Li Weina, Liu Gang
Institute of Reproduction and Stem Cell Engineering, Central South University, Reproductive and Genetic Hospital of CITIC-Xiangya, Changsha 410078, China.
BMC Dev Biol. 2014 Sep 19;14:38. doi: 10.1186/s12861-014-0038-5.
Hundreds of HSP40s derived from various species have been identified, of which several proteins are involved in spermatogenesis. DNAJB13 is a type II HSP40/DnaJ protein. In a previous study, we cloned mouse Dnajb13, which is up-regulated in cryptorchidism. To date, however, little is known about the localization and functions of DNAJB13 during spermatogenesis. This study intends to identify the expression pattern of DNAJB13 during mammalian spermatogenesis.
RT-PCR and western blot revealed that the Dnajb13 gene and DNAJB13 protein were expressed not only in the testis but also in several other ciliated cell-containing tissues like the trachea, lung and oviduct. Quantitative PCR showed that the expression of Dnajb13 mRNA in testis was detectable as early as postnatal week 1, and sharply increased from postnatal week 3. Western blotting and immunohistochemistry determined that the DNAJB13 protein, which was located in the cytoplasm of spermatids and the sperm flagellum, was detectable from postnatal week 4.
Based on the spatiotemporal expression observed in the cytoplasm of spermatids and sperm flagella, we suggest that DNAJB13 participates in spermiogenesis and the motility of mature spermatozoa.
已鉴定出数百种源自不同物种的热休克蛋白40(HSP40),其中几种蛋白质参与精子发生过程。DNAJB13是一种II型HSP40/DnaJ蛋白。在先前的研究中,我们克隆了小鼠Dnajb13,其在隐睾症中上调。然而,迄今为止,关于DNAJB13在精子发生过程中的定位和功能知之甚少。本研究旨在确定DNAJB13在哺乳动物精子发生过程中的表达模式。
逆转录-聚合酶链反应(RT-PCR)和蛋白质免疫印迹法显示,Dnajb13基因和DNAJB13蛋白不仅在睾丸中表达,还在其他一些含有纤毛细胞的组织中表达,如气管、肺和输卵管。定量PCR显示,睾丸中Dnajb13 mRNA的表达早在出生后第1周即可检测到,并从出生后第3周开始急剧增加。蛋白质免疫印迹法和免疫组织化学检测确定,DNAJB13蛋白位于精子细胞的细胞质和精子鞭毛中,从出生后第4周开始即可检测到。
基于在精子细胞的细胞质和精子鞭毛中观察到的时空表达,我们认为DNAJB13参与精子形成和成熟精子的运动。