DNA methylation is a prevalent epigenetic modification in vertebrates, and it has been shown to be involved the regulation of gene expression and embryo development. However, it remains unclear how DNA methylation regulates sexual development, especially in species without sex chromosomes. To determine this, we utilized zebrafish to investigate DNA methylation reprogramming during juvenile germ cell development and adult female-to-male sex transition. We reveal that primordial germ cells (PGCs) undergo significant DNA methylation reprogramming during germ cell development, and the methylome of PGCs is reset to an oocyte/ovary-like pattern at 9 days post fertilization (9 dpf). When DNA methyltransferase (DNMT) activity in juveniles was blocked after 9 dpf, the zebrafish developed into females. We also show that Tet3 is involved in PGC development. Notably, we find that DNA methylome reprogramming during adult zebrafish sex transition is similar to the reprogramming during the sex differentiation from 9 dpf PGCs to sperm. Furthermore, inhibiting DNMT activity can prevent the female-to-male sex transition, suggesting that methylation reprogramming is required for zebrafish sex transition. In summary, DNA methylation plays important roles in zebrafish germ cell development and sexual plasticity.
研究问题:斑马鱼生殖细胞的DNA甲基化是如何重编程的?生殖细胞的DNA甲基化重编程是否与斑马鱼的性别决定有关? 成年雌性斑马鱼可以改变性别成为雄性,其背后的分子机制是什么? 研究方法:在本研究中,利用流式分选技术和显微操作分离斑马鱼生殖细胞,通过微量细胞转录组和DNA甲基化测序,绘制了全面的斑马鱼幼年生殖细胞发育和性别反转过程中的DNA甲基化图谱。利用基因编辑技术,证明了Tet3蛋白参与斑马鱼的生殖细胞发育。并通过抑制DNA甲基化转移酶 (DNMTs)活性的方法,揭示了DNA甲基化在斑马鱼性别决定和性别反转过程中的作用。主要结果1:斑马鱼生殖细胞在发育过程中经历了显著的重编程,但与哺乳类动物生殖细胞中情况完全不同。主要结果2:DNA甲基化重编程与斑马鱼生殖发育和性别决定有关。主要结果3:Tet3蛋白参与斑马鱼生殖细胞的发育。主要结果4:在成年斑马鱼性别转换过程中,DNA甲基化可以从雌性模式重置为雄性模式。通过抑制DNMTs活性,证明DNA甲基化对成年斑马鱼的性别可塑性至关重要。
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The Role of DNA Methylation Reprogramming During Sex Determination and Transition in Zebrafish
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