Inhibition of carbachol-induced inositol phosphate accumulation in the embryonic retina promoted by kainate and veratridine

In the present study, we report that low concentrations of the glutamate ionotropic agonist kainate decreased the turnover of [3H]-phosphoinositides ([3H]-InsPs) induced by muscarinic receptors in the chick embryonic retina. When 100 muM carbachol was used, the estimated IC50 value for kainate was 0.2 muM and the maximal inhibition of ~50 percent was obtained with 1 muM or higher concentrations of the glutamatergic agonist. Our data also show that veratridine, a neurotoxin that increases the permeability of voltage-sensitive sodium channels, had no effect on [3H]-InsPs levels of the embryonic retina. However, 50 muM veratridine, but not 50 mM KCl, inhibited ~65 percent of the retinal response to carbachol. While carbachol increased [3H]-InsPs levels from 241.2 + 38.0 to 2044.5 + 299.9 cpm/mg protein, retinal response decreased to 861.6 + 113.9 cpm/mg protein when tissues were incubated with carbachol plus veratridine. These results suggest that the accumulation of phosphoinositides induced by activation of muscarinic receptors can be inhibited by the influx of Na+ ions triggered by activation of kainate receptors or opening of voltage-sensitive sodium channels in the chick embryonic retina.

Veratridine increases the survival of retinal ganglion cells in vitro

Neuronal cell death is an important phenomenon involving many biochemical pathways. This degenerative event has been studied to understand how the cells activate the mechanisms that lead to self-destruction. Target cells and afferent cells play a relevant role in the regulation of natural cell death. We studied the effect of veratridine (1.5, 3.0, 4.5 and 6.0 muM) on the survival of neonatal rat retinal ganglion cells in vitro. Veratridine (3.0 muM), a well-known depolarizing agent that opens the Na+ channel, promoted a two-fold increase in the survival of retinal ganglion cells kept in culture for 48 h. This effect was dose-dependent and was blocked by 1.0 muM tetrodotoxin (a classical voltage-dependent Na+ channel blocker) and 3.0 muM flunarizine (a Na+ and Ca2+ channel blocker). These results indicate that electrical activity is also important for the maintenance of retinal ganglion cell survival in vitro.

Evidence for two independent mechanisms of gaba release induced by veratridine and glutamate in monolayer cultures of chick embryo retinal cells

GABA is a major inhibitory neurotransmitter in the central nervous system, including the retina. In the present paper we present evidence for the existence of two independent mechanisms for GABA release in cultured retina cells. Eight-day-old chick embryo retinas were dissociated and plated in 35-mm plastic dishes and cultured for 3 or 7 days at 37 grade C. An increase of 3 to 5-fold in GABA release was observed in cultures of 3 or 7 days in vitro preloaded with 0.5 uCi[3H} GABA and stimulated with glutamate (100 uM) or veratridine (100 uM). Tetrodotoxin (1 uM) blocked the release induced by veratridine but not by glutamate. In contrast, the non-N-methyl-D-aspartate (NMDA)glutamate antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, 100 uM) was able to inhibit GABA release promoted by glutamate but not by veratridine. These results indicate that depolarization of retinal cells byopening of voltage-dependent sodium channels or activation of non-NMDA glutamate receptors can trigger intracellular events that lead to calcium-independent GABA release