Effects of Excitatory Amino Acids on Phosphoinositide Metabolism in Frog Retina

The effects of excitatory amino acid receptor agonists on the hydrolysis of phosphoinositides were examined using frog retinal membranes prelabeled in vitro with either 32PO4 or [3H]inositol. Glutamate stimulated release of [3H]inositol phosphates (IPs) from the retinas and altered the 32P-labeling pattern of phosphatidylinositol (PI) cycle intermediates. This indicates that glutamate affects not only the hydrolysis of phosphoinositides but possibly other steps involved in the PI cycle. Among glutamate analogs, kainate (KA), quisqualate (QA), and, to a lesser extent, N-methyl-D-aspartate (NMDA) mimicked the glutamate effect, whereas L-2-amino-4-phosphonobutyrate (L-AP4) was not effective in causing either the accumulation of [3H]IPs or the alteration of the 32P-labeling pattern of PI cycle intermediates. Among QA specific agonists, alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA), but not ibotenate (IBO) or trans-1-aminocyclopentane-1,3-dicarboxylate (ACPD) was active in stimulating IPs formation. KA effect on IPs formation may be due to indirect (polysynaptic) activation of receptor(s) other than L-AP4, IBO, or ACPD specific QA receptors. To avoid activating polysynaptic pathways, retinal synaptoneurosomes prelabeled with [3H]inositol were used to examine the hydrolysis of phosphoinositides. As in whole retinas, KA, carbachol (CARB), and NMDA stimulated the release of IPs while L-AP4 had minimal effect. Glycine (GLY) had no effect. Our results show CARB and KA to be the most effective in stimulating the production of IPs. Their effects were exerted directly through separate receptors and not through polysynaptic pathways. ACPD and IBO were the least effective in eliciting the release of IPs. Our studies provide evidence that ionotropic and not metabotropic glutamate receptors are involved in PI metabolism in the retina.