Department of Obstetrics and Gynecology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
National Clinical Research Center for Obstetrical and Gynecological Diseases, Wuhan, China.
Mol Hum Reprod. 2023 Apr 29;29(5). doi: 10.1093/molehr/gaad010.
Semaphorins are a family of evolutionarily conserved morphogenetic molecules that were initially found to be associated with axonal guidance. Semaphorin 4C (Sema4C), a member of the fourth subfamily of semaphorins, has been demonstrated to play multifaceted and important roles in organ development, immune regulation, tumor growth, and metastasis. However, it is completely unknown whether Sema4C is involved in the regulation of ovarian function. We found that Sema4C was widely expressed in the stroma, follicles, and corpus luteum of mouse ovaries, and its expression was decreased at distinct foci in ovaries of mice of mid-to-advanced reproductive age. Inhibition of Sema4C by the ovarian intrabursal administration of recombinant adeno-associated virus-shRNA significantly reduced oestradiol, progesterone, and testosterone levels in vivo. Transcriptome sequencing analysis showed changes in pathways related to ovarian steroidogenesis and the actin cytoskeleton. Similarly, knockdown of Sema4C by siRNA interference in mouse primary ovarian granulosa cells or thecal interstitial cells significantly suppressed ovarian steroidogenesis and led to actin cytoskeleton disorganization. Importantly, the cytoskeleton-related pathway RHOA/ROCK1 was simultaneously inhibited after the downregulation of Sema4C. Furthermore, treatment with a ROCK1 agonist after siRNA interference stabilized the actin cytoskeleton and reversed the inhibitory effect on steroid hormones described above. In conclusion, Sema4C may play an important role in ovarian steroidogenesis through regulation of the actin cytoskeleton via the RHOA/ROCK1 signaling pathway. These findings shed new light on the identification of dominant factors involved in the endocrine physiology of female reproduction.
信号蛋白是一个进化上保守的形态发生分子家族，最初被发现与轴突导向有关。信号蛋白 4C（Sema4C）是信号蛋白的第四个亚家族的成员，它在器官发育、免疫调节、肿瘤生长和转移中发挥着多方面的重要作用。然而，Sema4C 是否参与调节卵巢功能完全未知。我们发现 Sema4C 在小鼠卵巢的基质、卵泡和黄体中广泛表达，并且在处于中晚期生殖年龄的小鼠卵巢中，其表达在特定部位减少。通过卵巢腔内给予重组腺相关病毒-shRNA 抑制 Sema4C，可显著降低体内雌二醇、孕酮和睾酮水平。转录组测序分析显示，与卵巢甾体生成和肌动蛋白细胞骨架相关的途径发生变化。同样，通过 siRNA 干扰在小鼠原代卵巢颗粒细胞或间质细胞中敲低 Sema4C，可显著抑制卵巢甾体生成，并导致肌动蛋白细胞骨架解聚。重要的是，Sema4C 下调后，与细胞骨架相关的途径 RHOA/ROCK1 也同时被抑制。此外，siRNA 干扰后 ROCK1 激动剂的处理稳定了肌动蛋白细胞骨架，并逆转了上述对甾体激素的抑制作用。总之，Sema4C 可能通过 RHOA/ROCK1 信号通路调节肌动蛋白细胞骨架在卵巢甾体生成中发挥重要作用。这些发现为鉴定女性生殖内分泌生理学中涉及的主导因素提供了新的视角。
