金乌贼生长期DNA甲基化水平和模式

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%,在水产无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。

DNA methylation level and pattern in growing Sepia esculenta Hoyle

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.