AN Shiyu,ZHANG Guomin,YOU Peihua,et al.Research on the change of PPARGC1A/NRF1/TFAM pathway genes expression in the development of Hu sheep testis[J].Journal of Nanjing Agricultural University,2019,42(2):345-351.[doi:10.7685/jnau.201806041]





Research on the change of PPARGC1A/NRF1/TFAM pathway genes expression in the development of Hu sheep testis
安世钰1 张国敏1 尤佩华2 孟繁星3 刘建玲1 张婷婷1 杨花1 樊懿萱1 王锋1
1. 南京农业大学江苏省家畜胚胎工程实验室, 江苏 南京 210095;
2. 江苏波杜农牧股份有限公司, 江苏 南京 211803;
3. 南京农业大学动物科学类国家级实验教学示范中心, 江苏 南京 210095
AN Shiyu1 ZHANG Guomin1 YOU Peihua2 MENG Fanxing3 LIU Jianling1 ZHANG Tingting1 YANG Hua1 FAN Yixuan1 WANG Feng1
1. Jiangsu Livestock Embryo Engineering Laboratory, Nanjing Agricultural University, Nanjing 210095, China;
2. Jiangsu AgriPortal Company Limited, Nanjing 211803, China;
3. National Experimental Teaching Demonstration Center for Animal Science, Nanjing Agricultural University, Nanjing 210095, China
Hu sheeptestisprepubertal and postpubertalPPARGC1A/RNF1/TFAM pathway
[目的] 本试验旨在探究PPARGC1A/NRF1/TFAM通路核心基因在湖羊性成熟前后的表达变化。[方法] 选取体况良好和系谱清楚的雄性湖羊18只(3和9月龄,各9只),颈静脉采血后进行阉割处理。ELISA检测血样中睾酮(T)、瘦素、胰岛素及胰岛素样生长因子Ⅰ(IGF-Ⅰ)的浓度;免疫组化法检测睾丸组织中PPARGC1A/NRF1/TFAM通路相关蛋白的表达定位;RT-qPCR和Western blot检测睾丸组织中PPARGC1A/NRF1/TFAM通路相关基因与蛋白的表达变化。[结果] 与3月龄湖羊相比,9月龄湖羊外周血中T、瘦素、胰岛素及IGF-Ⅰ的浓度显著升高(P<0.05)。免疫组化法检测发现PPARGC1A、NRF1和TFAM蛋白在3月龄和9月龄湖羊睾丸组织的多种细胞内均呈时空特异性表达。此外,与3月龄湖羊相比,9月龄湖羊睾丸组织中PPARGC1A、NRF1TFAM mRNA和蛋白表达水平均显著升高(P<0.05)。[结论] PPARGC1A/RNF1/TFAM通路相关基因可能与湖羊睾丸发育和性成熟过程密切相关。
[Objectives] The objective of this study was to elucidate the localization and expression of the key PPARGC1A/NRF1/TFAM pathway genes in the testis of prepubertal(3-month-old) and postpubertal(9-month-old) Hu sheep.[Methods] Eighteen male Hu sheeps with good body conditions and clear pedigree were selected for this experiment(3-and 9-month-old,n=9). The testis was collected after blood collection. The concentrations of testosterone(T),leptin,insulin and insulin-like growth factorⅠ(IGF-Ⅰ) in the blood samples was detected by ELISA;cellular localization of PPARGC1A/NEF1/TFAM pathway key proteins in testis was detected by immunohistochemistry;and the expression patterns of the PPARGC1A/NRF1/TFAM pathway core genes in testis tissue were detected by RT-qPCR and Western blot.[Results] Compared with the 3-month-old Hu sheep,the concentrations of the T,leptin,insulin,and IGF-Ⅰ in the peripheral blood of 9-month-old Hu sheep significantly increased(P<0.05,respectively). Immunohistochemistry results showed an age-related and tissue-specific expressions of PPARGC1A/RNF1/TFAM pathway key proteins in the testis of 3-month-old and 9-month-old Hu sheep. In addition,the mRNA of PPARGC1A,NRF1 and TFAM and its protein expressions in the testis of 9-month-old Hu sheep were significantly higher than those in 3-month-old Hu sheep(P<0.05).[Conclusions] The PPARGC1A/RNF1/TFAM pathway may be closely associated with testicular development and sexual maturation of Hu sheep.


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