DU Ping, Yao Yu-Wei, Shi Yan, Gu Zheng, Wang Jian, Sun Zhao-Gui, Zuo Jia-Ke
Shanghai Medical College, Fudan University, Shanghai 200032, China; Key Laboratory of Contraceptive Drugs and Devices of National Population and Family Planning Commission of China, Shanghai Institute of Planned Parenthood Research, Shanghai 200032, China.
Sheng Li Xue Bao. 2014 Oct 25;66(5):528-36.
Uchl1 was found to be involved in spermatocyte apoptosis. The aim of the present study was to test whether Uchl1 and its associated proteins Jab1 and p27(kip1) were involved in spermatogenic damages in response to heat-stress in cryptorchidism. Hematoxylin and eosin (HE) staining and DNA end labeling (TUNEL) were used to observe morphological and apoptotic characteristics of spermatogenic cells; Immunohistochemical analysis was used to detect changes of Uchl1 and its associated proteins Jab1 and p27(kip1) in response to heat-stress from cryptorchidism leading to spermatocyte losses; And protein affinity analysis (pull-down) and immunofluorescence co-localization were used to verify the relevance among the three proteins in spermatocytes. The results showed that, Jab1 and p27(kip1), in parallel to Uchl1, increased in spermatocytes of apoptotic appearances in response to heat-stress, but not in multinucleated giant cells; Jab1 bound to Uchl1 in testis protein extracts, and co-localized with Uchl1 and p27(kip1) specifically in spermatocytes with apoptotic appearances. These results suggest that the accumulation of Uchl1 protein is involved in the heat-stress-induced spermatocyte apoptosis through a new pathway related with Jab1 and p27(kip1), but not the formation of multinucleated giant cells.
研究发现泛素羧基末端水解酶L1(Uchl1)参与精母细胞凋亡。本研究旨在检测Uchl1及其相关蛋白Jab1和p27(kip1)是否参与隐睾症热应激所致的生精损伤。采用苏木精-伊红(HE)染色和DNA末端标记(TUNEL)法观察生精细胞的形态和凋亡特征;采用免疫组织化学分析法检测Uchl1及其相关蛋白Jab1和p27(kip1)在隐睾症热应激导致精母细胞丢失时的变化;采用蛋白质亲和分析(下拉法)和免疫荧光共定位法验证三种蛋白在精母细胞中的相关性。结果显示,与Uchl1一样,Jab1和p27(kip1)在热应激诱导出现凋亡的精母细胞中表达增加,但在多核巨细胞中未增加;在睾丸蛋白提取物中Jab1与Uchl1结合,并与Uchl1和p27(kip1)特异性共定位于出现凋亡的精母细胞中。这些结果表明,Uchl1蛋白的积累通过一条与Jab1和p27(kip1)相关的新途径参与热应激诱导的精母细胞凋亡,而不参与多核巨细胞的形成。