Metabolic incorporation of unsaturated fatty acids into boar spermatozoa lipids and de novo formation of diacylglycerols

Abstract

Lipids play an important role in the maturation, viability and function of sperm cells. In this study, we examined the neutral and polar lipid composition of boar spermatozoa by thin-layer chromatography/mass spectrometry. Main representatives of the neutral lipid classes were diacylglycerols containing saturated (myristoyl, palmitoyl and stearoyl) fatty acyl residues. Glycerophosphatidylcholine and glycerophosphatidylethanolamine with alk(en)yl ether residues in the sn-1 position and unsaturated long chained fatty acyl residues in sn-2 position were identified as the most prominent polar lipids. The only glycoglycerolipid was sulfogalactosylglycerolipid carrying 16:0-alkyl- and 16:0-acyl chains. Using stable isotope-labelling, the metabolic incorporation of exogenously supplied fatty acids was analysed. Boar spermatozoa incorporated hexadecenoic (16:1), octadecenoic (18:1), octadecadienoic (18:2) and octadecatrienoic (18:3) acids primarily in the diacylglycerols and glycerophosphatidylcholines. In contrast, incorporation of eicosapentaenoic acid (20:5) was not detected. The analysis of molecular species composition subsequent to the incorporation of exogenous [(14)C]-octadecadienoic acid suggests two pathways for incorporation of exogenous fatty acids into glycerophosphatidylcholine: (1) de novo synthesis of glycerophosphatidylcholine via the CDP-choline pathway and (2) reacylation of lysophosphatidylcholine via an acyltransferase.

Keywords: AA; BTS; Beltsville Thawing Solution; Boar spermatozoa; DAG; DHB; Diacylglycerol; FFA; Fatty acid; GC; GPA; GPC; GPE; GPI; GPL; GPS; Glycerophospholipid; LPC; MAG; MS; PLA(2); PLC; SGG; TLC; aminoacridine; diacylglycerol; dihydroxybenzoic acid; free fatty acids; gas chromatography; glycerophosphatidic acid; glycerophosphatidylcholine; glycerophosphatidylethanolamine; glycerophosphatidylinositol; glycerophosphatidylserine; glycerophospholipid; lyso-glycerophosphatidylcholine (monoacylglycerophosphatidylcholine); mass spectrometry; monoacylglycerol; phospholipase A(2); phospholipase C; sulfogalactosylglycerolipid; thin-layer chromatography.