Bisaha J G, Bader D
Department of Cell Biology and Anatomy, Cornell University Medical College, New York, New York 10021.
Dev Biol. 1991 Nov;148(1):355-64. doi: 10.1016/0012-1606(91)90343-2.
To investigate the initial differentiative processes of avian cardiac and skeletal myogenesis, we have isolated and characterized a molecular marker of the cardiac myocyte cell lineage, ventricular myosin heavy chain 1 (VMHC1). Our goal in this initial study was to use a gene-specific probe to analyze the expression pattern of VMHC1 RNA during development. DNA sequence analysis confirmed that VMHC1 represented a novel member of the MHC gene family. PCR analysis using gene-specific primers determined that the VMHC1 RNA is first expressed in the stage 7 cardiac primordia, much earlier than the appearance of a tubular beating heart. RNA blot analyses determined that the VMHC1 message was present in the embryonic and adult ventricles but not in the embryonic or adult atria or skeletal muscle tissues of either the fast or slow type after definitive muscle structures were formed. Still, PCR and in situ hybridization analyses of the initial phases of cardiac and skeletal myogenic differentiation determined that VMHC1 was expressed in both progenitor populations at the initiation of myogenesis regardless of the source of myoblast or site of initial differentiation. The transient expression in skeletal muscle precursors coincided with the onset of differentiation in these cells. These data suggest that the differentiative programs of cardiac and skeletal myocytes overlap during their initial phases, then quickly become distinct. The VMHC1 gene should provide a model for identification of transcription factors involved in cardiac myocyte differentiation.
为了研究禽类心脏和骨骼肌生成的初始分化过程,我们分离并鉴定了心肌细胞谱系的一个分子标志物——心室肌球蛋白重链1(VMHC1)。我们这项初步研究的目标是使用基因特异性探针来分析VMHC1 RNA在发育过程中的表达模式。DNA序列分析证实,VMHC1代表了MHC基因家族的一个新成员。使用基因特异性引物进行的PCR分析确定,VMHC1 RNA首先在第7阶段的心脏原基中表达,比管状跳动心脏的出现要早得多。RNA印迹分析确定,在确定的肌肉结构形成后,VMHC1信息存在于胚胎和成年心室中,但不存在于胚胎或成年心房或快肌或慢肌类型的骨骼肌组织中。然而,对心脏和骨骼肌生成分化初始阶段的PCR和原位杂交分析确定,无论成肌细胞的来源或初始分化的部位如何,VMHC1在成肌起始时的两个祖细胞群体中均有表达。在骨骼肌前体细胞中的瞬时表达与这些细胞分化的开始相吻合。这些数据表明,心肌细胞和骨骼肌细胞的分化程序在其初始阶段重叠,然后迅速变得不同。VMHC1基因应该为鉴定参与心肌细胞分化的转录因子提供一个模型。