Specific interaction of glutamate with membranes from cultured retinal pigment epithelium.
Excitatory amino acids (EAA) have been shown to induce phagocytosis in retinal pigment epithelial (RPE) cells. In order to explore if this action is receptor-mediated, we have identified and characterized receptors for L-glutamate through the binding of [3H]L-glutamate to membranes from chick RPE cells in primary culture. Specific binding was found saturable, with KB = 333nM and Bmax = 3.2 pmol/mg protein in frozen/thawed membranes. Na(+)-independent binding was present in cultures of 16 and 25 days in vitro, and was not affected by temperature. Pharmacological profile of analogues of EAA at different receptor types suggests the presence of a metabotropic type receptor (L-glutamate > S-2-amino-3-phosphonopropionate > 2-amino-4-phosphonobutyrate = trans-(1S,3R)-1-aminocyclopentane-1,3-dicarboxylate > quisqualate). Excitatory amino acid analogues acting at the NMDA-receptor also displaced bound L-glutamate, and a noticeable stimulation of specific binding of this ligand by glycine was shown; this effect was mimicked by D-serine and 1-hydroxy-3-aminopyrrolidone-2 (HA-966) but not by 7-chlorokynurenate, and was not inhibited by strychnine. Since taurine and GABA also increased specific binding, it is likely that modulation of EAA receptors in RPE differs from that in neurons.
López-Colomé
A M
AM
Instituto de Fisiología Cellular, U.N.A.M. México, D.F. México.
Salceda
R
R
Fragoso
G
G
D000596:Amino Acids / Q000494:pharmacology; D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D002642:Chick Embryo; D005971:Glutamates / Q000378:metabolism*; D018698:Glutamic Acid; D007700:Kinetics; D010587:Phagocytosis / Q000187:drug effects; D010857:Pigment Epithelium of Eye / Q000378:metabolism*; D011485:Protein Binding; D017470:Receptors, Glutamate / Q000187:drug effects / Q000378:metabolism; D012160:Retina / Q000378:metabolism*; D012964:Sodium / Q000494:pharmacology; D013696:Temperature
D000596:Amino Acids; D005971:Glutamates; D017470:Receptors, Glutamate; D018698:Glutamic Acid; D012964:Sodium