GPx5基因在绵羊附睾上的时空表达特性

Temporal and spatial expression characteristics of GPx5 in epididymis of sheep

【目的】研究谷胱甘肽过氧化酶5(GPx5)基因在绵羊附睾及附睾不同发育时期的表达特性。【方法】采用荧光定量PCR技术、免疫印迹技术及免疫荧光等方法,分别检测GPx5基因在绵羊附睾不同发育时期、不同部位上mRNA和蛋白的表达及分布情况。【结果】荧光定量PCR结果表明,青年羊附睾GPx5基因在头部的mRNA表达量显著高于体部和尾部(P<0.05);青年羊附睾头部GPx5基因mRNA表达量显著高于羔羊和老年羊附睾头部(P<0.05)。免疫印迹结果表明,羔羊附睾无GPx5蛋白表达;青年羊附睾各部位均有表达,而老年羊附睾头部和体部有表达、尾部无表达。灰度分析结果表明,青年羊附睾头部GPx5相对表达量显著高于其他部位(P<0.05),且青年羊附睾头部GPx5蛋白相对表达量显著高于老年羊附睾头部(P<0.05)。免疫荧光结果表明,在羔羊附睾未检测到GPx5蛋白表达信号;青年羊附睾各部位均有荧光信号表达,且附睾头部和体部荧光强度明显高于尾部,并在附睾管液体中也发现有荧光信号;老年羊附睾头部和体部有荧光信号,荧光遍布于附睾管假复层上皮细胞的细胞核及细胞质中。【结论】GPx5基因mRNA在绵羊附睾不同发育时期均有表达,但在不同部位具有表达差异性;GPx5蛋白于附睾管假复层上皮细胞表达,且在性成熟前羔羊附睾及老年羊附睾尾部未表达,因此推测GPx5蛋白的表达与绵羊性发育时期有密切关联。

【Objective】The study was conducted to explore the expression of glutathione peroxidase 5（GPx5）in sheep epididymis at different developmental stages.【Method】The mRNA and protein expression levels of GPx5 in different parts of sheep epididymis at different developmental periods were determined using fluorescence quantitative PCR,immunoblotting and immunofluorescence methods.【Result】The quantitative real-time PCR results showed that the mRNA expression level of GPx5 at epididymis caput of young sheep was significantly higher than at other parts（P〈0.05）.The mRNA expression level of GPx5 in adult sheep epididymis caput was significantly higher than that in lamb and elder sheep（P〈0.05）.Western blot results showed that GPx5 protein was not expressed in epididymis of lamb.GPx5 was expressed in all parts of adult sheep epididymis and it was not expressed in epididymis cauda of elder sheep.The protein expression level of GPx5 was significantly higher at epididymis caput of adult sheep than at other parts（P〈0.05）.The protein expression of GPx5 in epididymis caput of adult sheep was significantly higher than the expression in elder sheep（P〈0.05）.Immunofluorescence results displayed that GPx5 protein positive signal was not detected in lamb epididymis.All parts of adult sheep epididymis had positive fluorescent signals,and the fluorescence intensities at epididymis caput and corpus were significantly higher than at epididymis cauda.Positive fluorescent signal was also found in liquid of epididymis tube.The epididymis caput and corpus in elder sheep had positive fluorescent signals and fluorescence was widely distributed in nucleus and cytoplasm of epithelium cell in epididymis duct.【Conclusion】The mRNA expression of GPx5 was detected in all developmental stages of sheep with significant difference in different parts.GPx5 protein distributed at the epithelium cell in epididymis duct but it was not expressed in epididymis of lamb and epididymis cauda of elder sheep.It was indicated that the protein expression of GPx5 was closely related with sexual development stages of sheep.