Modulation of glycine receptor subunits and gephyrin expression in the rat facial nucleus after axotomy

In the last decade, numerous studies have investigated molecular changes in excitatory glutamatergic receptors in axotomized motoneurons, but few data are available concerning the modulation of inhibitory amino acid receptors. We report here the effect of axotomy on the expression of glycine receptors, gephyrin, vesicular inhibitory amino acid transporter (VIAAT) and synapsin I in rat facial motor neurons as demonstrated by in situ hybridization and immunohistochemistry. The facial nerve trunk was sectioned unilaterally and rats were killed 1, 3, 8, 30 or 60 days after surgery. We investigated the mechanisms underlying the changes in production of these proteins following axotomy by perfusing the facial nerve with colchicine or tetrodotoxin, and injecting cardiotoxin or botulinum toxin independently and unilaterally into the whisker pads of normal rats. Animals were killed 8 days later and processed for immunohistochemistry. The abundance of GlyR subunits and gephyrin fell sharply in the axotomized facial nucleus. This decrease began 1 day after axotomy and was lowest at 8 days, with protein levels returning to normal by day 60. Abnormal synapsin immunolabelling was also observed between days 8 and 60 after axotomy but we detected no change in VIAAT immunoreactivity. The effect of colchicine was similar to, but weaker than, that of axotomy. In contrast, tetrodotoxin, cardiotoxin and botulinum toxin had no significant effect. Thus, axotomy‐induced changes probably resulted from a loss of trophic factor transported from the periphery or a positive injury signal, or both. They did not seem to depend on the disruption of activity.

[1]  P. Vidal,et al.  Modulation of GABA receptor subunits in rat facial motoneurons after axotomy. , 2005, Brain research. Molecular brain research.

[2]  L. Moran,et al.  The facial nerve axotomy model , 2004, Brain Research Reviews.

[3]  A. Burlingame,et al.  From snails to sciatic nerve: Retrograde injury signaling from axon to soma in lesioned neurons. , 2004, Journal of neurobiology.

[4]  P. Vidal,et al.  An in situ hybridization and immunofluorescence study of glycinergic receptors and gephyrin in the vestibular nuclei of the intact and unilaterally labyrinthectomized rat , 2004, Experimental Brain Research.

[5]  M. Kirsch,et al.  Ciliary neurotrophic factor is an early lesion-induced retrograde signal for axotomized facial motoneurons , 2003, Molecular and Cellular Neuroscience.

[6]  P. Vidal,et al.  Modulation of the voltage-gated sodium- and calcium-dependent potassium channels in rat vestibular and facial nuclei after unilateral labyrinthectomy and facial nerve transsection: an in situ hybridization study , 2003, Neuroscience.

[7]  L. Arnold,et al.  Acute reductions in GABAA receptor binding in layer IV of adult primate somatosensory cortex after peripheral nerve injury , 2002, Brain Research.

[8]  G. Franchi Time course of motor cortex reorganization following botulinum toxin injection into the vibrissal pad of the adult rat , 2002, The European journal of neuroscience.

[9]  N. Akaike,et al.  Reduced NR2A expression and prolonged decay of NMDA receptor‐mediated synaptic current in rat vagal motoneurons following axotomy , 2002, The Journal of physiology.

[10]  P. Klosen,et al.  In vivo expression of the intermediate filament peripherin in rat motoneurons: modulation by inhibitory and stimulatory signals , 2000, Neuroscience.

[11]  V. Haftel,et al.  Factors regulating AMPA‐type glutamate receptor subunit changes induced by sciatic nerve injury in rats , 2000, The Journal of comparative neurology.

[12]  J. Wrathall,et al.  The role of activity blockade on glutamate receptor subunit expression in the spinal cord , 2000, Brain Research.

[13]  J. Meier,et al.  Formation of glycine receptor clusters and their accumulation at synapses. , 2000, Journal of cell science.

[14]  L. Martı́nez-Millán,et al.  Ionotropic glutamate receptor subunits are differentially regulated in the motoneuronal pools of the rat hypoglossal nucleus in response to axotomy , 2000, Journal of neurocytology.

[15]  W. Zieglgänsberger,et al.  Axotomy of the sciatic nerve differentially affects expression of metabotropic glutamate receptor mRNA in adult rat motoneurons , 2000, Brain Research.

[16]  B. Gasnier The loading of neurotransmitters into synaptic vesicles. , 2000, Biochimie.

[17]  M. Tohyama,et al.  Changes in mRNA for post-synaptic density-95 (PSD-95) and carboxy-terminal PDZ ligand of neuronal nitric oxide synthase following facial nerve transection. , 2000, Brain research. Molecular brain research.

[18]  N. Akaike,et al.  Reduction of voltage-dependent magnesium block of N-methyl-d-aspartate receptor-mediated current by in vivo axonal injury , 2000, Neuroscience.

[19]  T. Cope,et al.  Neurotrophin expression by spinal motoneurons in adult and developing rats , 2000, The Journal of comparative neurology.

[20]  A. Windebank,et al.  Differential Expression of Brain-Derived Neurotrophic Factor, Neurotrophin-3, and Neurotrophin-4/5 in the Adult Rat Spinal Cord: Regulation by the Glutamate Receptor Agonist Kainic Acid , 1999, The Journal of Neuroscience.

[21]  Georg W. Kreutzberg,et al.  Neuroglial activation repertoire in the injured brain: graded response, molecular mechanisms and cues to physiological function , 1999, Brain Research Reviews.

[22]  M. Sofroniew,et al.  Re-expression of p75NTR by adult motor neurons after axotomy is triggered by retrograde transport of a positive signal from axons regrowing through damaged or denervated peripheral nerve tissue , 1999, Neuroscience.

[23]  Guido Stoll,et al.  Nerve Injury, Axonal Degeneration and Neural Regeneration: Basic Insights , 1999, Brain pathology.

[24]  K. Osen,et al.  The Vesicular GABA Transporter, VGAT, Localizes to Synaptic Vesicles in Sets of Glycinergic as Well as GABAergic Neurons , 1998, The Journal of Neuroscience.

[25]  A. Pastor,et al.  Effects of botulinum neurotoxin type A on the expression of gephyrin in cat abducens motoneurons , 1998, The Journal of comparative neurology.

[26]  P. Rostaing,et al.  Localization of components of glycinergic synapses during rat spinal cord development , 1998, The Journal of comparative neurology.

[27]  H. Aldskogius,et al.  Central neuron–glial and glial–glial interactions following axon injury , 1998, Progress in Neurobiology.

[28]  F. Tang,et al.  Expression of glutamate receptor subunits 2/3 and 4 in the hypoglossal nucleus of the rat after neurectomy , 1997, Experimental Brain Research.

[29]  B. Giros,et al.  Cloning of a functional vesicular GABA and glycine transporter by screening of genome databases , 1997, FEBS letters.

[30]  T. Cope,et al.  Low-frequency depression of the monosynaptic reflex is not altered by tetrodotoxin-induced nerve conduction blockade. , 1997, Journal of neurophysiology.

[31]  Y Q Li,et al.  Distribution of GABAergic and glycinergic premotor neurons projecting to the facial and hypoglossal nuclei in the rat , 1997, The Journal of comparative neurology.

[32]  M. Jacob,et al.  Changes in the Regulatory Effects of Cell–Cell Interactions on Neuronal AChR Subunit Transcript Levels after Synapse Formation , 1996, The Journal of Neuroscience.

[33]  R. Banati,et al.  Microglial Turnover in the Injured CNS: Activated Microglia Undergo Delayed DNA Fragmentation Following Peripheral Nerve Injury , 1995, Journal of neuropathology and experimental neurology.

[34]  J. Kirsch,et al.  The postsynaptic localization of the glycine receptor-associated protein gephyrin is regulated by the cytoskeleton , 1995, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[35]  Q. Yan,et al.  The biological responses of axotomized adult motoneurons to brain- derived neurotrophic factor , 1994, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[36]  S. Leong,et al.  Glial reaction after facial nerve compression in the facial canal of the albino rat. , 1994, Acta oto-laryngologica.

[37]  P. Vidal,et al.  Distribution of Glutamatergic Receptors and GAD mRNA‐Containing Neurons in the Vestibular Nuclei of Normal and Hemilabyrinthectomized Rats , 1994, The European journal of neuroscience.

[38]  H. Takagi,et al.  Distinct regulation of sodium channel types I, II and III following nerve transection. , 1994, Brain research. Molecular brain research.

[39]  A. Y. Chiu,et al.  Distinct neurotrophic responses of axotomized motor neurons to BDNF and CNTF in adult rats. , 1994, Neuroreport.

[40]  D. W. Vaughan Effects of peripheral axotomy on presynaptic axon terminals with GABA‐like immunoreactivity , 1994, The Anatomical record.

[41]  T. Hökfelt,et al.  GAP‐43, aFGF, CCK and α‐ and β‐CGRP in Rat Spinal Motoneurons Subjected to Axotomy and/or Dorsal Root Severance , 1993 .

[42]  J. Kirsch,et al.  Widespread expression of gephyrin, a putative glycine receptor-tubulin linker protein, in rat brain , 1993, Brain Research.

[43]  J. Kirsch,et al.  Distribution of Gephyrin Transcripts in the Adult and Developing Rat Brain , 1993, The European journal of neuroscience.

[44]  F. Miller,et al.  Evidence that the loss of homeostatic signals induces regeneration-associated alterations in neuronal gene expression. , 1993, Developmental biology.

[45]  M. Svensson,et al.  Regeneration of Hypoglossal Nerve Axons Following Blockade of the Axotomy‐induced Microglial Cell Reaction in the Rat , 1993, The European journal of neuroscience.

[46]  Tetsuro Yamamoto,et al.  Observations on morphology and electrophysiological properties of the normal and axotomized facial motoneurons in the cat , 1992, Brain Research.

[47]  M. Tohyama,et al.  Regional distribution of cells expressing glycine receptor α 2 subunit mRNA in the rat brain , 1992, Brain Research.

[48]  藤田 昌宏 Regional distribution of the cells expressing glycine receptor β subunit mRNA in the rat brain , 1992 .

[49]  H. Korn,et al.  Partial glycinergic denervation induces transient changes in the distribution of a glycine receptor-associated protein in a central neuron , 1992, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[50]  M. Tohyama,et al.  Localization of glycine receptor α1 subunit mRNA-containing neurons in the rat brain: An analysis using in situ hybridization histochemistry , 1991, Neuroscience.

[51]  Makoto Sato,et al.  Regional distribution of the cells expressing glycine receptor β subunit mRNA in the rat brain , 1991, Brain Research.

[52]  B. Marquèze-Pouey,et al.  Widespread expression of glycine receptor subunit mRNAs in the adult and developing rat brain. , 1991, The EMBO journal.

[53]  C. Becker,et al.  The inhibitory glycine receptor: a ligand-gated chloride channel of the central nervous system. , 1990, European journal of biochemistry.

[54]  K. Barron,et al.  Perineuronal glial responses after axotomy of central and peripheral axons. A comparison , 1990, Brain Research.

[55]  M. Tohyama,et al.  Localization of glycine receptors in the rat central nervous system: An immunocytochemical analysis using monoclonal antibody , 1988, Neuroscience.

[56]  M. Graeber,et al.  Delayed astrocyte reaction following facial nerve axotomy , 1988, Journal of neurocytology.

[57]  Georg W. Kreutzberg,et al.  Astrocytes increase in glial fibrillary acidic protein during retrograde changes of facial motor neurons , 1986, Journal of neurocytology.

[58]  H. Korn,et al.  Distribution of glycine receptors at central synapses: an immunoelectron microscopy study , 1985, The Journal of cell biology.

[59]  A. Rotter,et al.  Regulation of glycine receptor binding in the mouse hypoglossal nucleus in response to axotomy , 1984, Brain Research Bulletin.

[60]  P. Greengard,et al.  Synapsin I (protein I), a nerve terminal-specific phosphoprotein. I. Its general distribution in synapses of the central and peripheral nervous system demonstrated by immunofluorescence in frozen and plastic sections , 1983, The Journal of cell biology.

[61]  B. Sumner A quantitative analysis of boutons with different types of synapse in normal and injured hypoglossal nuclei , 1975, Experimental Neurology.

[62]  B. Grafstein The nerve cell body response to axotomy , 1975, Experimental Neurology.

[63]  L. Landmesser,et al.  Axotomy Mimicked by Localized Colchicine Application , 1972, Science.

[64]  B. Barres,et al.  The relationship between neuronal survival and regeneration. , 2000, Annual review of neuroscience.

[65]  S. Cullheim,et al.  Ultrastructural evidence for a preferential elimination of glutamate-immunoreactive synaptic terminals from spinal motoneurons after intramedullary axotomy. , 2000, The Journal of comparative neurology.

[66]  G. Kreutzberg Principles of neuronal regeneration. , 1996, Acta neurochirurgica. Supplement.

[67]  S. Cullheim,et al.  GAP-43, aFGF, CCK and alpha- and beta-CGRP in rat spinal motoneurons subjected to axotomy and/or dorsal root severance. , 1993, The European journal of neuroscience.

[68]  M. Tohyama,et al.  Regional distribution of cells expressing glycine receptor alpha 2 subunit mRNA in the rat brain. , 1992, Brain research.

[69]  J. Kaas,et al.  Injury-induced reorganization of somatosensory cortex is accompanied by reductions in GABA staining. , 1991, Somatosensory & motor research.

[70]  M. Jouvet,et al.  Neurones immunoréactifs à la glycine dans le bulbe rachidien du chat. , 1990 .

[71]  M. Jouvet,et al.  [Glycine immunoreactive neurons in the medulla oblongata in cats]. , 1990, Comptes rendus de l'Academie des sciences. Serie III, Sciences de la vie.

[72]  Lieberman Ar The Axon Reaction: A Review of the Principal Features of Perikaryal Responses to Axon Injury , 1971 .

[73]  A. Lieberman The axon reaction: a review of the principal features of perikaryal responses to axon injury. , 1971, International review of neurobiology.