The properties of glycine receptors (GlyRs) depend upon their subunit composition. While the prevalent adult forms of GlyRs are heteromers, previous reports suggested functional α homomeric receptors in mature nervous tissues. Here we show two functionally different GlyRs populations in the rat medial nucleus of trapezoid body (MNTB). Postsynaptic receptors formed α1/β-containing clusters on somatodendritic domains of MNTB principal neurons, colocalizing with glycinergic nerve endings to mediate fast, phasic IPSCs. In contrast, presynaptic receptors on glutamatergic calyx of Held terminals were composed of dispersed, homomeric α1 receptors. Interestingly, the parent cell bodies of the calyces of Held, the globular bushy cells of the cochlear nucleus, expressed somatodendritic receptors (α1/β heteromers) and showed similar clustering and pharmacological profile as GlyRs on MNTB principal cells. These results suggest that specific targeting of GlyR β-subunit produces segregation of GlyR subtypes involved in two different mechanisms of modulation of synaptic strength.

Department of Auditory Neuroscience Institute of Experimental Medicine ASCR 14220 Prague Czech Republic

Zobrazit více v PubMed

Alvarez FJ, Dewey DE, Harrington DA, Fyffe RE. Cell-type specific organization of glycine receptor clusters in the mammalian spinal cord. J Comp Neurol. 1997;379:150–170. PubMed

Awatramani GB, Turecek R, Trussell LO. Inhibitory control at a synaptic relay. J Neurosci. 2004;24:2643–2647. PubMed PMC

Awatramani GB, Price GD, Trussell LO. Modulation of transmitter release by presynaptic resting potential and background calcium levels. Neuron. 2005a;48:109–121. PubMed

Awatramani GB, Turecek R, Trussell LO. Staggered development of GABAergic and glycinergic transmission in the MNTB. J Neurophysiol. 2005b;93:819–828. PubMed

Banks MI, Smith PH. Intracellular recordings from neurobiotin-labeled cells in brain slices of the rat medial nucleus of the trapezoid body. J Neurosci. 1992;12:2819–2837. PubMed PMC

Bausen M, Weltzien F, Betz H, O'Sullivan GA. Regulation of postsynaptic gephyrin cluster size by protein phosphatase 1. Mol Cell Neurosci. 2010;44:201–209. PubMed

Beato M. The time course of transmitter at glycinergic synapses onto motoneurons. J Neurosci. 2008;28:7412–7425. PubMed PMC

Betz H, Laube B. Glycine receptors: recent insights into their structural organization and functional diversity. J Neurochem. 2006;97:1600–1610. PubMed

Billups B. Colocalization of vesicular glutamate transporters in the rat superior olivary complex. Neurosci Lett. 2005;382:66–70. PubMed

Bormann J, Rundström N, Betz H, Langosch D. Residues within transmembrane segment M2 determine chloride conductance of glycine receptor homo- and hetero-oligomers. EMBO J. 1993;12:3729–3737. PubMed PMC

Borst JG, Soria van Hoeve J. The calyx of held synapse: from model synapse to auditory relay. Annu Rev Physiol. 2012;74:199–224. PubMed

Burzomato V, Beato M, Groot-Kormelink PJ, Colquhoun D, Sivilotti LG. Single-channel behavior of heteromeric alpha1beta glycine receptors: an attempt to detect a conformational change before the channel opens. J Neurosci. 2004;24:10924–10940. PubMed PMC

Charrier C, Machado P, Tweedie-Cullen RY, Rutishauser D, Mansuy IM, Triller A. A crosstalk between beta1 and beta3 integrins controls glycine receptor and gephyrin trafficking at synapses. Nat Neurosci. 2010;13:1388–1395. PubMed

Chesnoy-Marchais D. Potentiation of chloride responses to glycine by three 5-HT3 antagonists in rat spinal neurones. Br J Pharmacol. 1996;118:2115–2125. PubMed PMC

Deleuze C, Runquist M, Orcel H, Rabié A, Dayanithi G, Alonso G, Hussy N. Structural difference between heteromeric somatic and homomeric axonal glycine receptors in the hypothalamo-neurohypophysial system. Neuroscience. 2005;135:475–483. PubMed

Dumoulin A, Rostaing P, Bedet C, Lévi S, Isambert MF, Henry JP, Triller A, Gasnier B. Presence of the vesicular inhibitory amino acid transporter in GABAergic and glycinergic synaptic terminal boutons. J Cell Sci. 1999;112:811–823. PubMed

Dumoulin A, Triller A, Kneussel M. Cellular transport and membrane dynamics of the glycine receptor. Front Mol Neurosci. 2009;2:28. PubMed PMC

Faber DS, Korn H. Synergism at central synapses due to lateral diffusion of transmitter. Proc Natl Acad Sci U S A. 1988;85:8708–8712. PubMed PMC

Felmy F, Schneggenburger R. Developmental expression of the Ca2+-binding proteins calretinin and parvalbumin at the calyx of held of rats and mice. Eur J Neurosci. 2004;20:1473–1482. PubMed

Feng G, Tintrup H, Kirsch J, Nichol MC, Kuhse J, Betz H, Sanes JR. Dual requirement for gephyrin in glycine receptor clustering and molybdoenzyme activity. Science. 1998;282:1321–1324. PubMed

Flint AC, Liu X, Kriegstein AR. Nonsynaptic glycine receptor activation during early neocortical development. Neuron. 1998;20:43–53. PubMed

Forsythe ID. Direct patch recording from identified presynaptic terminals mediating glutamatergic EPSCs in the rat CNS, in vitro. J Physiol. 1994;479:381–387. PubMed PMC

Friauf E. Transient appearance of calbindin-D28k-positive neurons in the superior olivary complex of developing rats. J Comp Neurol. 1993;334:59–74. PubMed

Friauf E, Hammerschmidt B, Kirsch J. Development of adult-type inhibitory glycine receptors in the central auditory system of rats. J Comp Neurol. 1997;385:117–134. PubMed

Fritschy JM, Harvey RJ, Schwarz G. Gephyrin: where do we stand, where do we go? Trends Neurosci. 2008;31:257–264. PubMed

Grothe B, Pecka M, McAlpine D. Mechanisms of sound localization in mammals. Physiol Rev. 2010;90:983–1012. PubMed

Grudzinska J, Schemm R, Haeger S, Nicke A, Schmalzing G, Betz H, Laube B. The beta subunit determines the ligand binding properties of synaptic glycine receptors. Neuron. 2005;45:727–739. PubMed

Harty TP, Manis PB. Glycine-evoked currents in acutely dissociated neurons of the guinea pig ventral cochlear nucleus. J Neurophysiol. 1996;75:2300–2311. PubMed

Hori T, Takahashi T. Mechanisms underlying short-term modulation of transmitter release by presynaptic depolarization. J Physiol. 2009;587:2987–3000. PubMed PMC

Jeong HJ, Jang IS, Moorhouse AJ, Akaike N. Activation of presynaptic glycine receptors facilitates glycine release from presynaptic terminals synapsing onto rat spinal sacral dorsal commissural nucleus neurons. J Physiol. 2003;550:373–383. PubMed PMC

Kandler K, Gillespie DC. Developmental refinement of inhibitory sound-localization circuits. Trends Neurosci. 2005;28:290–296. PubMed PMC

Karnani MM, Venner A, Jensen LT, Fugger L, Burdakov D. Direct and indirect control of orexin/hypocretin neurons by glycine receptors. J Physiol. 2011;589:639–651. PubMed PMC

Kawa K. Glycine facilitates transmitter release at developing synapses: a patch clamp study from Purkinje neurons of the newborn rat. Brain Res Dev Brain Res. 2003;144:57–71. PubMed

Kirsch J, Langosch D, Prior P, Littauer UZ, Schmitt B, Betz H. The 93-kDa glycine receptor-associated protein binds to tubulin. J Biol Chem. 1991;266:22242–22245. PubMed

Kneussel M, Betz H. Clustering of inhibitory neurotransmitter receptors at developing postsynaptic sites: the membrane activation model. Trends Neurosci. 2000a;23:429–435. PubMed

Kneussel M, Betz H. Receptors, gephyrin and gephyrin-associated proteins: novel insights into the assembly of inhibitory postsynaptic membrane specializations. J Physiol. 2000b;525:1–9. PubMed PMC

Kneussel M, Loebrich S. Trafficking and synaptic anchoring of ionotropic inhibitory neurotransmitter receptors. Biol Cell. 2007;99:297–309. PubMed

Kopp-Scheinpflug C, Dehmel S, Tolnai S, Dietz B, Milenkovic I, Rübsamen R. Glycine-mediated changes of onset reliability at a mammalian central synapse. Neuroscience. 2008;157:432–445. PubMed

Kopp-Scheinpflug C, Steinert JR, Forsythe ID. Modulation and control of synaptic transmission across the MNTB. Hear Res. 2011;279:22–31. PubMed

Korada S, Schwartz IR. Development of GABA, glycine, and their receptors in the auditory brainstem of gerbil: a light and electron microscopic study. J Comp Neurol. 1999;409:664–681. PubMed

Kubota H, Alle H, Betz H, Geiger JR. Presynaptic glycine receptors on hippocampal mossy fibers. Biochem Biophys Res Commun. 2010;393:587–591. PubMed

Kulik A, Vida I, Luján R, Haas CA, López-Bendito G, Shigemoto R, Frotscher M. Subcellular localization of metabotropic GABA(B) receptor subunits GABA(B1a/b) and GABA(B2) in the rat hippocampus. J Neurosci. 2003;23:11026–11035. PubMed PMC

Kungel M, Friauf E. Physiology and pharmacology of native glycine receptors in developing rat auditory brainstem neurons. Brain Res Dev Brain Res. 1997;102:157–165. PubMed

Laube B. Potentiation of inhibitory glycinergic neurotransmission by Zn2+: a synergistic interplay between presynaptic P2X2 and postsynaptic glycine receptors. Eur J Neurosci. 2002;16:1025–1036. PubMed

Laube B, Maksay G, Schemm R, Betz H. Modulation of glycine receptor function: a novel approach for therapeutic intervention at inhibitory synapses? Trends Pharmacol Sci. 2002;23:519–527. PubMed

Leao RN, Oleskevich S, Sun H, Bautista M, Fyffe RE, Walmsley B. Differences in glycinergic mIPSCs in the auditory brain stem of normal and congenitally deaf neonatal mice. J Neurophysiol. 2004;91:1006–1012. PubMed

Lee EA, Cho JH, Choi IS, Nakamura M, Park HM, Lee JJ, Lee MG, Choi BJ, Jang IS. Presynaptic glycine receptors facilitate spontaneous glutamate release onto hilar neurons in the rat hippocampus. J Neurochem. 2009;109:275–286. PubMed

Legendre P. Pharmacological evidence for two types of postsynaptic glycinergic receptors on the Mauthner cell of 52-h-old zebrafish larvae. J Neurophysiol. 1997;77:2400–2415. PubMed

Legendre P. The glycinergic inhibitory synapse. Cell Mol Life Sci. 2001;58:760–793. PubMed PMC

Legendre P, Muller E, Badiu CI, Meier J, Vannier C, Triller A. Desensitization of homomeric alpha1 glycine receptor increases with receptor density. Mol Pharmacol. 2002;62:817–827. PubMed

Legendre P, Förstera B, Jüttner R, Meier JC. Glycine receptors caught between genome and proteome-functional implications of RNA editing and splicing. Front Mol Neurosci. 2009;2:23. PubMed PMC

Lim R, Alvarez FJ, Walmsley B. Quantal size is correlated with receptor cluster area at glycinergic synapses in the rat brainstem. J Physiol. 1999;516:505–512. PubMed PMC

Lim R, Alvarez FJ, Walmsley B. GABA mediates presynaptic inhibition at glycinergic synapses in a rat auditory brainstem nucleus. J Physiol. 2000;525:447–459. PubMed PMC

Lim R, Oleskevich S, Few AP, Leao RN, Walmsley B. Glycinergic mIPSCs in mouse and rat brainstem auditory nuclei: modulation by ruthenium red and the role of calcium stores. J Physiol. 2003;546:691–699. PubMed PMC

Lohmann C, Friauf E. Distribution of the calcium-binding proteins parvalbumin and calretinin in the auditory brainstem of adult and developing rats. J Comp Neurol. 1996;367:90–109. PubMed

Lu T, Rubio ME, Trussell LO. Glycinergic transmission shaped by the corelease of GABA in a mammalian auditory synapse. Neuron. 2008;57:524–535. PubMed

Lynch JW. Molecular structure and function of the glycine receptor chloride channel. Physiol Rev. 2004;84:1051–1095. PubMed

Lynch JW. Native glycine receptor subtypes and their physiological roles. Neuropharmacology. 2009;56:303–309. PubMed

Lynch JW, Rajendra S, Barry PH, Schofield PR. Mutations affecting the glycine receptor agonist transduction mechanism convert the competitive antagonist, picrotoxin, into an allosteric potentiator. J Biol Chem. 1995;270:13799–13806. PubMed

Maas C, Belgardt D, Lee HK, Heisler FF, Lappe-Siefke C, Magiera MM, van Dijk J, Hausrat TJ, Janke C, Kneussel M. Synaptic activation modifies microtubules underlying transport of postsynaptic cargo. Proc Natl Acad Sci U S A. 2009;106:8731–8736. PubMed PMC

Mohammadi B, Krampfl K, Cetinkaya C, Moschref H, Grosskreutz J, Dengler R, Bufler J. Kinetic analysis of recombinant mammalian alpha(1) and alpha(1)beta glycine receptor channels. Eur Biophys J. 2003;32:529–536. PubMed

Mørkve SH, Hartveit E. Properties of glycine receptors underlying synaptic currents in presynaptic axon terminals of rod bipolar cells in the rat retina. J Physiol. 2009;587:3813–3830. PubMed PMC

Moss SJ, Smart TG. Constructing inhibitory synapses. Nat Rev Neurosci. 2001;2:240–250. PubMed

Muller E, Le-Corronc H, Legendre P. Extrasynaptic and postsynaptic receptors in glycinergic and GABAergic neurotransmission: a division of labor? Front Mol Neurosci. 2008;1:3. PubMed PMC

Oertel D. The role of timing in the brain stem auditory nuclei of vertebrates. Annu Rev Physiol. 1999;61:497–519. PubMed

Pecka M, Brand A, Behrend O, Grothe B. Interaural time difference processing in the mammalian medial superior olive: the role of glycinergic inhibition. J Neurosci. 2008;28:6914–6925. PubMed PMC

Pfeiffer F, Simler R, Grenningloh G, Betz H. Monoclonal antibodies and peptide mapping reveal structural similarities between the subunits of the glycine receptor of rat spinal cord. Proc Natl Acad Sci U S A. 1984;81:7224–7227. PubMed PMC

Piechotta K, Weth F, Harvey RJ, Friauf E. Localization of rat glycine receptor alpha1 and alpha2 subunit transcripts in the developing auditory brainstem. J Comp Neurol. 2001;438:336–352. PubMed

Pribilla I, Takagi T, Langosch D, Bormann J, Betz H. The atypical M2 segment of the beta subunit confers picrotoxinin resistance to inhibitory glycine receptor channels. EMBO J. 1992;11:4305–4311. PubMed PMC

Price GD, Trussell LO. Estimate of the chloride concentration in a central glutamatergic terminal: a gramicidin perforated-patch study on the calyx of Held. J Neurosci. 2006;26:11432–11436. PubMed PMC

Rigo JM, Badiu CI, Legendre P. Heterogeneity of postsynaptic receptor occupancy fluctuations among glycinergic inhibitory synapses in the zebrafish hindbrain. J Physiol. 2003;553:819–832. PubMed PMC

Rundström N, Schmieden V, Betz H, Bormann J, Langosch D. Cyanotriphenylborate: subtype-specific blocker of glycine receptor chloride channels. Proc Natl Acad Sci U S A. 1994;91:8950–8954. PubMed PMC

Sato K, Kuriyama H, Altschuler RA. Expression of glycine receptor subunits in the cochlear nucleus and superior olivary complex using non-radioactive in-situ hybridization. Hear Res. 1995;91:7–18. PubMed

Sato K, Shiraishi S, Nakagawa H, Kuriyama H, Altschuler RA. Diversity and plasticity in amino acid receptor subunits in the rat auditory brain stem. Hear Res. 2000;147:137–144. PubMed

Schmieden V, Grenningloh G, Schofield PR, Betz H. Functional expression in Xenopus oocytes of the strychnine binding 48 kd subunit of the glycine receptor. EMBO J. 1989;8:695–700. PubMed PMC

Schneggenburger R, Forsythe ID. The calyx of Held. Cell Tissue Res. 2006;326:311–337. PubMed

Schofield PR, Shivers BD, Seeburg PH. The role of receptor subtype diversity in the CNS. Trends Neurosci. 1990;13:8–11. PubMed

Singer JH, Berger AJ. Development of inhibitory synaptic transmission to motoneurons. Brain Res Bull. 2000;53:553–560. PubMed

Smith PH, Joris PX, Yin TC. Anatomy and physiology of principal cells of the medial nucleus of the trapezoid body (MNTB) of the cat. J Neurophysiol. 1998;79:3127–3142. PubMed

Specht CG, Triller A. The dynamics of synaptic scaffolds. Bioessays. 2008;30:1062–1074. PubMed

Supplisson S, Chesnoy-Marchais D. Glycine receptor beta subunits play a critical role in potentiation of glycine responses by ICS-205,930. Mol Pharmacol. 2000;58:763–770. PubMed

Suwa H, Saint-Amant L, Triller A, Drapeau P, Legendre P. High-affinity zinc potentiation of inhibitory postsynaptic glycinergic currents in the zebrafish hindbrain. J Neurophysiol. 2001;85:912–925. PubMed

Takahashi T, Momiyama A, Hirai K, Hishinuma F, Akagi H. Functional correlation of fetal and adult forms of glycine receptors with developmental changes in inhibitory synaptic receptor channels. Neuron. 1992;9:1155–1161. PubMed

Telgkamp P, Raman IM. Depression of inhibitory synaptic transmission between Purkinje cells and neurons of the cerebellar nuclei. J Neurosci. 2002;22:8447–8457. PubMed PMC

Triller A, Cluzeaud F, Pfeiffer F, Betz H, Korn H. Distribution of glycine receptors at central synapses: an immunoelectron microscopy study. J Cell Biol. 1985;101:683–688. PubMed PMC

Turecek R, Trussell LO. Presynaptic glycine receptors enhance transmitter release at a mammalian central synapse. Nature. 2001;411:587–590. PubMed

Turecek R, Trussell LO. Reciprocal developmental regulation of presynaptic ionotropic receptors. Proc Natl Acad Sci U S A. 2002;99:13884–13889. PubMed PMC

Tyagarajan SK, Ghosh H, Yévenes GE, Nikonenko I, Ebeling C, Schwerdel C, Sidler C, Zeilhofer HU, Gerrits B, Muller D, Fritschy JM. Regulation of GABAergic synapse formation and plasticity by GSK3beta-dependent phosphorylation of gephyrin. Proc Natl Acad Sci U S A. 2011;108:379–384. PubMed PMC

Veenman CL, Reiner A, Honig MG. Biotinylated dextran amine as an anterograde tracer for single- and double-labeling studies. J Neurosci Methods. 1992;41:239–254. PubMed

von Gersdorff H, Borst JG. Short-term plasticity at the calyx of held. Nat Rev Neurosci. 2002;3:53–64. PubMed

Walmsley B, Alvarez FJ, Fyffe RE. Diversity of structure and function at mammalian central synapses. Trends Neurosci. 1998;21:81–88. PubMed

Wang DS, Mangin JM, Moonen G, Rigo JM, Legendre P. Mechanisms for picrotoxin block of alpha2 homomeric glycine receptors. J Biol Chem. 2006;281:3841–3855. PubMed

Wang DS, Buckinx R, Lecorronc H, Mangin JM, Rigo JM, Legendre P. Mechanisms for picrotoxinin and picrotin blocks of alpha2 homomeric glycine receptors. J Biol Chem. 2007;282:16016–16035. PubMed

Webb TI, Lynch JW. Molecular pharmacology of the glycine receptor chloride channel. Curr Pharm Des. 2007;13:2350–2367. PubMed

Wojcik SM, Katsurabayashi S, Guillemin I, Friauf E, Rosenmund C, Brose N, Rhee JS. A shared vesicular carrier allows synaptic corelease of GABA and glycine. Neuron. 2006;50:575–587. PubMed

Wu SH, Kelly JB. Inhibition in the superior olivary complex: pharmacological evidence from mouse brain slice. J Neurophysiol. 1995;73:256–269. PubMed

Wu SH, Oertel D. Inhibitory circuitry in the ventral cochlear nucleus is probably mediated by glycine. J Neurosci. 1986;6:2691–2706. PubMed PMC

Yang Z, Cromer BA, Harvey RJ, Parker MW, Lynch JW. A proposed structural basis for picrotoxinin and picrotin binding in the glycine receptor pore. J Neurochem. 2007a;103:580–589. PubMed

Yang Z, Ney A, Cromer BA, Ng HL, Parker MW, Lynch JW. Tropisetron modulation of the glycine receptor: femtomolar potentiation and a molecular determinant of inhibition. J Neurochem. 2007b;100:758–769. PubMed

Ye JH, Wang F, Krnjevic K, Wang W, Xiong ZG, Zhang J. Presynaptic glycine receptors on GABAergic terminals facilitate discharge of dopaminergic neurons in ventral tegmental area. J Neurosci. 2004;24:8961–8974. PubMed PMC

Zarbin MA, Wamsley JK, Kuhar MJ. Glycine receptor: light microscopic autoradiographic localization with [3H]strychnine. J Neurosci. 1981;1:532–547. PubMed PMC

Zhang XB, Sun GC, Liu LY, Yu F, Xu TL. Alpha2 subunit specificity of cyclothiazide inhibition on glycine receptors. Mol Pharmacol. 2008;73:1195–1202. PubMed

Zhorov BS, Bregestovski PD. Chloride channels of glycine and GABA receptors with blockers: Monte Carlo minimization and structure-activity relationships. Biophys J. 2000;78:1786–1803. PubMed PMC

The role of GABAB receptors in the subcortical pathways of the mammalian auditory system

Distribution of glycine receptors on the surface of the mature calyx of Held nerve terminal