Cox W G, Neff A W
Medical Sciences Program, Indiana University, School of Medicine, Bloomington 47405, USA.
Differentiation. 1995 Apr;58(4):269-80. doi: 10.1046/j.1432-0436.1995.5840269.x.
Muscle-specific gene expression in the heart during Xenopus development was investigated using reverse transcription-polymerase chain reaction (RT-PCR) and whole-mount in situ hybridization to detect transcripts of the gene for the cardiac myosin heavy chain (CMHC). RT-PCR analysis determined that CMHC transcripts are present in the cardiac mesoderm at state 13, demonstrating that muscle-specific gene expression in the primitive myocardium has begun by the early neurula stage, approximately 30 h before the heart beat begins. Xenopus, therefore, is similar to amniotes and mammals in that cardiac precursor cells begin to express muscle-specific gene transcripts soon after commitment to the cardiac myocyte lineage. The earliest CMHC gene transcripts can be detected in the heart using whole-mount in situ hybridization is early tailbud stage 28, which coincides with the onset of heart tube morphogenesis. CMHC gene expression was also detected in skeletal muscle: RT-PCR analysis determined that CMHC transcripts are transiently expressed in the somite during the initial phases of skeletal muscle differentiation. Furthermore, CMHC mRNAs are expressed in a subset of head muscles of the feeding tadpole. CMHC gene expression is induced in ectodermal cells of the animal cap in blastula-stage embryos injected with synthetic MyoD or Myf5 RNA, suggesting that the CMHC gene contains regulatory elements that are responsive to the activity of those skeletal-muscle-specific transcription factors.
利用逆转录聚合酶链反应(RT-PCR)和全胚胎原位杂交技术,检测心肌肌球蛋白重链(CMHC)基因的转录本,对非洲爪蟾发育过程中心脏的肌肉特异性基因表达进行了研究。RT-PCR分析确定,在13期的心脏中胚层中存在CMHC转录本,这表明在原心肌中,肌肉特异性基因表达在神经胚早期就已开始,大约在心跳开始前30小时。因此,非洲爪蟾与羊膜动物和哺乳动物相似,即心脏前体细胞在确定为心肌细胞谱系后不久就开始表达肌肉特异性基因转录本。使用全胚胎原位杂交技术在心脏中最早检测到CMHC基因转录本是在尾芽早期28期,这与心管形态发生的开始时间一致。在骨骼肌中也检测到了CMHC基因表达:RT-PCR分析确定,在骨骼肌分化的初始阶段,CMHC转录本在体节中短暂表达。此外,CMHC mRNA在摄食期蝌蚪的一部分头部肌肉中表达。在注射了合成MyoD或Myf5 RNA的囊胚期胚胎的动物帽外胚层细胞中诱导了CMHC基因表达,这表明CMHC基因含有对那些骨骼肌特异性转录因子的活性有反应的调控元件。