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2025年4月6日 星期日
农业生物技术学报  2021, Vol. 29 Issue (9): 1741-1751    DOI: 10.3969/j.issn.1674-7968.2021.09.009
  研究论文与报告 本期目录 | 过刊浏览 | 高级检索 |
从江香猪发情期卵巢网格蛋白介导的内吞作用相关基因表达研究
唐靓婷1,2, 黄世会1, 王嘉福1,*, 冉雪琴1,*, 李升1, 牛熙1
1 贵州大学 农业生物工程研究院/动物科学学院,贵阳 550025;
2 贵州开放大学,贵阳 550023
The Expression of Genes Related to Clathrin-mediated Endocytosis in Estrus Ovary of Congjiang Xiang Pig (Sus scrofa)
TANG Liang-Ting1,2, HUANG Shi-Hui1, WANG Jia-Fu1,*, RAN Xue-Qin1,*, LI Sheng1, NIU Xi1
1 Institute of Agri-bioengineering / College of Animal Science, Guizhou University, Guiyang 550025, China;
2 Guizhou Open University, Guiyang 550023, China
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摘要 在卵巢卵泡发育过程中,卵母细胞与周围颗粒细胞之间形成缝隙连接通道,介导胞间通讯和物质转运,缝隙连接内在化可导致缝隙连接降解,是由网格蛋白介导的内吞过程(clathrin-mediated endocytosis, CME)。为了揭示CME对从江香猪(Sus scrofa)卵巢功能的调节作用,本研究以未发情期为对照,利用转录组测序技术,通过Limma、DESeq2和rMATS等软件以及逆转录PCR等技术,分析从江香猪发情期卵巢CME相关基因的表达和转录本发生的可变剪接。从江香猪卵巢中检测到8个CME基因表达,包括DAB衔接蛋白2 (DAB adaptor protein 2, DAB2),衔接蛋白复合体2亚基μ1 (adaptor related protein complex 2 subunit μ1, AP2M1), 网格蛋白轻链(clathrin light chain, CLT) A (CLTA)和B (CLTB), 动力蛋白1 (dynamin-1, DNM1), 肌球蛋白Ⅵ (myosin Ⅵ, MYO6),动力蛋白2 (dynamin-2, DNM2)和网格蛋白重链(clathrin heavy chain, CLTC),其中差异表达的基因有DAB2AP2M1CLTB;除了CLTB外,其余7个基因在发情和未发情卵巢之间发生差异的可变剪接,这些可变剪接产生的7个转录本是目前Ensembl数据库暂未收录的新转录本。说明可变剪接是从江香猪发情过程中卵巢功能重要的调控方式,以CME过程中产生新的转录本和蛋白的方式组建蛋白互作网络。结果表明,通过转录和转录后水平调控CME相关基因的表达和转录本的剪接,来影响缝隙连接的内在化,从而调节从江香猪卵巢的功能。本研究有助于提高对猪转录后调控过程的认识,对从转录组水平解析猪卵巢功能的分子调节机制具有重要意义。
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唐靓婷
黄世会
王嘉福
冉雪琴
李升
牛熙
关键词 从江香猪网格蛋白介导的内吞作用(CME)缝隙连接(GJ)可变剪接卵巢    
Abstract:During the development of ovarian follicles, gap junctions between oocytes and surrounding granulosa cells mediate intercellular communication and material transport. Gap junction internalization refers to the clathrin-mediated endocytosis (CME) process. In order to study the effect of CME-related genes on ovary function of Congjiang Xiang pig (Sus scrofa), transcriptome sequencing technology was used to analyze the expression of CME-related genes and the alternative splicing of transcripts in Congjiang Xiang pigs ovaries at estrus stage by Limma, DESeq2, rMATs softwares and reverse transcription PCR (RT-PCR) method. Transcriptome changes at diestrus stage were used as controls. As a result, 8 CME genes were detected in the ovaries of Congjiang Xiang pigs, including DAB adaptor protein 2 (DAB2), adaptor related protein complex 2 subunit μ1 (AP2M1), clathrin light chain B (CLTB), clathrin light chain A (CLTA), dynamin-1 (DNM1), myosin Ⅵ (MYO6), dynamin-2 (DNM2) and clathrin heavy chain (CLTC). Among them, the differentially expressed genes were DAB2, AP2M1 and CLTB. Except for CLTB, the other 7 genes produced different alternative splicing events between estrous and diestrous ovaries, and the 7 transcripts produced by these alternative splicing events were the new transcripts not included in the Ensembl database up to now. It showed that alternative splicing was an important regulation mode of ovary in Congjiang Xiang pigs at estrus stage, and a specific network of protein interaction was formed by producing new transcripts and proteins during CME. The results of this study indicated that the internalization of gap junction was regulated by the expression of CME-related genes and splicing of transcripts at transcription and post-transcription levels, thus regulating the ovarian function of Congjiang Xiang pig. These researches would help to understand of the post-transcriptional regulation process and the molecular regulation mechanism of porcine ovarian function.
Key wordsCongjiang Xiang pig    Clathrin-mediated endocytosis (CME)    Gap junction (GJ)    Alternative splicing    Ovary
收稿日期: 2021-03-22     
ZTFLH:  S828  
基金资助:国家自然科学基金(31960641; 31672390); 贵州省“百”层次创新型人才培养项目(黔科合人才(2016)4012号); 贵州省科技创新人才团队项目(黔科合平台人才[2019]5615); 贵州省科技支撑计划(黔科合支撑[2017]2585)
通讯作者: * jfwang@gzu.edu.cn;xqran@gzu.edu.cn   
引用本文:   
唐靓婷, 黄世会, 王嘉福, 冉雪琴, 李升, 牛熙. 从江香猪发情期卵巢网格蛋白介导的内吞作用相关基因表达研究[J]. 农业生物技术学报, 2021, 29(9): 1741-1751.
TANG Liang-Ting, HUANG Shi-Hui, WANG Jia-Fu, RAN Xue-Qin, LI Sheng, NIU Xi. The Expression of Genes Related to Clathrin-mediated Endocytosis in Estrus Ovary of Congjiang Xiang Pig (Sus scrofa). 农业生物技术学报, 2021, 29(9): 1741-1751.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/10.3969/j.issn.1674-7968.2021.09.009     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2021/V29/I9/1741
 
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