The cardiac ventricular myosin phenotype is developmentally and hormonally regulated. The genes coding for the two myosin heavy chains ( MHCs ), alpha and beta, have been recently isolated and characterized. In this study, we establish the precise temporal expression of these MHC genes in correlation with the myosin phenotype both during cardiac development and in response to different thyroid hormone levels and also document their expression in other muscle tissues. The close correlation observed between the relative abundance of the alpha- and beta-MHC mRNAs and corresponding isozymes demonstrates that the MHC phenotype is produced by the expression of the alpha- and beta-MHC genes and is regulated by changes in the level of their respective mRNAs. The opposite effect of thyroid hormone on the expression of the alpha- and beta-MHC genes in the ventricular myocardium indicates that these genes are regulated in an antithetic fashion. Finally, the MHC mRNAs encoded by the alpha- and beta-MHC genes are also present in the atrial myocardium and in the soleus, respectively.