Correlation between ubiquitination and defects of bull spermatozoa and removal of defective spermatozoa using anti-ubiquitin antibody-coated magnetized beads

Jian Zhang; Jie Su; Shuxiang Hu; Jindun Zhang; Rui Ding; Jitong Guo; Guifang Cao; Rongfeng Li; Qing-Yuan Sun; Xihe Li

doi:10.1016/j.anireprosci.2018.01.018

Correlation between ubiquitination and defects of bull spermatozoa and removal of defective spermatozoa using anti-ubiquitin antibody-coated magnetized beads

Anim Reprod Sci. 2018 May:192:44-52. doi: 10.1016/j.anireprosci.2018.01.018. Epub 2018 Feb 12.

Authors

Jian Zhang¹, Jie Su², Shuxiang Hu³, Jindun Zhang², Rui Ding², Jitong Guo³, Guifang Cao², Rongfeng Li⁴, Qing-Yuan Sun⁵, Xihe Li⁶

Affiliations

¹ Research Center for Animal Genetic Resources of Mongolia Plateau, Inner Mongolia University, Huhhot 010021, China.
² Inner Mongolia Saikexing Institute of Breeding and Reproductive Biotechnology in Domestic Animal, Huhhot 011517, China.
³ Research Center for Animal Genetic Resources of Mongolia Plateau, Inner Mongolia University, Huhhot 010021, China; Inner Mongolia Saikexing Institute of Breeding and Reproductive Biotechnology in Domestic Animal, Huhhot 011517, China.
⁴ Key Laboratory of Xenotransplantation of Jiangsu Province, Nanjing Medical University, Nanjing 210029, China.
⁵ State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China.
⁶ Research Center for Animal Genetic Resources of Mongolia Plateau, Inner Mongolia University, Huhhot 010021, China; Inner Mongolia Saikexing Institute of Breeding and Reproductive Biotechnology in Domestic Animal, Huhhot 011517, China. Electronic address: Lixh@life.imu.edu.cn.

PMID: 29526513
DOI: 10.1016/j.anireprosci.2018.01.018

Abstract

Ubiquitination is an important cellular process in spermatogenesis and involves the regulation of spermatid differentiation and spermiogenesis. In the current study, the correlation between bull sperm ubiquitination and sperm defects was analyzed, and the feasibility using anti-ubiquitin specific antibody immobilized magnetic beads to remove the spermatozoa with defects was assessed. A total of nine bulls were examined, and the amount of sperm ubiquitination ranged from 55 to 151. Correspondingly, the percentage of sperm deformity ranged from 9.3% to 28.1%. The coefficient of correlation was r = 0.92, indicating a significant correlation between the percentage of sperm deformity and the amount of ubiquitination (P < 0.05). The results from use of fluorescence staining and single-channel flow cytometry indicated there was a significant correlation between the sperm deformity and amount of ubiquitination (r = 0.86, P < 0.05). Results gained by use of the TUNEL and ubiquitination assays by double-channel flow cytometry indicated that the proportion of genetically defective spermatozoa with ubiquitination in Q3 and Q2 quartiles was markedly greater than that of spermatozoa with ubiquitination in Q1 and Q4 quartiles (82.1% compared with 17.9%). All these results confirmed that sperm ubiquitination is associated with genetic DNA defects (P < 0.01). Furthermore, nine semen samples with sperm motility of less than 50% (minimal motility), 50% to 70% (moderate motility) and greater than 70% (greatest motility) were selected for sorting defective spermatozoa using anti-ubiquitin specific antibody-coated magnetic beads. Strikingly, the percentage of sperm deformity significantly decreased from 18.8%, 19.0% and 17.1% to 11.7%, 11.0% and 11.0%, respectively (P < 0.05), suggesting that this method might be a feasible technology to improve the productivity via removal of the defective spermatozoa from bull semen.

Keywords: Bull sperm; Defective sperm; Magnetic-activated cell sorting; Ubiquitination.

MeSH terms

Animals
Antibodies*
Cattle
DNA Damage
Male
Semen Preservation / methods
Semen Preservation / veterinary
Specimen Handling / veterinary*
Sperm Count
Spermatozoa / abnormalities*
Spermatozoa / physiology
Ubiquitin
Ubiquitination

Substances

Antibodies
Ubiquitin