Journal of Andrology
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Published-Ahead-of-Print December 27, 2006, DOI:10.2164/jandrol.106.001289

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Acrosome reaction induced by recombinant human zona pellucida 3 peptides rhuZP3a22~176 and rhuZP3b177~348 and their mechanism

Ya Ni , Kun Li , Wanxiang Xu , Liwen Song , Kangshou Yao , Xinzong Zhang , Hefeng Huang , Yanling Zhang , and Qi-Xian Shi *

* To whom correspondence should be addressed. E-mail: qxshi{at}mail.hz.zj.cn.

To explore the biological characteristics of the recombinant zona pellucida 3 (ZP3) peptides of rhuZP3a22~176 and rhuZP3b177~348, we examined whether rhuZP3a22~176 or rhuZP3b177~348 may trigger acrosome reaction of human spermatozoa and the possible mechanism. The assessment of acrosome reaction was performed using chlortetracycline staining. Intracellular free calcium concentration ([Ca2+]i) in Fura-2/AM-loaded human sperm was monitored with spectrofluorophotometer. We found that the peptides rhuZP3a22~176 and rhuZP3b177~348 were significantly capable of eliciting acrosome reaction at different concentrations, respectively. With an addition of either peptide, [Ca2+]i level was raised like a peak with or without a plateau. The acrosome reaction could be inhibited by G protein sensitivity pertussis toxin (PTX), EGTA and pimozide (a T-type calcium channel blocker), whereas verapamil was less effective. This study suggests that the peptides rhuZP3a22~176 and rhuZP3b177~348 have a role similar to human ZP3, the mechanism in response to the peptides may involve in influx of calcium, G protein pathway and T-type calcium channel.


Key words: Semen • Sperm • G protein • calcium channel • chlortetracycline • intracellular calcium • pertussis toxin




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