Acrosome reaction induced by recombinant human zona pellucida 3 peptides rhuZP3a22 approximately 176 and rhuZP3b177 approximately 348 and their mechanism.
To explore the biological characteristics of the recombinant zona pellucida 3 (ZP3) peptides of rhuZP3a22 approximately 176 and rhuZP3b177 approximately 348, we examined whether rhuZP3a22 approximately 176 or rhuZP3b177 approximately 348 trigger the acrosome reaction (AR) of human spermatozoa and we investigated the underlying mechanism. The assessment of AR was performed using chlortetracycline staining. The intracellular free calcium concentration ([Ca2+]i) in Fura-2/AM-loaded human sperm was monitored with a spectrofluorophotometer. We found that rhuZP3a22 approximately 176 and rhuZP3b177 approximately 348 were capable of eliciting AR at different concentrations. With the addition of either peptide, the [Ca2+]i level was raised to a peak with or without a plateau. The AR could be inhibited by pertussis toxin (PTX), EGTA, and pimozide (a T-type calcium channel blocker), whereas verapamil was less effective in this regard. The results of the present study suggest that peptides rhuZP3a22 approximately 176 and rhuZP3b177 approximately 348 have a role similar to human ZP3, and that the mechanism of the response to the peptides involves influx of calcium, the G protein pathway, and a T-type calcium channel.
Ni
Ya
Y
Department of Reproductive Physiology, Zhejiang Academy of Medical Sciences, 182 Tian Mu Shan Road, Hangzhou, and Women's Hospital, School of Medicine, Zhejiang University, China.
Li
Kun
K
Xu
Wanxiang
W
Song
Liwen
L
Yao
Kangshou
K
Zhang
Xinzong
X
Huang
Hefeng
H
Zhang
Yanling
Y
Shi
Qi-Xian
QX
D020101:Acrosome Reaction / Q000502:physiology*; D002118:Calcium / Q000378:metabolism*; D004527:Egg Proteins / Q000502:physiology*; D004533:Egtazic Acid; D006801:Humans; D008297:Male; D008562:Membrane Glycoproteins / Q000502:physiology*; D010455:Peptides / Q000502:physiology; D037342:Pertussis Toxin; D010868:Pimozide; D011956:Receptors, Cell Surface / Q000502:physiology*; D011994:Recombinant Proteins; D013094:Spermatozoa / Q000378:metabolism / Q000502:physiology*; D000072216:Zona Pellucida Glycoproteins
D004527:Egg Proteins; D008562:Membrane Glycoproteins; D010455:Peptides; D011956:Receptors, Cell Surface; D011994:Recombinant Proteins; C000606517:ZP3 protein, human; D000072216:Zona Pellucida Glycoproteins; D010868:Pimozide; D004533:Egtazic Acid; D037342:Pertussis Toxin; D002118:Calcium