A comparison of the rates of incorporation of fatty acids during the rapid synthesis in vitro of endogenous triacylglycerols by neuronal nuclei

Abstract

The incorporation of radioactive palmitate, oleate, linoleate, and arachidonate into endogenous triacylglycerols was followed in vitro using neuronal nuclei (N1) isolated from cerebral cortices of 15-day-old rabbits. Specific rates of incorporation of fatty acids into N1 triacylglycerols were 33–42 times and more than 100 times the corresponding values for cerebral cortex homogenates and microsomal fractions (P3), respectively. Acyl-CoA synthetase specific activities in N1 were 2.2 to 3.2 times the specific rates for fatty acid incorporation into N1 triacylglycerols. Using single fatty acids, N1 acyl-CoA synthetase showed a preference for linoleate which was more highly marked in linoleate–palmitate and linoleate–arachidonate competitions. In fatty acid incorporation into N1 triacylglycerols a preference for linoleate in competition with palmitate was noted; however, there was also a relatively higher utilization of arachidonate shown competitively than was noted in acyl-CoA synthesis. The data suggested that N1 diacylglycerol acyl transferase shows a selectivity for arachidonoyl-CoA in comparison with CoA esters of palmitate or oleate. Molecular class analyses of radioactive triacylglycerol products indicated that native endogenous N1 diacylglycerols bearing arachidonate or fatty acids of equal or higher unsaturation were used preferentially in N1 triacylglycerol synthesis. This preference was significantly decreased when higher levels of endogenous diacylglycerols were produced in N1 following a phospholipase C preincubation.