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通过RNA干扰利用比较性染色质免疫沉淀测序和转录组分析研究对卵巢功能的调控

Study of Regulation on Ovarian Function in Through Comparative ChIP-Seq and Transcriptome Analysis Using RNA Interference.

作者信息

Liu Xiaoling, Yun Han, Xing Yan, Wang Shuo, Zhou Xueying, Zhang Jianbai

机构信息

College of life Science, Yantai University, Yantai 264006, China.

Marine Economic Research Institute, Yantai 264006, China.

出版信息

Biology (Basel). 2025 Sep 12;14(9):1259. doi: 10.3390/biology14091259.

Abstract

(forkhead box protein L2) is a transcription factor, its function and regulatory mechanism have been mainly studied in mammals; related research on marine invertebrates is still insufficient. It was found that oogenesis was affected, and even a small number of cells resembling spermatogonial morphology appeared in ovaries after the was knocked down through RNA interference (RNAi) technology in our laboratory previously. Based on previous research, this paper conducted transcriptome sequencing and differential expression analysis on the ovarian tissues between the experimental group (post-RNAi) and the control group (pre-RNAi) of , and used recombinant protein for antibody production in Chromatin Immunoprecipitation Sequencing (ChIP seq) experiments to comprehensively analyze the pathways and key genes regulated by during oogenesis. The results showed that in the RNAi experimental group, 389 genes were upregulated, and 1615 genes were downregulated. Among the differentially expressed genes (DEGs), the differential genes related to gender or gonadal development are relatively concentrated in physiological processes such as steroid hormone synthesis, spermatogenesis, gonadal development, and ovarian function maintenance, as well as the FoxO and estrogen signaling pathways. Combining transcriptome and ChIP-seq data, it was found that there were some genes related to sex gonadal development among genes which were directly regulated by FOXL2, such as , , , , , , , , , , , and . This study lays the foundation for a deeper understanding of the FOXL2's specific regulatory mechanism during oogenesis in scallops as a transcription factor.

摘要

(叉头框蛋白L2)是一种转录因子,其功能和调控机制主要在哺乳动物中得到研究;海洋无脊椎动物的相关研究仍不充分。此前在我们实验室通过RNA干扰(RNAi)技术敲低该蛋白后发现,卵子发生受到影响,甚至在卵巢中出现了少量类似精原细胞形态的细胞。基于前期研究,本文对该蛋白的实验组(RNAi后)和对照组(RNAi前)的卵巢组织进行了转录组测序和差异表达分析,并在染色质免疫沉淀测序(ChIP seq)实验中使用重组蛋白制备抗体,以全面分析该蛋白在卵子发生过程中调控的通路和关键基因。结果显示,在RNAi实验组中,389个基因上调,1615个基因下调。在差异表达基因(DEGs)中,与性别或性腺发育相关的差异基因相对集中在类固醇激素合成、精子发生、性腺发育和卵巢功能维持等生理过程,以及FoxO和雌激素信号通路中。结合转录组和ChIP-seq数据发现,在直接受FOXL2调控的基因中存在一些与性性腺发育相关的基因,如、、、、、、、、、、和。本研究为深入了解扇贝中作为转录因子的FOXL2在卵子发生过程中的具体调控机制奠定了基础。

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