College of Animal Science and Technology, Sichuan Agricultural University, Chengdu 611130, China.
Int J Mol Sci. 2022 Sep 12;23(18):10555. doi: 10.3390/ijms231810555.
Apoptosis plays a key role in the effective removal of excessive and defective germ cells, which is essential for sequential hermaphroditism and sex change in vertebrates. The ricefield eel, is a protogynous hermaphroditic fish that undergoes a sequential sex change from female to male. Previous studies have demonstrated that apoptosis is involved in sex change in . However, the apoptotic signaling pathway is unclear. In the current study, we explored the underlying mechanism of apoptosis during gonadal development and focused on the role of the mitochondrial apoptosis signaling pathway in sex change in . Flow cytometry was performed to detect apoptosis in gonads at five sexual stages and ovary tissues exposed to hydrogen peroxide (HO) in vitro. Then the expression patterns of key genes and proteins in the mitochondrial pathway, death receptor pathway and endoplasmic reticulum (ER) pathway were examined. The results showed that the apoptosis rate was significantly increased in the early intersexual stage and then decreased with the natural sex change from female to male. Quantitative real-time PCR revealed that , , and were mainly expressed in the five stages. ELISA demonstrated that the relative content of cytochrome-c (cyt-c) in the mitochondrial pathway was significantly higher than that of caspase8 and caspase12, with a peak in the early intersexual stage, while the levels of caspase8 and caspase12 peaked in the late intersexual stage. Interestingly, the Pearson's coefficient between cyt-c and the apoptosis rate was 0.705, which suggests that these factors are closely related during the gonadal development of Furthermore, the cyt-c signal was found to be increased in the intersexual stage by immunohistochemistry. After incubation with HO, the mRNA expression of mitochondrial pathway molecules such as , , and increased in ovary tissues. In conclusion, the present results suggest that the mitochondrial apoptotic pathway may play a more important role than the other apoptotic pathways in sex change in
细胞凋亡在有效清除多余和有缺陷的生殖细胞中起着关键作用，这对于脊椎动物的连续雌雄同体和性别转变至关重要。 日本鳗鲡是一种雌雄同体的鱼类，经历从雌性到雄性的连续性别转变。先前的研究表明，细胞凋亡参与了 的性别转变。然而，细胞凋亡信号通路尚不清楚。在本研究中，我们探讨了细胞凋亡在性腺发育过程中的潜在机制，并重点研究了线粒体凋亡信号通路在 的性别转变中的作用。通过流式细胞术检测了五个性阶段的性腺和体外暴露于过氧化氢（HO）的卵巢组织中的细胞凋亡。然后，检测了线粒体通路、死亡受体通路和内质网（ER）通路中的关键基因和蛋白的表达模式。结果表明，在早期两性阶段，凋亡率显著增加，然后随着从雌性到雄性的自然性别转变而降低。定量实时 PCR 显示 、 和 在五个阶段中主要表达。ELISA 表明线粒体通路中的细胞色素-c（cyt-c）相对含量明显高于 caspase8 和 caspase12，在早期两性阶段达到峰值，而 caspase8 和 caspase12 的水平在晚期两性阶段达到峰值。有趣的是，cyt-c 与凋亡率之间的 Pearson 相关系数为 0.705，这表明这些因素在 的性腺发育过程中密切相关。此外，免疫组织化学显示 cyt-c 信号在两性阶段增加。用 HO 孵育后，卵巢组织中线粒体通路分子如 、 、 和 的 mRNA 表达增加。综上所述，本研究结果表明，在线粒体凋亡途径在 的性别转变中可能比其他凋亡途径发挥更重要的作用。
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