获能处理对猪精子质量及泛素对顶体酶抑制剂水平调控的影响

doi:10.16431/j.cnki.1671-7236.2023.01.018

Abstract

Abstract: 【Objective】 The aim of this study was to investigate the changes of acrosin inhibitor (AI) expression and AI ubiquitination level before and after boar sperm capacitation in vitro, and to understand the relationship between AI and ubiquitin-proteasome system (UPS), so as to provide a reference for further study of the role of ubiquitin-proteasome pathway (UPP) in boar sperm capacitation.【Method】 Adult healthy Landwhite boars aged 18-24 months were selected, and boar semen was collected by hand-holding method, a part of sperm was capacitated, and a part of sperm was as control (fresh sperm).The kinetic parameters, quality parameters, tyrosine phosphorylation level and zinc ions (Zn²⁺) content of sperm were detected by computer assisted sperm analysis system (CASA), hypotonic swelling method (HOST), Coomassie brilliant blue staining, Western blotting and Zn²⁺ labeling, respectively.Western blotting was used to detect the expression of AI and ubiquitin (Ub) in sperm before and after capacitation.The localization of AI and Ub in sperm was detected by immunofluorescence.The binding of AI and Ub was analyzed by immunoprecipitation.【Result】 Compared with fresh sperm, the kinetic parameters VSL and BCF of capacitated sperm were extremely significantly increased (P<0.01), and the motility, plasma membrane integrity, acrosomal membrane integrity and viability of capacitated sperm were extremely significantly decreased (P<0.01).The tyrosine phosphorylation level of capacitated sperm protein was extremely significantly increased (P<0.01), and Zn²⁺ efflux was extremely significantly increased (P<0.01).The results of Western blotting showed that compared with fresh sperm, the expression of AI in capacitated sperm was extremely significantly decreased (P<0.01), and the expression of Ub was significantly increased (P<0.05).Immunofluorescence results showed that both AI and Ub were located in the acrosome region of sperm head.The results of immunoprecipitation analysis showed that AI was ubiquitinated in capacitated sperm, and the ubiquitinated AI was extremely decreased (P<0.01).【Conclusion】 During in vitro capacitation of boar sperm, the tyrosine phosphorylation level of sperm protein was extremely significantly increased and the ubiquitination level was enhanced, and AI was degraded, thereby releasing active acrosomal enzymes, in order to prepare for the subsequent sperm penetration through the zona pellucida (ZP) of oocytes to complete fertilization.

Key words: pigs; sperm; capacitation; acrosin inhibitor (AI); ubiquitination

CLC Number:

S828

YIN Yi, LYU Yanqiu, CHEN Xuan, CAO Lipeng, ZHANG Junzheng, JIN Yi. Effects of Capacitation on Sperm Quality and Regulation of Acrosin Inhibitor Levels by Ubiquitin in Pigs[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 174-185.

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Effects of Capacitation on Sperm Quality and Regulation of Acrosin Inhibitor Levels by Ubiquitin in Pigs

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