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Group I metabotropic glutamate receptor-mediated calcium signalling and immediate early gene expression in cultured rat striatal neurons



Group I metabotropic glutamate receptor-mediated calcium signalling and immediate early gene expression in cultured rat striatal neurons



European Journal of Neuroscience 17(4): 741-750



Group I metabotropic glutamate receptors (mGluRs) are positively coupled to phospholipase C (PLC) via Galphaq-proteins and are expressed in the medium-sized projection neurons of striatum. To characterize the group I mGluR/PLC-sensitive modulation of intracellular Ca2+ ([Ca2+]i) signalling, primary neuronal cultures were prepared from the striatum of E19 rat embryos or neonatal day-1 rat pups. Cytoplasmic Ca2+ signals were examined with fura-2/AM at a signal cell level. After 17-18 days in culture, a profound Ca2+ response consisting of two phases was induced in cultured striatal neurons following bath application of the selective group I agonist, 3,5-dihydroxyphenylglycine (DHPG). The [Ca2+]i elevation was concentration- and time-dependent, and was blocked by coexposure to the group I antagonist, N-phenyl-7-(hydroxyimino)cyclopropa[b]chromen-1a-carboxamide (PHCCC), or the PLC inhibitor, U-73122, but not to the group II/III antagonist (RS)-alpha-methylserine-O-phosphate monophenyl ester (MSOPPE). A series of further pharmacological studies demonstrated that the initial spike-like transient was dependent on intracellular Ca2+ mobilization through 1,4,5-triphosphate-sensitive stores, and the second long-lasting rise was dependent on extracellular Ca2+ influx through N-methyl-d-aspartate (NMDA) receptors and especially L-type voltage-operated Ca2+ channels. Lastly, using an immediate early gene c-fos as a report of inducible gene expression, the resultant [Ca2+]i elevation contributes to DHPG-stimulated c-fos mRNA and Fos protein expression in striatal neurons as revealed by quantitative in situ hybridization and immunocytochemistry, respectively. These results demonstrate that group I mGluRs are able to affect Ca2+ homeostasis at multiple levels and trigger Ca2+-sensitive gene transcription in striatal neurons.

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Accession: 010725129

Download citation: RISBibTeXText

PMID: 12603264

DOI: 10.1046/j.1460-9568.2003.02495.x


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