Ubiquitin-mediated downregulation of the proteasome subunit ADRM1 mediates in vitro capacitation of bovine spermatozoa

Lipeng Cao; Chunyu Li; Yue Han; Yanqiu Lv; Jie Wang; Xinglin Qu; Yuyang Zhang; Weiyu Gu; Qinglong Xu; Shunfa Yao; Jinwei Lu; Aihui Lv; Xuan Chen; Yi Jin

doi:10.1016/j.anireprosci.2026.108210

Ubiquitin-mediated downregulation of the proteasome subunit ADRM1 mediates in vitro capacitation of bovine spermatozoa

Anim Reprod Sci. 2026 Aug:291:108210. doi: 10.1016/j.anireprosci.2026.108210. Epub 2026 May 2.

Authors

Lipeng Cao¹, Chunyu Li¹, Yue Han¹, Yanqiu Lv¹, Jie Wang¹, Xinglin Qu¹, Yuyang Zhang¹, Weiyu Gu¹, Qinglong Xu¹, Shunfa Yao¹, Jinwei Lu¹, Aihui Lv², Xuan Chen³, Yi Jin⁴

Affiliations

¹ Department of Animal Science, College of Agriculture, Yanbian University, Yanji 133000, China.
² Yanbian Livestock Development Corporation, Yanbian University, Yanji, Jilin Province 133000, China.
³ Department of Animal Science, College of Agriculture, Yanbian University, Yanji 133000, China. Electronic address: 0000008862@ybu.edu.cn.
⁴ Department of Animal Science, College of Agriculture, Yanbian University, Yanji 133000, China. Electronic address: yijin@ybu.edu.cn.

PMID: 42140061
DOI: 10.1016/j.anireprosci.2026.108210

Abstract

Sperm capacitation is a complex process that induces biological and biochemical changes in spermatozoa, leading to successful oocyte fertilisation. In this study, we investigated the role of the ubiquitin-proteasome system (UPS) in the remodelling of proteasomal subunits during in vitro capacitation of bovine spermatozoa. We selectively separated proteins from non-capacitated and in vitro-capacitated (IVC) bovine spermatozoa and analysed the purified proteins using four liquid chromatography-mass spectrometry (LC-MS) replicates. This analysis indicated the downregulation of 26S proteasome components, specifically the proteasome 26S subunit, non-ATPase 1 (PSMD1), and adhesion regulating molecule 1 (ADRM1), during bovine spermatozoa capacitation. Immunofluorescence localisation, image-based flow cytometry, and western blotting were used to demonstrate changes in PSMD1, ADRM1, and ubiquitination (UB) during in vitro capacitation. The results showed a significant increase in protein tyrosine phosphorylation and ubiquitination levels, along with a significant decrease in PSMD1 and ADRM1 expression. The binding levels of ADRM1 to PSMD1 and UB were also significantly enhanced. Additionally, inhibiting ADRM1 binding to PSMD1 resulted in significant increases in PSMD1 and ADRM1 expression, and a significant decrease in ADRM1 binding to UB. These findings confirmed that the UPS plays a role in proteasomal subunit changes during bovine sperm capacitation.

Keywords: ADRM1; Bovine spermatozoa; PSMD1; Ubiquitin; capacitation.