Docosahexaenoic acid is taken up by the inner segment of frog photoreceptors leading to an active synthesis of docosahexaenoyl inositol lipids: similarities in metabolismin vivoandin vitro

Abstract

Retinal uptake and metabolism of docosahexaenoic acid (DHA) was studied in vivo in frogs 1, 2, and 6 hours after dorsal lymph sac injections of [3H]-DHA (50 microCi/g). Light microscope autoradiography and biochemical techniques were used to compare the profiles of cellular uptake and lipid labeling with those obtained from 6 hour [3H]-DHA retinal incubations (final DHA concentration, 0.11 and 25 microM). Light microscope autoradiography demonstrated that rod photoreceptor ellipsoids and synaptic terminals preferentially labeled both in vivo and in vitro conditions. Also, the cytoplasm and oil droplets of retinal pigment epithelial cells became very heavily labeled after 6 hours of in vivo labeling. Phosphatidic acid showed the highest labeling in one hour, while other phospholipids accumulated label throughout the 6 hours. At that time point, most label was recovered in phosphatidyl-ethanolamine (37%), phosphatidylcholine (27%), and phosphatidylinositol (16%), the latter displaying 1.6-fold higher labeling than phosphatidylserine. The profile of labeled lipids was similar to that obtained in vitro when the concentration of DHA was in the nanomolar range. Our results suggest that de novo lipid synthesis is a major route for esterification of [3H]-DHA into retinal lipids, giving rise to an early and rapid labeling of DHA-phosphatidylinositol, both in vivo and in vitro, when DHA is present at low concentrations. Furthermore, the profile of labeled retinal cells under in vivo conditions closely resembles in vitro DHA labeling.