Stimulation-evoked release of purines from the rabbit retina

The evoked release of purines from rabbit retinae preloaded with [(3)H]adenosine was studied in vitro. Potassium (8.6-43.6 mM) and ouabain (1 or 10 microM) increased the release of radioactivity in a concentration-dependent manner. The K(+)-evoked release was significantly reduced when the superfusion was carried out at 2-4 degrees C. The effect of K(+) (8.6, 13.6 and 23.6 mM) and of ouabain (1 microM) were completely abolished when the retinae were superfused with a Ca(2+)-free medium containing 0.1 mM EGTA. Calcium removal only partially reduced the effect of higher K(+) and ouabain concentrations (43.6 mM and 10 microM, respectively). Further, the effect of K(+) was found to be independent of extracellular Ca(2+) when retinae were pretreated with ouabain for 30 min. Stimulation of the retina with light flashes induced a small, persistent increase in the release of radioactivity observable for several minutes after the end of stimulation. The superfusate contained mainly hypoxanthine and inosine. There were no significant changes in the relative proportions of the different purine compounds released before or in response to either K(+) (23.6 mM) or ouabain (10 microM) stimulation. Potassium stimulation significantly increased the release of adenosine, inosine and hypoxanthine. Addition of the adenosine deaminase inhibitor, erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA), significantly increased the relative proportions of released endogenous adenosine and inosine. The results indicate that K(+) stimulation induces the release of purines from the rabbit retina by a Ca(2+)- and energy-dependent process. Light flashes also induce a purine release. The results suggest an active role for adenosine in retinal neurotransmission.