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. Author manuscript; available in PMC: 2007 Jun 15.
Published in final edited form as: Dev Biol. 2006 Dec 15;304(1):116–126. doi: 10.1016/j.ydbio.2006.12.023

Figure 2.

Figure 2

Zona pellucida agonists activate phosphoinositide signaling pathways in sperm. (A) Examples of the early effects of ZP3 on sperm phosphoinositides are shown. ZP3 (5 μg/ml) stimulated labeling of PI(3,4,5)P3 but not of PI(3)P. Enhanced labeling was inhibited by 100 nM wortmannin. Relative migration of sperm phosphoinositides and other phospholipids were identified using purified standards. Abbreviations: PC, phosphatidylcholine; PA, phosphatidic acid; phosphatidylinositides were defined previously. The spot migrating beneath PI(3)P could not be identified but did not change during treatment with ZPse or ZP3. (B) Time courses of the effects of ZP3 or of ZPse (5 and 20 μg/ml, respectively) on sperm phosphoinositides are shown. Data (mean ± SD, 7-10 experiments) were obtained from chromatograms of response time courses, similar to those in panel A, and expressed as the ratio of labeling at the indicated time, Pn, to the initial activity prior to addition of agonist or control proteins, Pi. ZPse and ZP3 had similar effects and results are pooled for presentation (ZP3 and ZPse, Inline graphic). PI3 kinase antagonists (100 nM wortmannin and 10 uM LY294002) had similar inhibitory effects on the ZP3/ZPse-evoked responses and data are pooled for presentation (Inline graphic). Similarly, control proteins (10 ug/ml ZP1 or ZP2; 20 ug/ml BSA, a1-acid glycoprotein) failed to evoke a phosphoinositide response and these results are pooled for presentation (Inline graphic).