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Potentiating and depressant effects of metabotropic glutamate receptor agonists on high-voltage-activated calcium currents in cultured retinal ganglion neurons from postnatal mice

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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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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

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