National Engineering Laboratory for Druggable Gene and Protein Screening, Northeast Normal University, Changchun, 130024, China; Institute of Genetics and Cytology, Northeast Normal University, Changchun, 130024, China.
Department of Hand Surgery, China-Japan Union Hospital, Jilin University, Changchun, 130033, China.
Biochem Biophys Res Commun. 2018 Jul 12;502(2):283-288. doi: 10.1016/j.bbrc.2018.05.169. Epub 2018 May 28.
TSP50, a testis-specific gene encoding a serine protease-like protein, was specifically expressed in the spermatocytes of testes but abnormally activated and expressed in many different kinds of cancers. Here, we aimed to analyze the expression of TSP50 in mouse embryo and its function in early embryonic development. Firstly, the distribution of TSP50 in oocytes and embryonic development was characterized by immunofluorescence, RT-PCR and western blotting, and the results showed that TSP50 was detected at all studied stages with a dynamic expression pattern. When overexpressed TSP50 in zygotes by microinjection, the zygotes development was highly accelerated. On the contrary, knocking down TSP50 expression by RNA interference greatly retarded the zygote development. Furthermore, TSP50 expression at embryonic day 6.5 (E6.5), day 8.5 (E8.5) and day 10.5 (E10.5) were increasingly enhanced, However, the expression of TSP50 decreased gradually in the development and differentiation of cardiac myocyte from E12.5 to postnatal (P0). Additionally, we found that TSP50 expression was decreased during cardiac myocyte differentiation of P19 cells. Overexpression of TSP50 could decrease the expression of GATA-4, and knockdown of TSP50 markedly increase the expression of GATA-4. Taken together, our data indicate that TSP50 may play an important role during the process of mouse embryonic development as well as myocardial cell differentiation.
TSP50 是一种睾丸特异性基因，编码一种丝氨酸蛋白酶样蛋白，它特异性地在精母细胞中表达，但在许多不同类型的癌症中异常激活和表达。在这里，我们旨在分析 TSP50 在小鼠胚胎中的表达及其在早期胚胎发育中的功能。首先，通过免疫荧光、RT-PCR 和 Western blot 分析了 TSP50 在卵母细胞和胚胎发育中的分布，结果表明 TSP50 在所有研究阶段均有检测到，且表达模式具有动态性。当通过微注射将 TSP50 过表达于受精卵时，受精卵的发育被高度加速。相反，通过 RNA 干扰敲低 TSP50 的表达则极大地阻碍了受精卵的发育。此外，TSP50 在胚胎第 6.5 天（E6.5）、第 8.5 天（E8.5）和第 10.5 天（E10.5）的表达逐渐增强，然而，在 E12.5 至出生后（P0）的心肌细胞发育和分化过程中，TSP50 的表达逐渐降低。此外，我们发现 TSP50 的表达在 P19 细胞的心肌细胞分化过程中降低。TSP50 的过表达可降低 GATA-4 的表达，而 TSP50 的敲低则显著增加 GATA-4 的表达。总之，我们的数据表明，TSP50 可能在小鼠胚胎发育以及心肌细胞分化过程中发挥重要作用。
