Yang Tong, Wei Bang-Hong, Hao Shuang-Li, Wei Ya-Lan, Yang Wan-Xi
The Sperm Laboratory, College of Life Sciences, Zhejiang University, Hangzhou 310058, China.
The Sperm Laboratory, College of Life Sciences, Zhejiang University, Hangzhou 310058, China.
Gene. 2020 Sep 5;754:144848. doi: 10.1016/j.gene.2020.144848. Epub 2020 Jun 6.
The TGF-beta superfamily is widely involved in cell events such as cell division and differentiation, while bone morphogenetic proteins (BMPs) belong to one of the subgroups. Their functions in crustacean spermatogenesis are still unknown. In this study, we first identified the bone morphogenetic protein 2 (bmp2) from Eriocheir sinensis (E. sinensis) testis. The es-BMP2 shows high expression in E. sinensis testis. We found that es-BMP2 is expressed in spermatids. The successfully knockdown of es-BMP2 through in vivo RNAi are used for functional analysis. Compared with the control group, the proportion of abnormal nuclear cup morphology in mature spermatozoa increased significantly after es-bmp2 RNAi, suggesting that es-BMP2 plays an important role in mature sperm morphogenesis. Immunofluorescence results confirm this finding. In order to study the specific mechanism of es-BMP2 involved in spermiogenesis, we tested kinesin-14 KIFC1, which functions in the nucleus formation of spermatozoa in E. sinensis. The results showed that knockdown of es-BMP2 caused a significant decrease of es-KIFC1 expression. We further performed es-bmp2 knockdown in vitro in primary cultured testis cells. es-KIFC1 expression was significantly reduced after es-bmp2 RNAi. The above results indicate that es-BMP2 participates in maintaining the spermiogenesis of E. sinensis by regulating es-KIFC1 expression.
转化生长因子-β(TGF-β)超家族广泛参与细胞分裂和分化等细胞事件,而骨形态发生蛋白(BMPs)属于其中一个亚组。它们在甲壳动物精子发生中的功能仍不清楚。在本研究中,我们首先从中华绒螯蟹(E. sinensis)睾丸中鉴定出骨形态发生蛋白2(bmp2)。中华绒螯蟹BMP2(es-BMP2)在中华绒螯蟹睾丸中高表达。我们发现es-BMP2在精子细胞中表达。通过体内RNA干扰成功敲低es-BMP2用于功能分析。与对照组相比,es-bmp2 RNA干扰后成熟精子中核杯形态异常的比例显著增加,表明es-BMP2在成熟精子形态发生中起重要作用。免疫荧光结果证实了这一发现。为了研究es-BMP2参与精子发生的具体机制,我们检测了在中华绒螯蟹精子细胞核形成中起作用的驱动蛋白-14 KIFC1。结果表明,敲低es-BMP2导致es-KIFC1表达显著下降。我们进一步在原代培养的睾丸细胞中进行体外es-bmp2敲低。es-bmp2 RNA干扰后es-KIFC1表达显著降低。上述结果表明,es-BMP2通过调节es-KIFC1表达参与维持中华绒螯蟹的精子发生。