GPx6在大白猪附睾细胞中的表达及其与繁殖性能的相关性

本试验旨在研究谷胱甘肽过氧化物酶6（glutathione peroxidase 6，GPx6）在大白公猪附睾细胞中的表达和定位，并探究其与公猪繁殖性能的相关性。选取15月龄的公猪和母猪各3头，取睾丸、附睾、前列腺、尿道球腺、精囊腺、卵巢和输卵管，提取蛋白，通过蛋白免疫印迹和免疫组化（IHC）检测组织和细胞中GPx6蛋白的表达和定位；根据评定大白公猪受精能力的数学模型，按照公猪配种胎次≥20胎、3次配种公猪为同一头的标准，筛选并采集符合要求的20头公猪精液，同时，统计相对应的1 279头母猪的生产成绩，计算得到公猪繁殖性能指标（包括窝产总仔数、窝产活仔数、分娩率和繁殖力）。提取精子蛋白和精清蛋白，通过BCA和ELISA检测精子和精清中GPx6蛋白的含量。使用SPSS软件的独立样本t检验及单因素方差分析（one-way ANOVA），进行差异显著性分析，用双变量Pearson进行相关性分析，P<0.05表示差异或相关显著。结果表明，GPx6蛋白在附睾中高表达，IHC结果显示，GPx6蛋白在附睾的顶细胞、基底细胞、晕细胞、主细胞和精子中表达，在肌样细胞中不表达；精清蛋白中GPx6的含量是精子蛋白的7倍，精液中GPx6蛋白的含量与窝产活仔数、窝产总仔数呈负相关关系。结果提示，GPx6在附睾的顶细胞、基底细胞、晕细胞、主细胞和精子中表达，且其在精液中的含量会影响公猪的繁殖性能，这为GPx6对公猪受精能力影响的研究奠定了理论和试验基础。

Expression of GPx6 in Epididymal Cells of Large White Boar and Its Correlation with Reproductive Performance

This study aimed to investigate the location and expression of glutathione peroxidase 6(GPx6) in epididymal cells of Large White boars as well to explore their correlation with reproductive performance of boars. Three female and male Large White pigs at 15 months old were selected, respectively. Testis, epididymis, prostate, urethral bulb gland, seminal vesicle, ovary and fallopian tube were collected for protein extraction. Expression and localization of GPx6 protein in different tissues and cells were detected by Western blot and immunohistochemistry (IHC). Boar fertilization ability was evaluated based on a mathematical mode, according to the standard of breeding parity ≥ 20 births and 3 times of breeding semen from the same boar, 20 Large White boars were selected along with their semen collected. Further, productive performance data of the 1 279 boar-paired sows were collected for evaluating the reproductive performance of those 20 breeding boars including total number born per litter (TNB), number born alive per litter (NBA), farrowing rate (FR) and fecundity. Protein from semen and sperm samples were extracted, BCA and ELISA were carried on to detect GPx6 content in sperm and seminal plasma. SPSS software was used for data statistics, while independent sample t-Test and one-way ANOVA were carried out for significant difference analysis, bivariate Pearson was used for correlation analysis, and P<0.05 indicated significant difference or correlation. The results indicated that GPx6 protein was highly expressed in epididymis, and IHC results showed GPx6 was expressed in apical cells, basal cells, halo cells, principal cells and spermatozoa from epididymis whereas was not expressed in myoid cells. The concentration of GPx6 protein in seminal plasma was as 7 times higher than that in sperm, meanwhile, the concentration of GPx6 protein in semen was negatively correlated with NBA and TNB. This study suggests that GPx6 is expressed in apical cells, basal cells, halo cells, principal cells and spermatozoa of epididymis, and its content in semen affects reproductive performance of boars, which provides a theoretical and experimental foundation for the research of function of GPx6 protein in boar fertility.