GPx6 is involved in the in vitro induced capacitation and acrosome reaction in porcine sperm

Yun Chen; Kai Wang; Delong Zhang; Zhihong Zhao; Jianhao Huang; Lele Zhou; Meiying Feng; Junsong Shi; Hengxi Wei; Li Li; Zhenfang Wu; Shouquan Zhang

doi:10.1016/j.theriogenology.2020.06.020

GPx6 is involved in the in vitro induced capacitation and acrosome reaction in porcine sperm

Theriogenology. 2020 Oct 15:156:107-115. doi: 10.1016/j.theriogenology.2020.06.020. Epub 2020 Jun 29.

Authors

Yun Chen¹, Kai Wang², Delong Zhang³, Zhihong Zhao⁴, Jianhao Huang⁵, Lele Zhou⁶, Meiying Feng⁷, Junsong Shi⁸, Hengxi Wei⁹, Li Li¹⁰, Zhenfang Wu¹¹, Shouquan Zhang¹²

Affiliations

¹ National Engineering Research Center for Breeding Swine Industry, Guangdong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou, 510642, China; Henry Fok College of Biology and Agriculture, Shaoguan University. Electronic address: luxixiworld@163.com.
² National Engineering Research Center for Breeding Swine Industry, Guangdong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou, 510642, China. Electronic address: 13556084553@163.com.
³ College of Animal Science, Xinjiang Agricultural University, Urumqi City, 830052, China. Electronic address: 1055325498@qq.com.
⁴ National Engineering Research Center for Breeding Swine Industry, Guangdong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou, 510642, China. Electronic address: zzh7797896@126.com.
⁵ National Engineering Research Center for Breeding Swine Industry, Guangdong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou, 510642, China. Electronic address: 839556615@qq.com.
⁶ National Engineering Research Center for Breeding Swine Industry, Guangdong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou, 510642, China. Electronic address: 823223094@qq.com.
⁷ National Engineering Research Center for Breeding Swine Industry, Guangdong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou, 510642, China; College of Life Sciences, Zhaoqing University, Zhaoqing, Guangdong, 526061, China. Electronic address: jony.ya@163.com.
⁸ Guangdong Wen's Breeding Swine Company, Yunfu, Guangdong, 527400, China. Electronic address: junsongstone@126.com.
⁹ National Engineering Research Center for Breeding Swine Industry, Guangdong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou, 510642, China. Electronic address: weihengxi@scau.edu.cn.
¹⁰ National Engineering Research Center for Breeding Swine Industry, Guangdong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou, 510642, China. Electronic address: 371530228@qq.com.
¹¹ National Engineering Research Center for Breeding Swine Industry, Guangdong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou, 510642, China. Electronic address: wzfemail@163.com.
¹² National Engineering Research Center for Breeding Swine Industry, Guangdong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou, 510642, China. Electronic address: sqzhang@scau.edu.cn.

PMID: 32698036
DOI: 10.1016/j.theriogenology.2020.06.020

Abstract

Glutathione peroxidases (GPxs) are regarded as important protectors against oxidative stress. Some members of this protein family were reported to play key roles in protecting sperm against oxidative stress. Whether GPx6 a member of the GPx family also plays a role in protection against oxidative stress is not known to date. The objective of the present study was to evaluate the localization and function of glutathione peroxidase 6 (GPx6) in boar accessory sex glands, seminal plasma, and sperm, as well as the effect of GPx6 on vitality and capacitation in boar sperm. qPCR and Western blot analysis demonstrated the presence of GPx6 in testis, epididymis, bulbourethral glands, prostate, seminal vesicle, sperm and seminal plasma. Incubation of sperm with an GPx6 antibody had no significant effect on the viability of boar sperm prior to capacitation. Surprisingly, when capacitated sperm was incubated with the GPx6 antibody for 240 min, sperm vitality was significantly improved. Western blotting showed that in capacitated sperm without prior pretreatment, GPx6 protein content was reduced compared to sperm before capacitation. To further confirm a role for GPx6 in sperm capacitation, we tested sperm acrosome reaction by ACR.2 and FITC-PSA. The results showed that treatment of sperm with the GPx6 antibody significantly increased sperm capacitation and acrosome reaction. Furthermore, we examined the concentration of cAMP in sperm after capacitation. ELISA demonstrated that the cAMP concentration in the sperm exposed to the GPx6 antibody was significantly higher than that of the control group. In addition, the exposure of sperm to the GPx6 antibody significantly increased the concentration of H₂O₂, while the expression of SOD3 and CAT were decreased. Based on these observations we would like to postulate that in the boar reproductive tract the GPx6 protein becomes attached to the sperm head preventing the sperm to undergo premature capacitation by affecting components of the antioxidant pathway. How GPx6 expression following ejaculation becomes suppressed to allow sperm capacitation to take place needs further investigation.

Keywords: Acrosome integrity; Capacitation; Semen; Seminal plasma; cAMP pathway.

MeSH terms

Acrosome
Acrosome Reaction*
Animals
Hydrogen Peroxide*
Male
Semen
Sperm Capacitation
Spermatozoa
Swine

Substances

Hydrogen Peroxide