Phosphoinositide 3-kinase is involved in the induction of the human sperm acrosome reaction downstream of tyrosine phosphorylation.
In somatic cells phosphoinositide 3-kinase (PI 3-kinase) is activated upon interaction with both receptor tyrosine kinases (RTK) and G-proteins resulting in the production of moieties involved in the inositol phospholipid signalling pathway. As G proteins, RTK and the inositol phospholipids have all been implicated in the human sperm acrosome reaction, experiments were carried out to determine whether PI 3-kinase was also involved in this phenomenon. Wortmannin is a selective inhibitor of PI 3-kinase and was shown to significantly inhibit the acrosome reaction induced by both mannose-bovine serum albumin (mannose-BSA) (10, 50 and 100 nM) and a polyclonal antibody raised against an extracellular region of the sperm zona receptor kinase (ZRK, at 100 nM only). Wortmannin did not inhibit the A23187- or progesterone-induced acrosome reaction. These results suggest that PI 3-kinase is involved in the human sperm acrosome reaction. The levels of tyrosine phosphorylation of sperm proteins as detected by Western blotting using antiphosphotyrosine antibodies was not affected by wortmannin in agonist (A23187 and mannose-BSA)-stimulated spermatozoa. This indicated that PI 3-kinase operates downstream of tyrosine phosphorylation in the signal transduction cascade which leads to the human sperm acrosome reaction.
Fisher
H M
HM
Department of Obstetrics and Gynaecology, University of Sheffield, Jessop Hospital for Women, UK.
Brewis
I A
IA
Barratt
C L
CL
Cooke
I D
ID
Moore
H D
HD
D020101:Acrosome Reaction / Q000187:drug effects / Q000502:physiology*; D000730:Androstadienes / Q000494:pharmacology; D015153:Blotting, Western; D000001:Calcimycin / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D066298:In Vitro Techniques; D007476:Ionophores / Q000494:pharmacology; D008297:Male; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D000081082:Phosphoinositide-3 Kinase Inhibitors; D010766:Phosphorylation; D011374:Progesterone / Q000494:pharmacology; D015398:Signal Transduction; D013094:Spermatozoa / Q000187:drug effects / Q000201:enzymology* / Q000378:metabolism; D014443:Tyrosine / Q000378:metabolism*; D000077191:Wortmannin
D000730:Androstadienes; D000001:Calcimycin; D004791:Enzyme Inhibitors; D007476:Ionophores; D019869:Phosphatidylinositol 3-Kinases; D000081082:Phosphoinositide-3 Kinase Inhibitors; D011374:Progesterone; D014443:Tyrosine; D000077191:Wortmannin