PHARMACOLOGICAL CHARACTERIZATION OF GLUTAMATE BINDING-SITES IN CULTURED CEREBELLAR GRANULE CELLS AND CORTICAL ASTROCYTES

Membranes prepared from cerebellar granule cells and cortical astrocytes exhibited specific, saturable binding of L-[H-3]glutamate. The apparent binding constant K-D was 135 nM and 393 nM and the maximal binding capacity B-max 42 and 34 mu mol/kg in granule cells and astrocytes, respectively. In granule cells the binding was strongly inhibited by the glutamate receptor agonists kainate, quisqualate, N-methyl-D-aspartate (NMDA), L-homocysteate and ibotenate, and the antagonist DL-5-aminophosphonovalerate. In astrocytes, only quisqualate among these was effective. L-Aspartate, L-cysteate, L-cysteinesulphinate and gamma-D-glutamylglycine were inhibitors in both cell types. The binding was totally displaced in both cell types by L-cysteinesulphinate with IC50 in the micromolar range. In astrocytes the binding was also totally displaced by quisqualate, but in granule cells only partially by NMDA, kainate and quisqualate in turn. It is concluded from the relative potencies of agonists and antagonists in [H-3]glutamate binding that cerebellar granule cells express the NMDA, kainate and quisqualate types of the glutamate receptor, while only the quisqualate-sensitive binding seems to be present in cortical astrocytes.