Key Laboratory of Freshwater Fish Reproduction and Development (Ministry of Education), Key Laboratory of Aquatic Science of Chongqing, School of Life Sciences, Southwest University, Chongqing 400715, China.
Int J Mol Sci. 2021 Sep 21;22(18):10170. doi: 10.3390/ijms221810170.
, a de novo methyltransferase, is essential for mammalian germ line DNA methylation. Only one is identified in mammals, and homozygous mutants of are lethal, while two paralogs, and , are identified in teleosts due to the third round of genome duplication, and homozygous mutants of and are viable in zebrafish. The expression patterns and roles of and in gonadal development remain poorly understood in teleosts. In this study, we elucidated the precise expression patterns of and in tilapia gonads. was highly expressed in oogonia, phase I and II oocytes and granulosa cells in ovaries and spermatogonia and spermatocytes in testes, while was mainly expressed in ovarian granulosa cells and testicular spermatocytes. The mutation of and was achieved by CRISPR/Cas9 in tilapia. Lower gonadosomatic index (GSI), increased apoptosis of oocytes and spermatocytes and significantly reduced sperm quality were observed in mutants, while normal gonadal development was observed in mutants. Consistently, the expression of apoptotic genes was significantly increased in mutants. In addition, the 5-methylcytosine (5-mC) level in gonads was decreased significantly, compared with that of and wild type (WT) gonads. Taken together, our results suggest that , not , plays important roles in maintaining gametogenesis in teleosts.
一个新的甲基转移酶,是哺乳动物生殖细胞 DNA 甲基化所必需的。哺乳动物中只鉴定出一个,而 的纯合突变体是致命的,而由于第三轮基因组复制,在硬骨鱼中鉴定出两个 同源物 和 ,并且 和 的纯合突变体在斑马鱼中是可行的。在硬骨鱼中, 和 在性腺发育中的表达模式和作用仍知之甚少。在这项研究中,我们阐明了 和 在罗非鱼性腺中的精确表达模式。 在卵巢中的卵原细胞、I 期和 II 期卵母细胞和颗粒细胞以及睾丸中的精原细胞和精母细胞中高度表达,而 主要在卵巢颗粒细胞和睾丸精母细胞中表达。通过 CRISPR/Cas9 在罗非鱼中实现了 和 的突变。 在 突变体中观察到性腺体指数 (GSI) 降低、卵母细胞和精母细胞凋亡增加以及精子质量显著降低,而 突变体中观察到正常的性腺发育。一致地,在 突变体中,凋亡基因的表达显著增加。此外,与 和野生型 (WT) 性腺相比, 性腺中的 5-甲基胞嘧啶 (5-mC) 水平显著降低。总之,我们的结果表明,在硬骨鱼中, 而不是 ,在维持配子发生中起重要作用。