摘要
[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、NRF1和TFAM m RNA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 PPARGC1 A/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 PPARGC1 A/NEF1/TFAM pathway key proteins in testis was detected by immunohistochemistry; and the expression patterns of the PPARGC1 A/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 PPARGC1 A/RNF1/TFAM pathway key proteins in the testis of 3-month-old and 9-month-old Hu sheep. In addition,the m RNA of PPARGC1 A,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 PPARGC1 A/RNF1/TFAM pathway may be closely associated with testicular development and sexual maturation of Hu sheep.
引文
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