Effect of stimulation with light on synthesis and release of acetylcholine by an isolated mammalian retina

Abstract

1. Acetylcholine synthesis and release were studied in rabbit retinas isolated from the eye and incubated under conditions in which their electrophysiological function is maintained. ACh synthesized from exogenous [14C] choline appeared in the retina at an initial rate of 16 nmol/g wet wt-h. Incorporation of labeled choline into ACh was accelerated by stimulation of the retina with light. 2. Retinas incubated for 40 min in the presence of labeled choline and then superfused with a medium containing an anticholinesterase released radioactive ACh into the perfusate. The rate of release increased approximately fourfold during stimulation with light. 3. When retinas were incubated with labeled choline and then superfused with medium containing no pharmacological agents, stimulation with light caused an increased release of choline into the perfusate. The recovery of labeled choline following stimulation was enhanced by hemicholinium 3. 4. Neither the light-induced release of ACh (in perfusate containing anticholinesterase) nor the light-induced release of choline (in perfusate containing no anticholinesterase) occurred if the perfusate contained 20 mM Mg2+ and 0.2 mM Ca2+. 5. Synthesis of ACh by the retina at a high rate, acceleration of choline incorporation by stimulation, and Ca2+-dependent release of ACh by stimulation are each presumptive evidence that the retina contains a cholinergic synapse. If this presumption is correct, one such synapse may be of an amacrine or bipolar cell since these cells can depolarize during illumination, whereas the predominant response of receptor and horizontal cells is hyperpolarization.