Human sperm degradation of zona pellucida proteins contributes to fertilization

Reproductive Biology and Endocrinology : RB&E
Analilia Saldívar-HernándezMayel Chirinos

Abstract

The mammalian oocyte extracellular matrix known as the zona pellucida (ZP) acts as a barrier to accomplish sperm fusion with the female gamete. Although penetration of the ZP is a limiting event to achieve fertilization, this is one of the least comprehended stages of gamete interaction. Even though previous studies suggest that proteases of sperm origin contribute to facilitate the passage of sperm through the ZP, in human this process is not yet fully understood. The aim of this study was to determine the ability of human sperm to degrade recombinant human ZP (rhZPs) proteins and to characterize the proteases involved in this process. Purified rhZP2, rhZP3 and rhZP4 proteins were incubated with capacitated sperm and the proteolytic activity was determined by Western blot analysis. To further characterize the proteases involved, parallel incubations were performed in the presence of the protease inhibitors o-phenanthroline, benzamidine and MG-132 meant to block the activity of metalloproteases, serine proteases and the proteasome, respectively. Additionally, protease inhibitors effect on sperm-ZP binding was evaluated by hemizona assay. The results showed that rhZPs were hydrolyzed in the presence of capacitated sperm. O-phena...Continue Reading

References

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Apr 12, 2011·Reproductive Sciences·Mayel ChirinosFernando Larrea

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Citations

Jan 20, 2018·Journal of Assisted Reproduction and Genetics·Alessandra AlteriPatrizia Rubino
Apr 14, 2016·Reproductive Sciences·Aideé Saray López-Torres, Mayel Chirinos
Sep 10, 2017·Reproduction : the Official Journal of the Society for the Study of Fertility·Mayel ChirinosFernando Larrea
Feb 1, 2017·Reproductive Sciences·Aideé Saray López-Torres, Mayel Chirinos

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Methods Mentioned

BETA
ubiquitination
fluorescence microscopy
affinity protein purification
phase contrast microscopy
affinity purification

Software Mentioned

GraphPad Prism
ImageJ

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