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.