Bonner TI


Full name : Bonner Tom I

First name : Tom I

Mail : Section on Functional Neuroscience, National Institute of Mental Health, Bethesda, Maryland

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References (30)

Title : International Union of Basic and Clinical Pharmacology. LXXXVIII. G protein-coupled receptor list: recommendations for new pairings with cognate ligands - Davenport_2013_Pharmacol.Rev_65_967
Author(s) : Davenport AP , Alexander SP , Sharman JL , Pawson AJ , Benson HE , Monaghan AE , Liew WC , Mpamhanga CP , Bonner TI , Neubig RR , Pin JP , Spedding M , Harmar AJ
Ref : Pharmacol Rev , 65 :967 , 2013
Abstract : In 2005, the International Union of Basic and Clinical Pharmacology Committee on Receptor Nomenclature and Drug Classification (NC-IUPHAR) published a catalog of all of the human gene sequences known or predicted to encode G protein-coupled receptors (GPCRs), excluding sensory receptors. This review updates the list of orphan GPCRs and describes the criteria used by NC-IUPHAR to recommend the pairing of an orphan receptor with its cognate ligand(s). The following recommendations are made for new receptor names based on 11 pairings for class A GPCRs: hydroxycarboxylic acid receptors [HCA(1) (GPR81) with lactate, HCA(2) (GPR109A) with 3-hydroxybutyric acid, HCA(3) (GPR109B) with 3-hydroxyoctanoic acid]; lysophosphatidic acid receptors [LPA(4) (GPR23), LPA(5) (GPR92), LPA(6) (P2Y5)]; free fatty acid receptors [FFA4 (GPR120) with omega-3 fatty acids]; chemerin receptor (CMKLR1; ChemR23) with chemerin; CXCR7 (CMKOR1) with chemokines CXCL12 (SDF-1) and CXCL11 (ITAC); succinate receptor (SUCNR1) with succinate; and oxoglutarate receptor [OXGR1 with 2-oxoglutarate]. Pairings are highlighted for an additional 30 receptors in class A where further input is needed from the scientific community to validate these findings. Fifty-seven human class A receptors (excluding pseudogenes) are still considered orphans; information has been provided where there is a significant phenotype in genetically modified animals. In class B, six pairings have been reported by a single publication, with 28 (excluding pseudogenes) still classified as orphans. Seven orphan receptors remain in class C, with one pairing described by a single paper. The objective is to stimulate research into confirming pairings of orphan receptors where there is currently limited information and to identify cognate ligands for the remaining GPCRs. Further information can be found on the IUPHAR Database website (http:\/\/
ESTHER : Davenport_2013_Pharmacol.Rev_65_967
PubMedSearch : Davenport_2013_Pharmacol.Rev_65_967
PubMedID: 23686350

Title : The status, quality, and expansion of the NIH full-length cDNA project: the Mammalian Gene Collection (MGC) - Gerhard_2004_Genome.Res_14_2121
Author(s) : Gerhard DS , Wagner L , Feingold EA , Shenmen CM , Grouse LH , Schuler G , Klein SL , Old S , Rasooly R , Good P , Guyer M , Peck AM , Derge JG , Lipman D , Collins FS , Jang W , Sherry S , Feolo M , Misquitta L , Lee E , Rotmistrovsky K , Greenhut SF , Schaefer CF , Buetow K , Bonner TI , Haussler D , Kent J , Kiekhaus M , Furey T , Brent M , Prange C , Schreiber K , Shapiro N , Bhat NK , Hopkins RF , Hsie F , Driscoll T , Soares MB , Casavant TL , Scheetz TE , Brown-stein MJ , Usdin TB , Toshiyuki S , Carninci P , Piao Y , Dudekula DB , Ko MS , Kawakami K , Suzuki Y , Sugano S , Gruber CE , Smith MR , Simmons B , Moore T , Waterman R , Johnson SL , Ruan Y , Wei CL , Mathavan S , Gunaratne PH , Wu J , Garcia AM , Hulyk SW , Fuh E , Yuan Y , Sneed A , Kowis C , Hodgson A , Muzny DM , McPherson J , Gibbs RA , Fahey J , Helton E , Ketteman M , Madan A , Rodrigues S , Sanchez A , Whiting M , Madari A , Young AC , Wetherby KD , Granite SJ , Kwong PN , Brinkley CP , Pearson RL , Bouffard GG , Blakesly RW , Green ED , Dickson MC , Rodriguez AC , Grimwood J , Schmutz J , Myers RM , Butterfield YS , Griffith M , Griffith OL , Krzywinski MI , Liao N , Morin R , Palmquist D , Petrescu AS , Skalska U , Smailus DE , Stott JM , Schnerch A , Schein JE , Jones SJ , Holt RA , Baross A , Marra MA , Clifton S , Makowski KA , Bosak S , Malek J
Ref : Genome Res , 14 :2121 , 2004
Abstract : The National Institutes of Health's Mammalian Gene Collection (MGC) project was designed to generate and sequence a publicly accessible cDNA resource containing a complete open reading frame (ORF) for every human and mouse gene. The project initially used a random strategy to select clones from a large number of cDNA libraries from diverse tissues. Candidate clones were chosen based on 5'-EST sequences, and then fully sequenced to high accuracy and analyzed by algorithms developed for this project. Currently, more than 11,000 human and 10,000 mouse genes are represented in MGC by at least one clone with a full ORF. The random selection approach is now reaching a saturation point, and a transition to protocols targeted at the missing transcripts is now required to complete the mouse and human collections. Comparison of the sequence of the MGC clones to reference genome sequences reveals that most cDNA clones are of very high sequence quality, although it is likely that some cDNAs may carry missense variants as a consequence of experimental artifact, such as PCR, cloning, or reverse transcriptase errors. Recently, a rat cDNA component was added to the project, and ongoing frog (Xenopus) and zebrafish (Danio) cDNA projects were expanded to take advantage of the high-throughput MGC pipeline.
ESTHER : Gerhard_2004_Genome.Res_14_2121
PubMedSearch : Gerhard_2004_Genome.Res_14_2121
PubMedID: 15489334
Gene_locus related to this paper: human-AFMID , human-CES4A , human-CES5A , human-NOTUM , human-SERAC1 , human-SERHL2 , human-TMEM53 , mouse-acot1 , mouse-adcl4 , mouse-Ces2f , mouse-Ces4a , mouse-notum , mouse-q6wqj1 , mouse-Q9DAI6 , mouse-rbbp9 , mouse-SERHL , mouse-srac1 , mouse-tmm53 , rat-abhd6 , rat-abhda , rat-abhea , rat-abheb , rat-Ldah , rat-cd029 , rat-estd , rat-Kansl3 , rat-nceh1 , ratno-acph , ratno-CMBL , mouse-b2rwd2 , rat-b5den3 , rat-ab17c

Title : Identification and characterization of the rat M1 muscarinic receptor promoter - Klett_1999_J.Neurochem_72_900
Author(s) : Klett CP , Bonner TI
Ref : Journal of Neurochemistry , 72 :900 , 1999
Abstract : Five subtypes of the muscarinic receptor have been cloned from both the rat and human genomes. Although all five genes have the coding sequences in a single exon, their structures 5' of the initiation codon are largely uncharacterized, except for the M4 receptor. In the brain, muscarinic receptors mediate motor and memory function by interaction with their ligand acetylcholine. In addition, the M1 muscarinic subtype has been implicated in behavior, stress-adaptive cardiovascular reflexes, and blood pressure regulation. In the current study the M1 muscarinic receptor noncoding 5'-flanking region has been identified and characterized, including the promoter and two 5' noncoding exons located approximately 13-14 kb from the coding exon. Similar to the M4 muscarinic receptor gene the M1 promoter is GC-rich, contains no TATA box, but has two potential CAAT boxes and several putative binding sites for transcription factors such as SP1 and AP-1-3. The transcription initiation site was identified by RNase protection and primer extension. Promoter activity was confirmed in transient expression assays, using luciferase reporter constructs. A 0.89-kb fragment consisting of 480 bp of the promoter, exon 1, and part of intron 1 expressed luciferase activity in two M1 receptor-expressing cell lines (CCL-107 and CCL-147), whereas a longer fragment (1.5 kb) that extends into intron 2 demonstrated significantly increased luciferase activity. The constructs exhibited responses indicating the presence of functional glucocorticoid-, acute-phase-, and heat shock-responsive elements.
ESTHER : Klett_1999_J.Neurochem_72_900
PubMedSearch : Klett_1999_J.Neurochem_72_900
PubMedID: 10037460

Title : Localization of the rat M1 muscarinic receptor gene to chromosome 1q43-51 -
Author(s) : Klett CP , Zimonjic DB , Keck CL , Bonner TI
Ref : Mamm Genome , 9 :476 , 1998
PubMedID: 9585439

Title : Upstream sequencing and functional characterization of the human cholinergic gene locus - Hahm_1997_J.Mol.Neurosci_9_223
Author(s) : Hahm SH , Chen L , Patel C , Erickson J , Bonner TI , Weihe E , Schafer MK , Eiden LE
Ref : Journal of Molecular Neuroscience , 9 :223 , 1997
Abstract : The 5' flanking region of the human VAChT gene was sequenced to approx 5350 bases upstream of the initiating methionine codon of the VAChT open reading frame (orf). The 5' flanks of the human and rat cholinergic gene loci were compared to identify regions of local sequence conservation, and therefore of potential regulatory importance. Several discrete domains of high homology, including a cluster of far-upstream cis-active consensus motifs, a neuronally restrictive silencer element consensus sequence, and additional conserved sequences within the putative nerve growth factor response domain of the locus, were identified. The probable start of transcription of the VAChT gene was deduced from mapping of sequences of rat and human VAChT cDNAs onto the 5' flanking regions of the human and rat cholinergic gene loci. The actual utilization of a putative 5' VAChT exon in rat central nervous system (CNS) tissue was assessed by in situ hybridization histochemistry. RNA transcripts containing both VAChT and ChAT protein-coding sequences were abundant in spinal cord motoneurons, sympathetic preganglionic cells, basal forebrain, striatum, and cranial motor nuclei. R-exon-containing transcripts could be detected only at low levels in these cell groups, implying that most transcription of VAChT proceeds from a promoter downstream of the R-exon. To assess the structural requirements for expression of the VAChT gene without bias regarding the actual start of transcription, a 5' fragment of the human gene corresponding to approximately 3 kb of sequence extending upstream from within the presumed 5' untranslated region of VAChT itself was fused to a luciferase-encoding reporter and transfected into VAChT-expressing and nonexpressing human and rat cell lines. This portion of the VAChT gene provided strong promoter expression in both cholinergic and noncholinergic cell lines. Deletion of the putative neuronally restrictive silencer element (NRSE) resulted in enhanced transcription in all cell lines. Lack of differential expression of VAChT transcription in VAChT-expressing vs non-VAChT-expressing cell lines suggested that additional enhancer elements controlling cell-specific expression of the VAChT gene exist further upstream in the cholinergic locus 5' flank. Conservation of potential cis-active elements within a 1.4 kb sequence immediately upstream of the NRSE in both rat and human cholinergic gene loci suggests that this domain is required for cholinergic-specific regulation of VAChT and ChAT gene transcription.
ESTHER : Hahm_1997_J.Mol.Neurosci_9_223
PubMedSearch : Hahm_1997_J.Mol.Neurosci_9_223
PubMedID: 9481623

Title : The VAChT\/ChAT cholinergic gene locus: new aspects of genetic and vesicular regulation of cholinergic function -
Author(s) : Erickson JD , Weihe E , Schafer MK , Neale E , Williamson L , Bonner TI , Tao-Cheng JH , Eiden LE
Ref : Prog Brain Res , 109 :69 , 1996
PubMedID: 9009694

Title : Molecular biology of the vesicular ACh transporter - Usdin_1995_Trends.Neurosci_18_218
Author(s) : Usdin TB , Eiden LE , Bonner TI , Erickson JD
Ref : Trends in Neurosciences , 18 :218 , 1995
Abstract : The cholinergic synapse has long been a model for biochemical studies of neurotransmission. The molecules that are responsible for synaptic transmission are being identified rapidly. The vesicular transporter for ACh, which is responsible for the concentration of ACh within synaptic vesicles, has been characterized recently, both at the molecular and functional level. Definitive identification of the cloned gene involved genetics of Caenorhabditis elegans, the specialized Torpedo electromotor system, and expression in mammalian tissue culture. Comparison of the vesicular transporter for ACh with the vesicular transporters for monoamines demonstrates a new gene family. Gene mapping has demonstrated a unique relationship between the genes for the vesicular ACh transporter and for choline acetyltransferase.
ESTHER : Usdin_1995_Trends.Neurosci_18_218
PubMedSearch : Usdin_1995_Trends.Neurosci_18_218
PubMedID: 7610492

Title : Multipoint genetic linkage analysis of the m2 human muscarinic receptor gene -
Author(s) : Badner JA , Yoon SW , Turner G , Bonner TI , Detera-Wadleigh SD
Ref : Mamm Genome , 6 :489 , 1995
PubMedID: 7579899

Title : Cloning and expression of the vesamicol binding protein from the marine ray Torpedo. Homology with the putative vesicular acetylcholine transporter UNC-17 from Caenorhabditis elegans - Varoqui_1994_FEBS.Lett_342_97
Author(s) : Varoqui H , Diebler MF , Meunier FM , Rand JB , Usdin TB , Bonner TI , Eiden LE , Erickson JD
Ref : FEBS Letters , 342 :97 , 1994
Abstract : Complementary DNA clones corresponding to a messenger RNA encoding a 56 kDa polypeptide have been obtained from Torpedo marmorata and Torpedo ocellata electric lobe libraries, by homology screening with a probe obtained from the putative acetylcholine transporter from the nematode Caenorhabditis elegans. The Torpedo proteins display approximately 50% overall identity to the C. elegans unc-17 protein and 43% identity to the two vesicle monoamine transporters (VMAT1 and VMAT2). This family of proteins is highly conserved within 12 domains which potentially span the vesicle membrane, with little similarity within the putative intraluminal glycosylated loop and at the N- and C-termini. The approximately 3.0 kb mRNA species is specifically expressed in the brain and highly enriched in the electric lobe of Torpedo. The Torpedo protein, expressed in CV-1 fibroblast cells, possesses a high-affinity binding site for vesamicol (Kd = 6 nM), a drug which blocks in vitro and in vivo acetylcholine accumulation in cholinergic vesicles.
ESTHER : Varoqui_1994_FEBS.Lett_342_97
PubMedSearch : Varoqui_1994_FEBS.Lett_342_97
PubMedID: 8143858

Title : Functional identification of a vesicular acetylcholine transporter and its expression from a cholinergic gene locus - Erickson_1994_J.Biol.Chem_269_21929
Author(s) : Erickson JD , Varoqui H , Schafer MK , Modi W , Diebler MF , Weihe E , Rand J , Eiden LE , Bonner TI , Usdin TB
Ref : Journal of Biological Chemistry , 269 :21929 , 1994
Abstract : The vesicular acetylcholine transporter (VAChT) has been identified and characterized based on the acquisition of high affinity vesamicol binding and proton-dependent, vesamicol-sensitive acetylcholine accumulation by a fibroblast cell line transfected with a clone from a rat pheochromocytoma cDNA library encoding this protein. The distribution of VAChT mRNA coincides with that reported for choline acetyltransferase (ChAT), the enzyme required for acetylcholine biosynthesis, in the peripheral and central cholinergic nervous systems. A human VAChT cDNA was used to localize the VAChT gene to chromosome 10q11.2, which is also the location of the ChAT gene. The entire sequence of the human VAChT cDNA is contained uninterrupted within the first intron of the ChAT gene locus. Transcription of VAChT and ChAT mRNA from the same or contiguous promoters within a single regulatory locus provides a previously undescribed genetic mechanism for coordinate regulation of two proteins whose expression is required to establish a mammalian neuronal phenotype.
ESTHER : Erickson_1994_J.Biol.Chem_269_21929
PubMedSearch : Erickson_1994_J.Biol.Chem_269_21929
PubMedID: 8071310

Title : Differential activation of inositol 1,4,5-trisphosphate-sensitive calcium pools by muscarinic receptors in Xenopus laevis oocytes - Ji_1993_Cell.Calcium_14_649
Author(s) : Ji H , Sandberg K , Bonner TI , Catt KJ
Ref : Cell Calcium , 14 :649 , 1993
Abstract : Muscarinic acetylcholine (ACh) receptors activate the phospholipase C signal transduction pathway to promote the formation of inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] and the consequent elevation of cytoplasmic calcium (Ca2+). The inositol phosphate and Ca(2+)-mobilization responses to ACh were analyzed in Xenopus oocytes possessing endogenous receptors, and in oocytes expressing exogenous receptors from injected muscarinic RNA transcripts, to evaluate the patterns of signal transduction mediated by native and expressed receptors. Activation of native ACh receptors elicited dose- and time-dependent increases in Ins(1,4,5)P3 and inositol bisphosphate (InsP2) production. ACh-induced Ins(1,4,5)P3 production increased rapidly within the first 2 min and continued to rise over the next 20 min. ACh was a much more effective stimulus of inositol phosphate production at native (up to 35-fold) than at expressed receptors (less than 2-fold). In contrast, measurements of Ca(2+)-mobilization in oocytes injected with the Ca(2+)-specific photoprotein, aequorin, revealed that ACh stimulation of expressed receptors evoked up to 200-fold increase in light emission, whereas ACh stimulation of native receptors elicited less than a 2-fold response. These observations indicate that the oocyte possesses functionally distinct agonist-sensitive Ca2+ pools which differ markedly in their sensitivity to Ins(1,4,5)P3 production and suggest that these pools are mobilized by different effector mechanisms. The finding that the magnitude of the intra-oocyte Ca2+ response is not necessarily determined by the degree of Ins(1,4,5)P3 production, but rather by another aspect of the signal transduction pathway (e.g. the nature and/or location of the Ins(1,4,5)P3 releasable Ca2+ pool), reveals an additional level of complexity in the transduction mechanisms responsible for intracellular Ca2+ signaling.
ESTHER : Ji_1993_Cell.Calcium_14_649
PubMedSearch : Ji_1993_Cell.Calcium_14_649
PubMedID: 8242720

Title : Genetic linkage mapping of the m4 human muscarinic receptor (CHRM4) -
Author(s) : Grewal RP , Martinez M , Hoehe M , Bonner TI , Gershon ES , Detera-Wadleigh S
Ref : Genomics , 13 :239 , 1992
PubMedID: 1577490

Title : Domains of muscarinic acetylcholine receptors that confer specificity of G protein coupling -
Author(s) : Bonner TI
Ref : Trends in Pharmacological Sciences , 13 :48 , 1992
PubMedID: 1561711

Title : Chimeric m2\/m3 muscarinic receptors: role of carboxyl terminal receptor domains in selectivity of ligand binding and coupling to phosphoinositide hydrolysis - Wess_1990_Mol.Pharmacol_38_872
Author(s) : Wess J , Bonner TI , Brann MR
Ref : Molecular Pharmacology , 38 :872 , 1990
Abstract : The cloning and expression of five mammalian muscarinic receptor genes (m1-m5) have shown that the individual receptor subtypes differ in their functional and ligand-binding properties. To study the role of the carboxyl terminal receptor domains in this pharmacological diversity, we constructed chimeric m2/m3 receptors in which a region comprising part of transmembrane domain VI, the third extracellular loop, transmembrane region VII, and the cytoplasmic tail (collectively referred to as C-terminal domains) was exchanged between the human m2 and the rat m3 receptor. The ability of the cloned receptors to mediate stimulation of phosphoinositide hydrolysis and to bind subtype-selective muscarinic ligands was studied after their transient expression in COS-7 cells. Whereas wild-type m3 strongly stimulated phosphoinositide breakdown, wild-type m2 gave only a poor response. Exchange of the C-terminal domains between m2 and m3 had no significant effect on the magnitude of these responses. In N-[3H]methylscopolamine competition binding studies, the muscarinic antagonists AF-DX 116 and methoctramine showed 11- and 23-fold higher affinities, respectively, for m2 than for m3, whereas hexahydro-silad-ifenidol (HHSiD) and 4-diphenylacetoxy-N-methylpiperidine methiodide (4-DAMP) displayed the reverse selectivity profile, having approximately 10-fold higher affinities for m3. In comparison with wild-type m3, the mutant m3 receptor containing the C-terminal domains of m2 displayed 2.5- and 8-fold higher affinities for AF-DX 116 and methoctramine but 7- and 3-fold lower affinities for HHSiD and 4-DAMP, respectively. The mutant m2 receptor with the C-terminal domains of m3 showed 2-3-fold lower affinities for AF-DX 116 and methoctramine but 2-3-fold higher affinities for HHSiD and 4-DAMP, as compared with wild-type m2. These data suggest that the C-terminal domains of the muscarinic receptors are not involved in conferring selectivity of coupling to phosphoinositide hydrolysis but contain major structural determinants of antagonist binding selectivity.
ESTHER : Wess_1990_Mol.Pharmacol_38_872
PubMedSearch : Wess_1990_Mol.Pharmacol_38_872
PubMedID: 2174507

Title : Delineation of muscarinic receptor domains conferring selectivity of coupling to guanine nucleotide-binding proteins and second messengers - Wess_1990_Mol.Pharmacol_38_517
Author(s) : Wess J , Bonner TI , Dorje F , Brann MR
Ref : Molecular Pharmacology , 38 :517 , 1990
Abstract : The cloning and functional expression of five mammalian muscarinic acetylcholine receptor genes (m1-m5) has revealed that m1, m3, and m5 primarily couple to stimulation of phosphoinositide (PI) turnover, whereas m2 and m4 are strongly linked to inhibition of adenylate cyclase, albeit not exclusively. To identify the structural domains responsible for this functional specificity, cDNAs encoding chimeric m2/m3 receptors were constructed. The abilities of these receptors to mediate stimulation of PI hydrolysis and inhibition of prostaglandin E2-stimulated cAMP accumulation, as well as the pertussis toxin (PTX) sensitivity of these responses, were examined after stable expression in mouse A9 L cells. Substitution of the putative third cytoplasmic loop (i3) of m2 with the corresponding m3 sequence resulted in a chimeric receptor that, similar to m3, stimulated PI breakdown by a PTX-insensitive mechanism but did not inhibit adenylate cyclase. Conversely, a chimeric m3 receptor containing the i3 domain of m2 showed the same functional profile as m2 (i.e., inhibition of adenylate cyclase and weak stimulation of PI turnover by a PTX-sensitive mechanism), indicating that the i3 loop is sufficient to determine coupling selectivity. Similarly, exchange of a short N-terminal segment of i3 (16 or 17 amino acids) between m2 and m3 resulted in chimeric receptors that gained the ability to mediate the functional responses of the wild-type receptor from which the segment was derived, although with substantially reduced efficiency. However, the chimeric m2 receptor containing the 17-amino acid m3 sequence in the N-terminal portion of i3 retained its ability to inhibit adenylate cyclase. Carbachol binding studies involving the use of the GTP analog 5'-guanylyl imidodiphosphate and PTX-pretreated cells generally correlated well with the functional findings. Our data indicate that the N-terminal portion of i3 is a sufficient but not the exclusive determinant of coupling selectivity displayed by the various muscarinic receptors.
ESTHER : Wess_1990_Mol.Pharmacol_38_517
PubMedSearch : Wess_1990_Mol.Pharmacol_38_517
PubMedID: 2172767

Title : An SstI polymorphism for the human muscarinic acetylcholine receptor gene, m4 (CHRM 4) -
Author(s) : Detera-Wadleigh SD , Wiesch D , Bonner TI
Ref : Nucleic Acids Research , 17 :6431 , 1989
PubMedID: 2570410

Title : Poster: Identification of a small intracellular region of the rat m3 receptor responsible for selective coupling to PI turnover -
Author(s) : Wess J , Brann MR , Bonner TI
Ref : Trends in Pharmacological Sciences , Suppl :115 , 1989

Title : New subtypes of muscarinic acetylcholine receptors - Bonner_1989_Trends.Pharmacol.Sci_Suppl_11
Author(s) : Bonner TI
Ref : Trends in Pharmacological Sciences , Suppl :11 , 1989
Abstract : Human and rat genes and/or cDNAs for five different mAChRs have been cloned. The m1, m2 and m3 receptors correspond most closely to the pharmacologically defined M1, M2 and M3 receptors and are expressed in both brain and peripheral tissues. The m4 and m5 receptors are previously unrecognized pharmacological subtypes whose mRNAs are found predominantly in brain. Other less related but uncharacterized genes could represent additional subtypes. The properties of the five receptors and their genes are reviewed and their implications for future research are discussed.
ESTHER : Bonner_1989_Trends.Pharmacol.Sci_Suppl_11
PubMedSearch : Bonner_1989_Trends.Pharmacol.Sci_Suppl_11
PubMedID: 2694516

Title : Poster: Carbachol stimulation causes inhibition of mitogenesis and cell elongation in CHO cells transfected with muscarinrc receptor genes -
Author(s) : Conklin BR , Brann MR , Buckley NJ , Bonner TI , Ma AL , Felder CC , Axelrod J
Ref : Trends in Pharmacological Sciences , Suppl :117 , 1989

Title : The molecular basis of muscarinic receptor diversity - Bonner_1989_Trends.Neurosci_12_148
Author(s) : Bonner TI
Ref : Trends in Neurosciences , 12 :148 , 1989
Abstract : The cloning of cDNAs and genes for five different muscarinic acetylcholine receptors provides a new basis for characterizing muscarinic receptor function. Studies of the cloned receptors when introduced into cells not expressing endogenous receptors have allowed the initial identification of two classes of functional response. The m1, m3 and m5 receptors belong to a class characterized by agonist-induced stimulation of phosphatidylinositol metabolism and are structurally more related to each other than they are to the m2 and m4 receptors, which belong to a class associated with agonist-induced inhibition of adenylate cyclase. While functional differences within these classes may yet be found, it appears likely that much of the difference between functionally similar receptors will be found to lie in their regulation.
ESTHER : Bonner_1989_Trends.Neurosci_12_148
PubMedSearch : Bonner_1989_Trends.Neurosci_12_148
PubMedID: 2470172

Title : Identification of a small intracellular region of the muscarinic m3 receptor as a determinant of selective coupling to PI turnover - Wess_1989_FEBS.Lett_258_133
Author(s) : Wess J , Brann MR , Bonner TI
Ref : FEBS Letters , 258 :133 , 1989
Abstract : Molecular cloning studies have demonstrated the existence of five different muscarinic receptors (m1-m5). While m1, m3 and m5 strongly couple to stimulation of phosphoinositide (PI) hydrolysis, m2 and m4 are more efficiently linked to inhibition of adenylate cyclase. The sequences of m1-m5 have a short segment at the N-terminal portion of the putative third cytoplasmic loop (i3) which is highly conserved among m1, m3 and m5, but different from the sequence which is well conserved among m2 and m4. To study the role of this region in conferring coupling selectivity, we constructed cDNAs encoding chimeric m2/m3 receptors. Transient expression of these receptor hybrids in COS-7 cells showed that a 17 amino acid segment at the N-terminal portion of i3 is a major determinant of how efficiently the different muscarinic receptors are coupled to PI hydrolysis.
ESTHER : Wess_1989_FEBS.Lett_258_133
PubMedSearch : Wess_1989_FEBS.Lett_258_133
PubMedID: 2556294

Title : Antagonist binding properties of five cloned muscarinic receptors expressed in CHO-K1 cells - Buckley_1989_Mol.Pharmacol_35_469
Author(s) : Buckley NJ , Bonner TI , Buckley CM , Brann MR
Ref : Molecular Pharmacology , 35 :469 , 1989
Abstract : A family of five cholinergic muscarinic receptor genes (m1, m2, m3, m4, and m5) has recently been identified and cloned. In order to investigate the pharmacological properties of the individual muscarinic receptors, we have transfected each of these genes into Chinese hamster ovary cells (CHO-K1) and have established stable cell lines expressing each receptor. In the present study we have examined the antagonist binding properties of each muscarinic receptor. Antagonists were chosen that had previously been proposed to be selective for muscarinic receptor subtypes and included pirenzepine, AF-DX 116, methoctramine, dicyclomine, hexohydrodifenidol, hexahydrosiladifenidol, hexocyclium, and silahexocyclium. m1, m2, and m3 receptors express binding properties similar to those expected of high affinity pirenzepine-type receptors of cerebral cortex ("M1"), low affinity pirenzepine-type receptors of atria ("M2 cardiac type"), and the intermediate affinity pirenzepine-type receptors found in exocrine glands ("M2 glandular type"), respectively. The M1/M2 schema cannot readily accommodate the binding properties of the m4 and m5 receptors. Pirenzepine, methoctramine, and hexahydrosiladifenidol were the most selective agents for the m1, m2, and m3 receptors, respectively. None of the antagonists used in this study were uniquely selective for either the m4 or m5 receptors. The diverse binding profiles of individual cloned receptors and the widespread distribution of m1-m4 mRNAs indicate that radioligand binding studies performed on primary tissues may actually be assessing the composite properties of a heterogeneous mixture of muscarinic receptor subtypes.
ESTHER : Buckley_1989_Mol.Pharmacol_35_469
PubMedSearch : Buckley_1989_Mol.Pharmacol_35_469
PubMedID: 2704370

Title : Cloning and expression of the human and rat m5 muscarinic acetylcholine receptor genes - Bonner_1988_Neuron_1_403
Author(s) : Bonner TI , Young AC , Brann MR , Buckley NJ
Ref : Neuron , 1 :403 , 1988
Abstract : The human and rat genes for a fifth muscarinic receptor have been cloned and expressed in mammalian cells. The 532 amino acid human protein has 89% sequence identity to the 531 amino acid rat protein and is most closely related to the m3 receptor. Both proteins are encoded by single exons. The receptor has intermediate affinity for pirenzepine and low affinity for AF-DX 116, and it increases metabolism of phosphatidylinositol when stimulated with carbachol. Expression of mRNA has yet to be observed in brain or selected peripheral tissues, suggesting that either it is substantially less abundant than m1-m4 or its distribution is quite different.
ESTHER : Bonner_1988_Neuron_1_403
PubMedSearch : Bonner_1988_Neuron_1_403
PubMedID: 3272174

Title : The striatum and cerebral cortex express different muscarinic receptor mRNAs - Brann_1988_FEBS.Lett_230_90
Author(s) : Brann MR , Buckley NJ , Bonner TI
Ref : FEBS Letters , 230 :90 , 1988
Abstract : The existence of four distinct muscarinic acetylcholine receptor genes (m1-mr) has recently been demonstrated. cDNAs for three of these receptors have been cloned from brain (m1, m3, m4) and one from heart (m2). To gain some understanding of the physiological role of the brain muscarinic receptors, we mapped the distribution of their mRNAs in rat brain by in situ hybridization. These mRNAs are barely detectable in the hindbrain and cerebellum. Within forebrain, each mRNA has a strikingly different pattern of distribution. The highest levels of m1 mRNA are in the cerebral cortex and hippocampus followed by the striatum. m3 mRNA is also prominent in the cerebral cortex, but has very low levels in the striatum. Conversely, the levels of m4 mRNA are highest in the striatum. Since the cognitive effects of muscarinic drugs have been localized to the cerebral cortex and hippocampus, and their psychomotor effects to the striatum, these data suggest that the muscarinic receptors which subserve these responses may be different gene products. Finally, we show that these muscarinic receptors can be distinguished pharmacologically, suggesting that it may be possible to develop drugs for the selective treatment of the psychomotor vs cognitive difficulties of Parkinson's and Alzheimer's disease, respectively.
ESTHER : Brann_1988_FEBS.Lett_230_90
PubMedSearch : Brann_1988_FEBS.Lett_230_90
PubMedID: 3350153

Title : Electrophysiological characterization of cloned m1 muscarinic receptors expressed in A9 L cells - Jones_1988_Proc.Natl.Acad.Sci.U.S.A_85_4056
Author(s) : Jones SV , Barker JL , Bonner TI , Buckley NJ , Brann MR
Ref : Proc Natl Acad Sci U S A , 85 :4056 , 1988
Abstract : The electrophysiological properties of A9 L cells stably transfected with m1 muscarinic receptor cDNA were examined by using the whole-cell patch-clamp technique. In current-clamp recordings, acetylcholine (AcCho) elicited a hyperpolarization of all transfected cells studied but had no effect on nontransfected A9 L cells. In voltage-clamp recordings, AcCho elicited an outward current at -50 mV accompanied by an increase in conductance. The onset of the current response was consistently delayed by several seconds with respect to the onset of the application of AcCho and could not be accounted for by diffusion. The AcCho-induced currents were reversibly inhibited by the muscarinic receptor antagonist atropine (1 microM) but were unaffected by the nicotinic receptor antagonist tubocurarine (50 microM). Ion-substitution experiments replacing K+ with N-methyl-D-glucamine and Cl- with methanesulfonate indicated that the current was carried mainly by K+, although a minor part appeared to be carried by Cl-. The AcCho-induced current could be blocked by the K+ channel blocking agents tetraethylammonium ion, 4-aminopyridine, apamin, and Ba2+ but not by Cs+. The AcCho-induced current was inhibited when 5 mM 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA) was included in the patch pipette or when extracellular Cd2+ or Co2+ was applied, indicating a role for intracellular Ca2+ in the generation of the response. Thus, these results show that cloned m1 muscarinic receptors expressed in A9 L cells can activate a Ca2+-dependent K+ conductance, possibly via a second-messenger system.
ESTHER : Jones_1988_Proc.Natl.Acad.Sci.U.S.A_85_4056
PubMedSearch : Jones_1988_Proc.Natl.Acad.Sci.U.S.A_85_4056
PubMedID: 2453885

Title : Localization of a family of muscarinic receptor mRNAs in rat brain - Buckley_1988_J.Neurosci_8_4646
Author(s) : Buckley NJ , Bonner TI , Brann MR
Ref : Journal of Neuroscience , 8 :4646 , 1988
Abstract : A family of 4 rat muscarinic receptors (m1, m2, m3, and m4) have recently been cloned and sequenced (Bonner et al., 1987). Since pharmacological probes that are presently available do not appear to distinguish among 3 of these muscarinic receptors, we constructed oligonucleotide probes corresponding to the N-terminal sequences of the muscarinic receptors and used them to specifically localize m1, m2, m3, and m4 mRNA in sections of rat brain using in situ hybridization histochemistry. Northern analysis demonstrated a 3.1 kilobase (kb) m 1 mRNA, a 4.5 kb m3 mRNA, and a 3.3 kb m4 mRNA in cerebral cortex, striatum, hippocampus, and cerebellum. In situ hybridization histochemistry indicated a prevalence of m1 mRNA in the pyramidal cell layer of the hippocampus, the granule cell layer of the dentate gyrus, the olfactory bulb, amygdala, olfactory tubercule, and piriform cortex. Caudate putamen and cerebral cortex showed moderate levels of labeling. m2 mRNA was detectable in the medial septum, diagonal band, olfactory bulb, and pontine nuclei. m3 and m4 mRNA were also prevalent in the olfactory bulb and pyramidal cell layer of the hippocampus but were present only in low levels in the dentate gyrus. m3 mRNA was present in superficial and deep layers of the cerebral cortex, whereas m4 mRNA was more evenly distributed with a slightly more intense labeling evident in the midcortical layer. In addition, m3 mRNA was present in a number of thalamic nuclei and brain-stem nuclei, while m4 mRNA predominated in the caudate putamen. These data offer a new basis on which to interpret the heterogeneity of muscarinic actions in the CNS.
ESTHER : Buckley_1988_J.Neurosci_8_4646
PubMedSearch : Buckley_1988_J.Neurosci_8_4646
PubMedID: 3199198

Title : Cloned muscarinic receptor subtypes expressed in A9 L cells differ in their coupling to electrical responses - Jones_1988_Mol.Pharmacol_34_421
Author(s) : Jones SV , Barker JL , Buckley NJ , Bonner TI , Collins RM , Brann MR
Ref : Molecular Pharmacology , 34 :421 , 1988
Abstract : The electrophysiological responses to cholinergic stimulation of four cloned muscarinic receptor subtypes (m1-m4) were studied in A9 L cells transfected with the expression plasmids of each of the different subtypes, using the tight-seal whole-cell recording technique. Cells transfected with m1 and m3 muscarinic receptor subtypes were hyperpolarized by acetylcholine (ACh), whereas m2- and m4-transfected cells did not respond to ACh concentrations of up to 1 mM. Stimulation of both m1 and m3 muscarinic receptor subtypes evoked outward currents in cells voltage-clamped at -50 mV, associated with an increase in membrane conductance. These outward currents were blocked by atropine but not by tubocurarine. The ACh-induced currents of m1- and m3-transfected cells primarily involved potassium ions, although chloride ions also contributed to a minor extent. The potassium and chloride conductances were blocked by barium or cobalt and by buffering the intracellular calcium to low levels with 5 mM 1,2-bis(2-aminophenoxy)ethane-N,N,N'N'-tetraacetic acid, showing a dependence of these conductances on calcium. Thus, m1- and m3-transfected cells respond to ACh in a manner that is qualitatively similar, evoking calcium-dependent potassium and chloride conductances, whereas m2- and m4-transfected cells are not coupled to electrically detectable responses in A9 L cells.
ESTHER : Jones_1988_Mol.Pharmacol_34_421
PubMedSearch : Jones_1988_Mol.Pharmacol_34_421
PubMedID: 3173332

Title : Stimulation of arachidonic acid release and inhibition of mitogenesis by cloned genes for muscarinic receptor subtypes stably expressed in A9 L cells - Conklin_1988_Proc.Natl.Acad.Sci.U.S.A_85_8698
Author(s) : Conklin BR , Brann MR , Buckley NJ , Ma AL , Bonner TI , Axelrod J
Ref : Proc Natl Acad Sci U S A , 85 :8698 , 1988
Abstract : A family of genes encoding four distinct muscarinic receptors (designated m1-m4) has been cloned and stably expressed in A9 L cells. When the m1 and m3 receptors were stimulated with carbachol, there was a rapid rise of liberated arachidonic acid, inositol phosphates, and cAMP, while m2 and m4 receptor stimulation had no detectable stimulation of these second messengers. Pretreatment with phorbol 12-myristate 13-acetate (PMA) caused a marked acceleration and amplification of m1 and m3 receptor-mediated arachidonic acid release. In contrast, m1- and m3-mediated inositol phosphate formation was inhibited by the same PMA pretreatment. Arachidonic acid release was unaffected by manipulations of cAMP levels. Arachidonic acid production was inhibited by calcium-free medium and 3,4,5-trimethoxybenzoic acid 8-(diethylamino)octyl ester (TMB-8; an inhibitor of cytosolic calcium mobilization) yet was unaffected by verapamil, a calcium-channel blocker. These experiments show that arachidonic acid release induced by the m1 and m3 receptors is regulated independently of phospholipase C and cAMP accumulation. Carbachol stimulation of the m1 and m3 cAMP accumulation. Carbachol stimulation of the m1 and m3 receptors also markedly decreased mitogenesis as measured by thymidine incorporation. The m1 receptor-mediated inhibition of mitogenesis could be partially blocked by indomethacin, a cyclooxygenase inhibitor. The inhibition of mitogenesis could be mimicked by cAMP elevation.
ESTHER : Conklin_1988_Proc.Natl.Acad.Sci.U.S.A_85_8698
PubMedSearch : Conklin_1988_Proc.Natl.Acad.Sci.U.S.A_85_8698
PubMedID: 2847172

Title : Identification of a family of muscarinic acetylcholine receptor genes - Bonner_1987_Science_237_527
Author(s) : Bonner TI , Buckley NJ , Young AC , Brann MR
Ref : Science , 237 :527 , 1987
Abstract : Complementary DNAs for three different muscarinic acetylcholine receptors were isolated from a rat cerebral cortex library, and the cloned receptors were expressed in mammalian cells. Analysis of human and rat genomic clones indicates that there are at least four functional muscarinic receptor genes and that these genes lack introns in the coding sequence. This gene family provides a new basis for evaluating the diversity of muscarinic mechanisms in the nervous system.
ESTHER : Bonner_1987_Science_237_527
PubMedSearch : Bonner_1987_Science_237_527
PubMedID: 3037705

Title : Expression of a cloned muscarinic receptor in A9 L cells - Brann_1987_Mol.Pharmacol_32_450
Author(s) : Brann MR , Buckley NJ , Jones SV , Bonner TI
Ref : Molecular Pharmacology , 32 :450 , 1987
Abstract : Using an oligonucleotide based on the sequence of a porcine brain muscarinic receptor cDNA, we recently cloned four distinct muscarinic receptors from the rat and human genomes. In the present study we transfected the rat homolog of the porcine brain muscarinic receptor cDNA into A9 L cells using a mammalian expression vector and a calcium phosphate precipitation procedure. Before transfection, A9 L cells do not bind muscarinic ligands and do not express muscarinic receptor mRNA. After transfection, A9 L cells expressed muscarinic receptor mRNA and saturable, high affinity binding sites for the muscarinic antagonists 3H-quinuclidinyl benzilate and 3H-pirenzepine. The muscarinic receptor antagonists AF DX-116 and pirenzepine displaced bound 3H-quinuclidinyl benzilate with inhibition curves suggestive of a single high affinity binding site. Competition of 3H-quinuclidinyl benzilate-labeled sites with the agonists acetylcholine and carbachol yielded broad inhibition curves, consistent with a heterogeneity of binding sites. In the presence of guanine nucleotide, the agonist inhibition curves were steeper, suggesting the presence of a single low affinity site. The effects of guanine nucleotides on agonist binding are consistent with coupling of these receptors to a guanine nucleotide-binding protein (G-protein) endogenous to A9 L cells. The electrical properties of the transfected A9 L cells were examined using the whole cell patch-clamp technique. Fifty microM acetylcholine induced a conductance which reversed in polarity at -60 mV. This conductance could be reversibly blocked by atropine. These data illustrate the utility of stable transfection of A9 L cells for the characterization of individual cloned muscarinic receptors, their G-protein coupling mechanisms, and resultant physiological responses.
ESTHER : Brann_1987_Mol.Pharmacol_32_450
PubMedSearch : Brann_1987_Mol.Pharmacol_32_450
PubMedID: 2444870