Xiong Zhaocheng, Zhang Haihang, Huang Ben, Liu Qingyou, Wang Yingqun, Shi Deshun, Li Xiangping
State Key Laboratory of Conservation and Utilization of Subtropical Agro-bioresources, Guangxi University, Nanning 530004, China.
Guangxi Livestock and Poultry Variety Reforming Station, Nanning 530001, China.
Asian-Australas J Anim Sci. 2018 Nov;31(11):1729-1737. doi: 10.5713/ajas.18.0025. Epub 2018 Apr 12.
The aim of the current study is to investigate the relationship between prohibitin (PHB), capping actin protein of muscle Z-line beta subunit (CAPZB), and tektin-2 (TEKT2) and sperm motility in Murrah buffalo.
We collected the high-motility and low-motility semen samples, testis, ovary, muscle, kidney, liver, brain and pituitary from Murrah buffalo, and analysed the expression of PHB, CAPZB, and TEKT2 in mRNA (message RNA) and protein level.
Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) result showed that the expression of PHB was higher and CAPZB, TEKT2 were specifically expressed in testis as compared to the other 6 tissues, and that in testis, the expression of TEKT2 was higher than that of CAPZB and PHB. Immunohistochemistry test revealed that all three genes were located on the convoluted seminiferous tubule and enriched in spermatogenic cells. Both qRT-PCR and Western Blot results showed that the expression levels of PHB, CAPZB, and TEKT2 were significantly lower in the low-motility semen group compared to the high-motility semen group (p<0.05).
The expression levels of PHB, CAPZB, and TEKT2 in Murrah buffalo sperm have a high positive correlation with sperm motility. And the three genes may be potential molecular markers for the decline of buffalo sperm motility.
本研究旨在探讨抑癌蛋白(PHB)、肌Z线肌动蛋白封端蛋白β亚基(CAPZB)和微管蛋白2(TEKT2)与摩拉水牛精子活力之间的关系。
我们采集了摩拉水牛的高活力和低活力精液样本、睾丸、卵巢、肌肉、肾脏、肝脏、大脑和垂体,并分析了PHB、CAPZB和TEKT2在mRNA(信使核糖核酸)和蛋白质水平的表达情况。
定量逆转录-聚合酶链反应(qRT-PCR)结果显示,与其他6个组织相比,PHB在睾丸中的表达较高,CAPZB和TEKT2在睾丸中特异性表达,且在睾丸中,TEKT2的表达高于CAPZB和PHB。免疫组织化学检测显示,这三个基因均位于曲细精管上,并在生精细胞中富集。qRT-PCR和蛋白质免疫印迹结果均显示,与高活力精液组相比,低活力精液组中PHB、CAPZB和TEKT2的表达水平显著降低(p<0.05)。
摩拉水牛精子中PHB、CAPZB和TEKT2的表达水平与精子活力呈高度正相关。这三个基因可能是水牛精子活力下降的潜在分子标志物。