High-affinity kainate binding sites in living slices of rat neocortex: characterization and regulation

We have characterized a high-affinity kainate binding site in in vitro living rat neocortical slices using [3H]kainate. [3H]Kainate labelled at least two binding sites, the higher affinity site with a Kd of 7.1 nM and a Bmax of 71.2 fmol/mg protein. This high-affinity binding site showed a pharmacology consistent with a kainate receptor with competition by kainate and domoic acid, as well as the (RS)-alpha-amino-3-hydroxy-5-methyl-isoxazole-4-propionate antagonist 6-cyano-2,3-dihydroxy-7-nitroquinoxaline. Increases in cellular depolarization induced by 2-h preincubations in veratridine and glutamate led to a significant 55% average decrease in [3H]kainate binding in adult cortex. Similarly, preincubation in kainate led to a significant average 26% decrease in binding. In both instances, Eadie-Hofstee analysis of saturation binding data revealed that the decreased binding reflected changes in receptor number. At different postnatal ages, increases in cellular depolarization significantly decreased binding (< 20 days postnatal age, -86%; > 60 days, -48%). Kainate treatment also significantly decreased binding at all ages (-64% at < 20 days; > 60 days, -18%), with significant differences noted between ages. These age-dependent effects are unlike those previously described for either N-methyl-D-aspartate [Lanius and Shaw (1992) Anat. Rec. 232, 54(A)] or (RS)-alpha-amino-3-hydroxy-5-methyl-isoxazole-4-propionate high affinity receptors [Shaw and Lanius (1992) Devl Brain Res. 68, 225-233].(ABSTRACT TRUNCATED AT 250 WORDS)