Activation of phospholipase D by the fucose-sulfate glycoconjugate that induces an acrosome reaction in spermatozoa.
We report for the first time that phospholipase D activity in sea urchin spermatozoa can be regulated by a component of egg jelly known to induce an acrosome reaction. The fucose-sulfate glycoconjugate (FSG) of egg jelly that induces an acrosome reaction in spermatozoa caused Ca2+-dependent increases in 1,2-diacylglycerol and phosphatidic acid. Diacylglycerol concentrations were increased 2-fold, and phosphatidic acid concentrations were elevated up to 10-fold 2 min after the addition of FSG to spermatozoa. FSG also caused increases in choline, but not in choline phosphate concentrations. Neither phosphorylation of diacylglycerol nor de novo synthesis from glycerol were significant routes of synthesis of phosphatidic acid during the acrosome reaction. When spermatozoa were incubated with FSG in the presence of ethanol, phosphatidylethanol was produced. As ethanol concentrations in the extracellular medium were increased from 0.1 to 2.5%, the amount of phosphatidylethanol increased, whereas phosphatidic acid concentrations decreased, suggesting a competitive transphosphatidylation reaction catalyzed by phospholipase D. Furthermore, when a phosphatidylcholine pool in spermatozoa was radiolabeled using [3H]1-O-alkyl-2-lyso-glycerol-3-phosphorylcholine, the subsequent addition of FSG caused a 4-fold accumulation of [3H]phosphatidic acid. FSG-induced elevations in [3H]phosphatidic acid were positively correlated with the percent of cells that had undergone an acrosome reaction.
Domino
S E
SE
Department of Pharmacology, Vanderbilt University Medical Center, Nashville, Tennessee 37232-0295.
Bocckino
S B
SB
Garbers
D L
DL
D000177:Acrosome / Q000201:enzymology* / Q000502:physiology; D000818:Animals; D002794:Choline / Q000378:metabolism; D004075:Diglycerides / Q000378:metabolism; D004789:Enzyme Activation / Q000187:drug effects; D005643:Fucose / Q000502:physiology* / Q000031:analogs & derivatives; D006001:Glycoconjugates / Q000502:physiology*; D008297:Male; D010712:Phosphatidic Acids / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000378:metabolism; D010766:Phosphorylation; D010767:Phosphorylcholine / Q000378:metabolism; D012617:Sea Urchins; D013094:Spermatozoa / Q000201:enzymology*; D013431:Sulfates / Q000502:physiology*; D013464:Sulfuric Acids; D014700:Verapamil / Q000494:pharmacology
D004075:Diglycerides; D006001:Glycoconjugates; D010712:Phosphatidic Acids; D013431:Sulfates; D013464:Sulfuric Acids; C061711:fucose-sulfate glycoconjugate; D010767:Phosphorylcholine; D005643:Fucose; D014700:Verapamil; D002794:Choline