Molecular cloning and expression analysis of the mstn and myod genes in the hybrids derived from Megalobrama amblycephala × Xenocypris davidi Bleeker

Siyu Fan , Ting Li , Zhong Tang , Zhifeng Zhou, Xin Deng, Lu Huang, Xinge Ouyang, Faxian Yu, Xiangqiong Yang, Liran Zhang, Min Tao
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Abstract

The hybrids (BY) derived from blunt snout bream (Megalobrama amblycephala, BSB, ♀) and Bleeker's yellow tai (Xenocypris davidi Bleeker, YT, ♂) have a fast growth rate and body thickness. However, the molecular mechanisms underlying the growth advantage of BY are still unclear. MSTN, a member of the TGF-β superfamily, is a negative regulator of muscle growth and development. MYOD, a member of the myogenic regulatory factor family (MRFs), is a positive regulator of muscle growth and development. MSTN and MYOD interact to co-regulate muscle growth and development. In this study, we cloned the full-length cDNAs of mstn and myod in BY, analyzed differential expression of the two genes in muscle of BY and its parents, and observed the localization of the two genes during embryonic development of BY. Multiple sequence alignment of amino acids and phylogenetic analysis showed that mstn and myod were highly conserved in BY, BSB, YT, and other carp family fishes. Quantitative Real-time PCR (qRT-PCR) showed that mstn and myod were primarily expressed in muscle of BY, and there were significant differences in the expression of the two genes in the red and white muscles of BY, BSB, and YT at different age. The results of in situ hybridization of embryos showed an overlap in the temporal and spatial expression of mstn and myod in BY embryo development, suggesting that the two genes might interact to co-regulate embryo growth and development. This study provides a theoretical basis for further research on the mechanisms of mstn and myod in muscle growth of hybrid fishes.

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驼背蛙(Megalobrama amblycephala)×大蛙(Xenocypris davidi Bleeker)杂交种中 mstn 和 myod 基因的分子克隆和表达分析
钝吻鳊(Megalobrama amblycephala, BSB, ♀)和布莱克黄台鱼(Xenocypris davidi Bleeker, YT, ♂)的杂交种(BY)生长速度快,体厚。然而,BY 生长优势的分子机制仍不清楚。TGF-β 超家族成员 MSTN 是肌肉生长和发育的负调控因子。MYOD是肌肉生长调节因子家族(MRFs)的成员,是肌肉生长和发育的正向调节因子。MSTN 和 MYOD 相互作用,共同调控肌肉的生长和发育。本研究克隆了BY中mstn和myod的全长cDNA,分析了这两个基因在BY及其亲本肌肉中的差异表达,并观察了这两个基因在BY胚胎发育过程中的定位。氨基酸多重序列比对和系统进化分析表明,mstn和myod在BY、BSB、YT和其他鲤科鱼类中高度保守。定量实时荧光定量PCR(qRT-PCR)显示,mstn和myod主要在BY的肌肉中表达,且在不同年龄的BY、BSB和YT的红白肌中,这两个基因的表达存在显著差异。胚胎原位杂交结果表明,在BY胚胎发育过程中,mstn和myod的表达在时间和空间上存在重叠,表明这两个基因可能相互作用,共同调控胚胎的生长发育。本研究为进一步研究mstn和myod在杂交鱼类肌肉生长中的作用机制提供了理论依据。
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