Kawashima K

General

Full name : Kawashima Koichiro

First name : Koichiro

Mail : Department of Molecular Pharmacology, Kitasato University, School of Pharmaceutical Sciences, Minato-ku, Tokyo 108-8641

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Country : Japan

Email : koichiro-jk@piano.ocn.ne.jp

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References (123)

Title : Phosphoproteomic of the acetylcholine pathway enables discovery of the PKC-beta-PIX-Rac1-PAK cascade as a stimulatory signal for aversive learning - Yamahashi_2022_Mol.Psychiatry__
Author(s) : Yamahashi Y , Lin YH , Mouri A , Iwanaga S , Kawashima K , Tokumoto Y , Watanabe Y , Faruk MO , Zhang X , Tsuboi D , Nakano T , Saito N , Nagai T , Yamada K , Kaibuchi K
Ref : Mol Psychiatry , : , 2022
Abstract : Acetylcholine is a neuromodulator critical for learning and memory. The cholinesterase inhibitor donepezil increases brain acetylcholine levels and improves Alzheimer's disease (AD)-associated learning disabilities. Acetylcholine activates striatal/nucleus accumbens dopamine receptor D2-expressing medium spiny neurons (D2R-MSNs), which regulate aversive learning through muscarinic receptor M1 (M1R). However, how acetylcholine stimulates learning beyond M1Rs remains unresolved. Here, we found that acetylcholine stimulated protein kinase C (PKC) in mouse striatal/nucleus accumbens. Our original kinase-oriented phosphoproteomic analysis revealed 116 PKC substrate candidates, including Rac1 activator beta-PIX. Acetylcholine induced beta-PIX phosphorylation and activation, thereby stimulating Rac1 effector p21-activated kinase (PAK). Aversive stimulus activated the M1R-PKC-PAK pathway in mouse D2R-MSNs. D2R-MSN-specific expression of PAK mutants by the Cre-Flex system regulated dendritic spine structural plasticity and aversive learning. Donepezil induced PAK activation in both accumbal D2R-MSNs and in the CA1 region of the hippocampus and enhanced D2R-MSN-mediated aversive learning. These findings demonstrate that acetylcholine stimulates M1R-PKC-beta-PIX-Rac1-PAK signaling in D2R-MSNs for aversive learning and imply the cascade's therapeutic potential for AD as aversive learning is used to preliminarily screen AD drugs.
ESTHER : Yamahashi_2022_Mol.Psychiatry__
PubMedSearch : Yamahashi_2022_Mol.Psychiatry__
PubMedID: 35665767

Title : Non-neuronal Cholinergic Muscarinic Acetylcholine Receptors in the Regulation of Immune Function - Mashimo_2022_Biol.Pharm.Bull_45_675
Author(s) : Mashimo M , Kawashima K , Fujii T
Ref : Biol Pharm Bull , 45 :675 , 2022
Abstract : Immune cells such as T and B cells, monocytes and macrophages all express most of the cholinergic components of the nervous system, including acetylcholine (ACh), choline acetyltransferase (ChAT), high affinity choline transporter, muscarinic and nicotinic ACh receptors (mAChRs and nAChRs, respectively), and acetylcholinesterase (AChE). Because of its efficient cleavage by AChE, ACh synthesized and released from immune cells acts only locally in an autocrine and/or paracrine fashion at mAChRs and nAChRs on themselves and other immune cells located in close proximity, leading to modification of immune function. Immune cells generally express all five mAChR subtypes (M(1)-M(5)) and neuron type nAChR subunits alpha2-alpha7, alpha9, alpha10, beta2-beta4. The expression pattern and levels of mAChR subtypes and nAChR subunits vary depending on the tissue involved and its immunological status. Immunological activation of T cells via T-cell receptor-mediated pathways and cell adhesion molecules upregulates ChAT expression, which facilitates the synthesis and release of ACh. At present, alpha7 nAChRs expressed in macrophages are receiving much attention because they play a central role in anti-inflammatory cholinergic pathways. However, it now appears that through modification of cytokine synthesis, G(q/11)-coupled mAChRs play a prominent role in regulation of T cell proliferation and differentiation and B cell immunoglobulin class switching. It is anticipated that greater understanding of G(q/11)-coupled mAChRs on immune cells will provide an opportunity to develop new and effective treatments for immunological disorders.
ESTHER : Mashimo_2022_Biol.Pharm.Bull_45_675
PubMedSearch : Mashimo_2022_Biol.Pharm.Bull_45_675
PubMedID: 35650095

Title : Regulation of Immune Functions by Non-Neuronal Acetylcholine (ACh) via Muscarinic and Nicotinic ACh Receptors - Mashimo_2021_Int.J.Mol.Sci_22_
Author(s) : Mashimo M , Moriwaki Y , Misawa H , Kawashima K , Fujii T
Ref : Int J Mol Sci , 22 : , 2021
Abstract : Acetylcholine (ACh) is the classical neurotransmitter in the cholinergic nervous system. However, ACh is now known to regulate various immune cell functions. In fact, T cells, B cells, and macrophages all express components of the cholinergic system, including ACh, muscarinic, and nicotinic ACh receptors (mAChRs and nAChRs), choline acetyltransferase, acetylcholinesterase, and choline transporters. In this review, we will discuss the actions of ACh in the immune system. We will first briefly describe the mechanisms by which ACh is stored in and released from immune cells. We will then address Ca(2+) signaling pathways activated via mAChRs and nAChRs on T cells and B cells, highlighting the importance of ACh for the function of T cells, B cells, and macrophages, as well as its impact on innate and acquired (cellular and humoral) immunity. Lastly, we will discuss the effects of two peptide ligands, secreted lymphocyte antigen-6/urokinase-type plasminogen activator receptor-related peptide-1 (SLURP-1) and hippocampal cholinergic neurostimulating peptide (HCNP), on cholinergic activity in T cells. Overall, we stress the fact that ACh does not function only as a neurotransmitter; it impacts immunity by exerting diverse effects on immune cells via mAChRs and nAChRs.
ESTHER : Mashimo_2021_Int.J.Mol.Sci_22_
PubMedSearch : Mashimo_2021_Int.J.Mol.Sci_22_
PubMedID: 34202925

Title : Physiological functions of the cholinergic system in immune cells - Fujii_2017_J.Pharmacol.Sci_134_1
Author(s) : Fujii T , Mashimo M , Moriwaki Y , Misawa H , Ono S , Horiguchi K , Kawashima K
Ref : J Pharmacol Sci , 134 :1 , 2017
Abstract : T and B cells, macrophages and dendritic cells (DCs) all express most of the components necessary for a functional cholinergic system. This includes choline acetyltransferase (ChAT), muscarinic and nicotinic acetylcholine (ACh) receptors (mAChRs and nAChRs, respectively) and acetylcholinesterase (AChE). Immunological activation of T cells up-regulates cholinergic activity, including ChAT and AChE expression. Moreover, toll-like receptor agonists induce ChAT expression in DCs and macrophages, suggesting cholinergic involvement in the regulation of immune function. Immune cells express all five M1-M5 mAChR subtypes and several nAChR subtypes, including alpha7. Modulation of antigen-specific antibody and pro-inflammatory cytokine production in M1/M5 mAChR gene-knockout (KO) and alpha7 nAChR-KO mice further support the idea of a non-neuronal cholinergic system contributing to the regulation of immune function. Evidence also suggests that alpha7 nAChRs are involved in suppressing DC and macrophage activity, leading to suppression of T cell differentiation into effector T cells. These findings suggest the possibility that immune function could be modulated by manipulating immune cell cholinergic activity using specific agonists and antagonists. Therefore, a fuller understanding of the immune cell cholinergic system should be useful for the development of drugs and therapeutic strategies for the treatment of inflammation-related diseases and cancers.
ESTHER : Fujii_2017_J.Pharmacol.Sci_134_1
PubMedSearch : Fujii_2017_J.Pharmacol.Sci_134_1
PubMedID: 28552584

Title : Expression and Function of the Cholinergic System in Immune Cells - Fujii_2017_Front.Immunol_8_1085
Author(s) : Fujii T , Mashimo M , Moriwaki Y , Misawa H , Ono S , Horiguchi K , Kawashima K
Ref : Front Immunol , 8 :1085 , 2017
Abstract : T and B cells express most cholinergic system components-e.g., acetylcholine (ACh), choline acetyltransferase (ChAT), acetylcholinesterase, and both muscarinic and nicotinic ACh receptors (mAChRs and nAChRs, respectively). Using ChATBAC-eGFP transgenic mice, ChAT expression has been confirmed in T and B cells, dendritic cells, and macrophages. Moreover, T cell activation via T-cell receptor/CD3-mediated pathways upregulates ChAT mRNA expression and ACh synthesis, suggesting that this lymphocytic cholinergic system contributes to the regulation of immune function. Immune cells express all five mAChRs (M1-M5). Combined M1/M5 mAChR-deficient (M1/M5-KO) mice produce less antigen-specific antibody than wild-type (WT) mice. Furthermore, spleen cells in M1/M5-KO mice produce less tumor necrosis factor (TNF)-alpha and interleukin (IL)-6, suggesting M1/M5 mAChRs are involved in regulating pro-inflammatory cytokine and antibody production. Immune cells also frequently express the alpha2, alpha5, alpha6, alpha7, alpha9, and alpha10 nAChR subunits. alpha7 nAChR-deficient (alpha7-KO) mice produce more antigen-specific antibody than WT mice, and spleen cells from alpha7-KO mice produce more TNF-alpha and IL-6 than WT cells. This suggests that alpha7 nAChRs are involved in regulating cytokine production and thus modulate antibody production. Evidence also indicates that nicotine modulates immune responses by altering cytokine production and that alpha7 nAChR signaling contributes to immunomodulation through modification of T cell differentiation. Together, these findings suggest the involvement of both mAChRs and nAChRs in the regulation of immune function. The observation that vagus nerve stimulation protects mice from lethal endotoxin shock led to the notion of a cholinergic anti-inflammatory reflex pathway, and the spleen is an essential component of this anti-inflammatory reflex. Because the spleen lacks direct vagus innervation, it has been postulated that ACh synthesized by a subset of CD4+ T cells relays vagal nerve signals to alpha7 nAChRs on splenic macrophages, which downregulates TNF-alpha synthesis and release, thereby modulating inflammatory responses. However, because the spleen is innervated solely by the noradrenergic splenic nerve, confirmation of an anti-inflammatory reflex pathway involving the spleen requires several more hypotheses to be addressed. We will review and discuss these issues in the context of the cholinergic system in immune cells.
ESTHER : Fujii_2017_Front.Immunol_8_1085
PubMedSearch : Fujii_2017_Front.Immunol_8_1085
PubMedID: 28932225

Title : Recent progress in revealing the biological and medical significance of the non-neuronal cholinergic system - Grando_2015_Int.Immunopharmacol_29(1)_1
Author(s) : Grando SA , Kawashima K , Kirkpatrick CJ , Kummer W , Wessler I
Ref : Int Immunopharmacol , 29 :1 , 2015
Abstract : This special issue of International Immunopharmacology is the proceedings of the Fourth International Symposium on Non-neuronal Acetylcholine that was held on August 28-30, 2014 at the Justus Liebig University of Giessen in Germany. It contains original contributions of meeting participants covering the significant progress in understanding of the biological and medical significance of the non-neuronal cholinergic system extending from exciting insights into molecular mechanisms regulating this system via miRNAs over the discovery of novel cholinergic cellular signaling circuitries to clinical implications in cancer, wound healing, immunity and inflammation, cardiovascular, respiratory and other diseases.
ESTHER : Grando_2015_Int.Immunopharmacol_29(1)_1
PubMedSearch : Grando_2015_Int.Immunopharmacol_29(1)_1
PubMedID: 26362206

Title : Non-neuronal cholinergic system in regulation of immune function with a focus on alpha7 nAChRs - Kawashima_2015_Int.Immunopharmacol_29(1)_127
Author(s) : Kawashima K , Fujii T , Moriwaki Y , Misawa H , Horiguchi K
Ref : Int Immunopharmacol , 29 :127 , 2015
Abstract : In 1929, Dale and Dudley described the first reported natural occurrence of acetylcholine (ACh) in an animal's body. They identified this ACh in the spleens of horses and oxen, which we now know suggests possible involvement of ACh in the regulation of lymphocyte activity and immune function. However, the source and function of splenic ACh were left unexplored for several decades. Recent studies on the source of ACh in the blood revealed ACh synthesis catalyzed by choline acetyltransferase (ChAT) in CD4(+) T cells. T and B cells, macrophages and dendritic cells (DCs) all express all five muscarinic ACh receptor subtypes (mAChRs) and several subtypes of nicotinic AChRs (nAChRs), including alpha7 nAChRs. Stimulation of these mAChRs and nAChRs by their respective agonists causes functional and biochemical changes in the cells. Using AChR knockout mice, we found that M(1)/M(5) mAChR signaling up-regulates IgG(1) and pro-inflammatory cytokine production, while alpha7 nAChR signaling has the opposite effect. These findings suggest that ACh synthesized by T cells acts in an autocrine/paracrine fashion at AChRs on various immune cells to modulate immune function. In addition, an endogenous allosteric and/or orthosteric alpha7 nAChR ligand, SLURP-1, facilitates functional development of T cells and increases ACh synthesis via up-regulation of ChAT mRNA expression. SLURP-1 is expressed in CD205(+) DCs residing in the tonsil in close proximity to T cells, macrophages and B cells. Collectively, these findings suggest that ACh released from T cells along with SLURP-1 regulates cytokine production by activating alpha7 nAChRs on various immune cells, thereby facilitating T cell development and/or differentiation, leading to immune modulation.
ESTHER : Kawashima_2015_Int.Immunopharmacol_29(1)_127
PubMedSearch : Kawashima_2015_Int.Immunopharmacol_29(1)_127
PubMedID: 25907239

Title : Transcriptional regulation of SLURP2, a psoriasis-associated gene, is under control of IL-22 in the skin: A special reference to the nested gene LYNX1 - Moriwaki_2015_Int.Immunopharmacol_29(1)_71
Author(s) : Moriwaki Y , Takada K , Tsuji S , Kawashima K , Misawa H
Ref : Int Immunopharmacol , 29 :71 , 2015
Abstract : A novel nicotinic acetylcholine (ACh) receptor (nAChR)-mediated transduction pathway, regulating keratinocyte function, has been elucidated in studies of secreted mammalian Ly6/urokinase plasminogen activator receptor-related protein (SLURP)-1 and -2. SLURPs are members of Ly6/neurotoxin superfamily (Ly6SF) of proteins containing the unique three-finger domain in their three-dimensional structure. Some endogenously expressed Ly6SF proteins (such as LYNX1, SLURP-1, and SLURP-2) modulate the function of nAChR, either as allosteric and/or orthosteric modulators, or as antagonists. Although the expression and functions of SLURP-1 and SLURP-2 in keratinocytes are well documented, the expression and the modes of action of LYNX1 in keratinocytes are unknown. Additionally, a particular hybrid transcript, LYNX1-SLURP2, which contains both LYNX1 and SLURP-2 sequences, with unknown function, has been reported. Furthermore, although SLURP2 is a gene strongly induced in psoriatic skin lesions, the mechanisms controlling SLURP2 expression are largely unknown. To better understand the function of nAChRs in keratinocytes, we investigated the expression profiles of LYNX1, LYNX1-SLURP-2, and SLURP-2 in keratinocytes under various inflammatory conditions. We found that keratinocytes express LYNX1 and SLURP2, but not LYNX1-SLURP2, at mRNA and protein levels. IL-22 treatment increased SLURP2 expression in keratinocytes, but this effect was completely abolished by IFN-gamma. Furthermore, the IL-22-induced up-regulation of SLURP2 was completely suppressed by the inhibitor or siRNA for STAT3, a major transcriptional factor downstream of IL-22. These findings provide new insights into the nAChR-mediated regulatory mechanism of SLURP-2 expression in keratinocytes.
ESTHER : Moriwaki_2015_Int.Immunopharmacol_29(1)_71
PubMedSearch : Moriwaki_2015_Int.Immunopharmacol_29(1)_71
PubMedID: 26033490

Title : SLURP-1, an endogenous alpha7 nicotinic acetylcholine receptor allosteric ligand, is expressed in CD205(+) dendritic cells in human tonsils and potentiates lymphocytic cholinergic activity - Fujii_2014_J.Neuroimmunol_267_43
Author(s) : Fujii T , Horiguchi K , Sunaga H , Moriwaki Y , Misawa H , Kasahara T , Tsuji S , Kawashima K
Ref : Journal of Neuroimmunology , 267 :43 , 2014
Abstract : Immune cells often express various nicotinic ACh receptor (nAChR) subtypes, including alpha7 nAChRs, as well as mRNA encoding secreted lymphocyte antigen-6/urokinase-type plasminogen activator receptor-related peptide (SLURP)-1, an endogenous alpha7 nAChR allosteric ligand. We detected SLURP-1 immunoreactivity in CD205(+) dendritic cells (DCs) residing in human tonsils. Phytohemagglutinin (PHA, 10mug/ml), a T cell activator, attenuated cell proliferation and increased the ACh content of MOLT-3 human leukemic T cells compared with the vehicle control. Methyllycaconitine (MLA, 100nM), a specific alpha7 nAChR antagonist, abolished all effects elicited by PHA. Recombinant (r)SLURP-1 (0.5mug/ml) attenuated peripheral blood mononuclear cell proliferation and increased ChAT gene expression and the ACh content in MOLT-3 cells compared with the control, all of which were abolished by MLA. This suggests SLURP-1 activates cholinergic transmission by potentiating ACh synthesis and its action at alpha7 nAChRs, thereby facilitating functional development of T cells. These findings support the notion that SLURP-1 acts as a key modulator of immune responses.
ESTHER : Fujii_2014_J.Neuroimmunol_267_43
PubMedSearch : Fujii_2014_J.Neuroimmunol_267_43
PubMedID: 24365495

Title : Effect of secreted lymphocyte antigen-6\/urokinase-type plasminogen activator receptor-related peptide-1 (SLURP-1) on airway epithelial cells - Narumoto_2013_Biochem.Biophys.Res.Commun_438_175
Author(s) : Narumoto O , Niikura Y , Ishii S , Morihara H , Okashiro S , Nakahari T , Nakano T , Matsumura H , Shimamoto C , Moriwaki Y , Misawa H , Yamashita N , Nagase T , Kawashima K
Ref : Biochemical & Biophysical Research Communications , 438 :175 , 2013
Abstract : Acetylcholine (ACh) exerts various anti-inflammatory effects through alpha7 nicotinic ACh receptors (nAChRs). We have previously shown that secreted lymphocyte antigen-6/urokinase-type plasminogen activator receptor-related peptide-1 (SLURP-1), a positive allosteric modulator of alpha7 nAChR signaling, is down-regulated both in an animal model of asthma and in human epithelial cells treated with an inflammatory cytokine related to asthma. Our aim of this study was to explore the effect of SLURP-1, signal through alpha7 nAChR, in the pathophysiology of airway inflammation. Cytokine production was examined using human epithelial cells. Ciliary beat frequency of murine trachea was measured using a high speed camera. The IL-6 and TNF-alpha production by human epithelial cells was augmented by siRNA of SLURP-1 and alpha7 nicotinic ACh receptor. The cytokine production was also dose-dependently suppressed by human recombinant SLURP-1 (rSLURP-1). The ciliary beat frequency and amplitude of murine epithelial cells were augmented by PNU282987, a selective alpha7 nAChR agonist. Those findings suggested that SLURP-1 and stimulus through alpha7 nicotinic ACh receptors actively controlled asthmatic condition by stimulating ciliary beating and also by suppressing airway inflammation.
ESTHER : Narumoto_2013_Biochem.Biophys.Res.Commun_438_175
PubMedSearch : Narumoto_2013_Biochem.Biophys.Res.Commun_438_175
PubMedID: 23876317

Title : The missing link between long-term stimulation of nicotinic receptors and the increases of acetylcholine release and vasodilation in the cerebral cortex of aged rats - Uchida_2013_J.Physiol.Sci_63_95
Author(s) : Uchida S , Hotta H , Misawa H , Kawashima K
Ref : J Physiol Sci , 63 :95 , 2013
Abstract : In adult rats (4-9 months), chronic nicotine infusion increases the basal level of acetylcholine (ACh) release in the cerebral cortex and enhances responses of cortical ACh release and cortical vasodilation elicited by nucleus basalis of Meynert (NBM) stimulation. In the present study, we examined whether these effects of nicotine are detected in aged rats. Aged rats (27-30 months) received sustained subcutaneous nicotine (100 mug/kg/h) or saline for 14 days. Under urethane anesthesia, ACh release and regional blood flow in the parietal cortex were measured. The basal level of ACh release in the cerebral cortex was not changed by chronic nicotine. In addition, the magnitudes of ACh release and vasodilation by NBM stimulation were similar between the saline-treated and nicotine-treated groups. The lack of an effect of chronic nicotine in aged rats may be due to a decrease in nicotinic receptors in the cerebral cortex during aging (Nordberg et al., J Neurosci Res 31:103-111, 1992).
ESTHER : Uchida_2013_J.Physiol.Sci_63_95
PubMedSearch : Uchida_2013_J.Physiol.Sci_63_95
PubMedID: 23086726

Title : Relationship between nicotine dependence and the endophenotype-related trait of cognitive function but not acoustic startle reponses in Japanese patients with schizophrenia - Kishi_2013_Hum.Psychopharmacol_28_220
Author(s) : Kishi T , Fukuo Y , Okochi T , Kawashima K , Moriwaki M , Furukawa O , Musso GM , Fujita K , Correll CU , Iwata N
Ref : Hum Psychopharmacol , 28 :220 , 2013
Abstract : OBJECTIVES: We investigated whether nicotine dependence affects these endophenotypes in Japanese schizophrenia patients and whether alpha4 and beta2 subunits of neuronal nicotinic acetylcholine receptor genes (alpha4 subunit of the nAChR gene (CHRNA4)/beta2 subunit of the nAChR gene (CHRNB2)) were associated with nicotine dependence in patients (n = 100) and healthy controls (n = 107).
METHODS: First, in patients, we evaluated cognitive function, using the Brief Assessment of Cognition in Schizophrenia, and acoustic startle responses. Second, we evaluated the severity of nicotine dependence, using the Tobacco Dependence Screener, the Fagerstrom Test for Nicotine Dependence, and the Brinkman index in current smokers in both groups. Third, we evaluated the relationship between acoustic startle responses, cognitive function, and severity of nicotine dependence. Finally, using 12 tagging single-nucleotide polymorphisms in each the CHRNA4/CHRNB2, we used multiple linear regression analysis to examine the association between nicotine dependence measures and each selected single-nucleotide polymorphism.
RESULTS: The presence and severity of nicotine dependence were associated with verbal memory and executive function in schizophrenia patients. However, nicotine dependence was not correlated with any acoustic startle response. In addition, rs755203 and rs1044397 in CHRNA4 were associated with nicotine dependence in healthy controls.
CONCLUSIONS: Nicotine dependence might influence the level of verbal memory and executive function in schizophrenia patients. In addition, rs755203 and rs1044397 in CHRNA4 might play a role in the pathophysiology of nicotine dependence in healthy controls in the Japanese population.
ESTHER : Kishi_2013_Hum.Psychopharmacol_28_220
PubMedSearch : Kishi_2013_Hum.Psychopharmacol_28_220
PubMedID: 23553665

Title : Mediatophore regulates acetylcholine release from T cells - Fujii_2012_J.Neuroimmunol_244_16
Author(s) : Fujii T , Takada-Takatori Y , Horiguchi K , Kawashima K
Ref : Journal of Neuroimmunology , 244 :16 , 2012
Abstract : Immunological stimulation of T cells by phytohemagglutinin (PHA) enhances the synthesis and release of acetylcholine (ACh), suggesting a role for the lymphocytic cholinergic system in the regulation of immune function. In the present study, we used two human leukemic T cell lines as models to investigate whether mediatophore, a homooligomer of a 16-kDa subunit homologous to the proteolipid subunit c of vacuolar H(+)-ATPase (V-ATPase), is involved in mediating ACh release from T cells. Immunohistochemical analysis revealed the presence of mediatophore in the cytoplasm and on the plasma membrane of both T cell lines. Mediatophore gene expression was up-regulated by immunological T cell activation by PHA. Transfection of anti-mediatophore small interference RNA down-regulated mediatophore gene expression and significantly reduced ACh release. These results suggest that T cells express mediatophore, which then plays a key role in mediating ACh release, and that mediatophore expression is regulated by immunological stimulation.
ESTHER : Fujii_2012_J.Neuroimmunol_244_16
PubMedSearch : Fujii_2012_J.Neuroimmunol_244_16
PubMedID: 22245286

Title : Reconciling neuronally and nonneuronally derived acetylcholine in the regulation of immune function - Kawashima_2012_Ann.N.Y.Acad.Sci_1261_7
Author(s) : Kawashima K , Fujii T , Moriwaki Y , Misawa H , Horiguchi K
Ref : Annals of the New York Academy of Sciences , 1261 :7 , 2012
Abstract : Immune cells, including lymphocytes, express muscarinic and nicotinic acetylcholine (ACh) receptors (mAChRs and nAChRs, respectively), and agonist stimulation of these AChRs causes functional and biochemical changes in the cells. The origin of the ACh that acts on immune cell AChRs has remained unclear until recently, however. In 1995, we identified choline acetyltransferase mRNA and protein in human T cells, and found that immunological T cell activation potentiated lymphocytic cholinergic transmission by increasing ACh synthesis and AChR expression. We also found that M(1) /M(5) mAChR signaling upregulates IgG(1) and proinflammatory cytokine production, whereas alpha7 nAChR signaling has the opposite effect. These findings suggest that ACh synthesized by T cells acts as an autocrine and/or paracrine factor via AChRs on immune cells to modulate immune function. In addition, a recently discovered endogenous allosteric alpha7 nAChR ligand, SLURP-1, also appears to be involved in modulating normal T cell function.
ESTHER : Kawashima_2012_Ann.N.Y.Acad.Sci_1261_7
PubMedSearch : Kawashima_2012_Ann.N.Y.Acad.Sci_1261_7
PubMedID: 22823388

Title : Regulatory mechanisms of acetylcholine synthesis and release by T cells - Fujii_2012_Life.Sci_91_981
Author(s) : Fujii T , Takada-Takatori Y , Kawashima K
Ref : Life Sciences , 91 :981 , 2012
Abstract : AIMS: Muscarinic and nicotinic acetylcholine (ACh) receptors are expressed in immune cells. ACh synthesized by choline acetyltransferase (ChAT) and released in T cells binds to these receptors. Furthermore, we have recently demonstrated the involvement of mediatophore, a homooligomer of a 16-kDa proteolipid subunit of vacuolar H(+)-ATPase, in ACh release from T cells. In this study, we investigated the effects of phorbol 12-myristate 13-acetate (PMA), dibutyryl cAMP (dbcAMP) and FK506, an immunosuppressant calcineurin inhibitor, on lymphocytic cholinergic activity in T cells. MAIN
METHODS: We determined the content and release of ACh in human leukemic T cell line MOLT-3 cells using a sensitive and specific radioimmunoassay for ACh. In addition, expression of ChAT mRNA and ChAT activity were investigated using reverse-transcription-polymerase chain reaction and Fonnum method, respectively. KEY FINDINGS: Phytohemagglutinin (PHA), a T-cell activator, up-regulated ChAT mRNA expression, synthesis and release of ACh. PMA, a protein kinase C (PKC) activator, and dbcAMP, a protein kinase A (PKA) activator, also increased ChAT activity and ACh synthesis by up-regulating ChAT gene expression. FK506 inhibited PHA-induced up-regulation of ChAT mRNA expression, suggesting the involvement of calcineurin-mediated pathways in ChAT gene transcription. SIGNIFICANCE: Activation of PKC and PKA up-regulates ACh synthesis in T cells, and immunological activation triggers ChAT gene transcription through calcineurin-mediated pathways.
ESTHER : Fujii_2012_Life.Sci_91_981
PubMedSearch : Fujii_2012_Life.Sci_91_981
PubMedID: 22569292

Title : The non-neuronal cholinergic system: basic science, therapeutic implications and new perspectives -
Author(s) : Grando SA , Kawashima K , Kirkpatrick CJ , Meurs H , Wessler I
Ref : Life Sciences , 91 :969 , 2012
PubMedID: 23141771

Title : Critical roles of acetylcholine and the muscarinic and nicotinic acetylcholine receptors in the regulation of immune function - Kawashima_2012_Life.Sci_91_1027
Author(s) : Kawashima K , Fujii T , Moriwaki Y , Misawa H
Ref : Life Sciences , 91 :1027 , 2012
Abstract : Lymphocytes express both muscarinic and nicotinic acetylcholine (ACh) receptors (mAChRs and nAChRs, respectively), and stimulation of mAChRs and nAChRs produces various biochemical and functional changes. Although it has been postulated that parasympathetic cholinergic nerves directly innervate immune cells, no evidence has supported this hypothesis. We measured ACh in the blood of various animal species and determined its localization in T cells using a sensitive and specific radioimmunoassay. Furthermore, we showed that T cells express choline acetyltransferase (ChAT), an ACh synthesizing enzyme. Immunological T cell activation enhances ACh synthesis through the up-regulation of ChAT expression, suggesting lymphocytic cholinergic activity is related to immunological activity. Most immune cells such as T cells, B cells, and monocytes express all five subtypes of mAChRs (M(1)-M(5)), and various subunits of the nAChR, such as alpha3, alpha5, alpha7, alpha9, and alpha10. Studies on serum antibody production in M(1) and M(5) combined mAChR gene knockout (KO) mice immunized with ovalbumin (OVA) revealed that M(1)/M(5) mAChRs up-regulate TNF-alpha, IFN-gamma and IL-6 production in spleen cells, leading to an elevation of serum anti-OVA specific IgG(1). In contrast, studies of nAChR alpha7 subunit gene KO mice immunized with OVA show that alpha7 nAChRs down-regulate these proinflammatory cytokines, thereby leading to a reduction of anti-OVA specific IgG(1). Taken together, these findings demonstrate that both mAChRs and nAChRs modulate production of cytokines, such as TNF-alpha, resulting in a modification of antibody production. These findings support the notion that a non-neuronal cholinergic system is involved in the regulation of immune cell function.
ESTHER : Kawashima_2012_Life.Sci_91_1027
PubMedSearch : Kawashima_2012_Life.Sci_91_1027
PubMedID: 22659391

Title : Localization of acetylcholine-related molecules in the retina: implication of the communication from photoreceptor to retinal pigment epithelium - Matsumoto_2012_PLoS.One_7_e42841
Author(s) : Matsumoto H , Shibasaki K , Uchigashima M , Koizumi A , Kurachi M , Moriwaki Y , Misawa H , Kawashima K , Watanabe M , Kishi S , Ishizaki Y
Ref : PLoS ONE , 7 :e42841 , 2012
Abstract : It has been long speculated that specific signals are transmitted from photoreceptors to the retinal pigment epithelium (RPE). However, such signals have not been identified. In this study, we examined the retinal expression and localization of acetylcholine-related molecules as putative candidates for these signals. Previous reports revealed that alpha7 nicotinic acetylcholine receptors (nAChRs) are present in the microvilli of RPE cells that envelope the tips of photoreceptor outer segments (OS). Secreted mammalian leukocyte antigen 6/urokinase-type plasminogen activator receptor-related protein-1 (SLURP-1) is a positive allosteric modulator of the alpha7 nAChR. Therefore, we first focused on the expression of SLURP-1. SLURP-1 mRNA was expressed in the outer nuclear layer, which is comprised of photoreceptor cell bodies. SLURP-1 immunoreactivity co-localized with rhodopsin and S-opsin in photoreceptor OS, while choline acetyltransferase (ChAT) and high affinity choline transporter (CHT-1) were also expressed in photoreceptor OS. Immunoelectron microscopy identified that the majority of SLURP-1 was localized to the plasma membranes of photoreceptor OS. These results provide evidence that SLURP-1 is synthesized in photoreceptor cell bodies and transported to photoreceptor OS, where SLURP-1 may also be secreted. Our findings suggest that photoreceptor OS communicate via neurotransmitters such as ACh and SLURP-1, while RPE cells might receive these signals through alpha7 nAChRs in their microvilli.
ESTHER : Matsumoto_2012_PLoS.One_7_e42841
PubMedSearch : Matsumoto_2012_PLoS.One_7_e42841
PubMedID: 22880119

Title : Sustained subcutaneous infusion of nicotine enhances cholinergic vasodilation in the cerebral cortex induced by stimulation of the nucleus basalis of Meynert in rats - Uchida_2011_Eur.J.Pharmacol_654_235
Author(s) : Uchida S , Hotta H , Misawa H , Kawashima K
Ref : European Journal of Pharmacology , 654 :235 , 2011
Abstract : The present study examined the effects of sustained nicotine exposure on the cholinergic vasodilative system originating in the nucleus basalis of Meynert (NBM) and projecting to the cerebral cortex in rats. Rats received sustained subcutaneous nicotine (100mug/kg/h) for 14 days. Under urethane anesthesia, the vasodilation response and acetylcholine release in the parietal cortex induced by electrical stimulation of the NBM (10-200muA) were measured. The basal level of acetylcholine release was significantly higher in nicotine-treated rats than in saline-treated control rats. In the control rats, both the acetylcholine release and blood flow were increased by NBM stimulation in a stimulus intensity-dependent manner, and a threshold of 50muA. In nicotine-treated rats, the threshold intensity of NBM stimulation producing increases in acetylcholine release and blood flow was reduced to 20muA. The stimulus intensity-dependent acetylcholine release and vasodilation by NBM stimulation were significantly larger in nicotine-treated rats than in control rats. We conclude that sustained subcutaneous infusion of nicotine enhances cholinergic vasodilative system in the cerebral cortex originating in the NBM.
ESTHER : Uchida_2011_Eur.J.Pharmacol_654_235
PubMedSearch : Uchida_2011_Eur.J.Pharmacol_654_235
PubMedID: 21237144

Title : Down-regulation of secreted lymphocyte antigen-6\/urokinase-type plasminogen activator receptor-related peptide-1 (SLURP-1), an endogenous allosteric alpha7 nicotinic acetylcholine receptor modulator, in murine and human asthmatic conditions - Narumoto_2010_Biochem.Biophys.Res.Commun_398_713
Author(s) : Narumoto O , Horiguchi K , Horiguchi S , Moriwaki Y , Takano-Ohmuro H , Shoji S , Misawa H , Yamashita N , Nagase T , Kawashima K
Ref : Biochemical & Biophysical Research Communications , 398 :713 , 2010
Abstract : Whereas acetylcholine (ACh) acts as a bronchoconstrictor and stimulator of mucus secretion from bronchial epithelium, it acts via alpha7 nicotinic Ach receptors (nAChRs) on macrophages in the airways to exert anti-inflammatory effects by reducing synthesis of pro-inflammatory cytokines, such as tumor necrosis factor-alpha (TNF-alpha). Moreover, the effects of ACh are modified by secreted ly-6/urokinase-type plasminogen activator receptor-related peptide-1 (SLURP-1), a positive allosteric modulator of alpha7 nAChR signaling. Our aim was to explore the roles played by SLURP-1 in the pathophysiology of asthma by assessing SLURP-1 expression in the OVA-sensitized murine asthma model and in cultured human bronchial epithelial cells. Using real-time PCR we found that expression of SLURP-1 mRNA is down-regulated in the lungs of asthmatic model mice, as compared to healthy mice. In addition, immunohistochemical studies confirmed the diminished expression of SLURP-1 in the bronchioles of asthmatic mice, and showed it was due to extensive metaplasia of mucus-secreting cells and the concomitant loss of ciliated epithelial cells. Expression of SLURP-1 mRNA and protein was also significantly down-regulated in human epithelial cells stimulated with the pro-inflammatory cytokine interleukin-13 (IL-13), which is related to asthmatic condition. Thus SLURP-1 appears to be down-regulated in both an animal model of asthma and human epithelial cells treated with an inflammatory cytokine related to asthma. Those findings suggest that diminished expression of SLURP-1 in asthma attenuates its negative regulation of airway inflammation, and that perhaps changes in SLURP-1 expression could serve as a marker of airway damage in asthma.
ESTHER : Narumoto_2010_Biochem.Biophys.Res.Commun_398_713
PubMedSearch : Narumoto_2010_Biochem.Biophys.Res.Commun_398_713
PubMedID: 20621062

Title : Complete genomic structure of the cultivated rice endophyte Azospirillum sp. B510 - Kaneko_2010_DNA.Res_17_37
Author(s) : Kaneko T , Minamisawa K , Isawa T , Nakatsukasa H , Mitsui H , Kawaharada Y , Nakamura Y , Watanabe A , Kawashima K , Ono A , Shimizu Y , Takahashi C , Minami C , Fujishiro T , Kohara M , Katoh M , Nakazaki N , Nakayama S , Yamada M , Tabata S , Sato S
Ref : DNA Research , 17 :37 , 2010
Abstract : We determined the nucleotide sequence of the entire genome of a diazotrophic endophyte, Azospirillum sp. B510. Strain B510 is an endophytic bacterium isolated from stems of rice plants (Oryza sativa cv. Nipponbare). The genome of B510 consisted of a single chromosome (3,311,395 bp) and six plasmids, designated as pAB510a (1,455,109 bp), pAB510b (723,779 bp), pAB510c (681,723 bp), pAB510d (628,837 bp), pAB510e (537,299 bp), and pAB510f (261,596 bp). The chromosome bears 2893 potential protein-encoding genes, two sets of rRNA gene clusters (rrns), and 45 tRNA genes representing 37 tRNA species. The genomes of the six plasmids contained a total of 3416 protein-encoding genes, seven sets of rrns, and 34 tRNAs representing 19 tRNA species. Eight genes for plasmid-specific tRNA species are located on either pAB510a or pAB510d. Two out of eight genomic islands are inserted in the plasmids, pAB510b and pAB510e, and one of the islands is inserted into trnfM-CAU in the rrn located on pAB510e. Genes other than the nif gene cluster that are involved in N(2) fixation and are homologues of Bradyrhizobium japonicum USDA110 include fixABCX, fixNOQP, fixHIS, fixG, and fixLJK. Three putative plant hormone-related genes encoding tryptophan 2-monooxytenase (iaaM) and indole-3-acetaldehyde hydrolase (iaaH), which are involved in IAA biosynthesis, and ACC deaminase (acdS), which reduces ethylene levels, were identified. Multiple gene-clusters for tripartite ATP-independent periplasmic-transport systems and a diverse set of malic enzymes were identified, suggesting that B510 utilizes C(4)-dicarboxylate during its symbiotic relationship with the host plant.
ESTHER : Kaneko_2010_DNA.Res_17_37
PubMedSearch : Kaneko_2010_DNA.Res_17_37
PubMedID: 20047946
Gene_locus related to this paper: azos1-d3nrk5 , azos1-d3ns85 , azos1-d3nsh5 , azos1-d3nt15 , azos1-d3ntk4 , azos1-d3nv04 , azos1-d3nx23 , azos1-d3ny33 , azos1-d3p0k9 , azos1-d3p0n8 , azos1-d3p1p1 , azos1-d3p2z0 , azos1-d3p4h2 , azos1-d3p4k9 , azos1-d3p5e7 , azos1-d3p281 , azos1-d3p626

Title : Acetylcholine synthesis and release in NIH3T3 cells coexpressing the high-affinity choline transporter and choline acetyltransferase - Fujii_2009_J.Neurosci.Res_87_3024
Author(s) : Fujii T , Masai M , Misawa H , Okuda T , Takada-Takatori Y , Moriwaki Y , Haga T , Kawashima K
Ref : Journal of Neuroscience Research , 87 :3024 , 2009
Abstract : Acetylcholine (ACh) is known to be a key neurotransmitter in the central and peripheral nervous systems, but it is also produced in a variety of non-neuronal tissues and cells, including lymphocytes, placenta, amniotic membrane, vascular endothelial cells, keratinocytes, and epithelial cells in the digestive and respiratory tracts. To investigate contribution made by the high-affinity choline transporter (CHT1) to ACh synthesis in both cholinergic neurons and nonneuronal cells, we transfected rat CHT1 cDNA into NIH3T3ChAT cells, a mouse fibroblast line expressing mouse choline acetyltransferase (ChAT), to establish the NIH3T3ChAT 112-1 cell line, which stably expresses both CHT1 and ChAT. NIH3T3ChAT 112-1 cells showed increased binding of the CHT1 inhibitor [(3)H]hemicholinium-3 (HC-3) and greater [(3)H]choline uptake and ACh synthesis than NIH3T3ChAT 103-1 cells, a CHT1-negative control cell line. HC-3 significantly inhibited ACh synthesis in NIH3T3ChAT 112-1 cells but did not affect synthesis in NIH3T3ChAT 103-1 cells. ACh synthesis in NIH3T3ChAT 112-1 cells was also reduced by amiloride, an inhibitor of organic cation transporters (OCTs) involved in low-affinity choline uptake, and by procaine and lidocaine, two local anesthetics that inhibit plasma membrane phospholipid metabolism. These results suggest that CHT1 plays a key role in ACh synthesis in NIH3T3ChAT 112-1 cells and that choline taken up by OCTs or derived from the plasma membrane is also utilized for ACh synthesis in both cholinergic neurons and nonneuronal cholinergic cells, such as lymphocytes.
ESTHER : Fujii_2009_J.Neurosci.Res_87_3024
PubMedSearch : Fujii_2009_J.Neurosci.Res_87_3024
PubMedID: 19405101

Title : Expression of SLURP-1, an endogenous alpha7 nicotinic acetylcholine receptor allosteric ligand, in murine bronchial epithelial cells - Horiguchi_2009_J.Neurosci.Res_87_2740
Author(s) : Horiguchi K , Horiguchi S , Yamashita N , Irie K , Masuda J , Takano-Ohmuro H , Himi T , Miyazawa M , Moriwaki Y , Okuda T , Misawa H , Ozaki H , Kawashima K
Ref : Journal of Neuroscience Research , 87 :2740 , 2009
Abstract : Mammalian secreted lymphocyte antigen-6/urokinase-type plasminogen activator receptor-related peptide-1 (SLURP-1) is a positive allosteric ligand for alpha7 nicotinic acetylcholine (ACh) receptors (alpha7 nAChRs) that potentiates responses to ACh and elicits proapoptotic activity in human keratinocytes. Mutations in the gene encoding SLURP-1 have been detected in patients with Mal de Meleda, a rare autosomal recessive skin disorder characterized by transgressive palmoplantar keratoderma. On the basis of these findings, SLURP-1 is postulated to be involved in regulating tumor necrosis factor-alpha (TNF-alpha) release from keratinocytes and macrophages via alpha7 nAChR-mediated pathways. In the present study, we assessed SLURP-1 expression in lung tissue from C57BL/6J mice to investigate the functions of SLURP-1 in pulmonary physiology and pathology. Immunohistochemical and in situ hybridization analyses revealed expression of SLURP-1 protein and mRNA, respectively, exclusively in ciliated bronchial epithelial cells. This was supported by Western blotting showing the presence of the 9.5-kDa SLURP-1 protein in whole-lung tissue and trachea. In addition, high-affinity choline transporter (CHT1) was detected in apical regions of bronchial epithelial cells and in neurons located in the lamina propria of the bronchus, suggesting that bronchial epithelial cells are able to synthesize both SLURP-1 and ACh. We also observed direct contact between F4/80-positive macrophages and bronchial epithelial cells and the presence of invading macrophages in close proximity to CHT1-positive nerve elements. Collectively, these results suggest that SLURP-1 contributes to the maintenance of bronchial epithelial cell homeostasis and to the regulation of TNF-alpha release from macrophages in bronchial tissue.
ESTHER : Horiguchi_2009_J.Neurosci.Res_87_2740
PubMedSearch : Horiguchi_2009_J.Neurosci.Res_87_2740
PubMedID: 19396877

Title : Long-term nicotine treatment reduces cerebral cortical vasodilation mediated by alpha4beta2-like nicotinic acetylcholine receptors in rats - Uchida_2009_Eur.J.Pharmacol_609_100
Author(s) : Uchida S , Hotta H , Kawashima K
Ref : European Journal of Pharmacology , 609 :100 , 2009
Abstract : Regional cortical cerebral blood flow is increased via activation of brain nicotinic acetylcholine receptors. Acute intravenous injection of nicotine increases cortical blood flow, without changing systemic blood pressure in anesthetized rats. Here, we examined whether the nicotine-induced cerebral cortical vasodilation is affected by chronic nicotine treatment. Rats received chronic subcutaneous nicotine (at a low or a high-dose) short-term (1 h) or long-term (14 days). Under urethane anesthesia, blood flow in the frontal cortex, before and after bolus injection of nicotine (0.3-30 microg/kg, i.v.) was measured by laser Doppler flowmetry. The threshold dose of nicotine (3 microg/kg, i.v.) producing vasodilation was not affected by chronic nicotine treatment. However, the vasodilation induced by nicotine at 30 microg/kg was reduced after long-term nicotine treatment (but not after short-term exposure). The degree of reduction was marked and was statistically significant with high-dose (100 microg/kg/h) nicotine; low-dose (33 microg/kg/h) nicotine had a small effect that was not statistically significant. In contrast, the vasodilation in the cortical vessels obtained by hypercapnia (inhalation of 10% CO2) was not changed by chronic nicotine treatment. The nicotine-induced cortical vasodilation was not influenced by methyllycaconitine, an alpha7-selective nicotinic antagonist, while it was completely abolished by dihydro-beta-erythroidine, an alpha4beta2-preferring nicotinic antagonist. We conclude that long-term nicotine treatment reduces the functional activity of alpha4beta2-like nicotinic receptors that mediate cortical vasodilation.
ESTHER : Uchida_2009_Eur.J.Pharmacol_609_100
PubMedSearch : Uchida_2009_Eur.J.Pharmacol_609_100
PubMedID: 19285493

Title : Primary sensory neuronal expression of SLURP-1, an endogenous nicotinic acetylcholine receptor ligand - Moriwaki_2009_Neurosci.Res_64_403
Author(s) : Moriwaki Y , Watanabe Y , Shinagawa T , Kai M , Miyazawa M , Okuda T , Kawashima K , Yabashi A , Waguri S , Misawa H
Ref : Neurosci Res , 64 :403 , 2009
Abstract : Secreted mammalian Ly6/urokinase plasminogen activator receptor-related protein-1 (SLURP-1) is a recently identified, endogenous ligand of the alpha7 subunit of nicotinic acetylcholine receptors. SLURP-1 is also the causative gene for an autosomal recessive palmoplantar keratoderma, Mal de Meleda. Although the function of SLURP-1 in keratinocyte development and differentiation has been extensively studied, little is known about its role in the nervous system. In the present study, we analyzed SLURP-1 expression in the spinal cord of rats, as a number of studies suggest spinal nicotinic acetylcholine receptors are important modulators of pain transmission. We detected intense SLURP-1 immunoreactivity in the dorsal horn of the spinal cord, especially in lamina I and outer II. In dorsal root ganglia, SLURP-1 immunoreactivity was detected in small- to medium-sized neurons, where in situ hybridization also revealed the presence of SLURP-1 mRNA. Fluorescent labeling of SLURP-1 partially overlapped that of calcitonin-gene related peptide (CGRP) or substance P (SP) in both the spinal cord dorsal horn and glabrous skin, and electron microscopic analysis revealed colocalization of SLURP-1 with SP or CGRP, in large synaptic vesicles in terminals within the superficial layer of the spinal cord. Finally, sciatic nerve axotomy reduced levels of SLURP-1 immunoreactivity in parallel with that of SP and CGRP in the ipsilateral superficial dorsal horn. These findings suggest that SLURP-1 is expressed in a subset of primary peptidergic sensory neurons.
ESTHER : Moriwaki_2009_Neurosci.Res_64_403
PubMedSearch : Moriwaki_2009_Neurosci.Res_64_403
PubMedID: 19409425

Title : Basic and clinical aspects of non-neuronal acetylcholine: expression of an independent, non-neuronal cholinergic system in lymphocytes and its clinical significance in immunotherapy - Fujii_2008_J.Pharmacol.Sci_106_186
Author(s) : Fujii T , Takada-Takatori Y , Kawashima K
Ref : J Pharmacol Sci , 106 :186 , 2008
Abstract : Lymphocytes possess all the components required to constitute an independent, non-neuronal cholinergic system. These include acetylcholine (ACh); choline acetyltransferase (ChAT), its synthesizing enzyme; and both muscarinic and nicotinic ACh receptors (mAChRs and nAChRs, respectively). ACh modifies T and B cell function via both mAChR- and nAChR-mediated pathways. Stimulation of lymphocytes with the T cell activator phytohemagglutinin, protein kinase C activator phorbol ester, or cell surface molecules enhances the synthesis and release of ACh and up-regulates ChAT and/or M(5) mAChR gene expression. Furthermore, animal models of immune disorders exhibit abnormal lymphocytic cholinergic activity. The cholesterol-lowering drug simvastatin attenuates the lymphocytic cholinergic activity of T cells by inhibiting LFA-1 signaling in a manner independent of its cholesterol-lowering activity. This suggests that simvastatin exerts its immunosuppressive effects in part by modifying lymphocytic cholinergic activity. Nicotine, an active ingredient of tobacco, ameliorates ulcerative colitis but exacerbates Crohn's disease. Expression of mRNAs encoding the nAChR alpha7 and alpha5 subunits are significantly diminished in peripheral mononuclear leukocytes from smokers, as compared with those from nonsmokers. In addition, long-term exposure of lymphocytes to nicotine reduces intracellular Ca(2+) signaling via alpha7 nAChR-mediated pathways. In fact, studies of humoral antibody production in M(1)/M(5) mAChR-deficient and alpha7 nAChR-deficient animals revealed the role of lymphocytic cholinergic activity in the regulation of immune function. These results provide clues to understanding the mechanisms underlying immune system regulation and could serve as the basis for the development of new immunomodulatory drugs.
ESTHER : Fujii_2008_J.Pharmacol.Sci_106_186
PubMedSearch : Fujii_2008_J.Pharmacol.Sci_106_186
PubMedID: 18285654

Title : Basic and clinical aspects of non-neuronal acetylcholine: overview of non-neuronal cholinergic systems and their biological significance - Kawashima_2008_J.Pharmacol.Sci_106_167
Author(s) : Kawashima K , Fujii T
Ref : J Pharmacol Sci , 106 :167 , 2008
Abstract : Acetylcholine (ACh) is a phylogenetically ancient molecule involved in cell-to-cell signaling in almost all life-forms on earth. Cholinergic components, including ACh, choline acetyltransferase, acetylcholinesterase, and muscarinic and nicotinic ACh receptors (mAChRs and nAChRs, respectively) have been identified in numerous non-neuronal cells and tissues, including keratinocytes, cancer cells, immune cells, urinary bladder, airway epithelial cells, vascular endothelial cells, and reproductive organs, among many others. Stimulation of the mAChRs and nAChRs elicits cell-specific functional and biochemical effects. These findings support the notion that non-neuronal cholinergic systems are expressed in certain cells and tissues and are involved in the regulation of their function and that cholinergic dysfunction is related to the pathophysiology of certain diseases. They also provide clues for development of drugs with novel mechanisms of action.
ESTHER : Kawashima_2008_J.Pharmacol.Sci_106_167
PubMedSearch : Kawashima_2008_J.Pharmacol.Sci_106_167
PubMedID: 18285657

Title : Alpha4 and beta2 subunits of neuronal nicotinic acetylcholine receptor genes are not associated with methamphetamine-use disorder in the Japanese population - Kishi_2008_Ann.N.Y.Acad.Sci_1139_70
Author(s) : Kishi T , Ikeda M , Kitajima T , Yamanouchi Y , Kinoshita Y , Kawashima K , Inada T , Harano M , Komiyama T , Hori T , Yamada M , Iyo M , Sora I , Sekine Y , Ozaki N , Ujike H , Iwata N
Ref : Annals of the New York Academy of Sciences , 1139 :70 , 2008
Abstract : The mesolimbic system is thought to be involved in the reinforcing action of many addictive drugs and the release of dopamine modulated by neuronal nicotine cholinergic receptors (nAChRs). Several investigations suggested that nAChRs on dopaminergic terminals play an important role in the development of some long-lasting adaptations associated with drug abuse. A majority of high-affinity nicotine binding sites in the brain have been showed in heteropentameric alpha4 (alpha4) and beta2 subunit (beta2) of nAChRs. Therefore, we conducted a genetic association analysis of the alpha4 gene (CHRNA4) and beta2 gene (CHRNB2) with methamphetamine (METH)-use disorder (191 cases and 753 controls). We first evaluated the linkage disequilibrium (LD) structure of these genes and selected 7 and 5 tagging SNPs (tag SNPs) on CHRNA4 and CHRNB2, respectively. Some tag SNPs were significantly associated with total METH-use disorder and METH-induced psychosis; however, these associations were no longer statistically significant after Bonferroni's correction for multiple testing. In conclusion, our results suggest that neither CHRNA4 nor CHRNB2 plays a major role in Japanese METH-use disorder.
ESTHER : Kishi_2008_Ann.N.Y.Acad.Sci_1139_70
PubMedSearch : Kishi_2008_Ann.N.Y.Acad.Sci_1139_70
PubMedID: 18991851

Title : Genetic association analysis of tagging SNPs in alpha4 and beta2 subunits of neuronal nicotinic acetylcholine receptor genes (CHRNA4 and CHRNB2) with schizophrenia in the Japanese population - Kishi_2008_J.Neural.Transm_115_1457
Author(s) : Kishi T , Ikeda M , Kitajima T , Yamanouchi Y , Kinoshita Y , Kawashima K , Okochi T , Inada T , Ozaki N , Iwata N
Ref : J Neural Transm , 115 :1457 , 2008
Abstract : Several lines of evidence suggest that nicotinic cholinergic dysfunction may contribute to the cognitive impairments in schizophrenia. The majority of high affinity nicotine binding sites in the human brain have been implicated in heteropentameric alpha4 and beta2 subunits of neuronal nicotinic acetylcholine receptors; therefore, these two neuronal nicotinic acetylcholine receptors genes (CHRNA4 and CHRNB2) are considered to be attractive candidate genes for the pathophysiology of schizophrenia. To represent these two genes in a gene-wide manner, we first evaluated the linkage disequilibrium structure using our own control samples. Thirteen SNPs (7 SNPs for CHRNA4 and 5 SNPs for CHRNB2) were selected as tagging SNPs. Using these tagging SNPs, we then conducted genetic association analysis of case-control samples (738 schizophrenia and 753 controls) in the Japanese population. No significant association was detected in the allele/genotype-wise or haplotype-wise analysis. Our results suggest that CHRNA4 and CHRNB2 do not play a major role in Japanese schizophrenia.
ESTHER : Kishi_2008_J.Neural.Transm_115_1457
PubMedSearch : Kishi_2008_J.Neural.Transm_115_1457
PubMedID: 18762859

Title : Aberrant trafficking of the high-affinity choline transporter in AP-3-deficient mice - Misawa_2008_Eur.J.Neurosci_27_3109
Author(s) : Misawa H , Fujigaya H , Nishimura T , Moriwaki Y , Okuda T , Kawashima K , Nakata K , Ruggiero AM , Blakely RD , Nakatsu F , Ohno H
Ref : European Journal of Neuroscience , 27 :3109 , 2008
Abstract : The high-affinity choline transporter (CHT) is expressed in cholinergic neurons and efficiently transported to axon terminals where it controls the rate-limiting step in acetylcholine synthesis. Recent studies have shown that the majority of CHT is unexpectedly localized on synaptic vesicles (SV) rather than the presynaptic plasma membrane, establishing vesicular CHT trafficking as a basis for activity-dependent CHT regulation. Here, we analyse the intracellular distribution of CHT in the adaptor protein-3 (AP-3)-deficient mouse model mocha. In the mocha mouse, granular structures in cell bodies are intensely labelled with CHT antibody, indicating possible deficits in CHT trafficking from the cell body to the axon terminal. Western blot analyses reveal that CHT on SV in mocha mice is decreased by 30% compared with wild-type mice. However, no significant difference in synaptosomal choline uptake activity is detected, consistent with the existence of a large reservoir pool for CHT. To further characterize CHT trafficking, we established a PC12D-CHT cell line. In this line, CHT is found associated with a subpopulation of synaptophysin-positive synaptic-like microvesicles (SLMV). The amounts of CHT detected on SLMV are greatly reduced by treating the cell with agents that halt AP-dependent membrane trafficking. These results demonstrate that APs have important functions for CHT trafficking in neuronal cells.
ESTHER : Misawa_2008_Eur.J.Neurosci_27_3109
PubMedSearch : Misawa_2008_Eur.J.Neurosci_27_3109
PubMedID: 18554297

Title : Immune system expression of SLURP-1 and SLURP-2, two endogenous nicotinic acetylcholine receptor ligands - Moriwaki_2007_Life.Sci_80_2365
Author(s) : Moriwaki Y , Yoshikawa K , Fukuda H , Fujii YX , Misawa H , Kawashima K
Ref : Life Sciences , 80 :2365 , 2007
Abstract : A novel transduction pathway via which apoptosis of keratinocytes is regulated through nicotinic acetylcholine (ACh) receptors (nAChRs) has emerged in studies of secreted mammalian Ly6/urokinase plasminogen-type activator receptor-related protein-1 and-2 (SLURP-1 and SLURP-2, respectively). SLURP-1 reportedly binds to alpha7 nAChRs and enhances the amplitude of macroscopic currents induced by ACh, leading to facilitation of apoptosis, whereas SLURP-2 binds to alpha3 nAChRs and prevents apoptosis. These observations prompted us to test whether SLURPs are expressed in immune cells and are involved in the regulation of immune function. We initially used reverse transcription-polymerase chain reaction analysis to characterize the expression profiles of SLURP mRNAs in several murine tissues and organs. Although SLURP-1 mRNA was not expressed in the pancreas, all other tissues and organs tested, including spleen and thymus, expressed both SLURP-1 and SLURP-2 mRNAs. Expression of both mRNAs also was detected in T and B cells, bone marrow-derived dendritic cells (DCs) and macrophages. Moreover, as in keratinocytes, stimulation of MOLT-3 human leukemic T cells with recombinant human SLURP-1 evoked intracellular Ca(2+) signaling. These results suggest that both SLURP-1 and SLURP-2 are expressed in various immune cells and organs, and that not only ACh but also SLURPs may be involved in regulating lymphocyte function via nAChR-mediated pathways.
ESTHER : Moriwaki_2007_Life.Sci_80_2365
PubMedSearch : Moriwaki_2007_Life.Sci_80_2365
PubMedID: 17286989

Title : Complete genomic structure of the bloom-forming toxic cyanobacterium Microcystis aeruginosa NIES-843 - Kaneko_2007_DNA.Res_14_247
Author(s) : Kaneko T , Nakajima N , Okamoto S , Suzuki I , Tanabe Y , Tamaoki M , Nakamura Y , Kasai F , Watanabe A , Kawashima K , Kishida Y , Ono A , Shimizu Y , Takahashi C , Minami C , Fujishiro T , Kohara M , Katoh M , Nakazaki N , Nakayama S , Yamada M , Tabata S , Watanabe MM
Ref : DNA Research , 14 :247 , 2007
Abstract : The nucleotide sequence of the complete genome of a cyanobacterium, Microcystis aeruginosa NIES-843, was determined. The genome of M. aeruginosa is a single, circular chromosome of 5,842,795 base pairs (bp) in length, with an average GC content of 42.3%. The chromosome comprises 6312 putative protein-encoding genes, two sets of rRNA genes, 42 tRNA genes representing 41 tRNA species, and genes for tmRNA, the B subunit of RNase P, SRP RNA, and 6Sa RNA. Forty-five percent of the putative protein-encoding sequences showed sequence similarity to genes of known function, 32% were similar to hypothetical genes, and the remaining 23% had no apparent similarity to reported genes. A total of 688 kb of the genome, equivalent to 11.8% of the entire genome, were composed of both insertion sequences and miniature inverted-repeat transposable elements. This is indicative of a plasticity of the M. aeruginosa genome, through a mechanism that involves homologous recombination mediated by repetitive DNA elements. In addition to known gene clusters related to the synthesis of microcystin and cyanopeptolin, novel gene clusters that may be involved in the synthesis and modification of toxic small polypeptides were identified. Compared with other cyanobacteria, a relatively small number of genes for two component systems and a large number of genes for restriction-modification systems were notable characteristics of the M. aeruginosa genome.
ESTHER : Kaneko_2007_DNA.Res_14_247
PubMedSearch : Kaneko_2007_DNA.Res_14_247
PubMedID: 18192279
Gene_locus related to this paper: micae-a8yde8 , micae-a8yen2 , micae-a8yma5 , micae-MCYC , mican-b0jqg0 , mican-b0jqq0 , mican-b0jsa2 , mican-b0jxh1 , mican-b0jux6 , mican-b0jyg0

Title : Enhanced serum antigen-specific IgG1 and proinflammatory cytokine production in nicotinic acetylcholine receptor alpha7 subunit gene knockout mice - Fujii_2007_J.Neuroimmunol_189_69
Author(s) : Fujii YX , Fujigaya H , Moriwaki Y , Misawa H , Kasahara T , Grando SA , Kawashima K
Ref : Journal of Neuroimmunology , 189 :69 , 2007
Abstract : Human and murine immune cells such as mononuclear leukocytes consisting of mainly T and B cells, bone marrow derived dendritic cells (DCs) and macrophages all express various nicotinic acetylcholine (ACh) receptor (nAChR) subunits. Activated T cells and DCs have the ability to synthesize ACh by choline acetyltransferase, suggesting the role of non-neuronal cholinergic system expressed in immune cells in the regulation of immune cell function. Stimulation of human leukemic T and B cell lines with nicotine causes a transient Ca(2+)-signaling that is antagonized by alpha-bungarotoxin, suggesting the involvement of alpha7 subunit. Furthermore, alpha7 nAChRs have been shown to negatively regulate synthesis and release of tumor necrosis factor (TNF)-alpha in macrophages. These findings suggest that immune cell function is regulated by its own non-neuronal cholinergic system, at least in part, via alpha7 nAChR-mediated pathways. In the present study, we tested the role of alpha7 nAChRs in the regulation of immune function by measuring total serum and antigen-specific IgG(1) and IgM, and production of TNF-alpha, gamma interferon (IFN-gamma) and interleukin (IL)-6 in activated spleen cells of nAChR alpha7 subunit gene knockout (alpha7 KO) and wild-type C57BL/6J mice immunized with ovalbumin (OVA). We found that serum levels of total and anti-OVA-specific IgG(1) were significantly elevated in alpha7 KO mice, though there were no significant differences in serum levels of total and anti-OVA-specific IgM between the two genotypes. Production of TNF-alpha, IFN-gamma and IL-6 in spleen cells was significantly facilitated in alpha7 KO mice. Expression of AChE mRNA was not different between the two genotypes. These results suggest that alpha7 nAChRs are involved in the regulation of cytokine production, through which modulates TNF-alpha, IFN-gamma and IL-6 productions, leading to modification of antibody production, but are not involved in expression of cholinergic components in immune cells.
ESTHER : Fujii_2007_J.Neuroimmunol_189_69
PubMedSearch : Fujii_2007_J.Neuroimmunol_189_69
PubMedID: 17675251

Title : Diminished antigen-specific IgG1 and interleukin-6 production and acetylcholinesterase expression in combined M1 and M5 muscarinic acetylcholine receptor knockout mice - Fujii_2007_J.Neuroimmunol_188_80
Author(s) : Fujii YX , Tashiro A , Arimoto K , Fujigaya H , Moriwaki Y , Misawa H , Fujii T , Matsui M , Kasahara T , Kawashima K
Ref : Journal of Neuroimmunology , 188 :80 , 2007
Abstract : Immunological activation of T cells enhances synthesis of acetylcholine (ACh) and transcription of choline acetyltransferase (ChAT), M5 muscarinic ACh receptor (mAChR) and acetylcholinesterase (AChE). Stimulation of mAChRs on T and B cells causes oscillating Ca(2+)-signaling and up-regulation of c-fos expression; moreover, M1 mAChRs play a crucial role in the differentiation of CD8(+) T cells into cytolytic T lymphocytes. Collectively, these findings suggest that immune cell function is regulated by its own cholinergic system. Bearing that in mind, we tested whether immune function can be regulated via mAChR-mediated pathways by immunizing combined M1 and M5 mAChR knockout (M1/M5 KO) and wild-type (WT) C57BL/6JJcl mice with ovalbumin (OVA) and measuring serum IgG1 and IgM 1 wk later. We found that serum levels of total and anti-OVA-specific IgG1 were significantly lower in M1/M5 KO than WT mice, though there was no difference in serum levels of total and anti-OVA-specific IgM between the two genotypes. Secretion of interleukin (IL)-6 from activated spleen cells was significantly reduced in M1/M5 KO mice, whereas there was no significant change in gamma interferon secretion. Expression of AChE mRNA was significantly reduced in activated spleen cells from M1/M5 KO mice. These results suggest that M1 and/or M5 mAChRs are involved in regulating cytokine (e.g., IL-6) production, leading to modulation of antibody class switching from IgM to IgG1, but are not involved in the initial generation of the antibody response. They also support the notion that a non-neuronal cholinergic system is involved in regulating immune cell function.
ESTHER : Fujii_2007_J.Neuroimmunol_188_80
PubMedSearch : Fujii_2007_J.Neuroimmunol_188_80
PubMedID: 17586055

Title : Roles played by lymphocyte function-associated antigen-1 in the regulation of lymphocytic cholinergic activity - Fujii_2007_Life.Sci_80_2320
Author(s) : Fujii T , Takada-Takatori Y , Kawashima K
Ref : Life Sciences , 80 :2320 , 2007
Abstract : Lymphocytes possess the essential components of a cholinergic system, including acetylcholine (ACh); choline acetyltransferase (ChAT), its synthesizing enzyme; and both muscarinic and nicotinic ACh receptors (mAChRs and nAChRs, respectively). Stimulation of lymphocytes with phytohemagglutinin, which activates T cells via the T cell receptor/CD3 complex, enhances the synthesis and release of ACh and up-regulates expression of ChAT and M(5) mAChR mRNAs. In addition, activation of protein kinase C and increases in intracellular cAMP also enhance cholinergic activity in T cells, and lymphocyte function associated antigen-1 (LFA-1; CD11a/CD18) is an important mediator of leukocyte migration and T cell activation. Anti-CD11a monoclonal antibody (mAb) as well as antithymocyte globulin containing antibodies against CD2, CD7 and CD11a all increase ChAT activity, ACh synthesis and release, and expression of ChAT and M(5) mAChR mRNAs in T cells. The cholesterol-lowering drug simvastatin inhibits LFA-1 signaling by binding to an allosteric site on CD11a (LFA-1 alpha chain), which leads to immunomodulation. We found that simvastatin abolishes anti-CD11a mAb-induced increases in lymphocytic cholinergic activity in a manner independent of its cholesterol-lowering activity. Collectively then, these results indicate that LFA-1 contributes to the regulation of lymphocytic cholinergic activity via CD11a-mediated pathways and suggest that simvastatin exerts its immunosuppressive effects in part via modification of lymphocytic cholinergic activity.
ESTHER : Fujii_2007_Life.Sci_80_2320
PubMedSearch : Fujii_2007_Life.Sci_80_2320
PubMedID: 17289088

Title : Recent progress in understanding the non-neuronal cholinergic system in humans -
Author(s) : Grando SA , Kawashima K , Kirkpatrick CJ , Wessler I
Ref : Life Sciences , 80 :2181 , 2007
PubMedID: 17467010

Title : The non-neuronal cholinergic system of human skin - Kurzen_2007_Horm.Metab.Res_39_125
Author(s) : Kurzen H , Wessler I , Kirkpatrick CJ , Kawashima K , Grando SA
Ref : Hormone & Metabolic Research , 39 :125 , 2007
Abstract : In human skin both resident and transiently residing cells are part of the extra- or non-neuronal cholinergic system, creating a highly complex and interconnected cosmos in which acetylcholine (ACh) and choline are the natural ligands of nicotinic and muscarinic receptors with regulatory function in both physiology and pathophysiology. ACh is produced in keratinocytes, endothelial cells and most notably in immune competent cells invading the skin at sites of inflammation. The cholinergic system is involved in basic functions of the skin through autocrine, paracrine, and endocrine mechanisms, like keratinocyte proliferation, differentiation, adhesion and migration, epidermal barrier formation, pigment-, sweat- and sebum production, blood circulation, angiogenesis, and a variety of immune reactions. The pathophysiological consequences of this complex cholinergic "concert" are only beginning to be understood. The present review aims at providing insight into basic mechanisms of this highly complex system.
ESTHER : Kurzen_2007_Horm.Metab.Res_39_125
PubMedSearch : Kurzen_2007_Horm.Metab.Res_39_125
PubMedID: 17326008

Title : Ubiquitous expression of acetylcholine and its biological functions in life forms without nervous systems - Kawashima_2007_Life.Sci_80_2206
Author(s) : Kawashima K , Misawa H , Moriwaki Y , Fujii YX , Fujii T , Horiuchi Y , Yamada T , Imanaka T , Kamekura M
Ref : Life Sciences , 80 :2206 , 2007
Abstract : Using a radioimmunoassay (RIA) with high specificity and sensitivity (1 pg/tube) for acetylcholine (ACh), we have been able to measure the ACh content in samples from the bacteria, archaea and eucarya domains of the universal phylogenetic tree. We found detectable levels of ACh to be ubiquitous in bacteria (e.g., Bacillus subtilis), archaea (e.g., Thermococcus kodakaraensis KOD1), fungi (e.g., shiitake mushroom and yeast), plants (e.g., bamboo shoot and fern) and animals (e.g., bloodworm and lugworm). The levels varied considerably, however, with the highest ACh content detected in the top portion of bamboo shoot (2.9 micromol/g), which contained about 80 times that found in rat brain. In addition, using the method of Fonnum, various levels of ACh-synthesizing activity also were detected, a fraction of which was catalyzed by a choline acetyltransferase (ChAT)-like enzyme (sensitive to bromoACh, a selective ChAT inhibitor) in T. kodakaraensis KOD1 (15%), bamboo shoot (91%) and shiitake mushroom (51%), bloodworm (91%) and lugworm (81%). Taken together, these findings demonstrate the ubiquitous expression of ACh and ACh-synthesizing activity among life forms without nervous systems, and support the notion that ACh has been expressed and may be active as a local mediator and modulator of physiological functions since the early beginning of life.
ESTHER : Kawashima_2007_Life.Sci_80_2206
PubMedSearch : Kawashima_2007_Life.Sci_80_2206
PubMedID: 17363003

Title : Expression and function of genes encoding cholinergic components in murine immune cells - Kawashima_2007_Life.Sci_80_2314
Author(s) : Kawashima K , Yoshikawa K , Fujii YX , Moriwaki Y , Misawa H
Ref : Life Sciences , 80 :2314 , 2007
Abstract : It is now evident that acetylcholine (ACh) synthesized by choline acetyltransferase (ChAT) and released from T cells during antigen presentation binds to muscarinic and nicotinic ACh receptors (mAChRs and nAChRs, respectively) on T and B cells or dendritic cells, leading to modulation of their function. In the present study, we used reverse transcription-polymerase chain reaction (RT-PCR) to investigate whether mononuclear leukocytes (MNLs), bone marrow-derived dendritic cells (DCs) and macrophages from C57BL/6J mice express components of the cholinergic system. Expression of ChAT mRNA was detected in MNLs activated with ConA and DCs stimulated with LPS, but not in resting MNLs and DCs or in resting and stimulated macrophages. MNLs, DCs and macrophages all expressed mRNAs encoding the five mAChR subtypes (M(1)-M(5)) and the nAChR alpha2, alpha5, alpha6, alpha7, alpha10 and beta2 subunits. Expression of VIP mRNA was detected in MNLs and macrophages, but not in DCs. MNLs, DCs and macrophages all expressed VIP receptor-1 (VPAC1) and -2 (VPAC2) mRNAs, as well as mRNAs encoding secreted mammalian Ly-6/urokinase-type plasminogen activator receptor-related protein (SLURP)-1 and SLURP-2, two endogenous nAChR ligands. These results suggest that the lymphocytic cholinergic system is activated by ACh via mAChR- and nAChR-mediated pathways during antigen presentation between T cells and DCs or macrophages, leading to modulation of immune cell function. Moreover, VIP released from both postganglionic cholinergic neurons and immune cells may play a role in the cholinergic anti-inflammatory reflex, acting via VPAC1 and VPAC2 on immune cells.
ESTHER : Kawashima_2007_Life.Sci_80_2314
PubMedSearch : Kawashima_2007_Life.Sci_80_2314
PubMedID: 17383684

Title : Simvastatin regulates non-neuronal cholinergic activity in T lymphocytes via CD11a-mediated pathways - Fujii_2006_J.Neuroimmunol_179_101
Author(s) : Fujii T , Masuyama K , Kawashima K
Ref : Journal of Neuroimmunology , 179 :101 , 2006
Abstract : Lymphocyte function associated antigen-1 (LFA-1; CD11a/CD18) is an important mediator of leukocyte migration and T cell activation. We previously showed that antithymocyte globulin stimulates an independent, non-neuronal cholinergic system in T cells via LFA-1-mediated pathways, as evidenced by increases in acetylcholine (ACh) synthesis and choline acetyltransferase (ChAT) mRNA expression. The cholesterol-lowering drug simvastatin inhibits LFA-1 signaling by binding to an allosteric site on CD11a (LFA-1 alpha chain), which leads to immunomodulation. In the present study, we investigated whether simvastatin modulates lymphocytic cholinergic activity in T cells. We found that anti-CD11a monoclonal antibody (mAb) increased ChAT activity, ACh synthesis and release, and expression of ChAT and M5 muscarinic ACh receptor mRNA in MOLT-3 cells, a human leukemic T cell line. Simvastatin abolished these anti-CD11a mAb-induced increases in lymphocytic cholinergic activity in a manner independent of its cholesterol-lowering activity. These results indicate that LFA-1 contributes to the regulation of lymphocytic cholinergic activity via CD11a-mediated pathways, and suggest that simvastatin exerts its immunosuppressive effects in part via modification of lymphocytic cholinergic activity.
ESTHER : Fujii_2006_J.Neuroimmunol_179_101
PubMedSearch : Fujii_2006_J.Neuroimmunol_179_101
PubMedID: 16828882

Title : [Expression of non-neuronal acetylcholine and its biological roles in mammalian species] -
Author(s) : Kawashima K
Ref : Nihon Yakurigaku Zasshi , 127 :368 , 2006
PubMedID: 16819242

Title : Expression of acetylcholine (ACh) and ACh-synthesizing activity in Archaea - Yamada_2005_Life.Sci_77_1935
Author(s) : Yamada T , Fujii T , Kanai T , Amo T , Imanaka T , Nishimasu H , Wakagi T , Shoun H , Kamekura M , Kamagata Y , Kato T , Kawashima K
Ref : Life Sciences , 77 :1935 , 2005
Abstract : Acetylcholine (ACh) is known generally as the neurotransmitter in the mammalian central and peripheral cholinergic nervous systems. However, ACh is also widely expressed in non-neuronal animal tissues and in plants, fungi and bacteria, where it is likely involved in the transport of water, electrolytes and nutrients, and in modulating various other cell functions. We have investigated the expression of ACh and ACh-synthesizing activity in various strains of Archaea, which are situated between Bacteria and Eucarya in the universal phylogenetic tree. Using a sensitive and specific radioimmunoassay, differing levels of ACh were detected in the Hyperthermophiles Thermococcus kodakaraensis KOD1, Sulfolobus tokodaii strain 7 and Pyrobaculum calidifontis VA1; the Methanogens Methanothermobacter thermautotrophicus deltaH and Methanosarcina barkeri; and the Halophiles Halobacterium sp. NRC-1 and Haloferax volcanii. T. kodakaraensis KOD1 expressed the highest levels of ACh among the Archaea tested; moreover, the substance expressed was verified to be ACh using high-performance liquid chromatography with electrochemical detection. Varying degrees of ACh-synthesizing activity were also identified in all of the strains, and the activity of bromoACh-sensitive choline acetyltransferase, an enzyme responsible for ACh synthesis in the nervous system, was detected in T. kodakaraensis KOD1. Our findings demonstrate that ACh and ACh-synthesizing activity are both expressed in evolutionally old Archaea. In the context of the recent discovery of non-neuronal ACh in bacteria, fungi, plants and animals, these findings support the notion that ACh has been expressed in organisms from the origin of life on the earth, functioning as a local mediator as well as a neurotransmitter.
ESTHER : Yamada_2005_Life.Sci_77_1935
PubMedSearch : Yamada_2005_Life.Sci_77_1935
PubMedID: 15936779

Title : Up-regulation of lymphocytic cholinergic activity by ONO-4819, a selective prostaglandin EP4 receptor agonist, in MOLT-3 human leukemic T cells - Suenaga_2004_Vascul.Pharmacol_41_51
Author(s) : Suenaga A , Fujii T , Ogawa H , Maruyama T , Ohuchida S , Katsube N , Obata T , Kawashima K
Ref : Vascul Pharmacol , 41 :51 , 2004
Abstract : We used a selective EP4 receptor agonist, ONO-4819, and a human leukemic T cell line MOLT-3 cells, which express all four prostaglandin E2 (PGE2) receptors (EP1-EP4), to investigate whether the EP4 PGE2 receptor subtype is involved in regulating lymphocytic cholinergic activity. Phytohemagglutinin (PHA), a T cell activator, significantly enhanced the expression of EP4 receptor mRNA during the first 3-6 h of exposure, after which, expression gradually declined. Furthermore, PHA stimulation slightly but significantly up-regulated the expression of EP2 mRNA after 12 h and of EP3 mRNA after 6 h. By contrast, expression level of EP1 receptor mRNA was not affected by PHA. ONO-4819 (1 microM), which was added to cultures after 3 h of PHA stimulation, significantly increased cellular ACh content and release, and up-regulated ChAT mRNA expression and activity but inhibited MOLT-3 cell proliferation. These findings suggest that the activation of T lymphocytes up-regulates EP4 receptor mRNA expression and, to a lesser extent, EP2 and EP3 receptors and that PGE2 enhances nonneuronal lymphocytic cholinergic transmission in activated T cells, at least in part, via EP4 receptor-mediated pathways.
ESTHER : Suenaga_2004_Vascul.Pharmacol_41_51
PubMedSearch : Suenaga_2004_Vascul.Pharmacol_41_51
PubMedID: 15196475

Title : Expression of non-neuronal acetylcholine in lymphocytes and its contribution to the regulation of immune function - Kawashima_2004_Front.Biosci_9_2063
Author(s) : Kawashima K , Fujii T
Ref : Front Biosci , 9 :2063 , 2004
Abstract : Lymphocytes express most components of the cholinergic system including acetylcholine (ACh), muscarinic and nicotinic ACh receptors (mAChRs and nAChRs, respectively), choline acetyltransferase (ChAT), high affinity choline transporter and acetylcholinesterase. ACh and mAChR agonists elicit intracellular Ca2+ signaling, up-regulation of c-fos expression and nitric oxide synthesis within T and B cells probably via M3 and M5 mAChRs. Stimulation of nAChRs with ACh or nicotine causes a rapid and transient Ca2+ signaling in T and B cells, probably via alpha7 nAChR subunit-mediated pathways. Phytohemagglutinin- or antigen-induced T cell activation via cell surface molecules (e.g., T cell receptor/CD3 complexes) enhances lymphocytic cholinergic transmission by up-regulating ChAT and M5 mAChR expression. It is thus likely that a local lymphocytic cholinergic system is involved in regulating immune function. This idea is supported by the findings that lymphocytic cholinergic activity is altered in animal models exhibiting immunological abnormalities. In addition, it appears likely that during interactions mediated by cell surface molecules T cells communicate via ACh with thymic epithelial cells and vascular endothelial cells, which also express ChAT and nAChRs or mAChRs. This interaction leads to T cell selection and maturation in the thymus and local vascular smooth muscle relaxation. Collectively, these data provide a compelling picture in which lymphocytes constitute a cholinergic system that is independent of cholinergic nerves, and which is involved in the regulation of immune function and local circulation.
ESTHER : Kawashima_2004_Front.Biosci_9_2063
PubMedSearch : Kawashima_2004_Front.Biosci_9_2063
PubMedID: 15353271

Title : Picrotoxin increased acetylcholine release from rat cultured embryonic septal neurons - Suzuki_2004_Neurosci.Lett_356_57
Author(s) : Suzuki T , Takagi R , Kawashima K
Ref : Neuroscience Letters , 356 :57 , 2004
Abstract : GABA is a major inhibitory neurotransmitter in the mature mammalian brain. In the early stages of brain development, it has been reported that GABA(A) receptor stimulation and the associated increase in Cl(-) conductance lead to membrane depolarization. In this study, we tested the effects of picrotoxin, a GABA(A) receptor Cl(-) channel blocker, on spontaneously released acetylcholine (ACh) from cultured rat embryonic septal cells. Picrotoxin increased spontaneously released ACh. These results indicate that blockade of GABA-activated Cl(-) channel increases neuronal excitability even in an early stage of the development.
ESTHER : Suzuki_2004_Neurosci.Lett_356_57
PubMedSearch : Suzuki_2004_Neurosci.Lett_356_57
PubMedID: 14746901

Title : The lymphocytic cholinergic system and its contribution to the regulation of immune activity - Kawashima_2003_Life.Sci_74_675
Author(s) : Kawashima K , Fujii T
Ref : Life Sciences , 74 :675 , 2003
Abstract : Lymphocytes express most of the cholinergic components found in the nervous system, including acetylcholine (ACh), choline acetyltransferase (ChAT), high affinity choline transporter, muscarinic and nicotinic ACh receptors (mAChRs and nAChRs, respectively), and acetylcholinesterase. Stimulation of T and B cells with ACh or another mAChR agonist elicits intracellular Ca2+ signaling, up-regulation of c-fos expression, increased nitric oxide synthesis and IL-2-induced signal transduction, probably via M3 and M5 mAChR-mediated pathways. Acute stimulation of nAChRs with ACh or nicotine causes rapid and transient Ca2+ signaling in T and B cells, probably via alpha7 nAChR subunit-mediated pathways. Chronic nicotine stimulation, by contrast, down-regulates nAChR expression and suppresses T cell activity. Activation of T cells with phytohemagglutinin or antibodies against cell surface molecules enhances lymphocytic cholinergic transmission by activating expression of ChAT and M5 mAChR, which is suggestive of local cholinergic regulation of immune system activity. This idea is supported by the facts that lymphocytic cholinergic activity reflects well the changes in immune system function seen in animal models of immune deficiency and immune acceleration. Collectively, these data provide a compelling picture in which lymphocytes constitute a cholinergic system that is independent of cholinergic nerves, and which is involved in the regulation of immune function.
ESTHER : Kawashima_2003_Life.Sci_74_675
PubMedSearch : Kawashima_2003_Life.Sci_74_675
PubMedID: 14654162

Title : Expression of multiple mRNA species for choline acetyltransferase in human T-lymphocytes - Ogawa_2003_Life.Sci_72_2127
Author(s) : Ogawa H , Fujii T , Watanabe Y , Kawashima K
Ref : Life Sciences , 72 :2127 , 2003
Abstract : Acetylcholine (ACh) is synthesized by choline acetyltransferase (ChAT) in cholinergic neurons. However, both ACh and mRNA for ChAT are expressed in mononuclear leukocytes and various human leukemic T-cell lines. Multiple ChAT mRNA species (R-, N0-, N1-, N2-, and M-types) having an identical coding region and different 5'-noncoding regions have been discovered in human brain and spinal cord. These mRNAs are transcribed by a combination of use of different promoter regions and alternative splicing. However, which types of ChAT mRNA species are expressed in T-lymphocytes remains to be elucidated. In the present study, we used two human leukemic T-cell lines, CCRF-CEM (CEM) and MOLT-3, which express the same ChAT mRNA as that in the nervous system. Major mRNA species in CEM were N2- and M-type, and to a lesser extent N1-type, while MOLT-3 expressed only N2-type. Neither CEM nor MOLT-3 expressed R-type mRNA. We previously found a lack of mRNA expression encoding vesicular acetylcholine transporter (VAChT) in CEM and MOLT-3, which mediates ACh transport to synaptic vesicles in cholinergic neurons. These findings suggest that the mechanisms regulating ChAT mRNA expression in T-lymphocytes differ from those in cholinergic neurons.
ESTHER : Ogawa_2003_Life.Sci_72_2127
PubMedSearch : Ogawa_2003_Life.Sci_72_2127
PubMedID: 12628468

Title : Upregulation of mRNA encoding the M5 muscarinic acetylcholine receptor in human T- and B-lymphocytes during immunological responses - Fujii_2003_Neurochem.Res_28_423
Author(s) : Fujii T , Watanabe Y , Inoue T , Kawashima K
Ref : Neurochem Res , 28 :423 , 2003
Abstract : Lymphocytes possess an independent, non-neuronal cholinergic system. Moreover, both T- and B-lymphocytes express multiple muscarinic acetylcholine receptors (mAChR). To obtain a better understanding of the regulatory mechanisms governing mAChR gene expression in the lymphocytic cholinergic system, we examined the effects of lymphocyte activation on expression of mAChR mRNA. Stimulation of T- and B-lymphocytes, respectively, with T-cell activator phytohemagglutinin and B-cell activator Staphylococcus aureus Cowan I upregulated M5 mAChR mRNA expression in the CEM human leukemic T-cell line and in the Daudi B-cell line, which served as models of lymphocytes. In striking contrast, M3 and M4 mAChR mRNA expression was not affected in either cell line. Nonetheless, stimulating lymphocytes with phorbol 12-myristate 13-acetate, a protein kinase C activator, plus ionomycin, a calcium ionophore, upregulated expression of both M3 and M5 mAChR mRNA. This represents the first demonstration that immunological stimulation leads to M5 mAChR gene expression in lymphocytes.
ESTHER : Fujii_2003_Neurochem.Res_28_423
PubMedSearch : Fujii_2003_Neurochem.Res_28_423
PubMedID: 12675126

Title : The endogenous, immunologically active peptide apelin inhibits lymphocytic cholinergic activity during immunological responses - Horiuchi_2003_J.Neuroimmunol_144_46
Author(s) : Horiuchi Y , Fujii T , Kamimura Y , Kawashima K
Ref : Journal of Neuroimmunology , 144 :46 , 2003
Abstract : We investigated the effects of apelin, an immunologically active peptide ligand for orphan receptor APJ, on acetylcholine (ACh) synthesis in MOLT-3 human leukemic T cells. We initially confirmed expression of APJ mRNA in several human T- and B-cell lines by reverse transcription-polymerase chain reaction (RT-PCR). We also found that in phytohemagglutinin (PHA)-stimulated MOLT-3 cells, an active apelin fragment, apelin-13, down-regulates expression of choline acetyltransferase (ChAT) mRNA and significantly reduces ChAT activity and cellular ACh content and release. It thus appears that apelin inhibits lymphocytic cholinergic activity via APJ during immunological responses.
ESTHER : Horiuchi_2003_J.Neuroimmunol_144_46
PubMedSearch : Horiuchi_2003_J.Neuroimmunol_144_46
PubMedID: 14597097

Title : Urodynamics in a rat neurogenic bladder model with a unilateral electrolytic lesion of the basal forebrain - Shimizu_2003_BJU.Int_91_861
Author(s) : Shimizu I , Kawashima K , Ishii D , Oka M
Ref : BJU Int , 91 :861 , 2003
Abstract : OBJECTIVE: To investigate the changes in bladder function in rats with an electrolytic lesion of the right basal forebrain (RBF) and to determine the effects of AH-9700, a novel sigma receptor ligand, on cystometry in RBF-lesioned rats. MATERIALS AND
METHODS: A lesion was made electrolytically in the RBF of male Wistar rats. At 7 or 8 days after the lesion or sham surgery, continuous cystometry was performed in awake rats. In addition, contractile responses to electrical field stimulation or carbachol were measured in isolated bladder strips, as were the forebrain contents of acetylcholine, monoamine neurotransmitters and their metabolites.
RESULTS: RBF-lesioned rats showed a remarkable increase in voiding frequency, with a decrease in voiding threshold pressure but no change in voiding pressure, compared with sham-operated rats. However, contractile responses in bladder strips isolated from RBF-lesioned rats were no different from those in strips isolated from sham-operated rats. In RBF-lesioned rats, the contents of acetylcholine, dopamine, 4-dihidroxyphenylacetic acid and homovanillic acid were significantly decreased in the right forebrain. AH-9700 dose-dependently decreased the voiding frequency and increased the threshold pressure in RBF-lesioned rats. Anti-muscarinic agents (oxybutynin and propiverine) also decreased the voiding frequency, but their effects were less potent than that of AH-9700.
CONCLUSIONS: The RBF-lesioned rat may be a useful model for the neurogenic bladder of supraspinal origin. Moreover, AH-9700 effectively improves bladder dysfunction in this model.
ESTHER : Shimizu_2003_BJU.Int_91_861
PubMedSearch : Shimizu_2003_BJU.Int_91_861
PubMedID: 12780849

Title : Evolutional study on acetylcholine expression - Horiuchi_2003_Life.Sci_72_1745
Author(s) : Horiuchi Y , Kimura R , Kato N , Fujii T , Seki M , Endo T , Kato T , Kawashima K
Ref : Life Sciences , 72 :1745 , 2003
Abstract : Acetylcholine (ACh) is a well-known neurotransmitter in the cholinergic nervous systems of vertebrates and insects; however, there is only indirect evidence for its presence in lower invertebrates, such as plants and fungi. We therefore investigated the expression of ACh in invertebrates (sea squirt, sea urchin, trepang, squid, abalone, nereis, sea anemone, coral and sponge), plants (arabidopsis, eggplant, bamboo shoot, cedar, hinoki, pine, podcarp, fern, horsetail and moss), fungi (yeast and mushroom) and bacteria by assaying ACh content and synthesis, focusing on the presence of two synthetic enzymes, choline acetyltransferase (ChAT) and carnitine acetyltransferase (CarAT). Using a specific radioimmunoassay, ACh was detected in all samples tested. The levels varied considerably, however, with the upper portion of bamboo shoots having the highest content (2.9 micromol/g). ACh synthesis was also detected in all samples tested; moreover, the activity in most samples from the animal kingdom, as well as bamboo shoots and the stem of the shiitake mushroom, were sensitive to both ChAT and CarAT inhibitors. Levels of ACh synthesis were lower in samples from other plants, fungi and bacteria and were insensitive to ChAT and CarAT inhibitors. These findings demonstrate the presence of ACh and ACh-synthesizing activity in evolutionally primitive life as well as in more complex multicellular organisms. In the context of the recent discovery of non-neuronal ACh in various mammalian species, these findings suggest that ACh been expressed in organisms from the beginning of life, functioning as a local mediator as well as a neurotransmitter.
ESTHER : Horiuchi_2003_Life.Sci_72_1745
PubMedSearch : Horiuchi_2003_Life.Sci_72_1745
PubMedID: 12559395

Title : Complete genome structure of Gloeobacter violaceus PCC 7421, a cyanobacterium that lacks thylakoids - Nakamura_2003_DNA.Res_10_137
Author(s) : Nakamura Y , Kaneko T , Sato S , Mimuro M , Miyashita H , Tsuchiya T , Sasamoto S , Watanabe A , Kawashima K , Kishida Y , Kiyokawa C , Kohara M , Matsumoto M , Matsuno A , Nakazaki N , Shimpo S , Takeuchi C , Yamada M , Tabata S
Ref : DNA Research , 10 :137 , 2003
Abstract : The nucleotide sequence of the entire genome of a cyanobacterium Gloeobacter violaceus PCC 7421 was determined. The genome of G. violaceus was a single circular chromosome 4,659,019 bp long with an average GC content of 62%. No plasmid was detected. The chromosome comprises 4430 potential protein-encoding genes, one set of rRNA genes, 45 tRNA genes representing 44 tRNA species and genes for tmRNA, B subunit of RNase P, SRP RNA and 6Sa RNA. Forty-one percent of the potential protein-encoding genes showed sequence similarity to genes of known function, 37% to hypothetical genes, and the remaining 22% had no apparent similarity to reported genes. Comparison of the assigned gene components with those of other cyanobacteria has unveiled distinctive features of the G. violaceus genome. Genes for PsaI, PsaJ, PsaK, and PsaX for Photosystem I and PsbY, PsbZ and Psb27 for Photosystem II were missing, and those for PsaF, PsbO, PsbU, and PsbV were poorly conserved. cpcG for a rod core linker peptide for phycobilisomes and nblA related to the degradation of phycobilisomes were also missing. Potential signal peptides of the presumptive products of petJ and petE for soluble electron transfer catalysts were less conserved than the remaining portions. These observations may be related to the fact that photosynthesis in G. violaceus takes place not in thylakoid membranes but in the cytoplasmic membrane. A large number of genes for sigma factors and transcription factors in the LuxR, LysR, PadR, TetR, and MarR families could be identified, while those for major elements for circadian clock, kaiABC were not found. These differences may reflect the phylogenetic distance between G. violaceus and other cyanobacteria.
ESTHER : Nakamura_2003_DNA.Res_10_137
PubMedSearch : Nakamura_2003_DNA.Res_10_137
PubMedID: 14621292
Gene_locus related to this paper: glovi-GLL0053 , glovi-GLL0778 , glovi-GLL1217 , glovi-GLL1281 , glovi-GLL1310 , glovi-GLL1435 , glovi-GLL2002 , glovi-gll2009 , glovi-GLL2335 , glovi-GLL2500 , glovi-GLL3208 , glovi-GLL3677 , glovi-GLL4259 , glovi-GLR0796 , glovi-GLR1368 , glovi-GLR1422 , glovi-GLR2241 , glovi-GLR2809 , glovi-GLR3058 , glovi-GLR3329 , glovi-GLR3546 , glovi-GLR3833 , glovi-q7ncx6 , glovi-q7nd10 , glovi-q7ndi8 , glovi-q7ndy7 , glovi-q7nek8 , glovi-q7net1 , glovi-q7new7 , glovi-q7ney7 , glovi-q7nfx3 , glovi-q7nga2 , glovi-q7ngw1 , glovi-q7nj78 , glovi-q7nj91 , glovi-q7nj98 , glovi-q7njx8 , glovi-q7njz2 , glovi-q7nkk7 , glovi-q7nly3 , glovi-q7nm82 , glovi-q7nmt4 , glovi-q7nmz0 , glovi-q7nn33 , glovi-q7nn46 , glovi-q7nn64 , glovi-q7nny4 , glovi-q7np81 , glovi-q7npc6

Title : Complete genome structure of Gloeobacter violaceus PCC 7421, a cyanobacterium that lacks thylakoids (supplement) -
Author(s) : Nakamura Y , Kaneko T , Sato S , Mimuro M , Miyashita H , Tsuchiya T , Sasamoto S , Watanabe A , Kawashima K , Kishida Y , Kiyokawa C , Kohara M , Matsumoto M , Matsuno A , Nakazaki N , Shimpo S , Takeuchi C , Yamada M , Tabata S
Ref : DNA Research , 10 :181 , 2003
PubMedID: 14621296
Gene_locus related to this paper: glovi-gll2009

Title : Detection of the high-affinity choline transporter in the MOLT-3 human leukemic T-cell line - Fujii_2003_Life.Sci_72_2131
Author(s) : Fujii T , Okuda T , Haga T , Kawashima K
Ref : Life Sciences , 72 :2131 , 2003
Abstract : We previously showed that lymphocytes possess the necessary components to constitute an independent, non-neuronal cholinergic system; these include acetylcholine (ACh) itself, choline acetyltransferase (the ACh-synthesizing enzyme), and both muscarinic and nicotinic ACh receptors (AChRs). In addition, we showed that stimulation of AChRs with their respective agonists elicits a variety of biochemical and functional effects, suggesting that lymphocytic cholinergic system is involved in the regulation of immune function. In nerve terminals, choline taken up via the high-affinity choline transporter (CHT1) is exclusively utilized for ACh synthesis. In the present study, therefore, we investigated the expression of CHT1 in T-lymphocytes. Reverse transcription-polymerase chain reaction analysis revealed that MOLT-3 cells, a human leukemic T-cell line used as a T-lymphocyte model, expressed CHT1 mRNA, but that the CEM and Jurkat T-cell lines did not. Consistent with that finding, specific binding of [3H]hemicholinium-3 (HC-3), an inhibitor of CHT1, and HC-3-sensitive [3H]choline uptake were also detected in MOLT-3 cells. These results suggest that CHT1 plays a role in mediating choline uptake in T-lymphocytes and provides further evidence for the presence of an independent lymphocytic cholinergic system.
ESTHER : Fujii_2003_Life.Sci_72_2131
PubMedSearch : Fujii_2003_Life.Sci_72_2131
PubMedID: 12628469

Title : Endogenous glutamatergic synaptic activity elicits acetylcholine release from rat cultured septal cells - Suzuki_2003_Neurosci.Res_47_341
Author(s) : Suzuki T , Matsugi T , Takagi R , Kawashima K
Ref : Neurosci Res , 47 :341 , 2003
Abstract : We tested the characteristics of acetylcholine (ACh) release from cultured rat septal cells. The spontaneous release was inhibited by treatment with tetrodotoxin (TTX) and omega-conotoxin (GVIA), indicating that the release was elicited by synaptic activity. The release was also inhibited by 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), an alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) receptor blocker, in both the absence and presence of nerve growth factor (NGF), suggesting that endogenously released glutamate produced the ACh release by stimulating AMPA receptors. This is the first report of detection of the release of ACh by endogenous spontaneous synaptic activity conducted by glutamate AMPA receptor activation in cultured septal cells. This in vitro experimental system is useful for the study of cholinergic functions.
ESTHER : Suzuki_2003_Neurosci.Res_47_341
PubMedSearch : Suzuki_2003_Neurosci.Res_47_341
PubMedID: 14568116

Title : Introduction: the non-neuronal cholinergic system in humans -
Author(s) : Grando SA , Kawashima K , Wessler I
Ref : Life Sciences , 72 :2009 , 2003
PubMedID: 12628450

Title : Acetylcholine increase in amniotic fluid of experimental rats for intrauterine growth retardation - Horikoshi_2003_Life.Sci_72_2145
Author(s) : Horikoshi T , Fujii T , Kawashima K , Sakuragawa N
Ref : Life Sciences , 72 :2145 , 2003
Abstract : Previous reports from this laboratory have demonstrated evidence for synthesis and release of acetylcholine (ACh) and catecholamines (CAs) by human amniotic epithelial cells (HAEC) and the presence of ACh and CAs in amniotic fluid. To study the physiological role of amniotic ACh, we used an experimental pregnant rat model for intrauterine growth retardation. Prior to this experiment, we confirmed the presence of choline acetyltransferase in the HAEC by immunocytochemical staining. Amniotic fluid was collected at 48 and 72 h after a transient ligation of the uterine vessels near the lower and upper ends of the right horn of the pregnant rat. The ACh concentration in the amniotic fluid from rats received intrauterine ischemia increased with time to a greater degree compared with the control rat, although the increase was not statistically significant. These results suggest that intrauterine hypoxic conditions cause a tendency to increase ACh concentrations in the amniotic fluid.
ESTHER : Horikoshi_2003_Life.Sci_72_2145
PubMedSearch : Horikoshi_2003_Life.Sci_72_2145
PubMedID: 12628471

Title : Nitric oxide (NO) synthase mRNA expression and NO production via muscarinic acetylcholine receptor-mediated pathways in the CEM, human leukemic T-cell line - Kamimura_2003_Life.Sci_72_2151
Author(s) : Kamimura Y , Fujii T , Kojima H , Nagano T , Kawashima K
Ref : Life Sciences , 72 :2151 , 2003
Abstract : Nitric oxide (NO) is synthesized from L-arginine by neuronal, endothelial and inducible isoforms of NO synthase (nNOS, eNOS and iNOS, respectively) and is involved in the regulation of a variety of physiological functions, including immune activity. In vascular endothelial cells, stimulation of M(3) subtype of muscarinic acetylcholine receptors (mAChRs) triggers NO synthesis by eNOS. Human lymphocytes express several mAChR subtypes and their stimulation increases the intracellular free Ca(2+) concentration and up-regulates c-fos gene expression. While the above findings suggest involvement of the lymphocytic cholinergic system in the regulation of immune function, little is known on NOS expression and NO synthesis in T-lymphocytes. In the present study, using reverse transcription-polymerase chain reaction, we found that CEM cells express mRNAs encoding iNOS and nNOS, but not for eNOS. In addition, using quantitative fluorescence microscopy and a novel NO-sensitive fluorescent indicator, DAF-2, we found that oxotremorine-M (Oxo-M) (100 microM), a non-selective mAChR agonist, enhances NO production in the cells. This effect of Oxo-M was antagonized by pirenzepine (10 microM), an antagonist acting preferentially at M(1) mAChR and by atropine (10 microM). Also 4-DAMP (10 microM), an antagonist acting preferentially at M(3) mAChR, reduced significantly the effect of Oxo-M, while AFDX-116 (10 microM), an antagonist acting preferentially at M(2) mAChR, was ineffective. These findings suggest that T-lymphocytes express functional mAChRs linked to NO synthesis by nNOS and/or iNOS.
ESTHER : Kamimura_2003_Life.Sci_72_2151
PubMedSearch : Kamimura_2003_Life.Sci_72_2151
PubMedID: 12628472

Title : The lymphocytic cholinergic system and its biological function - Kawashima_2003_Life.Sci_72_2101
Author(s) : Kawashima K , Fujii T
Ref : Life Sciences , 72 :2101 , 2003
Abstract : Lymphocytes are now known to possess the essential components for a non-neuronal cholinergic system. These include acetylcholine (ACh); choline acetyltransferase (ChAT), its synthesizing enzyme; and both muscarinic and nicotinic ACh receptors (mAChRs and nAChRs, respectively). Stimulating lymphocytes with phytohemagglutinin, a T-cell activator; Staphylococcus aureus Cowan I, a B-cell activator; or cell surface molecules enhances the synthesis and release of ACh and up-regulates expression of ChAT and M(5) mAChR mRNAs. Activation of mAChRs and nAChRs on lymphocytes elicits increases in the intracellular Ca(2+) concentration and stimulates c-fos gene expression and nitric oxide synthesis. On the other hand, long-term exposure to nicotine down-regulates expression of nAChR mRNA. Abnormalities in the lymphocytic cholinergic system have been detected in spontaneously hypertensive rats and MRL-lpr mice, two animal models of immune disorders. Taken together, these data present a compelling picture in which immune function is, at least in part, under the control of an independent non-neuronal lymphocytic cholinergic system.
ESTHER : Kawashima_2003_Life.Sci_72_2101
PubMedSearch : Kawashima_2003_Life.Sci_72_2101
PubMedID: 12628464

Title : Nicotine-induced Ca2+ signaling and down-regulation of nicotinic acetylcholine receptor subunit expression in the CEM human leukemic T-cell line - Kimura_2003_Life.Sci_72_2155
Author(s) : Kimura R , Ushiyama N , Fujii T , Kawashima K
Ref : Life Sciences , 72 :2155 , 2003
Abstract : We previously showed that T- and B-lymphocytes express both muscarinic and nicotinic acetylcholine (ACh) receptors (mAChR and nAChR, respectively), and that stimulation of M(3) mAChRs on lymphocytes increases the intracellular free Ca(2+) concentration ([Ca(2+)](i)) and up-regulates c-fos gene expression. Little is known about the effects of nicotinic stimulation on lymphocyte function, however. We therefore investigated the acute effect of nicotine on [Ca(2+)](i) in CEM cells, a model of T-lymphocytes, using confocal laser scanning microscopy with fluo-3, a Ca(2+)-sensitive fluorescent indicator. In addition, we examined the long-term effect of nicotine on the expression of selected nAChR subunits using semiquantitative reverse transcription-polymerase chain reaction analysis. In the presence of extracellular Ca(2+), nicotine (30 microM) evoked rapid, transient increases in [Ca(2+)](i). This effect was concentration-dependently inhibited by the alpha7 nAChR subunit antagonists, alpha-bungarotoxin (0.01-10 microM) and methyllycaconitine (0.01-10 mM), suggesting that the alpha7 nAChR subunit mediates Ca(2+) signaling in T-lymphocytes. Nicotine (0.01-10 microM) also concentration-dependently down-regulated expression of mRNAs for all the nAChR subunits tested: expression of the alpha6 and alpha7 subunits was down-regulated within 1 week, while expression of the alpha3 and alpha5 subunits declined gradually throughout the 8-week experimental period. These findings indicate that nicotine--and therefore likely smoking--affects immune function by suppressing expression of the neuronal nAChR subtype involved in Ca(2+) signaling in lymphocytes.
ESTHER : Kimura_2003_Life.Sci_72_2155
PubMedSearch : Kimura_2003_Life.Sci_72_2155
PubMedID: 12628473

Title : Complete genome structure of the thermophilic cyanobacterium Thermosynechococcus elongatus BP-1 - Nakamura_2002_DNA.Res_9_123
Author(s) : Nakamura Y , Kaneko T , Sato S , Ikeuchi M , Katoh H , Sasamoto S , Watanabe A , Iriguchi M , Kawashima K , Kimura T , Kishida Y , Kiyokawa C , Kohara M , Matsumoto M , Matsuno A , Nakazaki N , Shimpo S , Sugimoto M , Takeuchi C , Yamada M , Tabata S
Ref : DNA Research , 9 :123 , 2002
Abstract : The entire genome of a thermophilic unicellular cyanobacterium, Thermosynechococcus elongatus BP-1, was sequenced. The genome consisted of a circular chromosome 2,593,857 bp long, and no plasmid was detected. A total of 2475 potential protein-encoding genes, one set of rRNA genes, 42 tRNA genes representing 42 tRNA species and 4 genes for small structural RNAs were assigned to the chromosome by similarity search and computer prediction. The translated products of 56% of the potential protein-encoding genes showed sequence similarity to experimentally identified and predicted proteins of known function, and the products of 34% of these genes showed sequence similarity to the translated products of hypothetical genes. The remaining 10% lacked significant similarity to genes for predicted proteins in the public DNA databases. Sixty-three percent of the T. elongatus genes showed significant sequence similarity to those of both Synechocystis sp. PCC 6803 and Anabaena sp. PCC 7120, while 22% of the genes were unique to this species, indicating a high degree of divergence of the gene information among cyanobacterial strains. The lack of genes for typical fatty acid desaturases and the presence of more genes for heat-shock proteins in comparison with other mesophilic cyanobacteria may be genomic features of thermophilic strains. A remarkable feature of the genome is the presence of 28 copies of group II introns, 8 of which contained a presumptive gene for maturase/reverse transcriptase. A trace of genome rearrangement mediated by the group II introns was also observed.
ESTHER : Nakamura_2002_DNA.Res_9_123
PubMedSearch : Nakamura_2002_DNA.Res_9_123
PubMedID: 12240834
Gene_locus related to this paper: theeb-q8dg48 , theeb-TLL0292 , theeb-TLL0340 , theeb-TLL0918 , theeb-TLL0989 , theeb-TLL1717 , theeb-TLL2163 , theeb-TLR0492 , theeb-TLR1045 , theeb-TLR1157 , theeb-TLR1274 , theeb-TLR1393 , theeb-TLR1423 , theeb-TLR1685 , theeb-TLR1725 , theeb-TLR1892 , theeb-TLR1982 , theeb-TLR2038 , theeb-TLR2066

Title : Effects of human antithymocyte globulin on acetylcholine synthesis, its release and choline acetyltransferase transcription in a human leukemic T-cell line - Fujii_2002_J.Neuroimmunol_128_1
Author(s) : Fujii T , Ushiyama N , Hosonuma K , Suenaga A , Kawashima K
Ref : Journal of Neuroimmunology , 128 :1 , 2002
Abstract : Lymphocytes possess an independent, nonneuronal cholinergic system. In the present study, we investigated the short- and long-term effects of antithymocyte globulin (ATG)-Fresenius (ATG-F), a human antithymocyte globulin that binds to CD2, CD7 and CD11a, on acetylcholine (ACh) synthesis and transcription of choline acetyltransferase (ChAT) in CCRF-CEM cells, a human leukemic T-cell line. In the short-term (6 h), ATG-F enhanced ACh release, likely through transient increases in intracellular Ca(2+) ([Ca(2+)](i)) mediated by CD7, which led to declines in intracellular ACh content. By 48 h, however, the ACh content had increased as compared to control due to up-regulation of ChAT expression mediated by CD11a.
ESTHER : Fujii_2002_J.Neuroimmunol_128_1
PubMedSearch : Fujii_2002_J.Neuroimmunol_128_1
PubMedID: 12098504

Title : Complete genomic sequence of nitrogen-fixing symbiotic bacterium Bradyrhizobium japonicum USDA110 - Kaneko_2002_DNA.Res_9_189
Author(s) : Kaneko T , Nakamura Y , Sato S , Minamisawa K , Uchiumi T , Sasamoto S , Watanabe A , Idesawa K , Iriguchi M , Kawashima K , Kohara M , Matsumoto M , Shimpo S , Tsuruoka H , Wada T , Yamada M , Tabata S
Ref : DNA Research , 9 :189 , 2002
Abstract : The complete nucleotide sequence of the genome of a symbiotic bacterium Bradyrhizobium japonicum USDA110 was determined. The genome of B. japonicum was a single circular chromosome 9,105,828 bp in length with an average GC content of 64.1%. No plasmid was detected. The chromosome comprises 8317 potential protein-coding genes, one set of rRNA genes and 50 tRNA genes. Fifty-two percent of the potential protein genes showed sequence similarity to genes of known function and 30% to hypothetical genes. The remaining 18% had no apparent similarity to reported genes. Thirty-four percent of the B. japonicum genes showed significant sequence similarity to those of both Mesorhizobium loti and Sinorhizobium meliloti, while 23% were unique to this species. A presumptive symbiosis island 681 kb in length, which includes a 410-kb symbiotic region previously reported by Gottfert et al., was identified. Six hundred fifty-five putative protein-coding genes were assigned in this region, and the functions of 301 genes, including those related to symbiotic nitrogen fixation and DNA transmission, were deduced. A total of 167 genes for transposases/104 copies of insertion sequences were identified in the genome. It was remarkable that 100 out of 167 transposase genes are located in the presumptive symbiotic island. DNA segments of 4 to 97 kb inserted into tRNA genes were found at 14 locations in the genome, which generates partial duplication of the target tRNA genes. These observations suggest plasticity of the B. japonicum genome, which is probably due to complex genome rearrangements such as horizontal transfer and insertion of various DNA elements, and to homologous recombination.
ESTHER : Kaneko_2002_DNA.Res_9_189
PubMedSearch : Kaneko_2002_DNA.Res_9_189
PubMedID: 12597275
Gene_locus related to this paper: braja-BLL0118 , braja-BLL0272 , braja-BLL0546 , braja-BLL0558 , braja-BLL0837 , braja-BLL0839 , braja-BLL1016 , braja-BLL1128 , braja-BLL1234 , braja-BLL1350 , braja-BLL1401 , braja-BLL2323 , braja-BLL2387 , braja-BLL2443 , braja-BLL2527 , braja-BLL2884 , braja-BLL2902 , braja-BLL3246 , braja-BLL3359 , braja-BLL3416 , braja-BLL3418 , braja-BLL3470 , braja-BLL3759 , braja-BLL3777 , braja-BLL4001 , braja-BLL4189 , braja-BLL4284 , braja-BLL4335 , braja-BLL4360 , braja-BLL4548 , braja-BLL4985 , braja-BLL4989 , braja-BLL4997 , braja-BLL5160 , braja-BLL5514 , braja-BLL5588 , braja-BLL5740 , braja-bll6073 , braja-BLL6264 , braja-BLL6275 , braja-BLL6428 , braja-bll6463 , braja-BLL6574 , braja-BLL6577 , braja-BLL6614 , braja-bll6820 , braja-BLL6841 , braja-BLL6890 , braja-BLL6974 , braja-BLL7123 , braja-BLL7368 , braja-BLL7370 , braja-BLL7497 , braja-BLL7506 , braja-BLL7509 , braja-BLL7545 , braja-BLL7692 , braja-BLL7862 , braja-BLL8011 , braja-BLL8277 , braja-BLR0230 , braja-BLR0418 , braja-BLR0711 , braja-BLR0899 , braja-BLR0908 , braja-BLR1078 , braja-BLR1197 , braja-BLR1251 , braja-BLR2261 , braja-BLR2321 , braja-BLR2487 , braja-BLR2879 , braja-BLR2885 , braja-BLR2889 , braja-BLR2982 , braja-BLR3322 , braja-BLR3456 , braja-BLR3519 , braja-blr3732 , braja-BLR3878 , braja-BLR4157 , braja-BLR4181 , braja-BLR5346 , braja-BLR5359 , braja-BLR6083 , braja-BLR6127 , braja-BLR6186 , braja-BLR6271 , braja-BLR6465 , braja-BLR6576 , braja-BLR6677 , braja-BLR6703 , braja-BLR6960 , braja-BLR6965 , braja-BLR7068 , braja-BLR7144 , braja-BLR7443 , braja-BLR7556 , braja-BLR7622 , braja-BLR7741 , braja-BLR7809 , braja-BLR7810 , braja-BLR7894 , braja-BLR8188 , braja-dhaa , braja-EPHA , braja-EPHB , braja-ID587 , braja-ID930 , braja-METX , braja-pcaD , braja-PIP , braja-PLDB , braja-PTRB , braja-q89c18 , braja-q89c36 , braja-q89mj3 , braja-q89ql9 , braja-q89y23

Title : Complete genomic sequence of nitrogen-fixing symbiotic bacterium Bradyrhizobium japonicum USDA110 (supplement) -
Author(s) : Kaneko T , Nakamura Y , Sato S , Minamisawa K , Uchiumi T , Sasamoto S , Watanabe A , Idesawa K , Iriguchi M , Kawashima K , Kohara M , Matsumoto M , Shimpo S , Tsuruoka H , Wada T , Yamada M , Tabata S
Ref : DNA Research , 9 :225 , 2002
PubMedID: 12597279
Gene_locus related to this paper: braja-bll6463 , braja-bll6820 , braja-EPHB , braja-ID930 , braja-pcaD

Title : An independent non-neuronal cholinergic system in lymphocytes - Fujii_2001_Jpn.J.Pharmacol_85_11
Author(s) : Fujii T , Kawashima K
Ref : Japanese Journal of Pharmacology , 85 :11 , 2001
Abstract : Acetylcholine (ACh) is a well characterized neurotransmitter occurring throughout the animal kingdom. In addition, both muscarinic and nicotinic ACh receptors have been identified on lymphocytes of various origin, and their stimulation by muscarinic or nicotinic agonists elicits a variety of functional and biochemical effects. It was thus initially postulated that the parasympathetic nervous system may play a role in modulating immune system function. However, ACh in the blood has now been localized to lymphocytes; indeed expression of choline acetyltransferase (ChAT), an ACh synthesizing enzyme, has been shown in human blood mononuclear leukocytes, human leukemic T-cell lines and rat lymphocytes. Stimulation of T-lymphocytes with phytohemagglutinin activates the lymphoid cholinergic system, as evidenced by increased synthesis and release of ACh and increased expression of mRNAs encoding ChAT and ACh receptors. The observation that M3 muscarinic receptor stimulation by ACh and other agonists increases the intracellular free Ca2+ concentration and upregulates c-fos gene expression strongly argues that ACh, synthesized and released from T-lymphocytes, acts as an autocrine and/or paracrine factor regulating immune function. These findings present a compelling picture in which immune function is, at least in part, under the control of an independent lymphoid cholinergic system.
ESTHER : Fujii_2001_Jpn.J.Pharmacol_85_11
PubMedSearch : Fujii_2001_Jpn.J.Pharmacol_85_11
PubMedID: 11243565

Title : Non-neuronal neurotransmitters and neurotrophic factors in amniotic epithelial cells: expression and function in humans and monkey - Sakuragawa_2001_Jpn.J.Pharmacol_85_20
Author(s) : Sakuragawa N , Elwan MA , Uchida S , Fujii T , Kawashima K
Ref : Japanese Journal of Pharmacology , 85 :20 , 2001
Abstract : Human amniotic epithelial cells (HAEC) are formed from epiblasts on the 8th day after fertilization. Because they lack major histocompatibility complex (MHC) antigen, human amniotic tissue transplantation has been used for allotranplantation to treat patients with lysosomal diseases. We have provided evidence that HAEC have multiple functions such as synthesis and release of acetylcholine (ACh) and catecholamine (CA) as well as expressing mRNA coding for dopamine receptors and dopamine (DA) transporter (DAT). On the other hand, we showed that monkey amniotic epithelial cells (MAEC) synthesize and release CA and posses DA receptors and DAT. Detection of muscarinic actylcholine receptors indicates the presence of an autocrine mechanism in HAEC. Recently, we found that HAEC have neurotrophic function in conditioned medium from HAEC, indicating the presence of a novel neurotrohpic factor that is synthesized and released from HAEC. The amniotic membrane may have a significant role in supplying neurotrophic factors as well as neurotransmitters to the amniotic fluid, suggesting an important function in the early stages of neural development of the embryo. This review will focus on the neuropharmacological aspects of HAEC and MAEC in relation to the physiology of amniotic membrane.
ESTHER : Sakuragawa_2001_Jpn.J.Pharmacol_85_20
PubMedSearch : Sakuragawa_2001_Jpn.J.Pharmacol_85_20
PubMedID: 11243569

Title : Decreased acetylcholine content and choline acetyltransferase mRNA expression in circulating mononuclear leukocytes and lymphoid organs of the spontaneously hypertensive rat - Fujimoto_2001_Life.Sci_69_1629
Author(s) : Fujimoto K , Matsui M , Fujii T , Kawashima K
Ref : Life Sciences , 69 :1629 , 2001
Abstract : It has been confirmed that the neurotransmitter acetylcholine (ACh) is present in blood; it is synthesized in T-lymphocytes by choline acetyltransferase (ChAT) and released upon T-lymphocyte activation. Both muscarinic and nicotinic ACh receptors have been identified on lymphocytes isolated from thymus, lymph node, spleen and blood, and their stimulation by muscarinic and nicotinic agonists elicits a variety of functional and biochemical effects, providing a strong argument that ACh synthesized and released from T-lymphocytes acts as an autocrine and/or paracrine factor regulating immune function. In the present study, we compared ACh levels in the blood, circulating mononuclear leukocytes (MNLs), thymus and spleen of spontaneously hypertensive rats (SHRs), which exhibit immune deficiencies related to the emergence of natural thymocytotoxic autoantibody, age-related decline of T-cell function and morphological changes in immune organs, with ACh levels in age-matched, normotensive Wistar Kyoto rats. In each case, ACh levels in 5-, 10- and 20-week-old SHRs were significantly lower than in WKYs. ChAT mRNA expression in MNLs was also significantly depressed in the SHRs. These results suggest that diminished synthesis and release of ACh from MNLs into blood and lymphoid organs likely reflects an immune deficiency related to T-cell dysfunction.
ESTHER : Fujimoto_2001_Life.Sci_69_1629
PubMedSearch : Fujimoto_2001_Life.Sci_69_1629
PubMedID: 11589503

Title : Functional expression of alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptors precedes the development of cholinergic phenotype in embryonic rat septal cells in culture - Suzuki_2001_Neurosci.Lett_311_89
Author(s) : Suzuki T , Matsugi T , Takagi R , Kanagawa M , Hirata M , Nakamura T , Kudo Y , Kawashima K
Ref : Neuroscience Letters , 311 :89 , 2001
Abstract : We examined the development of cholinergic neuronal functions and alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) responses in cultured embryonic rat septal cells. Choline acetyltransferase activity was increased from 4 to 6 days in culture and reached a plateau at day 8. Acetylcholine release was increased from 6 to 8 days in culture. AMPA-induced increase in intracellular Ca(2+) level was observed at 3 days in culture and most of the AMPA-responsive cells coincided with high-K(+) responsive cells. These results suggest that cholinergic neurons develop their neuronal functions about 8 days under cultured conditions, and functional expression of AMPA receptors precedes the cholinergic functional development.
ESTHER : Suzuki_2001_Neurosci.Lett_311_89
PubMedSearch : Suzuki_2001_Neurosci.Lett_311_89
PubMedID: 11567785

Title : Glucocorticoid treatment increases inhibitory m(2) muscarinic receptor expression and function in the airways - Jacoby_2001_Am.J.Respir.Cell.Mol.Biol_24_485
Author(s) : Jacoby DB , Yost BL , Kumaravel B , Chan-Li Y , Xiao HQ , Kawashima K , Fryer AD
Ref : American Journal of Respiratory Cellular & Molecular Biology , 24 :485 , 2001
Abstract : M(2) muscarinic receptors on parasympathetic nerve endings inhibit acetylcholine release in the airways. In this study, the effects of dexamethasone on M(2) receptors in vivo and in primary cultures of airway parasympathetic neurons were tested. Treating guinea pigs with dexamethasone (0.1 mg/kg, daily for 2 d) substantially increased inhibitory M(2) muscarinic receptor function, decreasing airway responsiveness to electrical stimulation of the vagi. At the same time, dexamethasone decreased the response to acetylcholine but not to methacholine, suggesting that cholinesterase activity was increased. When both cholinesterase and M(2) receptors were blocked (using physostigmine and gallamine, respectively) vagally induced bronchoconstriction was increased to control values. In primary cultures of airway parasympathetic neurons, dexamethasone significantly decreased the release of acetylcholine in response to electrical stimulation. Blocking inhibitory M(2) receptors using atropine (10(-5) M) increased acetylcholine release. After the M(2) receptors were blocked there was no difference in acetylcholine release between control and dexamethasone-treated cultures. M(2) receptor gene expression was increased by more than fivefold in dexamethasone-treated cultures. Immunostaining of dexamethasone-treated neurons demonstrated more intense staining. Thus, decreased vagally mediated reflex bronchoconstriction after glucocorticoid treatment may be the result on increased M(2) receptor expression and function as well as increased degradation of acetylcholine by cholinesterase.
ESTHER : Jacoby_2001_Am.J.Respir.Cell.Mol.Biol_24_485
PubMedSearch : Jacoby_2001_Am.J.Respir.Cell.Mol.Biol_24_485
PubMedID: 11306443

Title : Complete genomic sequence of the filamentous nitrogen-fixing cyanobacterium Anabaena sp. strain PCC 7120 - Kaneko_2001_DNA.Res_8_205
Author(s) : Kaneko T , Nakamura Y , Wolk CP , Kuritz T , Sasamoto S , Watanabe A , Iriguchi M , Ishikawa A , Kawashima K , Kimura T , Kishida Y , Kohara M , Matsumoto M , Matsuno A , Muraki A , Nakazaki N , Shimpo S , Sugimoto M , Takazawa M , Yamada M , Yasuda M , Tabata S
Ref : DNA Research , 8 :205 , 2001
Abstract : The nucleotide sequence of the entire genome of a filamentous cyanobacterium, Anabaena sp. strain PCC 7120, was determined. The genome of Anabaena consisted of a single chromosome (6,413,771 bp) and six plasmids, designated pCC7120alpha (408,101 bp), pCC7120beta (186,614 bp), pCC7120gamma (101,965 bp), pCC7120delta (55,414 bp), pCC7120epsilon (40,340 bp), and pCC7120zeta (5,584 bp). The chromosome bears 5368 potential protein-encoding genes, four sets of rRNA genes, 48 tRNA genes representing 42 tRNA species, and 4 genes for small structural RNAs. The predicted products of 45% of the potential protein-encoding genes showed sequence similarity to known and predicted proteins of known function, and 27% to translated products of hypothetical genes. The remaining 28% lacked significant similarity to genes for known and predicted proteins in the public DNA databases. More than 60 genes involved in various processes of heterocyst formation and nitrogen fixation were assigned to the chromosome based on their similarity to the reported genes. One hundred and ninety-five genes coding for components of two-component signal transduction systems, nearly 2.5 times as many as those in Synechocystis sp. PCC 6803, were identified on the chromosome. Only 37% of the Anabaena genes showed significant sequence similarity to those of Synechocystis, indicating a high degree of divergence of the gene information between the two cyanobacterial strains.
ESTHER : Kaneko_2001_DNA.Res_8_205
PubMedSearch : Kaneko_2001_DNA.Res_8_205
PubMedID: 11759840
Gene_locus related to this paper: anasp-ALL0111 , anasp-ALL0193 , anasp-ALL0254 , anasp-ALL0316 , anasp-ALL0955 , anasp-ALL0969 , anasp-ALL1161 , anasp-ALL1205 , anasp-ALL1353 , anasp-ALL1695 , anasp-ALL2050 , anasp-ALL2056 , anasp-ALL2058 , anasp-ALL2068 , anasp-ALL2529 , anasp-ALL2533 , anasp-ALL2753 , anasp-ALL2761 , anasp-ALL3898 , anasp-ALL4221 , anasp-ALL4875 , anasp-ALL8511 , anasp-ALR0039 , anasp-ALR0079 , anasp-ALR0130 , anasp-ALR0235 , anasp-ALR0851 , anasp-ALR1077 , anasp-ALR1270 , anasp-ALR1352 , anasp-ALR1362 , anasp-ALR1709 , anasp-ALR2045 , noss1-ALR3140 , anasp-ALR3514 , anasp-ALR3685 , anasp-ALR3911 , anasp-ALR4625 , anasp-ALR5028 , anasp-AROE , anasp-q8ymv5 , anasp-q8yxx2 , anasp-y1448 , noss1-ALL3113 , noss1-ALL4967 , noss1-ALR4786 , noss1-q8yrg0 , noss1-y2406

Title : Extraneuronal cholinergic system in lymphocytes - Kawashima_2000_Pharmacol.Ther_86_29
Author(s) : Kawashima K , Fujii T
Ref : Pharmacol Ther , 86 :29 , 2000
Abstract : Acetylcholine (ACh) is well known as a neurotransmitter in both the central and peripheral nervous systems in mammalian species. Both muscarinic and nicotinic ACh receptors have been identified in lymphocytes isolated from thymus, lymph node, spleen, and peripheral blood, and their stimulation by muscarinic and nicotinic agonists elicits a variety of functional and biochemical effects. On the basis of these findings, it has been postulated that the parasympathetic nervous system may play a role in immune-neurohumoral crosstalk. However, ACh present in the blood of several species has been localized to lymphocytes from various origins using radioimmunoassay. Moreover, using Northern blots or reverse transcription-polymerase chain reaction, expression of choline acetyltransferase, an ACh synthesizing enzyme, has been identified in human blood mononuclear leukocytes, human leukemic T-cell lines, and rat lymphocytes. Stimulation of T-lymphocytes with phytohemagglutinin activates the lymphoid cholinergic system, as evidenced by increased synthesis and release of ACh, increased acetylcholinesterase activity, and the increased expression of mRNA encoding choline acetyltransferase and ACh receptors. The observation that muscarinic receptor stimulation by ACh or agonists increases in [Ca(2)+](i) and up-regulates c-fos expression strongly argues that ACh synthesized and released from T-lymphocytes acts as an autocrine and/or paracrine factor regulating immune function. In summary, these data present a compelling picture in which immune function is not only regulated by the cytokine system, but is also under the control of an independent, lymphoid cholinergic system.
ESTHER : Kawashima_2000_Pharmacol.Ther_86_29
PubMedSearch : Kawashima_2000_Pharmacol.Ther_86_29
PubMedID: 10760545

Title : Ca2+ oscillation and c-fos gene expression induced via muscarinic acetylcholine receptor in human T- and B-cell lines - Fujii_2000_Naunyn.Schmiedebergs.Arch.Pharmacol_362_14
Author(s) : Fujii T , Kawashima K
Ref : Naunyn Schmiedebergs Arch Pharmacol , 362 :14 , 2000
Abstract : We previously reported that blood acetylcholine (ACh) mainly originates from T-lymphocytes and that muscarinic (Ms) ACh receptor mRNA is expressed in both T- and B-lymphocytes. In the present study, we used confocal laser scanning microscopy and fluo-3, a calcium-sensitive indicator, to investigate the effects of Ms-ACh receptor agonists on the intracellular free Ca2+ concentration ([Ca2+]i) in single cells from human T-cell (CEM) and B-cell (Daudi) lines, which we used as models of lymphocytes. In both cell lines, stimulation of Ms-ACh receptors with ACh (0.1-100 microM), bethanechol (100 microM), car-bachol (100 microM) or oxotremorine-M (Oxo-M; 0.1-100 microM) induced [Ca2+]i-dependent increases in fluo-3 fluorescence, which in the presence of extracellular Ca2+ were followed by oscillations in [Ca2+]i that persisted for at least 10 min. All effects were completely blocked by atropine (1 microM), an Ms-ACh receptor antagonist. In both cell lines Oxo-M (100 microM) up-regulated expression of c-fos mRNA in an extracellular Ca2+-dependent manner. Again, the effect was blocked by 1 microM atropine. These results provide the first evidence that stimulation of Ms-ACh receptors induces Ca2+ oscillations and up-regulates c-fos gene expression in T- and B-lymphocytes, which is consistent with the notion that ACh released from T-lymphocytes triggers nuclear signaling via Ms-ACh receptors.
ESTHER : Fujii_2000_Naunyn.Schmiedebergs.Arch.Pharmacol_362_14
PubMedSearch : Fujii_2000_Naunyn.Schmiedebergs.Arch.Pharmacol_362_14
PubMedID: 10935528

Title : Sequence and analysis of chromosome 5 of the plant Arabidopsis thaliana - Tabata_2000_Nature_408_823
Author(s) : Tabata S , Kaneko T , Nakamura Y , Kotani H , Kato T , Asamizu E , Miyajima N , Sasamoto S , Kimura T , Hosouchi T , Kawashima K , Kohara M , Matsumoto M , Matsuno A , Muraki A , Nakayama S , Nakazaki N , Naruo K , Okumura S , Shinpo S , Takeuchi C , Wada T , Watanabe A , Yamada M , Yasuda M , Sato S , de la Bastide M , Huang E , Spiegel L , Gnoj L , O'Shaughnessy A , Preston R , Habermann K , Murray J , Johnson D , Rohlfing T , Nelson J , Stoneking T , Pepin K , Spieth J , Sekhon M , Armstrong J , Becker M , Belter E , Cordum H , Cordes M , Courtney L , Courtney W , Dante M , Du H , Edwards J , Fryman J , Haakensen B , Lamar E , Latreille P , Leonard S , Meyer R , Mulvaney E , Ozersky P , Riley A , Strowmatt C , Wagner-McPherson C , Wollam A , Yoakum M , Bell M , Dedhia N , Parnell L , Shah R , Rodriguez M , See LH , Vil D , Baker J , Kirchoff K , Toth K , King L , Bahret A , Miller B , Marra M , Martienssen R , McCombie WR , Wilson RK , Murphy G , Bancroft I , Volckaert G , Wambutt R , Dusterhoft A , Stiekema W , Pohl T , Entian KD , Terryn N , Hartley N , Bent E , Johnson S , Langham SA , McCullagh B , Robben J , Grymonprez B , Zimmermann W , Ramsperger U , Wedler H , Balke K , Wedler E , Peters S , van Staveren M , Dirkse W , Mooijman P , Lankhorst RK , Weitzenegger T , Bothe G , Rose M , Hauf J , Berneiser S , Hempel S , Feldpausch M , Lamberth S , Villarroel R , Gielen J , Ardiles W , Bents O , Lemcke K , Kolesov G , Mayer K , Rudd S , Schoof H , Schueller C , Zaccaria P , Mewes HW , Bevan M , Fransz P
Ref : Nature , 408 :823 , 2000
Abstract : The genome of the model plant Arabidopsis thaliana has been sequenced by an international collaboration, The Arabidopsis Genome Initiative. Here we report the complete sequence of chromosome 5. This chromosome is 26 megabases long; it is the second largest Arabidopsis chromosome and represents 21% of the sequenced regions of the genome. The sequence of chromosomes 2 and 4 have been reported previously and that of chromosomes 1 and 3, together with an analysis of the complete genome sequence, are reported in this issue. Analysis of the sequence of chromosome 5 yields further insights into centromere structure and the sequence determinants of heterochromatin condensation. The 5,874 genes encoded on chromosome 5 reveal several new functions in plants, and the patterns of gene organization provide insights into the mechanisms and extent of genome evolution in plants.
ESTHER : Tabata_2000_Nature_408_823
PubMedSearch : Tabata_2000_Nature_408_823
PubMedID: 11130714
Gene_locus related to this paper: arath-At5g11650 , arath-At5g16120 , arath-at5g18630 , arath-AT5G20520 , arath-At5g21950 , arath-AT5G27320 , arath-CXE15 , arath-F1N13.220 , arath-F14F8.240 , arath-q3e9e4 , arath-q8lae9 , arath-Q8LFB7 , arath-q9ffg7 , arath-q9fij5 , arath-Q9LVU7 , arath-q66gm8 , arath-SCPL34 , arath-B9DFR3 , arath-a0a1p8bcz0

Title : Calcium signaling and c-Fos gene expression via M3 muscarinic acetylcholine receptors in human T- and B-cells - Fujii_2000_Jpn.J.Pharmacol_84_124
Author(s) : Fujii T , Kawashima K
Ref : Japanese Journal of Pharmacology , 84 :124 , 2000
Abstract : We previously showed that blood acetylcholine (ACh) originates mainly from T-lymphocytes, and that stimulation of muscarinic ACh receptors (mAChRs) induces Ca2+ oscillations and up-regulates c-fos gene expression in both T- and B-lymphocytes. In the present study, we investigated which mAChR subtypes are involved in Ca2+ signaling and c-fos gene expression in human T- (CEM) and B- (Daudi) cells. Stimulation of mAChRs with 100 microM oxotremorine-M, an M1/M3 agonist, increased levels of intracellular free Ca2+ ([Ca2+]i) and c-fos mRNA expression in both cell lines. 4-DAMP, an M3 antagonist, more effectively blocked the oxotremorine-M-induced increase in [Ca2+]i than pirenzepine and telenzepine, M1-receptor antagonists; AF-DX 116, an M2 antagonist; hexahydrosiladifenidol, a weak M3 antagonist; or hexamethonium and d-tubocurarine, nicotinic receptor antagonists. McN-A-343 (100 microM), a partial M1-receptor agonist, had no apparent effect on [Ca2+]i in either cell line. The oxotremorine-M-induced up-regulation of c-fos transcription was inhibited by 4-DAMP, but not by pirenzepine or AF-DX 116. Our findings thus suggest that ACh released from T-lymphocytes acts as an autocrine/paracrine factor, transmitting a Ca2+-dependent signal to the nuclei of T- and B-lymphocytes via M3 receptors.
ESTHER : Fujii_2000_Jpn.J.Pharmacol_84_124
PubMedSearch : Fujii_2000_Jpn.J.Pharmacol_84_124
PubMedID: 11128034

Title : YM905, a novel M3 antagonist, inhibits Ca2+ signaling and c-fos gene expression mediated via muscarinic receptors in human T cells - Fujii_2000_Gen.Pharmacol_35_71
Author(s) : Fujii T , Kawashima K
Ref : General Pharmacology , 35 :71 , 2000
Abstract : Our earlier observations suggest that M3 muscarinic acetylcholine (ACh) receptors (mAChRs) are involved in Ca2+ signaling and regulation of c-fos gene expression in T lymphocytes. Here, we describe the effects of YM905, a novel M3 antagonist, on evoked Ca2+ signaling and c-fos gene expression in CEM human leukemic T cells. YM905 significantly inhibited increases in intracellular free Ca2+ evoked by 10 microM oxotremorine-M, an M1/M3 agonist (IC50=100 nM), and also inhibited 10 microM oxotremorine-M-induced upregulation of c-fos gene expression at 1 microM. These findings demonstrate that YM905 antagonizes the intracellular responses in T cells induced via mAChRs, possibly M3 receptors.
ESTHER : Fujii_2000_Gen.Pharmacol_35_71
PubMedSearch : Fujii_2000_Gen.Pharmacol_35_71
PubMedID: 11707312

Title : Enhancement of cerebral cortical acetylcholine release by intraperitoneal acetic acid and its suppression by analgesics in freely moving rats - Harada_2000_Neurosci.Lett_284_163
Author(s) : Harada H , Hosonuma K , Fujii T , Kawashima K
Ref : Neuroscience Letters , 284 :163 , 2000
Abstract : Several lines of evidence suggest that central cholinergic neurons play a key role in the perception and control of pain. We investigated the effects of analgesics on the increase in central cholinergic activity and writhing responses elicited by i.p. injection of acetic acid. ACh efflux from the rat cerebral cortex and hippocampus was measured in the absence of a cholinesterase inhibitor using an in vivo microdialysis technique and a highly sensitive and specific radioimmunoassay. ACh efflux from the cerebral cortex was significantly increased during the first 30 min after acetic acid injection and then returned to the control levels. In contrast, acetic acid-induced writhing responses, indicative of the perception of pain, persisted for almost the entire 120 min observation period. No changes in ACh efflux were observed in the hippocampus. The centrally-acting analgesic morphine and the peripherally-acting analgesic indomethacin each completely abolished the enhanced cerebral cortical ACh efflux and the writhing, whereas diazepam, a muscle relaxant, selectively suppressed only the writhing. These results demonstrate that peripheral nociceptive stimulation transiently increases cholinergic activity in the cerebral cortex, but not in the hippocampus, and that analgesics suppress both the enhanced ACh efflux and the writhing induced by acetic acid.
ESTHER : Harada_2000_Neurosci.Lett_284_163
PubMedSearch : Harada_2000_Neurosci.Lett_284_163
PubMedID: 10773424

Title : Effects of physostigmine and calcium on acetylcholine efflux from the hippocampus of freely moving rats as determined by in vivo microdialysis and a radioimmunoassay - Fujii_2000_Neurosci.Lett_289_181
Author(s) : Fujii T , Harada H , Koyama T , Nakajima Y , Kawashima K
Ref : Neuroscience Letters , 289 :181 , 2000
Abstract : The effects varying the concentration of Ca2+ in perfused artificial cerebrospinal fluid ([Ca2+]csf) on basal acetylcholine (ACh) efflux from the hippocampus of freely moving rats, in the presence and absence of the cholinesterase (ChE) inhibitor physostigmine, were investigated using in vivo microdialysis and a highly specific radioimmunoassay for ACh. In the absence of physostigmine, basal ACh efflux was 3.4+/-0.7 pg/30 min (mean +/- SEM) at [Ca2+]csf = 1.26 mM. Stepwise increases in [Ca2+]csf elicited a gradual increase in ACh efflux that was significant at [Ca2+]csf = 5.04 mM. Inhibition of ChE by addition of 10 microM physostigmine to the perfusate increased the efflux of ACh to 103.2+/-21.1 pg/30 min ([Ca2+]csf = 1.26 mM), and the efflux was augmented still further by increasing [Ca2+]csf, a change that became significant at [Ca2+]csf = 3.78. These results illustrate the sensitivity of basal ACh efflux from the hippocampus to changes in the extracellular Ca2+ concentration, and suggest that a more accurate picture of hippocampal cholinergic activity is obtained by microdialysis using normal artificial cerebrospinal fluid, under physiological conditions, rather than in the presence of a ChE inhibitor.
ESTHER : Fujii_2000_Neurosci.Lett_289_181
PubMedSearch : Fujii_2000_Neurosci.Lett_289_181
PubMedID: 10961659

Title : Enhancement of hippocampal cholinergic neurotransmission through 5-HT1A receptor-mediated pathways by repeated lithium treatment in rats - Fujii_2000_Can.J.Physiol.Pharmacol_78_392
Author(s) : Fujii T , Nakai K , Nakajima Y , Kawashima K
Ref : Canadian Journal of Physiology & Pharmacology , 78 :392 , 2000
Abstract : Hippocampal cholinergic neuronal activity is reported to be regulated, at least partly, through serotonin1A (5-HT1A) receptors. Chronic lithium treatment has been shown to alter both behavioral and neurochemical responses mediated by postsynaptic 5-HT1A receptors. We investigated whether long-term lithium treatment affects central cholinergic neurotransmission through 5-HT1A receptor-mediated pathways. Changes in acetylcholine (ACh) release induced by 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT), a 5-HT1A receptor agonist, in the rat hippocampus were measured using a microdialysis technique and a radioimmunoassay for ACh. Administration of lithium for 21 days resulted in a serum lithium concentration of 1.03 mM and caused little change in density or affinity of [3H]8-OH-DPAT binding sites in the hippocampus. The local application of 8-OH-DPAT into the hippocampus of lithium treated rats increased the ACh efflux in both the absence and the presence of physostigmine, a cholinesterase (ChE) inhibitor, in the perfusion fluid. The basal ACh efflux of lithium treated rats was not different from that of the control rats under normal conditions, but was significantly higher than that of the controls when ChE was inhibited. These results demonstrate that chronic lithium treatment increases spontaneous ACh release in the hippocampus under conditions of ChE inhibition, but not under normal conditions, and enhances cholinergic neurotransmission through 5-HT1A receptor-mediated pathways, and suggest that activation of 5-HT1A receptor function by lithium is related to the enhancement of hippocampal cholinergic neurotransmission.
ESTHER : Fujii_2000_Can.J.Physiol.Pharmacol_78_392
PubMedSearch : Fujii_2000_Can.J.Physiol.Pharmacol_78_392
PubMedID: 10841434

Title : Sequence and analysis of chromosome 3 of the plant Arabidopsis thaliana - Salanoubat_2000_Nature_408_820
Author(s) : Salanoubat M , Lemcke K , Rieger M , Ansorge W , Unseld M , Fartmann B , Valle G , Blocker H , Perez-Alonso M , Obermaier B , Delseny M , Boutry M , Grivell LA , Mache R , Puigdomenech P , de Simone V , Choisne N , Artiguenave F , Robert C , Brottier P , Wincker P , Cattolico L , Weissenbach J , Saurin W , Quetier F , Schafer M , Muller-Auer S , Gabel C , Fuchs M , Benes V , Wurmbach E , Drzonek H , Erfle H , Jordan N , Bangert S , Wiedelmann R , Kranz H , Voss H , Holland R , Brandt P , Nyakatura G , Vezzi A , D'Angelo M , Pallavicini A , Toppo S , Simionati B , Conrad A , Hornischer K , Kauer G , Lohnert TH , Nordsiek G , Reichelt J , Scharfe M , Schon O , Bargues M , Terol J , Climent J , Navarro P , Collado C , Perez-Perez A , Ottenwalder B , Duchemin D , Cooke R , Laudie M , Berger-Llauro C , Purnelle B , Masuy D , de Haan M , Maarse AC , Alcaraz JP , Cottet A , Casacuberta E , Monfort A , Argiriou A , Flores M , Liguori R , Vitale D , Mannhaupt G , Haase D , Schoof H , Rudd S , Zaccaria P , Mewes HW , Mayer KF , Kaul S , Town CD , Koo HL , Tallon LJ , Jenkins J , Rooney T , Rizzo M , Walts A , Utterback T , Fujii CY , Shea TP , Creasy TH , Haas B , Maiti R , Wu D , Peterson J , Van Aken S , Pai G , Militscher J , Sellers P , Gill JE , Feldblyum TV , Preuss D , Lin X , Nierman WC , Salzberg SL , White O , Venter JC , Fraser CM , Kaneko T , Nakamura Y , Sato S , Kato T , Asamizu E , Sasamoto S , Kimura T , Idesawa K , Kawashima K , Kishida Y , Kiyokawa C , Kohara M , Matsumoto M , Matsuno A , Muraki A , Nakayama S , Nakazaki N , Shinpo S , Takeuchi C , Wada T , Watanabe A , Yamada M , Yasuda M , Tabata S
Ref : Nature , 408 :820 , 2000
Abstract : Arabidopsis thaliana is an important model system for plant biologists. In 1996 an international collaboration (the Arabidopsis Genome Initiative) was formed to sequence the whole genome of Arabidopsis and in 1999 the sequence of the first two chromosomes was reported. The sequence of the last three chromosomes and an analysis of the whole genome are reported in this issue. Here we present the sequence of chromosome 3, organized into four sequence segments (contigs). The two largest (13.5 and 9.2 Mb) correspond to the top (long) and the bottom (short) arms of chromosome 3, and the two small contigs are located in the genetically defined centromere. This chromosome encodes 5,220 of the roughly 25,500 predicted protein-coding genes in the genome. About 20% of the predicted proteins have significant homology to proteins in eukaryotic genomes for which the complete sequence is available, pointing to important conserved cellular functions among eukaryotes.
ESTHER : Salanoubat_2000_Nature_408_820
PubMedSearch : Salanoubat_2000_Nature_408_820
PubMedID: 11130713
Gene_locus related to this paper: arath-MES17 , arath-AT3G12150 , arath-At3g61680 , arath-AT3g62590 , arath-CXE12 , arath-eds1 , arath-SCP25 , arath-F1P2.110 , arath-F1P2.140 , arath-F11F8.28 , arath-F14D17.80 , arath-F16B3.4 , arath-SCP27 , arath-At3g50790 , arath-At3g05600 , arath-PAD4 , arath-At3g51000 , arath-SCP16 , arath-gid1 , arath-GID1B , arath-Q9LUG8 , arath-Q84JS1 , arath-Q9SFF6 , arath-q9m236 , arath-q9sr22 , arath-q9sr23 , arath-SCP7 , arath-SCP14 , arath-SCP15 , arath-SCP17 , arath-SCP36 , arath-SCP37 , arath-SCP39 , arath-SCP40 , arath-SCP49 , arath-T19F11.2

Title : Complete genome structure of the nitrogen-fixing symbiotic bacterium Mesorhizobium loti - Kaneko_2000_DNA.Res_7_331
Author(s) : Kaneko T , Nakamura Y , Sato S , Asamizu E , Kato T , Sasamoto S , Watanabe A , Idesawa K , Ishikawa A , Kawashima K , Kimura T , Kishida Y , Kiyokawa C , Kohara M , Matsumoto M , Matsuno A , Mochizuki Y , Nakayama S , Nakazaki N , Shimpo S , Sugimoto M , Takeuchi C , Yamada M , Tabata S
Ref : DNA Research , 7 :331 , 2000
Abstract : The complete nucleotide sequence of the genome of a symbiotic bacterium Mesorhizobium loti strain MAFF303099 was determined. The genome of M. loti consisted of a single chromosome (7,036,071 bp) and two plasmids, designated as pMLa (351,911 bp) and pMLb (208, 315 bp). The chromosome comprises 6752 potential protein-coding genes, two sets of rRNA genes and 50 tRNA genes representing 47 tRNA species. Fifty-four percent of the potential protein genes showed sequence similarity to genes of known function, 21% to hypothetical genes, and the remaining 25% had no apparent similarity to reported genes. A 611-kb DNA segment, a highly probable candidate of a symbiotic island, was identified, and 30 genes for nitrogen fixation and 24 genes for nodulation were assigned in this region. Codon usage analysis suggested that the symbiotic island as well as the plasmids originated and were transmitted from other genetic systems. The genomes of two plasmids, pMLa and pMLb, contained 320 and 209 potential protein-coding genes, respectively, for a variety of biological functions. These include genes for the ABC-transporter system, phosphate assimilation, two-component system, DNA replication and conjugation, but only one gene for nodulation was identified.
ESTHER : Kaneko_2000_DNA.Res_7_331
PubMedSearch : Kaneko_2000_DNA.Res_7_331
PubMedID: 11214968
Gene_locus related to this paper: meslo-acoc , meslo-EphB , meslo-est , meslo-lipest , meslo-MLL0014 , meslo-MLL0351 , meslo-MLL0537 , meslo-mll0601 , meslo-MLL0618 , meslo-MLL1209 , meslo-MLL1226 , meslo-mll1328 , meslo-MLL1329 , meslo-MLL1495 , meslo-MLL1869 , meslo-mll1900 , meslo-MLL2018 , meslo-MLL2072 , meslo-mll2481 , meslo-mll2689 , meslo-MLL2788 , meslo-MLL3556 , meslo-MLL3568 , meslo-MLL3682 , meslo-mll3776 , meslo-MLL4497 , meslo-MLL4552 , meslo-MLL5128 , meslo-mll5179 , meslo-mll5392 , meslo-MLL5717 , meslo-mll5743 , meslo-MLL6746 , meslo-MLL6752 , meslo-mll6871 , meslo-MLL7643 , meslo-mll7742 , meslo-MLL9722 , meslo-MLR0094 , meslo-mlr0145 , meslo-mlr0170 , meslo-MLR0240 , meslo-mlr0493 , meslo-MLR0937 , meslo-mlr0978 , meslo-MLR0992 , meslo-MLR1612 , meslo-mlr1789 , meslo-mlr1864 , meslo-mlr2176 , meslo-MLR2262 , meslo-mlr2612 , meslo-mlr2710 , meslo-mlr3034 , meslo-MLR3538 , meslo-mlr3816 , meslo-mlr4436 , meslo-MLR4903 , meslo-MLR5045 , meslo-MLR5063 , rhilo-dhaa , meslo-MLR6087 , meslo-MLR6657 , meslo-mlr6682 , meslo-mlr6683 , meslo-MLR6684 , meslo-MLR6787 , meslo-MLR6993 , meslo-mlr6999 , meslo-mlr7206 , meslo-mlr7232 , meslo-mlr7803 , meslo-MLR9053 , meslo-mlr9622 , meslo-mlr9641 , rhilo-MLL0076 , rhilo-MLL1824 , rhilo-MLL7123 , rhilo-MLL8374 , rhilo-MLR1247 , rhilo-MLR2444 , rhilo-MLR4383 , rhilo-MLR8175 , rhilo-q98nf6 , rhilo-q98nf8 , rhilo-q988i9

Title : Nerve growth factor increases the synthesis and release of acetylcholine and the expression of vesicular acetylcholine transporter in primary cultured rat embryonic septal cells - Oosawa_1999_J.Neurosci.Res_57_381
Author(s) : Oosawa H , Fujii T , Kawashima K
Ref : Journal of Neuroscience Research , 57 :381 , 1999
Abstract : Acetylcholine (ACh) is synthesized by choline acetyltransferase (ChAT) in the cytoplasm of cholinergic nerve terminals and transported into synaptic vesicles by vesicular ACh transporter (VAChT). The genes encoding ChAT and VAChT are colocalized within the genome, and their products are known to be coregulated by various neurotrophic factors. In the present study, nerve growth factor (NGF; 100 ng/ml) was shown to enhance expression of VAChT and ChAT mRNA in primary cultured rat embryonic septal cells. By using a radioimmunoassay, we also found that NGF increased both neuronal content and spontaneous release of ACh, which were first detected on day 2 of culture and time-dependently increased up to day 10. Stimulated release of ACh elicited by high K+ (50 mM KCl) was also significantly greater in NGF-treated cells than in control cells. NGF enhanced immunoreactivity to antiserum against VAChT, indicating that the augmented responses were due to, at least in part, increased expression of VAChT protein. In contrast, the numbers of immunocytochemically positive cells were unaffected. Thus, NGF appears to augment ACh synthesis, its transport into synaptic vesicles, and its subsequent release. The data also suggest that NGF facilitates growth and development of cholinergic neurons, but not their survival.
ESTHER : Oosawa_1999_J.Neurosci.Res_57_381
PubMedSearch : Oosawa_1999_J.Neurosci.Res_57_381
PubMedID: 10412029

Title : Toluene inhalation induced epididymal sperm dysfunction in rats - Ono_1999_Toxicology_139_193
Author(s) : Ono A , Kawashima K , Sekita K , Hirose A , Ogawa Y , Saito M , Naito K , Yasuhara K , Kaneko T , Furuya T , Inoue T , Kurokawa Y
Ref : Toxicology , 139 :193 , 1999
Abstract : Toluene is a widely abused inhaled solvent. This study was designed to determine whether toluene abuse affects the reproductive functions or general health of males. Seven-week-old male Sprague-Dawley rats were exposed to toluene vapor inhalation (0, 4000, or 6000 ppm; 2 h/day) daily for 5 weeks. Exposure-related suppression of body weight gain and food consumption were observed. Salivation and lacrimation were observed during exposure periods and intensified with repeated exposure. Rats exposed to 6000 ppm toluene had decreased spleen and thymus weights, as well as suppressed lymphocyte counts. In 6000 ppm group, the epididymal sperm counts, sperm motility, sperm quality and in vitro penetrating ability to zona-free hamster eggs were significantly reduced, while no exposure-related changes in the testes weight or spermatogenesis within testes were detected. Tail-less sperm heads were seen within zona-free eggs incubated with sperm from rats exposed to 6000 ppm toluene, but not control rats. No significant changes were observed in serum luteinizing hormone, follicle-stimulating hormone, or testosterone levels following 1 month of exposure to 6000 ppm toluene. These results indicate that high concentrations of toluene may directly target sperm in the epididymis and disrupt sperm maturation.
ESTHER : Ono_1999_Toxicology_139_193
PubMedSearch : Ono_1999_Toxicology_139_193
PubMedID: 10647920

Title : A possible role for acetylcholine in the dialogue between thymocytes and thymic stroma - Rinner_1999_Neuroimmunomodulation_6_51
Author(s) : Rinner I , Globerson A , Kawashima K , Korsatko W , Schauenstein K
Ref : Neuroimmunomodulation , 6 :51 , 1999
Abstract : In this article we will review data suggesting that acetylcholine takes part in the mutual interplay between developing T cells and thymic epithelium, and thereby may influence the generation of the T-cell repertoire. In the first part we will recapitulate our findings according to which cholinergic agonists affect thymocyte apoptosis via a nicotinergic effect on thymic epithelial cells. In the second part we will present evidence that acetylcholine within the thymus is mainly derived from the thymocytes themselves, and that the production and release of this neurotransmitter is dependent on activation of thymic lymphocytes.
ESTHER : Rinner_1999_Neuroimmunomodulation_6_51
PubMedSearch : Rinner_1999_Neuroimmunomodulation_6_51
PubMedID: 9876235

Title : Enhancement of the serotonin-mediated acetylcholine release by repeated desmethylimipramine treatment in the hippocampus of freely moving rats - Fujii_1999_Jpn.J.Pharmacol_80_303
Author(s) : Fujii T , Ohba S , Nakai K , Fujimoto K , Suzuki T , Kawashima K
Ref : Japanese Journal of Pharmacology , 80 :303 , 1999
Abstract : A possible involvement of serotonin-mediated cholinergic activation in the antidepressant effect of desmethylimipramine (DMI) was investigated by determination of the effects of a single or repeated DMI administration on acetylcholine (ACh) release in the hippocampus using an in vivo microdialysis technique and a radioimmunoassay for ACh. Rats were administered DMI (10 mg/kg, i.p.) acutely or repeatedly for 21 days. A single or repeated DMI administration did not cause any significant effects on the basal ACh release compared with the respective controls. Atropine perfusion in the acutely DMI-treated or control rats increased the ACh release to the same degree. In repeatedly DMI-treated rats, serotonin (5-HT) (1 to 10 microM) perfusion enhanced significantly the ACh release. However, 5-HT in acutely DMI-treated rats enhanced significantly the ACh release only at 10 microM. 5-HT did not cause any changes in ACh release in control rats. Hippocampal 5-HT content of acutely DMI-treated rats was significantly higher than that of saline-treated control rats, while no difference was observed between the repeatedly DMI- and saline-treated rats. These findings suggest, for the first time, that DMI induced a facilitation of cholinergic neurotransmission in the rat hippocampus through the activation of 5-HT-receptor function.
ESTHER : Fujii_1999_Jpn.J.Pharmacol_80_303
PubMedSearch : Fujii_1999_Jpn.J.Pharmacol_80_303
PubMedID: 10496330

Title : Constitutive expression of mRNA for the same choline acetyltransferase as that in the nervous system, an acetylcholine-synthesizing enzyme, in human leukemic T-cell lines - Fujii_1999_Neurosci.Lett_259_71
Author(s) : Fujii T , Tajima S , Yamada S , Watanabe Y , Sato KZ , Matsui M , Misawa H , Kasahara T , Kawashima K
Ref : Neuroscience Letters , 259 :71 , 1999
Abstract : Both muscarinic and nicotinic acetylcholine (ACh) receptors are known to be present on the surface of lymphocytes. We have shown that variable amounts of ACh are detectable in the blood of various mammals including humans, and a major portion of blood ACh is localized in circulating mononuclear leukocytes in humans. In order to investigate which types of blood cell are the source of ACh in human blood, expression of mRNA for choline acetyltransferase (ChAT, EC 2.3.1.6), which catalyzes ACh synthesis, was analyzed using human leukemic cell lines as models of lymphocytes and the reverse transcription-polymerase chain reaction (RT-PCR) method. We observed that mRNA for the same ChAT as that in the nervous system is expressed constitutively in all the T-cell lines tested, but not in B-, pre-lymphoma or monocytic cell lines. Furthermore, only T-cell lines showed high ACh-synthesizing activities and intracellular ACh contents. These results suggest that the major portion of ACh in the circulating blood originates from T-lymphocytes.
ESTHER : Fujii_1999_Neurosci.Lett_259_71
PubMedSearch : Fujii_1999_Neurosci.Lett_259_71
PubMedID: 10025560

Title : Possible dynamic neurotransmitter metabolism surrounding the fetus - Sakuragawa_1999_J.Child.Neurol_14_265
Author(s) : Sakuragawa N , Elwan MA , Fujii T , Kawashima K
Ref : Journal of Child Neurology , 14 :265 , 1999
Abstract : Human amniotic epithelial cells express makers of glial and neural stem cells. These cells also synthesize and release acetylcholine and catecholamines. This study of amniotic fluid demonstrated that acetylcholine and catecholamines can be readily identified in the fluid. Norepinephrine was the major catecholamine present, although dopamine and DOPAC could also be detected. The physiologic role of these amniotic fluid neurotransmitters in fetal-placental interactions and nervous system development is currently under investigation.
ESTHER : Sakuragawa_1999_J.Child.Neurol_14_265
PubMedSearch : Sakuragawa_1999_J.Child.Neurol_14_265
PubMedID: 10334403

Title : Diversity of mRNA expression for muscarinic acetylcholine receptor subtypes and neuronal nicotinic acetylcholine receptor subunits in human mononuclear leukocytes and leukemic cell lines - Sato_1999_Neurosci.Lett_266_17
Author(s) : Sato KZ , Fujii T , Watanabe Y , Yamada S , Ando T , Kazuko F , Kawashima K
Ref : Neuroscience Letters , 266 :17 , 1999
Abstract : Previously, we reported that various levels of acetylcholine (ACh), currently known as a neurotransmitter, are detectable in the blood of several mammals including humans and that most blood ACh originates from T-lymphocytes. To investigate whether ACh in the blood acts on lymphocytes and participates in the modulation of immune responses, we have analyzed the expression of mRNA for muscarinic (Ms) ACh receptor subtypes and nicotinic (Nc) ACh receptor subunits using reverse transcription-polymerase chain reaction (RT-PCR) methods. The cells tested were human peripheral mononuclear leukocytes (MNLs) from seven healthy donors and eight leukemic cell lines, as models of lymphocytes. We detected mRNA expression for various neuronal Nc receptor subunits and Ms receptor subtypes in all of the MNL samples and in all of the cell lines tested. However, the expression pattern of mRNA for neuronal Nc receptor subunits (alpha2-alpha7 and beta2-beta4) and Ms receptor subtypes (m1-m5) varied among the individuals and cell lines. No expression of mRNA for three muscle-type Nc receptor subunits (alpha1, beta1 and epsilon) was observed in the MNLs and cell lines. These results indicate that both neuronal-type Nc and Ms ACh receptors are present on the surface of lymphocytes.
ESTHER : Sato_1999_Neurosci.Lett_266_17
PubMedSearch : Sato_1999_Neurosci.Lett_266_17
PubMedID: 10336173

Title : Enhancement of cortical and hippocampal cholinergic neurotransmission through 5-HT1A receptor-mediated pathways by BAY x 3702 in freely moving rats - Koyama_1999_Neurosci.Lett_265_33
Author(s) : Koyama T , Nakajima Y , Fujii T , Kawashima K
Ref : Neuroscience Letters , 265 :33 , 1999
Abstract : Involvement of serotonin (5-HT) in the regulation of cholinergic neuronal activity by modulation of acetylcholine (ACh) release has been reported for various regions of the brain. Cortical and hippocampal cholinergic neurotransmission is of particular importance in the mechanisms of attention as well as learning and memory. In the present study, we investigated the effect of R-(-)-2-[4-[(chroman-2-yl-methyl)-amino-butyl]-1,1-dioxo-benzo[d]++ +isothiazolone hydrochloride (BAY x 3702), a novel, high-affinity 5-HT1A receptor agonist, on ACh release in the cerebral cortex and hippocampus of freely moving rats using an in vivo microdialysis technique. Acetylcholine efflux from the cortex and hippocampus was measured every 30 m using a sensitive and specific radioimmunoassay and was stable for at least 5 h. The ACh efflux from the cortex and hippocampus was increased significantly by BAY x 3702 (0.3 mg/kg, i.p.) compared with saline administration. WAY-100635 (0.6 mg/kg, s.c.), a selective 5-HT1A receptor antagonist, eliminated the augmentation of ACh efflux induced by BAY x 3702 in both brain regions. These results demonstrate that stimulation by BAY x 3702 enhanced ACh release in both the cortex and hippocampus through 5-HT1A receptor-mediated pathways.
ESTHER : Koyama_1999_Neurosci.Lett_265_33
PubMedSearch : Koyama_1999_Neurosci.Lett_265_33
PubMedID: 10327199

Title : Effect of WAY-100135 on the hippocampal acetylcholine release potentiated by 8-OH-DPAT, a serotonin1A receptor agonist, in normal and p-chlorophenylalanine-treated rats as measured by in vivo microdialysis - Nakai_1998_Neurosci.Res_31_23
Author(s) : Nakai K , Fujii T , Fujimoto K , Suzuki T , Kawashima K
Ref : Neurosci Res , 31 :23 , 1998
Abstract : The mechanisms involved in the enhancement of acetylcholine (ACh) release in the rat hippocampus by 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT), a serotonin (5-HT)1A receptor agonist, were investigated using in vivo microdialysis. Administration of p-chlorophenylalanine (PCPA, 300 mg/kg, i.p.), a tryptophan hydroxylase inhibitor, 3 days before the dialysis experiments reduced the hippocampal 5-HT content to 30% of that in saline-treated rats, but did not affect basal ACh release in the hippocampus. 8-OH-DPAT administered systemically (0.5 mg/kg, s.c.) or applied locally (30 microM) into the hippocampus through the dialysis probe significantly enhanced the release of ACh in the hippocampus of PCPA-treated rats to the same degree as that in saline-treated rats. Pretreatment with (+)WAY-100135 (5 mg/kg, i.p.), a selective 5-HT1A receptor antagonist, completely eliminated the enhancement of ACh release induced by locally applied 8-OH-DPAT, but only partially reduced the effects induced by systemically administered 8-OH-DPAT, in both groups of rats. Systemically administered 8-OH-DPAT induced hyperlocomotion in the both saline- and PCPA-treated rats, but this was not eliminated by (+)WAY-100135. 8-OH-DPAT applied locally into the hippocampus did not elicit hyperlocomotion in either group of rats. These results suggest that the modification of endogenous 5-HT release via the 5-HT1A autoreceptor is not involved in the 8-OH-DPAT-induced increase of hippocampal ACh release, and that the increase of ACh release induced by locally applied 8-OH-DPAT involves mainly hippocampal postsynaptic 5-HT1A receptor stimulation. In addition, a possibility that subtypes of 5-HT receptors other than the 5-HT1A receptor, probably 5-HT7 receptor in the septum as well as postsynaptic 5-HT1A receptor in the hippocampus, are involved in the increased hippocampal ACh release induced by systemically administered 8-OH-DPAT is discussed.
ESTHER : Nakai_1998_Neurosci.Res_31_23
PubMedSearch : Nakai_1998_Neurosci.Res_31_23
PubMedID: 9704975

Title : Rat lymphocytes produce and secrete acetylcholine in dependence of differentiation and activation - Rinner_1998_J.Neuroimmunol_81_31
Author(s) : Rinner I , Kawashima K , Schauenstein K
Ref : Journal of Neuroimmunology , 81 :31 , 1998
Abstract : Previous data from this laboratory suggested for the first time that immune cells of the immune system of different species are capable to synthesize the neurotransmitter acetylcholine. In the present study we detected the RNA message for choline acetyltransferase in thymic, splenic and peripheral blood lymphocytes of rats using RT-PCR. Furthermore, using a sensitive radioimmunoassay, we measured acetylcholine in thymic, splenic and peripheral blood lymphocytes. T-cells were found to contain about three times the amount of acetylcholine as compared to B-cells, and CD4+ cells showed significantly higher levels as compared to CD8+ cells. Mitogenic stimulation with PHA increased the acetylcholine levels in lymphoid cells as well as the release into the supernatants.
ESTHER : Rinner_1998_J.Neuroimmunol_81_31
PubMedSearch : Rinner_1998_J.Neuroimmunol_81_31
PubMedID: 9521603

Title : Induction of choline acetyltransferase mRNA in human mononuclear leukocytes stimulated by phytohemagglutinin, a T-cell activator - Fujii_1998_J.Neuroimmunol_82_101
Author(s) : Fujii T , Yamada S , Watanabe Y , Misawa H , Tajima S , Fujimoto K , Kasahara T , Kawashima K
Ref : Journal of Neuroimmunology , 82 :101 , 1998
Abstract : The induction of mRNA for choline acetyltransferase (ChAT), which catalyzes acetylcholine (ACh) synthesis was investigated in human mononuclear leukocytes (MNL) stimulated by phytohemagglutinin (PHA), a T-cell activator, using the reverse transcription-polymerase chain reaction. Stimulation of MNL by PHA induced the expression of ChAT mRNA, and potentiated ACh synthesis. ChAT mRNA induction required more time than the induction of interleukin-2 mRNA. Expression of the gene encoding the vesicular ACh transporter, which mediates ACh transport in cholinergic neurons, was not observed in PHA-stimulated MNL, suggesting that the mechanisms controlling ACh release from T-lymphocytes differ from those in cholinergic neurons. These findings demonstrate that activation of T-lymphocytes up-regulates ACh synthesis in the blood, and suggest that ACh plays an important role as a neuroimmunomodulator besides its role as a neurotransmitter.
ESTHER : Fujii_1998_J.Neuroimmunol_82_101
PubMedSearch : Fujii_1998_J.Neuroimmunol_82_101
PubMedID: 9526852

Title : Acetylcholine synthesis and muscarinic receptor subtype mRNA expression in T-lymphocytes - Kawashima_1998_Life.Sci_62_1701
Author(s) : Kawashima K , Fujii T , Watanabe Y , Misawa H
Ref : Life Sciences , 62 :1701 , 1998
Abstract : We used a sensitive and specific radioimmunoassay for acetylcholine (ACh), and detected significant amounts of ACh in the blood of various mammals, including humans. About 60% of human blood ACh was localized in mononuclear leukocytes. Human leukemic T-cell lines, used as T-lymphocyte models, contained both ACh and choline acetyltransferase (ChAT) activity. Furthermore, ChAT mRNA and protein were detected in the T-cell line MOLT-3. Phytohemagglutinin, a T-cell activator, increased both synthesis and release of ACh by MOLT-3 cells. Muscarinic receptor subtype mRNA expression was confirmed in various T-cell lines. These findings indicate that ACh synthesized by ChAT in T-lymphocytes acts on the muscarinic receptors on lymphocytes in autocrine and/or paracrine pathways and suggest that ACh in blood functions as a modulator of T-cell-dependent immune responses.
ESTHER : Kawashima_1998_Life.Sci_62_1701
PubMedSearch : Kawashima_1998_Life.Sci_62_1701
PubMedID: 9585160

Title : Demonstration of the facilitatory role of 8-OH-DPAT on cholinergic transmission in the rat hippocampus using in vivo microdialysis - Fujii_1997_Brain.Res_761_244
Author(s) : Fujii T , Yoshizawa M , Nakai K , Fujimoto K , Suzuki T , Kawashima K
Ref : Brain Research , 761 :244 , 1997
Abstract : The role of the serotonin (5-HT)1A receptor in the regulation of acetylcholine (ACh) release in the hippocampus was investigated using an in vivo microdialysis technique and a sensitive radioimmunoassay specific for ACh. The mean (+/- S.E.M.) basal ACh contents in the hippocampal perfusate of conscious, freely moving rats was 60 +/- 4 (n = 29) and 3691 +/- 265 fmol/30 min (n = 31), respectively, in the absence and presence of physostigmine (Phy) in the perfusion fluid. Systemic administration of 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT, 0.5 mg/kg, s.c.), a 5-HT1A agonist, significantly enhanced ACh release both in the presence and absence of Phy. Local application of 8-OH-DPAT (3-30 microM) into the hippocampus through the microdialysis probe significantly potentiated ACh release only in the presence of Phy, whereas no significant effect was observed in its absence. Pretreatment with NAN-190 (3 mg/kg, i.p.), a 5-HT1A antagonist, eliminated the increasing effect of systemically applied 8-OH-DPAT on ACh release, while NAN-190 alone had no effect on basal ACh release either in the absence or presence of Phy. Consistent with the time course of ACh release, systemic administration of 8-OH-DPAT evoked hyperlocomotion, which was reversed by NAN-190. However, local hippocampal application of 8-OH-DPAT did not affect the locomotor activity of the rats. These findings suggest that at least two different sites are involved in the 8-OH-DPAT-induced increase in the release of ACh in the rat hippocampus in vivo.
ESTHER : Fujii_1997_Brain.Res_761_244
PubMedSearch : Fujii_1997_Brain.Res_761_244
PubMedID: 9252022

Title : Oral administration of KW-5092, a novel gastroprokinetic agent with acetylcholinesterase inhibitory and acetylcholine release enhancing activities, causes a dose-dependent increase in the blood acetylcholine content of beagle dogs - Yamada_1997_Neurosci.Lett_225_25
Author(s) : Yamada S , Fujii T , Kawashima K
Ref : Neuroscience Letters , 225 :25 , 1997
Abstract : Acetylcholine (ACh) was detected in the blood and plasma of beagle dogs using a specific, sensitive radioimmunoassay. The mean basal ACh contents in the blood and plasma of beagle dogs were 451 +/- 65 and 83.5 +/- 12.3 pg/ml (+/- SEM, n = 7), respectively, and were lower than the contents in humans reported previously by our laboratory. Oral administration of KW-5092 (10-30 mg/kg), a gastroprokinetic agent with acetylcholinesterase (AChE) inhibitory and ACh release enhancing activities, caused a dose-dependent increase in the ACh content of both the blood and plasma, as well as several behavioral side effects due to peripheral cholinergic stimulation. The size of the increase in the plasma ACh content at each dose of KW-5092 was greater than that in the blood, indicating that KW-5092 caused the increase in the blood ACh content through elevation of the plasma ACh content, by inhibition of AChE and facilitation of ACh release. These results demonstrate that the blood ACh of beagle dogs is present mainly in the blood cells and to a lesser degree in the plasma, and that KW-5092 increased the blood ACh content mainly by increasing the plasma ACh concentration.
ESTHER : Yamada_1997_Neurosci.Lett_225_25
PubMedSearch : Yamada_1997_Neurosci.Lett_225_25
PubMedID: 9143009

Title : Nitric oxide increases stimulation-evoked acetylcholine release from rat hippocampal slices by a cyclic GMP-independent mechanism - Suzuki_1997_Brain.Res_760_158
Author(s) : Suzuki T , Nakajima K , Fujimoto K , Fujii T , Kawashima K
Ref : Brain Research , 760 :158 , 1997
Abstract : Nitric oxide (NO) is an endothelium-derived relaxing factor and its main mechanism of action is activation of soluble guanylyl cyclase. NO and NO-related compounds have been reported to affect several neuronal functions in the central nervous system. In this study, we investigated the effects of NO donors (sodium nitroprusside (SNP) and (+/-)-(E)-4-ethyl-2-[(E)-hydroxyimino]-5-nitro-3-hexenamide (FK409)) on acetylcholine (ACh) release from rat hippocampal slices. SNP (10(-5) M) and FK409 (10(-4) M) increased electrical stimulation-evoked ACh release without affecting basal release. As dibutyryl cyclic GMP inhibited stimulation-evoked ACh release, the effects of these NO donors were not due to soluble guanylyl cyclase activation. Atropine increased stimulation-evoked ACh release by blocking presynaptic muscarinic autoreceptors, and SNP increased stimulation-evoked ACh release in the presence of atropine, suggesting that SNP and atropine increase stimulation-evoked ACh release by different mechanisms. The present results indicate that NO enhances some part of the excitation-secretion coupling pathway without inducing ACh release directly and these effects are mediated by cyclic GMP-independent mechanism.
ESTHER : Suzuki_1997_Brain.Res_760_158
PubMedSearch : Suzuki_1997_Brain.Res_760_158
PubMedID: 9237530

Title : Maintenance of constant blood acetylcholine content before and after feeding in young chimpanzees - Fujii_1997_Neurosci.Lett_227_21
Author(s) : Fujii T , Mori Y , Tominaga T , Hayasaka I , Kawashima K
Ref : Neuroscience Letters , 227 :21 , 1997
Abstract : We have shown that acetylcholine (ACh) is present in the blood of various species of mammals using a specific, sensitive radioimmunoassay. In the present study, the effect on blood and plasma ACh levels of feeding after overnight fasting was studied in one male and five female 4- to 7-year-old chimpanzees. The mean basal ACh concentrations of the blood and plasma were 3143 +/- 380 and 184 +/- 10 pg/ml (+/-SEM, n = 6), respectively. Feeding each chimpanzee 500 g boiled sweet potatoes as breakfast at 1000 h and tap water given ad libitum did not affect the ACh content of the blood and plasma, and constant values of the blood and plasma ACh contents were observed for 4 h after the feeding. Hematocrit and plasma acetylcholinesterase (AChE) activity were also insensitive to feeding. No correlation was observed between plasma AChE activity and either blood or plasma ACh content. The results of the present study indicate that the blood ACh of chimpanzees is distributed mainly in the blood cell fraction, and that the blood ACh content is not regulated directly by cholinergic nerve activity or by plasma AChE activity.
ESTHER : Fujii_1997_Neurosci.Lett_227_21
PubMedSearch : Fujii_1997_Neurosci.Lett_227_21
PubMedID: 9178849

Title : Determination of acetylcholine concentration in cerebrospinal fluid of patients with neurologic diseases - Yamada_1996_Acta.Neurol.Scand_93_76
Author(s) : Yamada H , Otsuka M , Fujimoto K , Kawashima K , Yoshida M
Ref : Acta Neurologica Scandinavica , 93 :76 , 1996
Abstract : INTRODUCTION: Acetylcholine (ACh) is a well known neurotransmitter in the central nervous system, but determining its level in cerebrospinal fluid (CSF) is very difficult and the origin of CSF ACh is still unknown. In this study, we attempted to measure CSF ACh by a specific and sensitive radioimmunoassay (RIA) from patients with neurologic diseases. MATERIAL AND
METHODS: Patients with cerebral infarction (n = 7), Parkinson's disease (n = 6), spinocerebellar degeneration (n = 6), Alzheimer's disease (n = 3), amyotrophic lateral sclerosis (n = 3) and disc herniation with no central nervous involvement (n = 8) participated to determine the CSF ACh levels.
RESULTS: Of these 33 patients, the mean ACh level in CSF was 282.2 +/- 61.7 fmol/ml (mean +/- SE, range 20-1505.8 fmol/ml). The mean ACh level of spinocerebellar degeneration group was lower than others, but not statistically significant. CONCLUSION: We conclude that an amount of ACh detectable by RIA is certainly present in CSF.
ESTHER : Yamada_1996_Acta.Neurol.Scand_93_76
PubMedSearch : Yamada_1996_Acta.Neurol.Scand_93_76
PubMedID: 8825278

Title : Localization and synthesis of acetylcholine in human leukemic T cell lines - Fujii_1996_J.Neurosci.Res_44_66
Author(s) : Fujii T , Tsuchiya T , Yamada S , Fujimoto K , Suzuki T , Kasahara T , Kawashima K
Ref : Journal of Neuroscience Research , 44 :66 , 1996
Abstract : In order to clarify the origin of acetylcholine (ACh) in human blood, we measured the content and synthesis activity of ACh in several human leukemic cell lines. The intracellular ACh content determined by a specific and sensitive radioimmunoassay in the human leukemic T cell lines, HSB-2, MOLT-3, and CEM, was 79.6, 36.2, and 9.5 pmol/10(6) cells, respectively. These values were 9-70-fold higher than those of other cell lines, including a helper T cell line, Jurkat. Stimulation of HSB-2 and MOLT-3 by phytohemagglutinin (PHA) increased both the intracellular content and release of ACh into the culture medium, but did not influence the intracellular content and release of ACh in CEM. ACh synthesis activity was found in all the T cell lines tested. Bromoacetylcholine (100 microM), a choline acetyltransferase (ChAT) inhibitor, and bromoacetyl-L-carnitine (100 microM), a carnitine acetyltransferase (CarAT) inhibitor, decreased ACh-synthesizing activity in MOLT-3, and HSB-2 and CEM, by about 50% and 30%, respectively, indicating that both ChAT, and to a lesser extent CarAt, are involved in ACh synthesis in T cells. These results suggest that T lymphocytes have the potential to synthesize and release ACh, which may play a role in regulating T cell-dependent immune responses.
ESTHER : Fujii_1996_J.Neurosci.Res_44_66
PubMedSearch : Fujii_1996_J.Neurosci.Res_44_66
PubMedID: 8926632

Title : Effect of TA-0910, a novel thyrotropin-releasing hormone analog, on in vivo acetylcholine release and turnover in rat brain - Kinoshita_1996_Jpn.J.Pharmacol_71_139
Author(s) : Kinoshita K , Kawashima K , Kawashima Y , Fukuchi I , Yamamura M , Matsuoka Y
Ref : Japanese Journal of Pharmacology , 71 :139 , 1996
Abstract : To examine the action of a novel thyrotropin-releasing hormone (TRH) analog, TA-0910 ((-)-N-[(S)-hexahydro-1-methyl-2,6-dioxo-4-pyrimidinylcarbonyl]-L- histidyl-L-prolinamide tetrahydrate), on the cerebral cholinergic systems, the release of acetylcholine (ACh) and choline in freely-moving rats and ACh accumulation in gamma-butyrolactone (GBL, a nerve impulse flow blocker)- and physostigmine-treated rats were examined. TA-0910 (0.1-1 mg/kg, i.p.) caused a marked dose-dependent increase in extracellular ACh levels and a decrease in choline levels in the hippocampus of freely moving rats. These effects were significantly stronger and longer-lasting than similar effects of TRH. TA-0910 (1, 3 mg/kg, i.p.) depressed the ACh accumulation in the cerebral cortex and hippocampus of GBL (1000 mg/kg, i.p.)-treated rats. Moreover, this analog (1, 3 mg/kg, i.p.) increased the accumulation rate of ACh in these regions in physostigmine (1 mg/kg, i.p.)-treated rats. TRH (30 mg/kg, i.p.) affected the ACh accumulation only in the hippocampus of the GBL-treated rats. These results suggest that TA-0910 not only enhances the release of ACh, but also accelerates the ACh turnover, i.e., ACh release and synthesis, at the cholinergic neuronal terminals in normal rats.
ESTHER : Kinoshita_1996_Jpn.J.Pharmacol_71_139
PubMedSearch : Kinoshita_1996_Jpn.J.Pharmacol_71_139
PubMedID: 8835640

Title : Species differences in the concentration of acetylcholine, a neurotransmitter, in whole blood and plasma - Fujii_1995_Neurosci.Lett_201_207
Author(s) : Fujii T , Yamada S , Yamaguchi N , Fujimoto K , Suzuki T , Kawashima K
Ref : Neuroscience Letters , 201 :207 , 1995
Abstract : Various concentrations of acetylcholine (ACh) were detected in samples of bovine, goat, horse, porcine, rat and sheep blood and plasma using a specific, sensitive radioimmunoassay. The ACh levels in whole blood in bovine and horse samples were about 40- and ten-fold higher, respectively, than in humans, but levels comparable to those in humans were measured in porcine samples. Goat, rat and sheep samples had lower whole blood ACh concentrations than those of humans. When plasma samples were assayed, the ACh contents of bovine and porcine plasma were found to be about two- to five-fold those of human. On the other hand, levels in horse, goat, rat and sheep samples were much lower than in humans. The ratio of the ACh contents of plasma to whole blood was high in porcine and rat samples, indicating that porcine and rat blood ACh is distributed mostly in the plasma, while in the other species tested most of the ACh is present in the blood cells. These results demonstrate that variable levels of ACh are present in the blood of different species, and that the distribution of ACh in the blood constituents varies according to species.
ESTHER : Fujii_1995_Neurosci.Lett_201_207
PubMedSearch : Fujii_1995_Neurosci.Lett_201_207
PubMedID: 8786841

Title : Effect of NIK-247 on basal concentrations of extracellular acetylcholine in the cerebral cortex of conscious, freely moving rats - Ishii_1994_Jpn.J.Pharmacol_66_289
Author(s) : Ishii Y , Kojima J , Ikeda N , Kawashima K
Ref : Japanese Journal of Pharmacology , 66 :289 , 1994
Abstract : We studied the effect of orally administered NIK-247 (9-amino-2,3,5,6,7,8-hexahydro-1H-cyclopenta[b]quinoline monohydrochloride monohydrate) on basal extracellular acetylcholine (ACh) concentrations in the rat cerebral cortex using microdialysis without the addition of cholinesterase inhibitor to the perfusion fluid and radioimmunoassay for ACh. In addition, the effect of oral administration of NIK-247 on acetylcholinesterase (AChE) activity in rat cerebral cortex was determined. The mean basal ACh content in the perfusate from the cerebral cortex of freely moving rats was 123.2 +/- 21.8 fmol/30 min (n = 7). NIK-247 (2.5-10.0 mg/kg, p.o.) increased the ACh content of the perfusate in a dose-dependent manner. NIK-247 at 10 mg/kg significantly increased the ACh content in the perfusate from 0.5 to 2.5 hr after administration, and the maximum increase was attained at 1 hr after administration. 9-Amino-1,2,3,4-tetrahydroacridine (5 mg/kg, p.o.) and physostigmine (0.5 mg/kg, i.p.) significantly increased the ACh content in the perfusate from 1 to 2 hr and from 0.5 to 1.5 hr after administration, respectively. AChE activities in the cerebral cortex were about 32% and 12% below the control value at 1 hr and 3 hr after administration of NIK-247 at 10 mg/kg, respectively. These findings demonstrate that NIK-247 increases extracellular ACh concentration and inhibits AChE activity in the cerebral cortex after oral administration, and they suggest that NIK-247 facilitates central cholinergic transmission.
ESTHER : Ishii_1994_Jpn.J.Pharmacol_66_289
PubMedSearch : Ishii_1994_Jpn.J.Pharmacol_66_289
PubMedID: 7869615

Title : Phorbol ester stimulates acetylcholine synthesis in cultured endothelial cells isolated from porcine cerebral microvessels - Ikeda_1994_Brain.Res_655_147
Author(s) : Ikeda C , Morita I , Mori A , Fujimoto K , Suzuki T , Kawashima K , Murota S
Ref : Brain Research , 655 :147 , 1994
Abstract : Acetylcholine (ACh) is one of the factor which induces vasodilation through the release of endothelium-derived relaxing factor. The aim of this study was to clarify whether endothelial cells can synthesize ACh and the types of substance which regulate the synthesis of ACh in endothelial cells. We determined the ACh content of endothelial cells isolated from porcine cerebral microvessels and of the culture medium. ACh was detected in the medium after 12 h incubation in the presence of diisopropylfluorophosphate, a non-specific cholinesterase inhibitor, and increased linearly up to 24 h. Phorbol 12-myristate 13-acetate (PMA, 10(-7) M) increased the ACh content of the medium in a dose-dependent manner. The effect of PMA was most apparent between 12 and 24 h after treatment, and was inhibited by cycloheximide. Calphostin C, a specific inhibitor of protein kinase C (PKC), did not inhibit the effect of PMA. Dioctanoyl glycerol, a specific activator of PKC, did not increase the intracellular ACh content or the amount released into the culture medium. ACh synthesis was not inhibited by bromoacetylcholine, a specific inhibitor of choline acetyltransferase (ChAT). PMA treatment did not affect the specific activity of ACh synthesis in endothelial cells. These data show that endothelial cells are able to synthesize ACh, and that ACh synthesis is up-regulated by PMA through the PKC independent mechanism via protein induction. The enzyme which synthesizes ACh in endothelial cells is not ChAT. The increase in ACh synthesis induced by PMA may not be due to induction of the ACh synthetic enzyme.
ESTHER : Ikeda_1994_Brain.Res_655_147
PubMedSearch : Ikeda_1994_Brain.Res_655_147
PubMedID: 7529125

Title : Nerve growth factor treatment induces high-potassium-evoked calcium-dependent acetylcholine release in cultured embryonic rat septal cells - Suzuki_1994_Brain.Res_665_311
Author(s) : Suzuki T , Kanagawa M , Takada Y , Fujimoto K , Kawashima K
Ref : Brain Research , 665 :311 , 1994
Abstract : The effects of dibutyryl cAMP (dbcAMP) and nerve growth factor (NGF) on acetylcholine (ACh) release from primary cultured cells of embryonic rat septum were studied. Both dbcAMP and NGF increased the activity of choline acetyltransferase and contents of intracellular and spontaneously released ACh without affecting the total protein content. High-potassium-evoked ACh release was observed in NGF-treated cells but not in untreated and dbcAMP-treated cells. These results indicate that NGF induces cholinergic maturation from induction of the ACh-synthetic enzyme to the excitation-secretion processes.
ESTHER : Suzuki_1994_Brain.Res_665_311
PubMedSearch : Suzuki_1994_Brain.Res_665_311
PubMedID: 7895068

Title : Effects of the centrally acting cholinesterase inhibitors tetrahydroaminoacridine and E2020 on the basal concentration of extracellular acetylcholine in the hippocampus of freely moving rats - Kawashima_1994_Naunyn.Schmiedebergs.Arch.Pharmacol_350_523
Author(s) : Kawashima K , Sato A , Yoshizawa M , Fujii T , Fujimoto K , Suzuki T
Ref : Naunyn Schmiedebergs Arch Pharmacol , 350 :523 , 1994
Abstract : The effects of the centrally acting cholinesterase (ChE) inhibitors, tetrahydroaminoacridine (THA) and E2020 (1-benzyl-4-[(5,6-dimethoxy-1-indanon)-2-yl] methylpiperidine hydrochloride), potential drugs for the treatment of senile dementia, on the basal extracellular acetylcholine (ACh) concentration in the hippocampus of freely moving rats, were determined using a microdialysis technique without the use of a ChE inhibitor in the perfusion fluid and a sensitive RIA. The mean (+/- SEM) basal ACh content in the perfusate was 103.1 +/- 3.6 fmol/sample collected over 30 min when microdialysis probes with a length of 3 mm dialysis membrane were used. The content of ACh decreased to an almost undetectable level upon perfusion of magnesium, suggesting that, in the present study, most of the ACh detected in the perfusates was due to cholinergic neuronal activity. THA (1.65 mg/kg, i.p.) produced an insignificant increase in the extracellular ACh concentration, but a dose of 5 mg/kg, i.p. caused a prolonged and significant 5.5-fold increase from the control value. E2020 (0.65 and 2 mg/kg, i.p.) produced significant, prolonged and dose-dependent increases (4 and 12 times the control value, respectively), the peak effect occurring within 1 h. Perfusion with 10 mumol/l physostigmine produced an about 30-fold increase of ACh output, suggesting that the basal extracellular ACh concentration is highly dependent on ChE activity. When ChE was inhibited locally by perfusion with physostigmine, THA (5 mg/kg) produced a transient and, at its maximum, a 1.42-fold increase in extracellular ACh concentration.
ESTHER : Kawashima_1994_Naunyn.Schmiedebergs.Arch.Pharmacol_350_523
PubMedSearch : Kawashima_1994_Naunyn.Schmiedebergs.Arch.Pharmacol_350_523
PubMedID: 7870192

Title : Tacrine increases stimulation-evoked acetylcholine release from rat hippocampal slices - Suzuki_1994_Jpn.J.Pharmacol_65_337
Author(s) : Suzuki T , Nonaka H , Fujimoto K , Kawashima K
Ref : Japanese Journal of Pharmacology , 65 :337 , 1994
Abstract : We examined the effects of tacrine (9-amino-1,2,3,4-tetrahydroacridine) on endogenous acetylcholine (ACh) release from rat hippocampal slices. Tacrine (more than 1 microM) increased the measurable amount of basal ACh release. On the other hand, in the presence of physostigmine (50 microM; under this condition, cholinesterase activity was inhibited), tacrine did not enhance the basal ACh release. Tacrine at more than 100 microM increased the submaximal electrical stimulation-evoked release of ACh in both the absence and presence of physostigmine (50 microM). This effect of tacrine was abolished by a combination of atropine (100 mM) and physostigmine. These results indicate that a high-dose of tacrine increases cholinergic neurotransmission not only by inhibition of cholinesterase but also by increasing ACh release through an atropine-like effect, perhaps by blockade of part of the process of muscarinic autoinhibition.
ESTHER : Suzuki_1994_Jpn.J.Pharmacol_65_337
PubMedSearch : Suzuki_1994_Jpn.J.Pharmacol_65_337
PubMedID: 7990271

Title : Effects of physostigmine and some nitric oxide-cyclic GMP-related compounds on muscarinic receptor-mediated autoinhibition of hippocampal acetylcholine release - Suzuki_1993_J.Neurochem_60_2285
Author(s) : Suzuki T , Nonaka H , Fujimoto K , Kawashima K
Ref : Journal of Neurochemistry , 60 :2285 , 1993
Abstract : We have investigated the effects of (a) the cholinesterase inhibitor physostigmine and (b) drugs that are known to change intracellular cyclic GMP levels on the autoinhibition of acetylcholine release from rat hippocampal slices. Autoinhibition was triggered by submaximal electrical stimulation in both the absence and presence of physostigmine. The results obtained indicate that an unusual increase in the extracellular acetylcholine content, such as that induced by cholinesterase inhibition, is not essential for autoinhibition triggering. Dibutyryl cyclic GMP reduced significantly the stimulation-evoked acetylcholine release in the presence, but not in the absence, of atropine. Neither sodium nitroprusside nor glyceryl trinitrate exerted a dibutyryl cyclic GMP-like effect. NG-Nitro-L-arginine did not lessen the autoinhibition. These results indicate that an increase in the intracellular cyclic GMP level reduces acetylcholine release, and that the muscarinic receptor stimulation-nitric oxide synthesis-(soluble) guanylyl cyclase activation pathway is not involved in the cholinergic autoinhibition process.
ESTHER : Suzuki_1993_J.Neurochem_60_2285
PubMedSearch : Suzuki_1993_J.Neurochem_60_2285
PubMedID: 8388037

Title : Acetylcholine synthesis is modulated by acetylcholine content of cytosolic fraction but not by that of releasable fraction - Suzuki_1992_Neurosci.Lett_144_127
Author(s) : Suzuki T , Kashima Y , Fujimoto K , Kawashima K
Ref : Neuroscience Letters , 144 :127 , 1992
Abstract : Synthesis and release of acetylcholine (ACh) in the rat hippocampal slices were examined to clarify the mechanism of modulation of ACh synthesis. Treatment with 2-(4-phenylpiperidino)cyclohexanol (AH5183, 50 microM), an inhibitor of ACh transport from cytosol to synaptic vesicles, inhibited the increase in ACh content of the membrane-bound fraction which is readily releasable, but did not affect the cytosolic ACh content. Under these conditions, the total ACh content reached a plateau value. These results indicate that ACh synthesis is modulated by cytosolic ACh content but not by the vesicular fraction.
ESTHER : Suzuki_1992_Neurosci.Lett_144_127
PubMedSearch : Suzuki_1992_Neurosci.Lett_144_127
PubMedID: 1436692

Title : Regional differences in extracellular choline dependency of acetylcholine synthesis in the rat brain - Suzuki_1991_Neurosci.Res_11_71
Author(s) : Suzuki T , Kashima Y , Fujimoto K , Oohata H , Kawashima K
Ref : Neurosci Res , 11 :71 , 1991
Abstract : Acetylcholine (ACh) synthesis under conditions of restricted extracellular choline uptake was investigated in order to clarify the procurement of choline for ACh synthesis using slices of several regions of the rat brain. Extracellular choline-independent ACh synthesis was observed in the hippocampus, frontal cortex and caudate putamen, which contain cholinergic nerve terminals, whereas little or no synthesis was observed in the medial septum or basal nucleus of Meynert, which contain cholinergic cell bodies. These results indicate that cholinergic nerve terminals, but not the cell bodies, may be able to synthesize choline for ACh biosynthesis. Extracellular choline-dependent ACh synthesis was observed in all regions examined. In the presence of 10 microM choline, the highest content of newly synthesized ACh and the proportionate increase in ACh compared with the unreleasable fraction (basal level) were observed in the caudate putamen. In the frontal cortex, although the level of synthesized ACh was low, the proportionate increase in ACh was high. In contrast, in the medial septum and basal nucleus of Meynert, high levels of ACh with a low proportionate increase compared with basal levels were observed. In the presence of hemicholinium-3, extracellular choline was also taken up for ACh synthesis in all regions examined, the level being especially high in the frontal cortex and medial septum. The present results indicate that the manner of choline procurement for ACh synthesis is heterogeneous among the various regions of the rat brain.
ESTHER : Suzuki_1991_Neurosci.Res_11_71
PubMedSearch : Suzuki_1991_Neurosci.Res_11_71
PubMedID: 1653922

Title : Determination of acetylcholine release in the striatum of anesthetized rats using in vivo microdialysis and a radioimmunoassay - Kawashima_1991_J.Neurochem_57_882
Author(s) : Kawashima K , Hayakawa T , Kashima Y , Suzuki T , Fujimoto K , Oohata H
Ref : Journal of Neurochemistry , 57 :882 , 1991
Abstract : A vertical-type in vivo microdialysis probe and a sensitive, specific radioimmunoassay (RIA) were used to study the mechanism of acetylcholine (ACh) release in the striatum of anesthetized rats. Without the use of physostigmine, a cholinesterase inhibitor, our RIA could still detect the amount of ACh present in the perfusate (5.6 +/- 0.6 fmol/min, n = 16). Tetrodotoxin (1 microM) produced a significant decrease in the amount of ACh collected in the perfusate, suggesting that basal ACh determined under the present experimental conditions was related to cholinergic neural activity. Atropine (0.1-1 microM) applied topically via the dialysis probe did not affect the amount of ACh recovered in the perfusate in the absence of physostigmine. Addition of physostigmine (10 microM) to the perfusion fluid produced about a 100-fold increase in the amount of ACh collected. In the presence of physostigmine, topical administration of atropine and pirenzepine (0.01-1 microM) through a dialysis probe produced a further three- to fourfold increase in ACh output, whereas a slight increase was produced by AF-DX 116 at the highest concentration (1 microM). These results indicate that presynaptic modulation of ACh release in the striatum does not occur under basal conditions, and that presynaptic M1 muscarinic receptors are involved in the modulation of ACh release when the ACh concentration is raised under certain conditions.
ESTHER : Kawashima_1991_J.Neurochem_57_882
PubMedSearch : Kawashima_1991_J.Neurochem_57_882
PubMedID: 1861156

Title : Substance P-evoked release of acetylcholine from isolated spinal cord of the newborn rat - Kobayashi_1991_Neurosci_45_331
Author(s) : Kobayashi N , Sakuma M , Yoshioka K , Onishi Y , Yanagisawa M , Kawashima K , Otsuka M
Ref : Neuroscience , 45 :331 , 1991
Abstract : Isolated spinal cords of newborn rats were perfused with artificial cerebrospinal fluid and the release of endogenous acetylcholine was measured using high-performance liquid chromatography with an electrochemical detection system. Application of high-K+ (90 mM) medium evoked about an eight-fold increase in the acetylcholine release, and the K(+)-evoked release was Ca2+ dependent. Veratridine (20 microM) also evoked about a four-fold increase in the acetylcholine release, and this increase was suppressed by 0.2 microM tetrodotoxin. Application of substance P at 0.3-3 microM evoked a concentration-dependent release of acetylcholine. The substance P-evoked acetylcholine release was Ca2+ dependent and abolished by tetrodotoxin. Neurokinin A, neurokinin B, acetyl-Arg6-septide and senktide (3 microM each) also evoked a release of acetylcholine. Electrophysiological experiments using isolated spinal cords of newborn rats showed that bath application of substance P induced a depolarization of motoneurons, which was enhanced by edrophonium. This enhancement of substance P-induced depolarization by edrophonium disappeared in a low-Ca2+ medium or in the presence of atropine and dihydro-beta-erythroidine. In the presence of edrophonium and dihydro-beta-erythroidine, substance P induced an inhibition of monosynaptic reflex, and this inhibition was abolished by atropine. These results suggest that substance P and other tachykinins induce a release of acetylcholine from the newborn rat spinal cord by exciting cholinergic neurons.
ESTHER : Kobayashi_1991_Neurosci_45_331
PubMedSearch : Kobayashi_1991_Neurosci_45_331
PubMedID: 1722289

Title : Pharmacological differentiation of presynaptic M1 muscarinic receptors modulating acetylcholine release from postsynaptic muscarinic receptors in guinea-pig ileum - Kawashima_1990_Gen.Pharmacol_21_17
Author(s) : Kawashima K , Fujimoto K , Suzuki T , Oohata H
Ref : General Pharmacology , 21 :17 , 1990
Abstract : 1. Effects of three muscarinic antagonists on electrically evoked ACh release and contractile response were investigated in longitudinal muscle strips of guinea-pig ileum suspended in an organ-bath and superfused with Krebs solution. ACh release was determined by a specific radioimmunoassay. 2. Telenzepine, a selective M1 muscarinic antagonist, increased the ACh release at a concentration of 100-fold less than that inhibiting the contractile response (10 vs 1000 nM). 3. AF-DX 116, a cardioselective M2 muscarinic antagonist, inhibited the contractile response at 10 microM, but did not affect the ACh release at this concentration. 4. (-)N-Methylscopolamine (NMS) did not affect the ACh release, but inhibited the contractile response at all concentrations tested (1-1000 nM), indicating (-)NMS can be used as an ileal specific postsynaptic muscarinic antagonist. 5. These data demonstrate that presynaptic muscarinic receptors modulating ACh release are distinct from postsynaptic ones involved in the contractile response and can be classified as M1 subtype.
ESTHER : Kawashima_1990_Gen.Pharmacol_21_17
PubMedSearch : Kawashima_1990_Gen.Pharmacol_21_17
PubMedID: 2298386

Title : Synthesis and release of acetylcholine by cultured bovine arterial endothelial cells - Kawashima_1990_Neurosci.Lett_119_156
Author(s) : Kawashima K , Watanabe N , Oohata H , Fujimoto K , Suzuki T , Ishizaki Y , Morita I , Murota S
Ref : Neuroscience Letters , 119 :156 , 1990
Abstract : Using cultured endothelial cells prepared from bovine carotid artery, and a specific radioimmunoassay for acetylcholine (ACh), the synthesis and release of ACh by vascular endothelial cells were investigated directly. ACh content in the culture medium after 24 h of incubation in the presence of isoflurophate, a nonspecific cholinesterase inhibitor, was about 16 times higher than that in endothelial cells, indicating that ACh synthesized inside the cells was released rapidly. The presence of ACh in the culture medium was further confirmed qualitatively using high-performance liquid chromatography with an electrocapture detection. These results represent the first direct evidence that endothelial cells can synthesize and release ACh, suggesting the possibility that ACh acts not only as a neurotransmitter but also as an autacoid under certain conditions.
ESTHER : Kawashima_1990_Neurosci.Lett_119_156
PubMedSearch : Kawashima_1990_Neurosci.Lett_119_156
PubMedID: 2280888

Title : Effects of TRH and DN-1417 on high potassium-evoked acetylcholine release from rat basal forebrain slices determined directly by radioimmunoassay - Suzuki_1989_Gen.Pharmacol_20_239
Author(s) : Suzuki T , Fujimoto K , Oohata H , Kawashima K
Ref : General Pharmacology , 20 :239 , 1989
Abstract : 1. High potassium (50 mM)-evoked acetylcholine (ACh) release from rat basal forebrain slices under conditions without an exogenous choline supply was determined using a radioimmunoassay for ACh. 2. A consistent amount of ACh release was observed at each repetitive stimulation and ACh content in brain slices was not altered by potassium stimulations. These results indicate the existence of a large intracellular releasable ACh store, which is independent of new synthesis from exogenous choline. 3. Atropine, even at a concentration of 10(-6) M, did not affect the potassium-evoked ACh release. Thus, modulation of ACh release by the muscarinic autoreceptor was not revealed under the conditions employed. 4. Thyrotropin-releasing hormone (TRH, 10(-4) M) caused a slight and statistically insignificant increase in potassium-evoked ACh release. DN-1417, a TRH analogue, at a concentration of 10(-4) M significantly increased potassium-evoked ACh release. These findings indicate that DN-1417 is able to enhance ACh output independently of ACh synthesis from exogenous choline.
ESTHER : Suzuki_1989_Gen.Pharmacol_20_239
PubMedSearch : Suzuki_1989_Gen.Pharmacol_20_239
PubMedID: 2565849

Title : Hemicholinium-3-resistant choline uptake system linked to acetylcholine synthesis in the rat hippocampus - Suzuki_1989_Neurosci.Lett_105_211
Author(s) : Suzuki T , Kashima Y , Kawashima K
Ref : Neuroscience Letters , 105 :211 , 1989
Abstract : Extracellular choline-dependency of acetylcholine (ACh) synthesis was examined in rat hippocampal slices. In the presence of hemicholinium-3 (HC-3, 5 microM), extracellular choline-dependent ACh synthesis appeared to consist of two different components. The first component (saturated at up to 50 microM choline) was dependent on the HC-3-resistant choline uptake system having a low capacity for choline in comparison with the high-affinity choline uptake system (HACU). The second component, observed in the presence of more than 50 microM choline, was considered due to competitive inhibition of HACU by HC-3. HACU-dependent ACh synthesis seemed to be saturated in the presence of up to 2 microM choline. These results indicate that both the HACU and HC-3-resistant choline uptake systems are linked to ACh synthesis in the hippocampus.
ESTHER : Suzuki_1989_Neurosci.Lett_105_211
PubMedSearch : Suzuki_1989_Neurosci.Lett_105_211
PubMedID: 2485880

Title : Extraneuronal localization of acetylcholine and its release upon nicotinic stimulation in rabbits - Kawashima_1989_Neurosci.Lett_104_336
Author(s) : Kawashima K , Oohata H , Fujimoto K , Suzuki T
Ref : Neuroscience Letters , 104 :336 , 1989
Abstract : A study was undertaken to examine the origin of plasma acetylcholine (ACh). The ACh content of blood cells was about 25 times higher than that of plasma in normal rabbits (3,722 vs 140 pg/ml blood, n = 7). Plasma ACh content in rabbits having antibody against ACh was about 80 times higher than in normal rabbits, while no difference was observed in the ACh content of blood cells between the groups. Nicotine (100 micrograms/kg, i.v.) produced a significant increase in plasma ACh content and a decrease in the ACh content of blood cells in normal rabbits. These data demonstrate that a large amount of ACh is localized in blood cells and that a considerable proportion of plasma ACh originates from blood cells, suggesting that ACh acts not only as a neurotransmitter but also as an autacoid.
ESTHER : Kawashima_1989_Neurosci.Lett_104_336
PubMedSearch : Kawashima_1989_Neurosci.Lett_104_336
PubMedID: 2812548

Title : Presynaptic M1 muscarinic receptor modulates spontaneous release of acetylcholine from rat basal forebrain slices - Suzuki_1988_Neurosci.Lett_84_209
Author(s) : Suzuki T , Fujimoto K , Oohata H , Kawashima K
Ref : Neuroscience Letters , 84 :209 , 1988
Abstract : Spontaneous release of acetylcholine (ACh) from rat basal forebrain slices in the presence of cholinesterase inhibitor was directly determined using a specific radioimmunoassay for ACh. The release was calcium dependent. A consistent amount of ACh release was observed throughout the experiment. Atropine (10(-8) to 10(-5) M) and pirenzepine (10(-7) to 10(-5) M) enhanced spontaneous ACh release. These findings indicate the presence of an M1 muscarinic autoreceptor that modulates spontaneous release of ACh in the rat basal forebrain.
ESTHER : Suzuki_1988_Neurosci.Lett_84_209
PubMedSearch : Suzuki_1988_Neurosci.Lett_84_209
PubMedID: 3340327

Title : Direct determination of acetylcholine release by radioimmunoassay and presence of presynaptic M1 muscarinic receptors in guinea pig ileum - Kawashima_1988_J.Pharmacol.Exp.Ther_244_1036
Author(s) : Kawashima K , Fujimoto K , Suzuki T , Oohata H
Ref : Journal of Pharmacology & Experimental Therapeutics , 244 :1036 , 1988
Abstract : Radioimmunoassay for acetylcholine (ACh) with a sensitivity of 10 pg/tube was applied to the direct determination of ACh output from the nerve endings in longitudinal muscle strips of guinea pig ileum. The strips were preincubated with an irreversible cholinesterase inhibitor and superfused with Krebs' solution under various experimental conditions. Pirenzepine (0.1-10 microM) and atropine (10-100 nM) produced an increase in electrically evoked ACh output through the inhibition of presynaptic muscarinic receptors. Contractile response to endogenous ACh released by electrical stimulation was enhanced by pirenzepine and atropine at lower concentrations, whereas the highest concentrations of pirenzepine (10 microM) and atropine (100 nM) caused a reduction in the enhanced contractile response and a significantly diminished response, respectively. These results demonstrate that the concentrations of pirenzepine and atropine, effective in inhibiting presynaptic muscarinic receptors, differ from those inhibiting postsynaptic muscarinic receptors and suggest the possibility that presynaptic M1 muscarinic receptors regulating ACh output may be present in the guinea pig ileum.
ESTHER : Kawashima_1988_J.Pharmacol.Exp.Ther_244_1036
PubMedSearch : Kawashima_1988_J.Pharmacol.Exp.Ther_244_1036
PubMedID: 3252021

Title : Plasma concentration of acetylcholine in young women - Kawashima_1987_Neurosci.Lett_80_339
Author(s) : Kawashima K , Oohata H , Fujimoto K , Suzuki T
Ref : Neuroscience Letters , 80 :339 , 1987
Abstract : A sensitive and specific radioimmunoassay was applied to the determination of acetylcholine (ACh) in plasma. The concentration of ACh in plasma sampled from 32 young women was 456.1 +/- 53.1 (mean +/- S.E.M.) pg/ml. No significant correlations were observed between plasma concentration of ACh and acetylcholinesterase (AChE) activity, or gonadal hormones. These data demonstrate that an amount of ACh measurable by radioimmunoassay is present in plasma and plasma ACh is not regulated by AChE activity and the menstrual cycle in young women. The origin and physiological as well as pathophysiological significance of ACh in plasma remain to be clarified.
ESTHER : Kawashima_1987_Neurosci.Lett_80_339
PubMedSearch : Kawashima_1987_Neurosci.Lett_80_339
PubMedID: 3683989

Title : Pharmacological properties of the novel antimuscarinic agent 4-[2-(1,2-benzisoxazol-3-yl)-2-(hexahydro-1H-azepin-1-yl)a cetoxy]-1-et hyl-1- methylpiperidinium iodide - Kawashima_1986_Arzneimittelforschung_36_927
Author(s) : Kawashima K , Yoshida N , Kadokawa T
Ref : Arzneimittelforschung , 36 :927 , 1986
Abstract : Pharmacological properties of 4-[2-(1, 2-benzisoxazol-3-yl)-2-(hexahydro-1H-azepin-1-yl)acetoxy]-1- ethyl-1- methylpiperidinium iodide (SX-810) were investigated and the following results were obtained. 1. In isolated guinea-pig ileum, SX-810 showed a competitive antagonistic effect against acetylcholine with a pA2 value of 7.93. 2. SX-810 (10-50 mg/kg p.o. or 10-50 micrograms/kg i.v.) inhibited the gastroduodenal contractions induced by bethanechol and carbachol in anaesthetized rats. 3. SX-810 (10-100 mg/kg p.o.) inhibited spontaneous gastric motility in conscious rats and rabbits. 4. SX-810 (10-100 mg/kg p.o.) inhibited gastric secretion in pylorus-ligated rats. 5. SX-810 (20-200 mg/kg p.o. or 1-10 mg/kg s.c.) inhibited the ulceration induced by pylorus ligation or by exposure to restrained and water-immersed stress in rats. 6. When administered orally to rats, SX-810 had no mydriatic effect even at 3000 mg/kg, but subcutaneously administered SX-810 (0.5-10 mg/kg) exhibited such action. 7. When administered orally to rats, SX-810 (100-500 mg/kg) caused no significant inhibition of the salivation induced by pilocarpine, but subcutaneously administered SX-810 (0.5-10 mg/kg) exhibited an inhibition of the salivation. In rabbits, SX-810 (200-500 mg/kg p.o.) also caused no significant inhibition of the salivation except at an extremely high dose of 1000 mg/kg p.o. 8. SX-810 (100-500 mg/kg p.o.) caused no significant effect on urine and electrolyte excretion in rats. These results indicate that oral use of SX-810 exhibits marked spasmolytic and antiulcerative activities without exerting systemic antimuscarinic side effects.
ESTHER : Kawashima_1986_Arzneimittelforschung_36_927
PubMedSearch : Kawashima_1986_Arzneimittelforschung_36_927
PubMedID: 3741526

Title : Effect of light on the acetylcholine concentrations of the suprachiasmatic nucleus in the rat - Murakami_1984_Brain.Res_311_358
Author(s) : Murakami N , Takahashi K , Kawashima K
Ref : Brain Research , 311 :358 , 1984
Abstract : Acetylcholine concentrations were determined in suprachiasmatic nucleus of rats sacrificed by irradiation of microwave, using radioimmunoassay. No significant rhythmicity was observed over a 24-h period in rats blinded for two weeks. When the light was given at 22.00 h (illumination schedule L:07.00-19.00 h) in intact rats, acetylcholine concentration increased 30 and 60 min after light on in suprachiasmatic nucleus, but not in the other control site. These results suggest that endogenous circadian rhythm of acetylcholine concentration is absent in suprachiasmatic nucleus, but light may affect acetylcholine concentration of this site.
ESTHER : Murakami_1984_Brain.Res_311_358
PubMedSearch : Murakami_1984_Brain.Res_311_358
PubMedID: 6541956

Title : A possible mechanism of a new antispasmodic drug, 2-(1,2-benzisoxazol-3-yl)-3-[2-(2-piperidinoethoxy) phenyl]acrylonitrile (SX-284) - Takayanagi_1982_Jpn.J.Pharmacol_32_973
Author(s) : Takayanagi I , Sone H , Kawashima K , Sohji Y , Kadokawa T
Ref : Japanese Journal of Pharmacology , 32 :973 , 1982
Abstract : A newly synthesized drug, SX-284, depressed the twitch response of the ileum from guinea pig to electrical stimulation at 0.1 Hz. SX-284 proved to be almost as active as atropine on electrically stimulated ileum. The responses of guinea pig ileum to nicotine and serotonin were also inhibited by SX-284. However, SX-284 did not influence the release of transmitters from the motor, sympathetic, nonadrenergic inhibitory, noncholinergic excitatory nerves, and responses of various smooth muscles mediated through drug receptors, and at these doses, SX-284 inhibited the release of acetylcholine from the vagus nerve. These facts indicate that SX-284 specifically inhibits the acetylcholine release from the vagus nerve. Furthermore, spontaneous movement of the guinea pig ileum was dose-dependently depressed by SX-284. The potency ratio for SX-284 relative to atropine was 1.7.
ESTHER : Takayanagi_1982_Jpn.J.Pharmacol_32_973
PubMedSearch : Takayanagi_1982_Jpn.J.Pharmacol_32_973
PubMedID: 7161971

Title : Radioimmunoassay for acetylcholine in the rat brain - Kawashima_1980_J.Pharmacol.Methods_3_115
Author(s) : Kawashima K , Ishikawa H , Mochizuki M
Ref : J Pharmacol Methods , 3 :115 , 1980
Abstract : Specific antiserum against acetylcholine (ACh) was produced in the rabbit immunized with a choline hemiglutarate-bovine serum albumin conjugate. The antiserum significantly cross-reacted with choline carboxylates, such as butyrylcholine, succinylcholine, and carbamylcholine. However, neither choline itself nor choline phosphates, such as phosphatidylcholine and phosphorylcholine, showed any significant cross-reaction. The antiserum was used to develop a radioimmunoassay for ACh. The assay can reliably determine as little as 170 pg of ACh. Acetylcholine concentrations in two regions of the rat brain were determined directly from an aqueous extract. After inhibition of acetylcholinesterase with physostigmine, ACh increased more in the forebrain than in the brainstem.
ESTHER : Kawashima_1980_J.Pharmacol.Methods_3_115
PubMedSearch : Kawashima_1980_J.Pharmacol.Methods_3_115
PubMedID: 7392652

Title : [Pharmacological studies of loperamide, an anti-diarrheal agent. III. Interaction between loperamide and various agonists in the guinea pig intestine (author's transl)] - Sohji_1978_Nihon.Yakurigaku.Zasshi_74_213
Author(s) : Sohji Y , Kawashima K , Shimizu M
Ref : Nihon Yakurigaku Zasshi , 74 :213 , 1978
Abstract : To clarify the mode of action of loperamide, the interaction with various agonists was investigated in guinea pig intestines. The following results were obtained. Prostaglandin E1 (0.1 microgram) or acetylcholine (1 approximately 3 microgram) injection into the mesenteric artery at 10 min intervals caused a constant rise of intraluminal pressure of the intestinal loop in situ. Loperamide (0.01 approximately 0.1 mg/kg i.v.) markedly suppressed the response induced by prostaglandin E1. Morphine (0.1 approximately 1.0 mg/kg i.v.) and atropine (0.01 mg/kg i.v.) also suppressed the response. The intestinal response induced by acetylcholine was inhibited markedly by atropine and slightly by loperamide (0.1 mg/kg i.v.) but not by morphine. Contractions of the isolated ileum induced by coaxial stimulation, BaCl2, nicotine and serotonin were suppressed by loperamide (10(-9) approximately 2 X 10(-7) g/ml). Morphine, methadone and codeine showed the same effect as loperamide but the activity was weaker than that of loperamide. Contractions of isolated ileum induced by acetylcholine, histamine and bradykinin, and Ca-contraction in the depolarized taenia coli were inhibited by relatively high concentrations of loperamide (2 X 10(-7) g/ml or above). These results suggest that loperamide suppresses the function of cholinergic neurons in the intestines.
ESTHER : Sohji_1978_Nihon.Yakurigaku.Zasshi_74_213
PubMedSearch : Sohji_1978_Nihon.Yakurigaku.Zasshi_74_213
PubMedID: 658837