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牙鲆胚胎和幼鱼骨骼肌发育过程中多种肌球蛋白重链基因的表达。

The expression of multiple myosin heavy chain genes during skeletal muscle development of torafugu Takifugu rubripes embryos and larvae.

机构信息

Department of Aquatic Bioscience, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Bunkyo, Tokyo 113-8657, Japan.

出版信息

Gene. 2013 Feb 15;515(1):144-54. doi: 10.1016/j.gene.2012.10.089. Epub 2012 Nov 28.

Abstract

In vertebrates, the development-dependent and tissue-specific expression of myosin heavy chain (MYH) genes (MYHs) contributes to the formation of diverged muscle fiber types. The expression patterns of developmentally regulated MYHs have been investigated in certain species of fish. However, the expression profiles of MYHs during torafugu Takifugu rubripes development, although extensively studied in adult tissues, have not been sufficiently studied, and also the expression orders of MYHs during development have remained unclear. In the present study, we comprehensively cloned four MYHs (MYH(M743-2), MYH(M86-2), MYH(M5) and MYH(M2126-1)) from embryos, and two MYHs (MYH(M2528-1) and MYH(M1034)) from larvae, and characterized their expression pattern in relation to developmental stages of torafugu by reverse transcription (RT)-PCR and in situ hybridization. The expression of MYHs from torafugu embryos and larvae appeared sequentially and varied largely in relation to the developmental stage-dependent and fibers-type-specific manners. The transcripts of MYH(M743-2) appeared first in embryos at 3 days post fertilization (dpf) and were localized in the epaxial and hypaxial domains of fast muscle fibers of larval myotome, whereas those of MYH(M5) and MYH(M86-2) in 3 dpf and 4 dpf, respectively, and both were localized in superficial slow and horizontal myoseptum regions. The expression of MYH(M1034) and MYH(M2126-1) was quite low and mostly undetectable. Different MYHs from torafugu embryos and larvae have also been found to be expressed differentially in pectoral fin and craniofacial muscles. Interestingly, the transcripts of MYH(M2528-1) first appeared at 6 dpf and were distinctly expressed at the dorsal and ventral extremes of larval myotome, suggesting its involvement in stratified hyperplasia. The novel involvement of MYH(M2528-1) in mosaic hyperplasia was further confirmed in juvenile torafugu, where the transcripts were expressed in fast fibers with small diameters as well as the inner part of superficial slow fibers.

摘要

在脊椎动物中,肌球蛋白重链(MYH)基因(MYHs)的发育依赖性和组织特异性表达有助于形成分化的肌肉纤维类型。在某些鱼类物种中,已经研究了发育调节的 MYH 的表达模式。然而,尽管在成年组织中广泛研究了斑点叉尾鮰 Takifugu rubripes 发育过程中 MYHs 的表达谱,但对其发育过程中的表达顺序仍不清楚。在本研究中,我们从胚胎中全面克隆了 4 种 MYHs(MYH(M743-2)、MYH(M86-2)、MYH(M5)和 MYH(M2126-1)),从幼虫中克隆了 2 种 MYHs(MYH(M2528-1)和 MYH(M1034)),并通过逆转录(RT)-PCR 和原位杂交技术,研究了它们在斑点叉尾鮰发育阶段的表达模式。斑点叉尾鮰胚胎和幼虫的 MYHs 的表达顺序不同,且与发育阶段和纤维类型特异性有关。MYH(M743-2)的转录本在受精后 3 天(dpf)的胚胎中最早出现,并定位于幼虫肌节的背侧和腹侧快肌纤维中,而 MYH(M5)和 MYH(M86-2)的转录本分别在 3 dpf 和 4 dpf 出现,均定位于浅层慢肌和水平肌隔区。MYH(M1034)和 MYH(M2126-1)的表达水平较低,大多无法检测到。来自斑点叉尾鮰胚胎和幼虫的不同 MYHs 也在胸鳍和颅面肌肉中表达不同。有趣的是,MYH(M2528-1)的转录本首先在 6 dpf 出现,并在幼虫肌节的背侧和腹侧极端明显表达,提示其参与分层增生。在幼年斑点叉尾鮰中,进一步证实了 MYH(M2528-1)在镶嵌增生中的新作用,该转录本在小直径的快肌纤维以及浅层慢肌的内部表达。

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