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转基因方法揭示的斑马鱼 Oryzias latipes 肌肉的早期发育,使用胚胎和幼虫类型的肌球蛋白重链基因。

Early development of medaka Oryzias latipes muscles as revealed by transgenic approaches using embryonic and larval types of myosin heavy chain genes.

机构信息

Department of Aquatic Bioscience, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Bunkyo, Tokyo, Japan.

出版信息

Dev Dyn. 2010 Jun;239(6):1807-17. doi: 10.1002/dvdy.22298.

Abstract

We cloned three full-length cDNAs encoding myosin heavy chains (MYHs) previously found to be expressed in embryos or larvae of medaka Oryzias latipes. Based on cDNA sequence information, the three medaka MYH genes, mMYH(emb1), mMYH(L1) and mMYH(L2), were localized on the chromosomes. In vivo promoter assay using the gene encoding green or red fluorescent protein and linked to the 5'-flanking region of mMYH demonstrated that the transcripts of fast-type mMYH(emb1), first expressed in embryos but belonging to the adult type in phylogenetic analysis, were located in the horizontal myoseptum. On the other hand, embryonic fast-type mMYH(L1) and mMYH(L2) were expressed in the whole myotomes. Interestingly, cells expressing mMYH(emb1) were localized together with engrailed, and cyclopamine, which blocks hedgehog signaling, inhibited mMYH(emb1) expression as well as the formation of the horizontal myoseptum, suggesting that muscle pioneer cells express mMYH(emb1) as a key protein in the formation of the horizontal myoseptum.

摘要

我们克隆了三个全长 cDNA,这些 cDNA 编码的肌球蛋白重链 (myosin heavy chains, MYHs) 之前在斑马鱼胚胎或幼虫中被发现有表达。基于 cDNA 序列信息,这三个斑马鱼 MYH 基因,mMYH(emb1)、mMYH(L1) 和 mMYH(L2),被定位在染色体上。使用编码绿色或红色荧光蛋白的基因进行体内启动子检测,并与 mMYH 的 5' 侧翼区相连,证明了快速型 mMYH(emb1)的转录物,最初在胚胎中表达,但在系统发育分析中属于成体类型,位于水平肌隔中。另一方面,胚胎快速型 mMYH(L1)和 mMYH(L2)在整个肌节中表达。有趣的是,表达 mMYH(emb1)的细胞与 engrailed 一起定位,hedgehog 信号阻断剂环巴胺不仅抑制了 mMYH(emb1)的表达,也抑制了水平肌隔的形成,表明肌肉先驱细胞表达 mMYH(emb1)作为形成水平肌隔的关键蛋白。

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