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肌动蛋白生成因子(myomaker)基因的甲基化状态和表达模式在日本牙鲆(Paralichthys olivaceus)的出生后发育中起着重要作用。

Methylation status and expression patterns of myomaker gene play important roles in postnatal development in the Japanese flounder (Paralichthys olivaceus).

机构信息

Key Laboratory of Mariculture (Ocean University of China), Ministry of Education, Qingdao 266003, China.

Key Laboratory of Mariculture (Ocean University of China), Ministry of Education, Qingdao 266003, China.

出版信息

Gen Comp Endocrinol. 2019 Sep 1;280:104-114. doi: 10.1016/j.ygcen.2019.04.017. Epub 2019 Apr 16.

Abstract

Myomaker is a membrane protein that plays a crucial role in the fusion of myoblasts during muscle growth. DNA methylation, a significant factor, regulates gene expression. The aim of this study was to examine the methylation and mRNA expression patterns of the myomaker gene during 8 different postnatal developmental stages in the Japanese flounder (L: 7 days post hatch (dph); M1: 21 dph; M2: 28 dph; M3: 35 dph; J1: 90 dph; J2: 180 dph; A1: 24 months; A2: 36 months). Muscle tissue samples were taken from Japanese flounder at different postnatal development stages to measure the extent of DNA methylation and gene expression. Methylation level in the promoter and exon 1 of myomaker was measured using bisulfite sequencing, and the relative expression of myomaker during each developmental stage was measured by quantitative PCR. The relative expression levels of myomaker were up-regulated from stages L to M2, M3 to J2, and methylation of myomaker was negatively correlated with mRNA expression. Furthermore, the CpG site located at -26 bp in the promoter was the lowest methylated region in all developmental stages. These results offer a basis for understanding the mechanism by which myomaker regulates muscle formation during postnatal development.

摘要

肌生成素是一种膜蛋白,在肌肉生长过程中肌细胞融合中起着至关重要的作用。DNA 甲基化是一个重要的调节因子,控制着基因的表达。本研究旨在研究肌生成素基因在日本牙鲆(L:孵化后 7 天(dph);M1:21 dph;M2:28 dph;M3:35 dph;J1:90 dph;J2:180 dph;A1:24 个月;A2:36 个月)的 8 个不同的出生后发育阶段的甲基化和 mRNA 表达模式。从日本牙鲆的不同出生后发育阶段采集肌肉组织样本,以测量 DNA 甲基化和基因表达的程度。使用亚硫酸氢盐测序测量肌生成素启动子和外显子 1 的甲基化水平,通过定量 PCR 测量每个发育阶段肌生成素的相对表达。肌生成素的相对表达水平从 L 期到 M2 期、M3 期到 J2 期上调,肌生成素的甲基化与 mRNA 表达呈负相关。此外,在所有发育阶段,位于启动子-26 bp 的 CpG 位点是甲基化程度最低的区域。这些结果为了解肌生成素在出生后发育过程中调节肌肉形成的机制提供了基础。

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