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Role of kainate receptor activation and desensitization on the [Ca 2+ ] i changes in cultured rat hippocampal neurons
Author(s) -
Silva Ana P.,
Malva João O.,
Ambrósio António F.,
Salgado António J.,
Carvalho Arsélio P.,
Carvalho Caetana M.
Publication year - 2001
Publication title -
journal of neuroscience research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.72
H-Index - 160
eISSN - 1097-4547
pISSN - 0360-4012
DOI - 10.1002/jnr.1164
Subject(s) - kainate receptor , ampa receptor , agonist , kainic acid , glutamate receptor , receptor , hippocampal formation , stimulation , biology , chemistry , medicine , endocrinology , biochemistry
Abstract We investigated the role of kainate (KA) receptor activation and desensitization in inducing the increase in the intracellular free Ca 2+ concentration ([Ca 2+ ] i ) in individual cultured rat hippocampal neurons. The rat hippocampal neurons in the cultures were shown to express kainate receptor subunits, KA2 and GluR6/7, either by immunocytochemistry or by immunoblot analysis. The effect of LY303070, an α‐amino‐3‐hydroxy‐5‐methyl‐isoxazole‐4‐propionate (AMPA) receptor antagonist, on the alterations in the [Ca 2+ ] i caused by kainate showed cell‐to‐cell variability. The [Ca 2+ ] i increase caused by kainate was mostly mediated by the activation of AMPA receptors because LY303070 inhibited the response to kainate in a high percentage of neurons. The response to kainate was potentiated by concanavalin A (Con A), which inhibits kainate receptor desensitization, in 82.1% of the neurons, and this potentiation was not reversed by LY303070 in about 38% of the neurons. Also, upon stimulation of the cells with 4‐methylglutamate (MGA), a selective kainate receptor agonist, in the presence of Con A, it was possible to observe [Ca 2+ ] i changes induced by kainate receptor activation, because LY303070 did not inhibit the response in all neurons analyzed. In toxicity studies, cultured rat hippocampal neurons were exposed to the drugs for 30 min, and the cell viability was evaluated at 24 hr using the 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT) assay. The selective activation of kainate receptors with MGA, in the presence of Con A, induced a toxic effect, which was not prevented by LY303070, revealing a contribution of a small subpopulation of neurons expressing kainate receptors that independently mediate cytotoxicity. Taken together, these results indicate that cultured hippocampal neurons express not only AMPA receptors, but also kainate receptors, which can modulate the [Ca 2+ ] i and toxicity. J. Neurosci. Res. 65:378–386, 2001. © 2001 Wiley‐Liss, Inc.