De-SUMO1ylated hexokinase 1 selectively degraded by ubiquitin-proteasome pathway promotes boar sperm capacitation in vitro

IF 2.2 2区农林科学 Q1 AGRICULTURE, DAIRY & ANIMAL SCIENCE

Animal Reproduction Science Pub Date : 2025-02-08 DOI:10.1016/j.anireprosci.2025.107794

Shunfa Yao , Qinglong Xu , Yanqiu Lv, Yuyang Zhang, Hao Li, Junzheng Zhang, Zhiwei Yao, Yang An, Xuan Chen, Yi Jin

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引用次数: 0

Abstract

This study investigates the effect of de-SUMOylated hexokinase 1 (HK1) degradation on sperm capacitation in vitro. Sperm from adult Landrace pigs was divided into three groups: non-capacitated group, capacitated group, and 16 mM 2-deoxy-D-glucose (2-DG) group. Immunoprecipitation was performed to detect the binding levels of HK1 and small ubiquitin-like modifier 1 (SUMO1) before and after capacitation. Western blotting was used to assess the degradation pathways of HK1 and tyrosine phosphorylation levels after capacitation. Immunofluorescence was employed to localize HK1 and SUMO1 in sperm. A kit was used to measure pyruvate and adenosine triphosphate (ATP) content. Fluorescent staining was used to detect reactive oxygen species (ROS) levels, zinc (Zn²⁺) levels, calcium (Ca²⁺) levels, and mitochondrial membrane potential (MMP) in sperm. Hoechst 33342 staining was used to visualize sperm nuclei and assess sperm-oocyte adhesion rates. The results showed that inhibition of HK1 activity significantly reduced pyruvate and ATP content, sperm capacitation markers (tyrosine phosphorylation, Zn²⁺), MMP, and sperm adhesion rates (P < 0.05), while significantly increasing ROS levels in capacitated sperm (P < 0.01). Immunoprecipitation and Western blotting results indicated that HK1 undergoes de-SUMOylation during capacitation and is selectively degraded via the ubiquitin-proteasome pathway (P < 0.01). Our findings suggest that de-SUMOylated HK1 activates the glycolytic pathway through ubiquitin-proteasome pathway (UPP) degradation, thereby promoting sperm capacitation. These results provide new theoretical insights into the molecular mechanisms of sperm capacitation.

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来源期刊

Animal Reproduction Science 农林科学-奶制品与动物科学

CiteScore

4.50

自引率

9.10%

发文量

136

审稿时长

54 days

期刊介绍： Animal Reproduction Science publishes results from studies relating to reproduction and fertility in animals. This includes both fundamental research and applied studies, including management practices that increase our understanding of the biology and manipulation of reproduction. Manuscripts should go into depth in the mechanisms involved in the research reported, rather than a give a mere description of findings. The focus is on animals that are useful to humans including food- and fibre-producing; companion/recreational; captive; and endangered species including zoo animals, but excluding laboratory animals unless the results of the study provide new information that impacts the basic understanding of the biology or manipulation of reproduction. The journal''s scope includes the study of reproductive physiology and endocrinology, reproductive cycles, natural and artificial control of reproduction, preservation and use of gametes and embryos, pregnancy and parturition, infertility and sterility, diagnostic and therapeutic techniques. The Editorial Board of Animal Reproduction Science has decided not to publish papers in which there is an exclusive examination of the in vitro development of oocytes and embryos; however, there will be consideration of papers that include in vitro studies where the source of the oocytes and/or development of the embryos beyond the blastocyst stage is part of the experimental design.