Zhang Haolin, Sheng Xia, Hu Xiao, Li Xiuwen, Xu Hui, Zhang Mengyuan, Li Ben, Xu Meiyu, Weng Qiang, Zhang Zhixiang, Taya Kazuyoshi
College of Biological Science and Technology, Beijing Forestry University, Beijing, China.
J Reprod Dev. 2010 Jun;56(3):297-302. doi: 10.1262/jrd.09-078t. Epub 2010 Mar 3.
The purpose of this study was to investigate seasonal changes of spermatogenesis and the cellular localization of P450c17 and P450arom in wild male ground squirrels during the breeding and non-breeding seasons. The testicular weight, testicular size and score count of spermatogenesis from April to September were measured, and histological and immunohistochemical observations of testicular tissues were performed in wild male ground squirrels. In addition, total proteins were extracted from testicular tissue in the breeding and non-breeding seasons and were used for Western blotting analysis for P450c17 and P450arom. There were marked variations in testicular weight, testicular size and score count of spermatogenesis from the breeding season (April) to the non-breeding season (September). Histologically, spermatogonia, primary spermatocytes, secondary spermatocytes and spermatozoa were identified in the breeding season (April). Immunolocalization of P450c17 was detected in Leydig cells and spermatozoa during the breeding season and was only found in Leydig cells during the non-breeding season. The positive signals of P450c17 by Western blotting were both observed in the breeding and non-breeding seasons. Immunolocalization of P450arom was observed in Leydig cells, Sertoli cells and all types of spermatogenic cells including mature-phase spermatozoa in the breeding season, while immunoreactivity for P450arom was not present in the testis of the non-breeding season. With P450arom antibody, a band was also only detected in the breeding season by Western blotting. These results suggest that the seasonal changes in testicular weight and size are correlated with spermatogenesis and immunolocalization of P450c17 and P450arom, and androgen and estrogen may play an important role in the spermatogenesis and testicular recrudescence and regression process.
本研究的目的是调查野生雄性地松鼠在繁殖季节和非繁殖季节精子发生的季节性变化以及细胞色素P450c17和细胞色素P450芳香化酶（P450arom）的细胞定位。测量了4月至9月野生雄性地松鼠的睾丸重量、睾丸大小和精子发生的评分计数，并对睾丸组织进行了组织学和免疫组织化学观察。此外，在繁殖季节和非繁殖季节从睾丸组织中提取总蛋白，用于细胞色素P450c17和细胞色素P450芳香化酶的蛋白质免疫印迹分析。从繁殖季节（4月）到非繁殖季节（9月），睾丸重量、睾丸大小和精子发生评分计数有明显变化。组织学上，在繁殖季节（4月）可识别出精原细胞、初级精母细胞、次级精母细胞和精子。在繁殖季节，细胞色素P450c17在睾丸间质细胞和精子中检测到免疫定位，而在非繁殖季节仅在睾丸间质细胞中发现。蛋白质免疫印迹法在繁殖季节和非繁殖季节均观察到细胞色素P450c17的阳性信号。在繁殖季节，细胞色素P450芳香化酶在睾丸间质细胞、支持细胞和包括成熟精子在内的所有类型生精细胞中观察到免疫定位，而非繁殖季节睾丸中未检测到细胞色素P450芳香化酶的免疫反应性。使用细胞色素P450芳香化酶抗体，蛋白质免疫印迹法也仅在繁殖季节检测到一条带。这些结果表明，睾丸重量和大小的季节性变化与精子发生以及细胞色素P450c17和细胞色素P450芳香化酶的免疫定位相关联，雄激素和雌激素可能在精子发生以及睾丸再生和退化过程中发挥重要作用。
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