Abstract
This study was aimed at clarifying the role of metabotropic glutamate receptors (mGluRs) in the regulation of intracellular Ca2+ concentration ([Ca2+]i) in postnatal mouse retinal ganglion neurons (RGNs). RGNs were maintained for 1–2 weeks in vitro by adding brain-derived neurotrophic factor (BDNF) and basic fibroblast growth factor (bFGF) to the culture medium. In order to select these cells for electrophysiological measurements, RGNs were vitally labelled with an antibody against Thy-1.2. Voltage-activated Ca2+ currents [I Ca(V)] were recorded with patch electrodes in the wholecell configuration. It was found that racemic ±-1-aminocyclopentane-trans-1, 3-dicarboxylic acid (t-ACPD) or its active enantiomer 1S,3R-ACPD rapidly and reversibly either enhanced or depressed I Ca(V). Quisqualate (QA), l-2-amino-4-phosphonobutyrate (l-AP4) and the endogenous transmitter glutamate induced similar effects when ionotropic glutamate receptors were blocked with d-2-amino-5-phosphonovalerate (d-APV) and 6,7-dinitroquinoxaline-2, 3-dione (DNQX). ω- Conotoxin GVIA (ω-CgTx GVIA), but not nifedipine prevented modulation of I Ca(V) by mGluR agonists. The depression of I Ca(V) by t-ACPD was irreversible when cells were dialysed with guanosine-5′-O-(3-thiotriphosphate) (GTP[γ-S]). Ratio measurements of fura-2 fluorescence in Thy-1+ cells showed that neither t-ACPD, QA nor l-AP4 affected [Ca2+]i by liberation of Ca2+ from intracellular stores. Our results suggest that cultured RGNs express mGluRs. These receptors cannot induce Ca2+ release from intracellular stores but regulate [Ca2+]i by a fast and reversible, G-protein-mediated action on a subpopulation of voltage-activated Ca2+ channels.
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Agopyan N, Miu P, Krnjevic K (1993) Modulation of highthreshold Ca current and spontaneous postsynaptic transient currents by phorbol 12,13-diacetate, 1-(5-isoquinolinesulfonyl)-2-methyl piperazine (H-7), and monosialoganglioside (GM1) in Ca1 pyramidal neurons of rat hippocampus in vitro. Hippocampus 3:67–76
Ayoub GS, Matthews G (1992) Substance P modulates calcium current in retinal bipolar neurons. Visual Neurosci 8:539–544
Bortolotto ZA, Collingridge GL (1993) Characterisation of LPT induced by the activation of glutamate metabotropic receptors in area CA1 of the hippocampus. Neuropharmacology 32:1–9
Bossu JL, Fagni L, Nooney J, Bockaert J, Feltz A (1993) Increased Ca channel activity due to metabotropic glutamate receptor stimulation in isolated rat cerebellar granule cells. J Physiol (Lond) 459:250P
De Waard M, Seagar M, Feltz A, Couraud F (1992) Inositol phosphate regulation of voltage-dependent calcium channels in cerebellar granule neurons. Neuron 9:497–503
Dolphin AC (1991) Regulation of calcium channel activity by GTP binding proteins and second messengers. Biochim Biophys Acta 1091:68–80
Dolphin AC (1991) Ca2+ channel currents in rat sensory neurones: interaction between guanine nucleotides, cyclic AMP and Ca2+ channel ligands. J Physiol (Lond) 432:23–43
Duvoisin R (1993) Isolation of a novel member of the metabotropic glutamate receptor gene family and its pattern of expression in mouse retina. Invest Ophthalmol Vis Sci 34:1381
Grantyn R, Korenbaum E (1992) Easy identification of dissociated rat retinal ganglion cells by a size criterion. In: Kettenmann H, Grantyn R (eds) Practical electrophysiological methods: a guide for in vitro studies in vertebrate neurobiology. Wiley-Liss, New York, pp 84–87
Grynkiewicz G, Poenie M, Tsien RY (1985) A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem 260:3440–3450
Guenther E, Rothe T, Taschenberger H, Grantyn R (1993) Identification of calcium currents in retinal ganglion cells from postnatal rat. Brain Res (in press)
Hamill OP, Marty A, Neher E, Sakmann B, Sigworth FJ (1981) Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflügers Arch 391:85–100
Hay M, Kunze DL (1993) Calcium current inhibition by L-Glu metabotropic receptor activation in nodose neurons. FASEB J 7:A99
Ishida AT (1991) Regenerative sodium and calcium currents in goldfish retinal ganglion cell somata. Vision Res 31:477–485
Kaneda M, Kaneko A (1991) Voltage-gated calcium currents in isolated retinal ganglion cells of the cat. Jpn J Physiol 41:35–48
Karschin A, Lipton SA (1989) Calcium channels in solitary retinal ganglion cells from post-natal rat. J Physiol (Lond) 418:379–396
Lasater EM, Witkovsky P (1990) Membrane currents of spiking cells isolated from turtle retina. J Comp Physiol [A] 167:11–21
Lester RAJ, Jahr CE (1990) Quisqualate receptor-mediated depression of calcium currents in hippocampal neurons. Neuron 4:741–749
Mayer ML, Miller RJ (1990) Excitatory amino acid receptors, second messengers and regulation of intracellular Ca2+ in mammalian neurons. Trends Pharmacol Sci 11:254–260
Mogul DJ, Adams ME, Fox AP (1993) Differential activation of adenosine receptors decreases N-type but potentiates P-type Ca2+ current in hippocampal CA3 neurons. Neuron 10:327–334
Moore EDW, Becker PL, Fogarty KE, Williams DA, Fay FS (1990) Ca2+ imaging in single living cells: theoretical and practical issues. Cell Calcium 11:157–179
Nakanishi S (1992) Molecular diversity of glutamate receptors and implications for brain function. Science 258:597–603
Nakashima I, Zhang Y, Rahman SMJ, Yoshida T, Isobe K, Ding L, Iwamoto T, Hamaguchi M, Ikezawa H, Taguchi R (1991) Evidence of synergy between Thy-1 and CD3/TCR complex in signal delivery to murine thymocytes for cell death. J Immunol 147:1153–1162
Ohishi H, Shigemoto R, Nakanishi S, Mizuno N (1993) Distribution of the messenger RNA for a metabotropic glutamate receptor, mGluR2, in the central nervous system of the rat. Neuroscience 53:1009–1018
Perouansky M, Grantyn R (1990) Is GABA release modulated by presynaptic excitatory amino acid receptors? Neurosci Lett 113:292–297
Plummer MR, Logothetis DE, Hess P (1989) Elementary properties and pharmacological sensitivities of calcium channels in mammalian peripheral neurons. Neuron 2:1453–1463
Rörig B, Grantyn R (1993) Rat retinal ganglion cells express Ca2+-permeable non-NMDA glutamate receptors during the period of histogenetic cell death. Neurosci Lett 153:32–36
Rörig B, Grantyn R (1993) Glutamatergic and GABAergic synaptic currents in ganglion cells from isolated retinae of pigmented rats during postnatal development. Dev Brain Res 74:98–110
Sahara Y, Westbrook GL (1991) Trans-ACPD blocks high threshold calcium currents on cultured hippocampal neurons. Soc Neurosci Abstr 17:1168
Sayer RJ, Schwindt PC, Crill WE (1992) Metabotropic glutamate receptor-mediated suppression of L-type calcium current in acutely isolated neocortical neurons. J Neurophysiol 68: 833–842
Schoepp DD, Conn PJ (1993) Metabotropic glutamate receptors in brain function and pathology. Trends Pharmacol Sci 14:13–20
Shapiro MS, Hille B (1993) Substance P and somatostatin inhibit calcium channels in rat sympathetic neurons via different G protein pathways. Neuron 10:11–20
Swandulla D, Carbone E, Lux HD (1991) Do calcium channel classifications account for neuronal calcium channel diversity? Trends Neurosci 14:46–51
Swartz KJ, Bean BP (1992) Inhibition of calcium channels in rat CA3 pyramidal neurons by a metabotropic glutamate receptor. J Neurosci 12:4358–4371
Swartz KJ, Merritt A, Bean BP, Lovinger DM (1993) Protein kinase C modulates glutamate receptor inhibition of Ca2+ channels and synaptic transmission. Nature 361:165–168
Tanabe Y, Nomura A, Masu M, Shigemoto R, Mizuno N, Nakanishi S (1993) Signal transduction, pharmacological properties, and expression patterns of two rat metabotropic glutamate receptors, mGluR3 and mGluR4. J Neurosci 13:1372–1378
Tanabe-Ohuchi T, Yoshimura N, Shigemoto R, Honda Y (1992) Localization of NMDA and metabotropic glutamate receptors in the rat retina by in situ hybridization technique. Invest Opththalmol Vis Sci 33:1030
Taschenberger H, Grantyn R (1993) Two types of spontaneous excitatory synaptic currents in cultures from the postnatal rat retina. In: Elsner N, Heisenberg M (eds) Gene — brain — behaviour. Proceedings of the 21st Göttingen Neurobiology Conference. Thieme, Stuttgart, p 407
Thomsen C, Kristensen P, Mulvihill E, Haldemann B, Suzdak PD (1992) L-2-Amino-4-phosphonobutyrate (L-AP4) is an agonist at the type IV metabotropic glutamate receptor which is negatively coupled to adenylate cyclase. Eur J Pharmacol 227:361–362
Trombley PQ, Westbrook GL (1992) L-AP4 inhibits calcium currents and synaptic transmission via a G-protein-coupled glutamate receptor. J Neurosci 12:2043–2050
Wang X, Treistman SN, Lemos JR (1992) Two types of highthreshold calcium currents inhibited by ω-conotoxin in nerve terminals of rat neurohypophysis. J Physiol (Lond) 445:181–199
Westenbroek RE, Hell JW, Warner C, Dubel SJ, Snutch TP, Catterall WA (1992) Biochemical properties and subcellular distribution of an N-type calcium channel α1 subunit. Neuron 9:1099–1115
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Rothe, T., Bigl, V. & Grantyn, R. Potentiating and depressant effects of metabotropic glutamate receptor agonists on high-voltage-activated calcium currents in cultured retinal ganglion neurons from postnatal mice. Pflugers Arch. 426, 161–170 (1994). https://doi.org/10.1007/BF00374684
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DOI: https://doi.org/10.1007/BF00374684