Protein phosphorylation regulates the mouse sperm acrosome reaction induced by the zona pellucida.
Recently, the ligand-receptor signal transduction mechanism has been implicated in mediating the zona pellucida (ZP)-induced acrosome reaction. Little is known about the role of protein phosphorylation in this specific event. We examine whether modification of protein phosphorylation and dephosphorylation affects the kinetics of the acid-solubilized ZP-induced acrosome reaction of mouse sperm. Mouse epididymal sperm were incubated in modified Krebs-Ringer bicarbonate medium for a period of 90 to 120 min and then treated with 2 acid-solubilized ZP/microliters for an additional 60 min. The chlortetracycline fluorescence assay was used to monitor the acrosome reaction. Capacitated sperm were inhibited from undergoing acid-solubilized ZP-induced acrosome reaction in the presence of an inhibitor of cyclic nucleotide-dependent protein kinase, H8; activators of the Ca(2+)- and phospholipid-dependent protein kinase (protein kinase C); an inhibitor of phosphatases 1 and 2A, okadaic acid; or an inhibitor of protein tyrosine kinases, genistein. The addition of inhibitors of protein kinase C, such as staurosporine, H7, and protein kinase C [19-36] pseudosubstrate, inhibited the phorbol ester-dependent inhibition of the acid-solubilized ZP-induced acrosome reaction. The present study suggests that protein phosphorylation and dephosphorylation play a regulatory role in the process of the ZP-induced acrosome reaction.
Furuya
S
S
Department of Obstetrics and Gynecology, Keio University School of Medicine, Tokyo, Japan.
Endo
Y
Y
Oba
M
M
Matsui
Y
Y
Nozawa
S
S
Suzuki
S
S
D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D000177:Acrosome / Q000187:drug effects / Q000502:physiology*; D000470:Alkaloids / Q000494:pharmacology; D000818:Animals; D004075:Diglycerides / Q000494:pharmacology; D004988:Ethers, Cyclic / Q000494:pharmacology; D005089:Exocytosis; D005260:Female; D019833:Genistein; D007529:Isoflavones / Q000494:pharmacology; D007546:Isoquinolines / Q000494:pharmacology; D008297:Male; D008561:Membrane Fusion; D051379:Mice; D019319:Okadaic Acid; D010703:Phorbol Esters / Q000494:pharmacology; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors / Q000502:physiology; D010766:Phosphorylation / Q000187:drug effects; D010879:Piperazines / Q000494:pharmacology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors; D047428:Protein Kinase Inhibitors; D011494:Protein Kinases / Q000502:physiology*; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D015398:Signal Transduction / Q000187:drug effects; D013084:Sperm-Ovum Interactions / Q000187:drug effects; D019311:Staurosporine; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D015044:Zona Pellucida / Q000187:drug effects / Q000502:physiology*
D000470:Alkaloids; D004075:Diglycerides; D004988:Ethers, Cyclic; D007529:Isoflavones; D007546:Isoquinolines; D010703:Phorbol Esters; D010879:Piperazines; D047428:Protein Kinase Inhibitors; C044295:1,2-dioctanoylglycerol; D019319:Okadaic Acid; C008876:phorbol-12,13-didecanoate; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; C053177:N-(2-(methylamino)ethyl)-5-isoquinolinesulfonamide; D019833:Genistein; D011494:Protein Kinases; D011493:Protein Kinase C; D010749:Phosphoprotein Phosphatases; D019311:Staurosporine; D013755:Tetradecanoylphorbol Acetate