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使用TUNEL和裂解的半胱天冬酶-3免疫组织化学检测野生埃及胡子鲶的睾丸细胞凋亡

Testicular apoptosis in feral Clarias gariepinus using TUNEL and cleaved caspase-3 immunohistochemistry.

作者信息

McClusky Leon M, Barnhoorn Irene E J, van Dyk Jacobus C, Bornman Maria S

机构信息

Department of Physiology, Faculty of Health Sciences, University of Pretoria, Pretoria, South Africa.

出版信息

Ecotoxicol Environ Saf. 2008 Sep;71(1):41-6. doi: 10.1016/j.ecoenv.2008.03.017. Epub 2008 May 23.

Abstract

This paper reports on the mechanistic basis of cellular death in the testis of Clarias gariepinus using the TUNEL and caspase-3 immunohistochemistry. It was also aimed to determine the testicular zone most suitable for the quantification of testicular apoptosis. The results showed that based on its immuno-expression patterns, activated caspase-3 has a clear and defined role in the progression of germ cell apoptosis in spermatogenically active catfish testis. Caspase-3 activation, and not TUNEL-detected DNA fragmentation, is associated with condensation of chromatin into a single mass. Testes of spermatogenically active catfish consist of spermatogenic, mature and spent tubules. Spermatogenic tubules were concluded to be the most suitable zone for the quantification of testicular cell death ratios as apoptotic events occurred predominantly in the secondary spermatocytes. The findings of this study will form the basis to link apoptotic events in the testes of C. gariepinus with the effects of EDC exposure in future studies.

摘要

本文利用TUNEL和半胱天冬酶-3免疫组织化学方法,报告了尖吻鲈睾丸细胞死亡的机制基础。研究还旨在确定最适合定量睾丸细胞凋亡的睾丸区域。结果表明,基于其免疫表达模式,活化的半胱天冬酶-3在生精活跃的鲶鱼睾丸生殖细胞凋亡进程中具有明确的作用。半胱天冬酶-3的激活而非TUNEL检测到的DNA片段化与染色质凝聚成单个团块有关。生精活跃的鲶鱼睾丸由生精小管、成熟小管和耗尽小管组成。由于凋亡事件主要发生在次级精母细胞中,因此得出结论,生精小管是定量睾丸细胞死亡率的最合适区域。本研究结果将为未来研究中尖吻鲈睾丸中的凋亡事件与环境内分泌干扰物暴露的影响之间建立联系奠定基础。

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