Kobiyama A, Nihei Y, Hirayama Y, Kikuchi K, Suetake H, Johnston IA, Watabe S
Laboratory of Aquatic Molecular Biology and Biotechnology, Laboratory of Aquatic Animal Physiology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Bunkyo, Tokyo 113-8657, Japan and Gatty Marine Laboratory, School of Environmental and Evolutionary Biology, University of St Andrews, St Andrews, Fife KY16 8LB, Scotland.
J Exp Biol. 1998 Sep 22;201(Pt 20):2801-2813.
cDNA clones encoding the myogenic regulatory factors (MRFs) myogenin, MyoD and myf-5 were isolated by reverse-transcription polymerase chain reaction from larvae and embryos of the common carp (Cyprinus carpio L.). Myocyte-specific enhancer factor 2 (MEF2) cDNAs were identified from a cDNA library from adult carp. Northern blot analysis showed that MyoD, myf-5 and MEF2C transcripts were present in three-somite embryos, whereas myogenin and MEF2A transcripts were not detected until the 15-somite stage. Intense signals of myogenin and MyoD transcripts were observed even in 1-month-old juveniles. Levels of MyoD, myogenin and MEF2A transcripts declined between 1 and 7 months after hatching, and myf-5 gave only a weak signal in the oldest fish. In contrast, levels of MEF2C transcripts were considerably higher in 7-month-old juveniles than in 1-month-old larvae. mRNAs encoding carp myosin heavy chain and -actin were first detected at approximately the time of the first heartbeat, and levels were maximal in juveniles 1 month post-hatching. The relatively high levels of MRF mRNA in juvenile fish probably reflect the recruitment of new muscle fibres from the satellite cell population. It was concluded that the relative importance of the different members of the MyoD and MEF2 families of transcription factors for muscle differentiation changes during ontogeny in the carp.
通过逆转录聚合酶链反应,从鲤鱼(Cyprinus carpio L.)的幼虫和胚胎中分离出编码生肌调节因子(MRF)肌细胞生成素、肌分化因子(MyoD)和肌原纤维蛋白-5(myf-5)的cDNA克隆。从成年鲤鱼的cDNA文库中鉴定出肌细胞特异性增强因子2(MEF2)的cDNA。Northern印迹分析表明,MyoD、myf-5和MEF2C转录本在三对体节胚胎中存在,而肌细胞生成素和MEF2A转录本直到15对体节阶段才被检测到。即使在1月龄的幼鱼中也观察到肌细胞生成素和MyoD转录本的强烈信号。孵化后1至7个月间,MyoD、肌细胞生成素和MEF2A转录本水平下降,而在最年长的鱼中myf-5仅给出微弱信号。相比之下,7月龄幼鱼中MEF2C转录本水平比1月龄幼虫中的高得多。编码鲤鱼肌球蛋白重链和肌动蛋白的mRNA大约在第一次心跳时首次被检测到,且在孵化后1个月的幼鱼中水平最高。幼鱼中相对较高水平的MRF mRNA可能反映了卫星细胞群体中新肌纤维的募集。得出结论,在鲤鱼个体发育过程中,MyoD和MEF2转录因子家族不同成员对肌肉分化的相对重要性发生变化。