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金乌贼生长期DNA甲基化水平和模式
何暮春1,2,3, 张金勇2,3,4, 刘长琳2, 项子龙2,3,5, 柳淑芳2,3, 庄志猛2, 薛同明6
1.浙江海洋大学水产学院 舟山 316003;2.农业农村部海洋渔业可持续发展重点实验室 中国水产科学研究院黄海水产研究所 青岛 266071;3.青岛海洋科学与技术试点国家实验室海洋渔业科学与食物产出过程功能实验室 青岛 266071;4.南京农业大学无锡渔业学院 无锡 214081;5.上海海洋大学水产与生命学院 上海 201306;6.青岛金沙滩水产开发有限公司 青岛 266000
摘要:
DNA甲基化在基因表达、细胞衰老、性状分化中发挥重要调控作用,为探讨生长期金乌贼(Sepia esculenta Hoyle)不同性别、不同组织DNA甲基化水平和模式的差异,本研究采用荧光标记的甲基化敏感扩增多态性(Fluorescence-labeled Methylation Sensitive Amplified Polymorphism, F-MSAP)技术,选取12对特异性引物,检测分析了雌、雄金乌贼肌肉、心脏、胰脏和性腺4种组织的基因组DNA甲基化。结果显示,生长期金乌贼基因组DNA总甲基化水平为23.97%~39.70%,在水产脊椎动物中处于较高水平;金乌贼4种组织中,肌肉的总甲基化水平最高,这可能与金乌贼存在异速生长现象且在生长期运动器官优先发育有关;金乌贼甲基化水平和模式存在性别差异,雌性金乌贼肌肉组织DNA总甲基化水平显著低于雄性,心脏和胰脏组织DNA总甲基化水平却显著高于雄性;此外,雌性金乌贼肌肉组织全甲基化水平对总甲基化水平贡献最大,与之不同的是,雌性其他组织、雄性金乌贼各组织中半甲基化水平和全甲基化水平差异不大,说明金乌贼DNA甲基化的水平和模式具有性别和组织差异。上述结果可为深入研究金乌贼生长发育、组织分化和衰老死亡等生命过程的表观遗传学调控提供基础数据。
关键词:  金乌贼  生长期  DNA甲基化  F-MSAP  组织差异性  性别差异性
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DNA methylation level and pattern in growing Sepia esculenta Hoyle
HE Muchun1,2,3, ZHANG Jinyong2,3,4, LIU Changlin2, XIANG Zilong2,3,5, LIU Shufang2,3, ZHUANG Zhimeng2, XUE Tongming6
1.School of Fishery, Zhejiang Ocean University, Zhoushan 316003;2.Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071;3.Laboratory for Marine Fisheries Science and Food Production Processes, Pilot National Laboratory for Marine Science and Technology (Qingdao), Qingdao 266071;4.Wuxi Fisheries College, Nanjing Agricultural University, Wuxi 214081;5.College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306;6.Qingdao Jinshatan Aquatic Product Development Co., Ltd., Qingdao 266000
Abstract:
DNA methylation plays an important role in regulating gene expression, cell aging and trait differentiation. In order to investigate the differences of DNA methylation level and pattern in different genders and tissues of Sepia esculenta Hoyle, we applied Fluorescence-labeled Methylation Sensitive Amplified Polymorphism (F-MSAP) technology and analyzed the level and pattern of methylation of genomic DNA in the muscle, the heart, the pancreas and the gonad of female and male S. esculenta. The total methylation rate of S. esculenta in the growing stage was 23.97%~39.70%, which was at a high level in aquatic invertebrates. Among the female and male's four tissues, the total methylation level in muscle was the highest, which may be related to the phenomenon of allometric growth and the preferential development of the muscle during the growing state. Moreover, both the methylation levels and patterns of S. esculenta were different in gender, the total methylation level of DNA in the muscle tissue of the female S. esculenta was significantly lower than that of the male, but that in the heart and pancreas tissue was significantly higher than that in the male. In addition, we also found that the fully methylation level in the muscle of female S. esculenta was the largest contributor to the total methylation level and was consistent with the level of total methylation in the other tissues of female S. esculenta and in the four tissues of male S. esculenta. The results showed that the level and pattern of DNA methylation of S. esculenta had gender and tissue differences. This study can provide basic data for epigenetic regulation of the life processes such as the growth, differentiation, and death of S. esculenta.
Key words:  Sepia esculenta Hoyle  Growing stage  DNA methylation  F-MSAP  Tissue differences  Gender differences