Molecular Characterization, Expression Pattern, DNA Methylation and Gene Disruption of Figla in Blotched Snakehead (Channa maculata)

文献类型： 外文期刊

第一作者： Zhang, Yang

作者： Zhang, Yang;Lu, Yuntao;Cui, Xiaojuan;Sun, Yuandong;Ou, Mi;Zhang, Yang;Lu, Yuntao;Zhang, Xiaotian;Wu, Yuxia;Zhao, Jian;Luo, Qing;Liu, Haiyang;Chen, Kunci;Fei, Shuzhan;Ou, Mi;Xu, Feng

作者机构：

关键词： Figla; ovarian development; gene expression; epigenetic regulation; CRISPR/Cas9 system

期刊名称：ANIMALS （ 影响因子：3.0； 五年影响因子：3.2 ）

ISSN： 2076-2615

年卷期： 2024 年 14 卷 3 期

页码：

收录情况： SCI

摘要： Simple Summary: There is a significant sexual dimorphism in the blotched snakehead (Channa maculata). However, studies on sex differentiation and gonadal development of C. maculata are limited. Factor in the germ line alpha (Figla) is an oocyte-specific basic helix-loop-helix (bHLH) transcription factor, involved in ovarian development. In the present research, the expression patterns and cellular location of Figla, as well as the CpG methylation of its promoter, demonstrated that Figla participated in ovarian differentiation and development in C. maculata. Meanwhile, the gene disruption of Figla was realized by the CRISPR/Cas9 system for further functional research. Figla is one of the earliest expressed genes in the oocyte during ovarian development. In this study, Figla was characterized in C. maculata, one of the main aquaculture species in China, and designated as CmFigla. The length of CmFigla cDNA was 1303 bp, encoding 197 amino acids that contained a conserved bHLH domain. CmFigla revealed a female-biased expression patterns in the gonads of adult fish, and CmFigla expression was far higher in ovaries than that in testes at all gonadal development stages, especially at 60 similar to 180 days post-fertilization (dpf). Furthermore, a noteworthy inverse relationship was observed between CmFigla expression and the methylation of its promoter in the adult gonads. Gonads at 90 dpf were used for in situ hybridization (ISH), and CmFigla transcripts were mainly concentrated in oogonia and the primary oocytes in ovaries, but undetectable in the testes. These results indicated that Figla would play vital roles in the ovarian development in C. maculata. Additionally, the frame-shift mutations of CmFigla were successfully constructed through the CRISPR/Cas9 system, which established a positive foundation for further investigation on the role of Figla in the ovarian development of C. maculata. Our study provides valuable clues for exploring the regulatory mechanism of Figla in the fish ovarian development and maintenance, which would be useful for the sex control and reproduction of fish in aquaculture.

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