Koulakoff A

References (19)

Title : Genetic and epigenetic control of the Na-G ion channel expression in glia - Gautron_2001_Glia_33_230
Author(s) : Gautron S , Gruszczynski C , Koulakoff A , Poiraud E , Lopez S , Cambier H , Dos Santos G , Berwald-Netter Y
Ref : Glia , 33 :230 , 2001
Abstract : The Na-G ion channel, previously cloned from a rat astroglia cDNA library, belongs to a new family of ion channels, related to but distinct from the predominant brain and muscle fast voltage-gated Na(+) channels. In vivo, the corresponding transcripts are widely expressed in peripheral nervous system neurons and glia, but only in selected subpopulations of neuronal and glia-like cells of the central nervous system. In the present report, we show that Na-G messenger RNA level in astrocyte and Schwann cell cultures is modulated in a cell-specific manner by several growth factors, hormones, and intracellular second messengers pathways. Striking changes in transcript level were observed in the two types of glia in response to protein-kinase A activation and to treatment with the neuregulin glial growth factor, indicating regulation of the Na-G gene by neuroglial signaling. By transient transfection of Na-G/reporter constructs into cultured cells, we show that a short genomic region, encompassing the first exon and 375 bp upstream, bears a high glial-specific transcriptional activity while part of the first intron behaves as a negative regulatory element. In vivo footprinting experiments revealed binding of glial-specific nuclear factors to several sites of the Na-G promoter region. Finally, Na-G/reporter constructs are shown to sustain a low but reproducible transcriptional response to cAMP, accounting in part for the elevation in mRNA level elicited by cAMP in Schwann cells and its reduction in astrocytes.
ESTHER : Gautron_2001_Glia_33_230
PubMedSearch : Gautron_2001_Glia_33_230
PubMedID: 11241741

Title : The Na-G ion channel is transcribed from a single promoter controlled by distinct neuron- and Schwann cell-specific DNA elements - Poiraud_1999_J.Neurochem_73_2575
Author(s) : Poiraud E , Gruszczynski C , Porteu A , Cambier H , Escurat M , Koulakoff A , Kahn A , Berwald-Netter Y , Gautron S
Ref : Journal of Neurochemistry , 73 :2575 , 1999
Abstract : Na-G is a putative sodium (or cationic) channel expressed in neurons and glia of the PNS, in restricted neuronal subpopulations of the brain, and in several tissues outside the nervous system, like lung and adrenal medulla. To analyze the mechanisms underlying tissue-specific expression of this channel, we isolated the 5' region of the corresponding gene and show that Na-G mRNA transcription proceeds from a single promoter with multiple initiation sites. By transgenic mice studies, we demonstrate that 600 bp containing the Na-G proximal promoter region and the first exon are sufficient to drive the expression of a beta-galactosidase reporter gene in neurons of both CNS and PNS, whereas expression in Schwann cells depends on more remote DNA elements lying in the region between -6,500 and -1,050 bp upstream of the main transcription initiation sites. Crucial elements for lung-specific expression seem to be located in the region between -1,050 and -375 bp upstream of the promoter. Using in vivo footprint experiments, we demonstrate that several sites of the Na-G proximal promoter region are bound specifically by nuclear proteins in dorsal root ganglion neurons, as compared with nonexpressing hepatoma cells.
ESTHER : Poiraud_1999_J.Neurochem_73_2575
PubMedSearch : Poiraud_1999_J.Neurochem_73_2575
PubMedID: 10582621

Title : Doublecortin is a developmentally regulated, microtubule-associated protein expressed in migrating and differentiating neurons - Francis_1999_Neuron_23_247
Author(s) : Francis F , Koulakoff A , Boucher D , Chafey P , Schaar B , Vinet MC , Friocourt G , McDonnell N , Reiner O , Kahn A , McConnell SK , Berwald-Netter Y , Denoulet P , Chelly J
Ref : Neuron , 23 :247 , 1999
Abstract : Recently, we and others reported that the doublecortin gene is responsible for X-linked lissencephaly and subcortical laminar heterotopia. Here, we show that Doublecortin is expressed in the brain throughout the period of corticogenesis in migrating and differentiating neurons. Immunohistochemical studies show its localization in the soma and leading processes of tangentially migrating neurons, and a strong axonal labeling is observed in differentiating neurons. In cultured neurons, Doublecortin expression is highest in the distal parts of developing processes. We demonstrate by sedimentation and microscopy studies that Doublecortin is associated with microtubules (MTs) and postulate that it is a novel MAP. Our data suggest that the cortical dysgeneses associated with the loss of Doublecortin function might result from abnormal cytoskeletal dynamics in neuronal cell development.
ESTHER : Francis_1999_Neuron_23_247
PubMedSearch : Francis_1999_Neuron_23_247
PubMedID: 10399932

Title : A novel CNS gene required for neuronal migration and involved in X-linked subcortical laminar heterotopia and lissencephaly syndrome - des Portes_1998_Cell_92_51
Author(s) : des Portes V , Pinard JM , Billuart P , Vinet MC , Koulakoff A , Carrie A , Gelot A , Dupuis E , Motte J , Berwald-Netter Y , Catala M , Kahn A , Beldjord C , Chelly J
Ref : Cell , 92 :51 , 1998
Abstract : X-SCLH/LIS syndrome is a neuronal migration disorder with disruption of the six-layered neocortex. It consists of subcortical laminar heterotopia (SCLH, band heterotopia, or double cortex) in females and lissencephaly (LIS) in males, leading to epilepsy and cognitive impairment. We report the characterization of a novel CNS gene encoding a 40 kDa predicted protein that we named Doublecortin and the identification of mutations in four unrelated X-SCLH/LIS cases. The predicted protein shares significant homology with the N-terminal segment of a protein containing a protein kinase domain at its C-terminal part. This novel gene is highly expressed during brain development, mainly in fetal neurons including precursors. The complete disorganization observed in lissencephaly and heterotopia thus seems to reflect a failure of early events associated with neuron dispersion.
ESTHER : des Portes_1998_Cell_92_51
PubMedSearch : des Portes_1998_Cell_92_51
PubMedID: 9489699

Title : Developmental regulation of polyglutamylated alpha- and beta-tubulin in mouse brain neurons - Audebert_1994_J.Cell.Sci_107 ( Pt 8)_2313
Author(s) : Audebert S , Koulakoff A , Berwald-Netter Y , Gros F , Denoulet P , Edde B
Ref : Journal of Cell Science , 107 ( Pt 8) :2313 , 1994
Abstract : Polyglutamylation is an important posttranslational modification of tubulin that is very active in nerve cells, where it accounts for the main factor responsible for tubulin heterogeneity. In the present work, we have analyzed quantitative and qualitative changes in glutamylated alpha- and beta-tubulin occurring during neuronal differentiation in culture. Glutamylated alpha- and beta-tubulin both markedly accumulate during this process with a time course remarkably similar to that observed in vivo during brain development. However, the characteristics of the glutamylation of the two subunits are not exactly the same. Glutamylated alpha-tubulin is already abundant in very young neurons and displays, at this stage, a wide range of its degree of glutamylation (1 to 6 glutamyl units present in the lateral polyglutamyl chain), which remains unchanged during the entire period of the culture. Glutamylated beta-tubulin is present at very low levels in young neurons and its accumulation during differentiation is accompanied by a progressive increase in its degree of glutamylation from 2 to 6 glutamyl units. Posttranslational incorporation of [3H]glutamate into alpha- and beta-tubulin decreases during differentiation, as well as the rate of the reverse deglutamylation reaction, suggesting that accumulation of glutamylated tubulin is accompanied by a decrease in the turnover of glutamyl units onto tubulin. Neuronal differentiation is also accompanied by an increase of other posttranslationally modified forms of tubulin, including acetylated and non-tyrosinatable alpha-tubulin, which can occur in combination with polyglutamylation and contributes to increase the complexity of tubulin in mature neurons.
ESTHER : Audebert_1994_J.Cell.Sci_107 ( Pt 8)_2313
PubMedSearch : Audebert_1994_J.Cell.Sci_107 ( Pt 8)_2313
PubMedID: 7527057

Title : Expression of the transcripts initiated in the 62nd intron of the dystrophin gene - Lambert_1993_Neuromuscul.Disord_3_519
Author(s) : Lambert M , Chafey P , Hugnot JP , Koulakoff A , Berwald-Netter Y , Billard C , Morris GE , Kahn A , Kaplan JC , Gilgenkrantz H
Ref : Neuromuscular Disorders , 3 :519 , 1993
Abstract : The pattern of expression of two distal transcripts initiated in the 62nd intron of the dystrophin gene was investigated under different circumstances; (i) during the development of different rat tissues these transcripts and Dp71, a protein encoded by one of them, increased with brain development and decreased with muscle development; (ii) in cultured glial and neuronal cells, the distal promoter was coactivated with tissue-specific upstream promoters, the muscle-type promoter in glial cells and the brain-type promoter in neuronal cells, which suggests that activity of the upstream promoter does not interfere with activity of the distal promoter; (iii) in lymphoblasts of DMD patients with various deletions of the dystrophin gene, the most distal of which included the 56th intron, the production of the distal transcript was not perturbed.
ESTHER : Lambert_1993_Neuromuscul.Disord_3_519
PubMedSearch : Lambert_1993_Neuromuscul.Disord_3_519
PubMedID: 8186704

Title : Adenoviral vector as a gene delivery system into cultured rat neuronal and glial cells - Caillaud_1993_Eur.J.Neurosci_5_1287
Author(s) : Caillaud C , Akli S , Vigne E , Koulakoff A , Perricaudet M , Poenaru L , Kahn A , Berwald-Netter Y
Ref : European Journal of Neuroscience , 5 :1287 , 1993
Abstract : Previous studies have demonstrated that a defective recombinant adenovirus can infect a wide range of postmitotic and slowly proliferating cell types such as hepatocytes, myotubes, pneumocytes and intestinal cells (Stratford-Perricaudet et al., Hum. Gene Ther., 1, 241-256, 1990; Quantin et al., Proc. Natl. Acad. Sci. USA, 89, 2581-2584, 1992; Jaffe et al., Nature Genetics, 1, 372-378, 1992). We have used a defective recombinant adenovirus, Ad.RSV beta gal, containing the Escherichia coli beta-galactosidase gene targeted to the nucleus under the transcriptional control of the Rous sarcoma virus long terminal repeat promoter (Stratford-Perricaudet et al., J. Clin. Invest., 90, 626-630, 1992) to infect non-dividing neural cells in primary culture. We show that 80-100% of neuronal and astroglial cells infected with a viral titre lower than 10(9) p.f.u./ml express beta-galactosidase for at least 1 month without cell damage. These results demonstrate the potential usefulness of recombinant adenovirus infection for the analysis of brain-specific gene regulation and for the transfer of genes into neural cells before their transplantation into the brain.
ESTHER : Caillaud_1993_Eur.J.Neurosci_5_1287
PubMedSearch : Caillaud_1993_Eur.J.Neurosci_5_1287
PubMedID: 8275231

Title : The glial voltage-gated sodium channel: cell- and tissue-specific mRNA expression - Gautron_1992_Proc.Natl.Acad.Sci.U.S.A_89_7272
Author(s) : Gautron S , Dos Santos G , Pinto-Henrique D , Koulakoff A , Gros F , Berwald-Netter Y
Ref : Proc Natl Acad Sci U S A , 89 :7272 , 1992
Abstract : Previous electrophysiological and pharmacological studies on central and peripheral glia revealed the presence of voltage-gated Na channels with properties that are similar but not identical to those of neuronal Na channels. Here we report the isolation and characterization of a cDNA encoding the C-terminal portion of a putative glial Na-channel (Na-G) alpha subunit. The amino acid sequence deduced from this cDNA indicates that the Na-G represents a separate molecular class within the mammalian Na-channel multigene family. By Northern blot, RNase protection, and in situ hybridization assays, we demonstrate that, in addition to brain astroglia, the Na-G mRNA is expressed in cultures of Schwann cells derived from dorsal root ganglia or from sciatic nerve. In vivo, the Na-G mRNA is detected not only in brain, dorsal root ganglia, and sciatic nerve, but also in tissues outside the nervous system including cardiac and skeletal muscle and lung. Its level varies according to the tissue and is developmentally regulated. The sequence and expression data concur in designating Na-G as an distinct type of Na channel, presumably with low sensitivity to tetrodotoxin.
ESTHER : Gautron_1992_Proc.Natl.Acad.Sci.U.S.A_89_7272
PubMedSearch : Gautron_1992_Proc.Natl.Acad.Sci.U.S.A_89_7272
PubMedID: 1379737

Title : Distal transcript of the dystrophin gene initiated from an alternative first exon and encoding a 75-kDa protein widely distributed in nonmuscle tissues - Hugnot_1992_Proc.Natl.Acad.Sci.U.S.A_89_7506
Author(s) : Hugnot JP , Gilgenkrantz H , Vincent N , Chafey P , Morris GE , Monaco AP , Berwald-Netter Y , Koulakoff A , Kaplan JC , Kahn A , et al.
Ref : Proc Natl Acad Sci U S A , 89 :7506 , 1992
Abstract : A transcript generated by the distal part of the Duchenne Muscular Dystrophy (DMD) gene was initially detected in cells where the full size 14-kilobase (kb) messenger RNA is not found at a significant level. This transcript, approximately 4.5 kb long, corresponds to the cysteine-rich and carboxyl-terminal domains of dystrophin. It begins with a novel 80- to 100-nucleotide exon containing an ATG start site for a new coding sequence of 17 nucleotides in-frame with the consecutive dystrophin cDNA sequence from exon 63. This result suggests the existence of a third promoter that would be localized about 8 kilobases upstream from exon 63 of the DMD gene. The distal transcript is widely distributed but is absent in adult skeletal and myometrial muscle. It is much more abundant in fetal tissues. With an antibody directed against the dystrophin carboxyl terminus, the protein corresponding to this transcript was detected as a 70- to 75-kDa entity on Western blots. It was found in all tissues analyzed except in skeletal muscle. It was not found in lymphoblastoid cells from a Duchenne patient with a complete deletion of the dystrophin gene. The role and subcellular localization of this protein is not known. It may explain extramuscular symptoms exhibited by some Duchenne patients.
ESTHER : Hugnot_1992_Proc.Natl.Acad.Sci.U.S.A_89_7506
PubMedSearch : Hugnot_1992_Proc.Natl.Acad.Sci.U.S.A_89_7506
PubMedID: 1380160

Title : A combination of posttranslational modifications is responsible for the production of neuronal alpha-tubulin heterogeneity - Edde_1991_J.Cell.Biochem_46_134
Author(s) : Edde B , Rossier J , Le Caer JP , Berwald-Netter Y , Koulakoff A , Gros F , Denoulet P
Ref : Journal of Cellular Biochemistry , 46 :134 , 1991
Abstract : We describe the presence of alpha-tubulin and MAP2 acetyltransferase activities in mouse brain. The enzyme(s) copurified with microtubules through two cycles of assembly-disassembly. Incubation of microtubule proteins with [3H]acetyl CoA resulted in a strong labeling of both alpha-tubulin and MAP2. To determine the site of the modification, tubulin was purified and digested with Glu-C endoproteinase. A unique radioactive peptide was detected and purified by HPLC. Edman degradation sequencing showed that this peptide contained epsilon N-acetyllysine at position 40 of the alpha-tubulin molecule. This result demonstrates that mouse brain alpha-tubulin was acetylated at the same site as in Chlamydomonas. Isoelectric focusing analysis showed that acetylated alpha-tubulin was resolved into five isoelectric variants, denoted alpha 3 and alpha 5 to alpha 8. This heterogeneity is not due to acetylation of other sites but results from a single acetylation of Lys40 of an heterogeneous population of alpha-tubulin isoforms. These isoforms are produced by posttranslational addition of one to five glutamyl units. Thus, neuronal alpha-tubulin is extensively modified by a combination of modifications including acetylation, glutamylation, tyrosylation, and other yet unknown modifications.
ESTHER : Edde_1991_J.Cell.Biochem_46_134
PubMedSearch : Edde_1991_J.Cell.Biochem_46_134
PubMedID: 1680872

Title : Dystrophin gene transcribed from different promoters in neuronal and glial cells - Chelly_1990_Nature_344_64
Author(s) : Chelly J , Hamard G , Koulakoff A , Kaplan JC , Kahn A , Berwald-Netter Y
Ref : Nature , 344 :64 , 1990
Abstract : It has been shown that the dystrophin gene, which is defective in patients with Duchenne and Becker muscular dystrophy (reviewed in ref. 1), is transcribed in brain from a specific promoter that is different from the one used in muscle, and so the two types of transcripts differ at least in their first exon. We recently found that the dystrophin gene is expressed at a higher level in primary cultures of neuronal cells than in astro-glial cells derived from adult mouse brain. Here we investigate the use of two different promoters in each cell type. Our results demonstrate that the brain-type promoter of the dystrophin gene is highly specific to neurons, in which there is a significant increase in the amount of brain-specific messenger RNA during the course of in vitro maturation. By contrast, the muscle-type promoter is active in a wider range of cell types, including not only striated and smooth muscle, but also glial cells to a lesser extent, and probably neurons.
ESTHER : Chelly_1990_Nature_344_64
PubMedSearch : Chelly_1990_Nature_344_64
PubMedID: 2406613

Title : Posttranslational modifications of tubulin in cultured mouse brain neurons and astroglia - Edde_1989_Biol.Cell_65_109
Author(s) : Edde B , Denoulet P , de Nechaud B , Koulakoff A , Berwald-Netter Y , Gros F
Ref : Biology of the cell , 65 :109 , 1989
Abstract : Posttranslational modifications of tubulin were analyzed in mouse brain neurons and glia developing in culture. Purified tubulin was resolved by isoelectric focusing. After 3 weeks of culture, neurons were shown to express a high degree of tubulin heterogeneity (8 alpha and 10 beta isoforms), similar to that found in the brain at the same developmental stage. Astroglial tubulin exhibits a less complex pattern consisting of 4 alpha and 4 beta isoforms. After incubation of neuronal and glial cells with 3H-acetate in the presence of cycloheximide, a major posttranslational label was found associated with alpha-tubulin and a minor one with beta-tubulin. The acetate-labeled isotubulins of neurons were resolved by isoelectric focusing into as many as 6 alpha and 7 beta isoforms, while those of astroglia were resolved into only 2 alpha and 2 beta isoforms. The same alpha isoforms were also shown to react with a monoclonal antibody recognizing selectively the acetylated form(s) of alpha-tubulin. Whether acetate-labeling of alpha-tubulin in these cells corresponds to the acetylation of Lys40, as reported for Chlamydomonas reinhardtii, is discussed according to very recent data obtained by protein sequence analysis. Tubulin phosphorylation was analyzed by incubation of cell cultures with 32PO4. No phosphorylation of alpha-tubulin isoforms was detected. A single beta-tubulin isoform (beta'2), expressed only in neurons, was found to be phosphorylated. This isoform is similar to that previously identified in differentiated mouse neuroblastoma cells.
ESTHER : Edde_1989_Biol.Cell_65_109
PubMedSearch : Edde_1989_Biol.Cell_65_109
PubMedID: 2736326

Title : Neurotoxin-sensitive sodium channels in neurons developing in vivo and in vitro - Couraud_1986_J.Neurosci_6_192
Author(s) : Couraud F , Martin-Moutot N , Koulakoff A , Berwald-Netter Y
Ref : Journal of Neuroscience , 6 :192 , 1986
Abstract : Fetal mouse brain cells were investigated by 22Na+ flux assays with the aim to determine the ontogenetic time course of appearance of functional voltage-sensitive sodium channels. Their pharmacological properties were assessed by measurement of the response to known neurotoxins, acting at site 1, 2, or 3 of the Na+ channel. Brain cell suspensions, prepared at 11-19 d of prenatal development in vivo, and fetal brain neurons in culture were explored. In vivo neurotoxin-sensitive Na+ influx becomes detectable at 12 d of gestation, in concordance with the time of appearance of saturable binding sites for alpha-scorpion toxin (alpha-ScTx) and saxitoxin. Progression in fetal age or in time in vitro is accompanied by an increase in the initial rate and in the amplitude of Na+ uptake stimulated by batrachotoxin or veratridine. The general pharmacological properties of developing Na+ channels are very similar to the known properties of voltage-dependent Na+ channels in adult nerve: Batrachotoxin acts as a full channel agonist and veratridine as a partial agonist. Their respective apparent affinities are increased in presence of alpha-ScTx, in agreement with the known positive cooperativity of toxins acting at sites 2 and 3 of the Na+ channel. alpha-ScTx alone induces a small increase in Na+ permeability; its effect is greatly amplified in the presence of batrachotoxin or veratridine. The apparent affinity of alpha-ScTx is reduced by cell depolarization. Tetrodotoxin and saxitoxin block the increase in Na+ permeability induced by batrachotoxin, veratridine, and alpha-ScTx.(ABSTRACT TRUNCATED AT 250 WORDS)
ESTHER : Couraud_1986_J.Neurosci_6_192
PubMedSearch : Couraud_1986_J.Neurosci_6_192
PubMedID: 2418173

Title : Ultrastructural visualization of Na+-channel associated [125I]alpha-scorpion toxin binding sites on fetal mouse nerve cells in culture - Boudier_1985_Brain.Res_352_137
Author(s) : Boudier JA , Berwald-Netter Y , Dellmann HD , Boudier JL , Couraud F , Koulakoff A , Cau P
Ref : Brain Research , 352 :137 , 1985
Abstract : Purified neurotoxin II from the scorpion Androctonus australis Hector (alpha-ScTx) has previously been shown to bind specifically to the voltage-sensitive Na+ channels of excitable cells. Recent studies, using high specific activity 125I-labeled alpha-ScTx, demonstrated specific binding to neuronal cells derived from fetal mouse brains. In the present study, 125I-labeled alpha-ScTx was used to localize the voltage-sensitive Na+ channels in cultured fetal mouse brain cells. By quantitative electron microscope autoradiography we demonstrate that specific alpha-ScTx binding sites are selectively located at the plasma membrane. Estimates of their density revealed that neurites at 13 days in vitro carry at least 6 X more specific alpha-ScTx sites than cell body membrane.
ESTHER : Boudier_1985_Brain.Res_352_137
PubMedSearch : Boudier_1985_Brain.Res_352_137
PubMedID: 2408712

Title : Neuronal acquisition of tetanus toxin binding sites: relationship with the last mitotic cycle - Koulakoff_1983_Dev.Biol_100_350
Author(s) : Koulakoff A , Bizzini B , Berwald-Netter Y
Ref : Developmental Biology , 100 :350 , 1983
Abstract : In an earlier study on the developing nervous system, the existence of a temporal correlation between the appearance of tetanus toxin-binding cells and neurogenesis was reported (A. Koulakoff, B. Bizzini, and Y. Berwald-Netter (1982). Dev. Brain Res. 5, 139-147). Using a combined approach of immunocytochemistry and [3H]thymidine autoradiography it is shown that, in the fetal mouse central nervous system, dividing cells do not express membrane binding sites for tetanus toxin. A time-course quantitative autoradiography revealed that the toxin-binding sites become apparent within 7 +/- 1 hr, following the last S phase, on cells undergoing the conversion from dividing to postmitotic state. The acquisition of surface binding sites for tetanus toxin may thus be an early property of nascent central neurons, marking the transition from cycling precursor neuroblasts to postmitotic neuronal cells. Parallel studies on in vivo-developing dorsal root ganglia disclosed that at least some peripheral nervous system cells are endowed with tetanus toxin-binding capacity while still capable of DNA synthesis and undergo one or more divisions.
ESTHER : Koulakoff_1983_Dev.Biol_100_350
PubMedSearch : Koulakoff_1983_Dev.Biol_100_350
PubMedID: 6653877

Title : Neurotropism of rabies virus. An in vitro study - Tsiang_1983_J.Neuropathol.Exp.Neurol_42_439
Author(s) : Tsiang H , Koulakoff A , Bizzini B , Berwald-Netter Y
Ref : J Neuropathol Experimental Neurology , 42 :439 , 1983
Abstract : The relative susceptibility of neurons and glia, grown as monolayers in vitro, to rabies virus infection was explored. Established cell lines of neuronal or glial phenotype and primary cultures of cells derived from mouse dorsal root ganglia (DRC) or brain were used as homologues of the targets of rabies virus in the nervous system. Fixed rabies virus (CVS) strain was used in most experiments; other fixed rabies strains (PV, HEP, ERA) and a street rabies virus isolate were used in some. Virus-cell tropism was determined by immunofluorescence assay for rabies nucleocapsid antigen and cell permissivity was assessed by titration of virus yields. Neuronal cells always exhibited a much greater susceptibility to infection and a greater propensity to sustain viral growth. By immunofluorescence, 90-100% of neurons commonly had viral inclusion bodies, while doses of the virus three to four orders of magnitude higher still left greater than 99% of astrocytes, in brain cell cultures and 90 +/- 5% of the non-neuronal cells in DRG cultures without any obvious signs of rabies virus. Neuroblastoma cells (95 +/- 5% with viral antigens) produced viral yields about four orders of magnitude higher than glioma cells (10 +/- 5% with viral antigens). Though the overall infectivity of street virus was lower than that of fixed virus strains, a significantly higher viral tropism for neurons than for glia was maintained. Thus, primary neuronal cultures offer a means of exploring molecular events in rabies virus infection and their role in pathogenesis.
ESTHER : Tsiang_1983_J.Neuropathol.Exp.Neurol_42_439
PubMedSearch : Tsiang_1983_J.Neuropathol.Exp.Neurol_42_439
PubMedID: 6864237

Title : A correlation between the appearance and the evolution of tetanus toxin binding cells and neurogenesis - Koulakoff_1982_Brain.Res_281_139
Author(s) : Koulakoff A , Bizzini B , Berwald-Netter Y
Ref : Brain Research , 281 :139 , 1982
Abstract : The ontogenesis of cells expressing surface membrane binding sites for tetanus toxin (Tt) was studied in the mouse nervous system. Cells were labeled shortly after the tissue dissociation and the toxin bound was revealed by immunofluorescence. In the brain, spinal cord and dorsal root ganglia the toxin binding cells (TBC) are found as of very early stages of nervous system organogenesis, i.e. at 10 days of gestation. There is a close temporal correlation between the pattern of emergence and accumulation of TBC and the known pattern of appearance of post-mitotic neurons in mouse cerebral cortex, cerebellum and spinal cord. The curves of TBC abundance as a function of fetal age in various nervous system areas are different. They show regional fluctuations in the proportion of TBC that reflect the cumulative changes in the dynamics of neuronal subpopulations. The results indicate that Tt can be used as an ontogenetically early marker of neuronal differentiation and that the acquisition of Tt receptors may represent one of the earliest detectable characteristics of the developing neurons.
ESTHER : Koulakoff_1982_Brain.Res_281_139
PubMedSearch : Koulakoff_1982_Brain.Res_281_139
PubMedID: 6754006

Title : Neurotoxins as probes in the study of neuronal development - Berwald-Netter_1982_Toxicon_20_129
Author(s) : Berwald-Netter Y , Couraud F , Koulakoff A , Martin-Moutot N
Ref : Toxicon , 20 :129 , 1982
Abstract : We have investigated the expression of surface membrane binding sites for tetanus toxin and alpha-scorpion toxin (AaHII) on cells of the in vivo developing mouse nervous system. There is a close temporal correlation in the pattern of emergence and accumulation of tetanus toxin binding cells (TBC) and that of post-mitotic neurons. In different nervous system areas, the fluctuations in relative TBC abundance reflect regional changes in the dynamics of neuronal subpopulations. The results indicate that the acquisition of membrane tetanus toxin binding sites may represent one of the earliest detectable characteristics of nascent neurons. The Na+ channel-associated scorpion toxin become detectable in fetal mouse brain two days after the appearance of TBC. Their density increases with fetal age without change in receptor properties. At all stages, scorpion toxin binds to a single class of noninteracting sites with a KD = 0.1 - 0.5 nM. The affinity of binding is voltage-dependent. Studies on brain cells and various cell lines grown in vitro suggest a selective association of the high affinity scorpion toxin receptors with neuronal phenotype. In culture, as in vivo, there is a time dependent increase in receptor density. These results indicate that both tetanus toxin and scorpion toxin can be used as qualitative markers of neuronal differentiation; moreover, estimates of the density of scorpion toxin binding sites provide a quantitative index of neuronal maturation.
ESTHER : Berwald-Netter_1982_Toxicon_20_129
PubMedSearch : Berwald-Netter_1982_Toxicon_20_129
PubMedID: 6281938

Title : Na+-channel-associated scorpion toxin receptor sites as probes for neuronal evolution in vivo and in vitro - Berwald-Netter_1981_Proc.Natl.Acad.Sci.U.S.A_78_1245
Author(s) : Berwald-Netter Y , Martin-Moutot N , Koulakoff A , Couraud F
Ref : Proc Natl Acad Sci U S A , 78 :1245 , 1981
Abstract : Purified neurotoxin II of the scorpion Androctonus australis Hector (ScTx) has previously been shown to bind specifically to the Na+-ionophore-associated, voltage-sensitive receptor sites of excitable cells. We have conducted binding studies, using high-specific-activity 125I-labeled ScTx, to detect and quantify the Na+-channel receptors on cells of the developing fetal mouse brain. In vivo, the onset of detectable specific binding is at 12 fetal days. The rate of receptor appearance is initially slow but increases sharply as of the 16th day of mouse ontogenesis. The mean number of receptors at 12 and 19 days is 120 and 20,000 per cell, respectively (i.e., 0.5 and 80 per square micrometer). When corrected for the fraction of cell population corresponding to putative neuroblasts and neurons, identified by immunofluorescence as tetanus toxin binding cells, these values are, respectively, 1040 and 33,900 ScTx receptors per tetanus toxin binding cell or 4.2 and 136 per square micrometer. At all stages, the toxin binds to a single class of noninteracting sites; Kd = 0.1-0.5 nM. Similar findings in terms of ScTx-receptor properties and quantitative evolution were obtained in vitro. Specific 125I-labeled ScTx binding the presence of tetanus toxin binding cells. In cultures of central nervous system glia without neurons, only nonspecific low-level ScTx binding was detected. These results suggest that the high-affinity scorpion toxin receptors may be used as quantitative markers of neuronal differentiation.
ESTHER : Berwald-Netter_1981_Proc.Natl.Acad.Sci.U.S.A_78_1245
PubMedSearch : Berwald-Netter_1981_Proc.Natl.Acad.Sci.U.S.A_78_1245
PubMedID: 6262759