Metabolic labeling of normal canine rod outer segment phospholipids in vivo and in vitro

Abstract

Twenty-four hours after the intravitreal injection of [ 3H]palmitate and [ 14C]docosahexaenoate in dogs, the rod outer segment phospholipids are highly labeled. Palmitate is found predominantly in phosphatidylcholine (PC), with lesser amounts in phosphatidylethanolamine (PE) and very little in either phosphatidylserine (PS) or phosphatidylinositol (PI). Docosahexaenoate most heavily labeled PE followed by PC, with lesser amounts in PS and very little in PI. Two-hour incubations of 3 mm trephine buttons removed from dog retinas produced very similar patterns of labeling with palmitate and docosahexaenoate. In vitro incubation of retina buttons with [ 3H]arachidonate produced heavy labeling of PI, with much less in PC and very little in either PS or PE. [ 3H]Glycerol labeled in PC, PI and PE in descending order but PS almost not at all, [ 3H]Serine labeled PS predominantly, but small amounts were found in PC, PE and PI. The trephine retina buttons can be utilized for multiple-precursor incubations and studies of differential metabolism in retinal regions, particularly when studying scarce tissue from mutant animals or humans with inherited retinal degenerations.