Although the small (22 Kd) heat shock protein/alpha B-crystallin functions as a major structural protein and molecular chaperone in the vertebrate lens, little is known about the protein's role in nonlenticular tissues such as the heart and skeletal muscle. Recent studies have demonstrated that alpha B-crystallin expression is uniquely regulated during myogenesis in vitro. We report here for the first time that the temporal and spatial expression of alpha B-crystallin is similarly regulated in vivo during mouse embryogenesis. Expression of alpha B-crystallin mRNA was detected by in situ hybridization in the primitive heart at 8.5 days postconception (p.c.) and in the myotome of the somites at 10.5 days p.c. This tissue-restricted pattern was corroborated by immunohistochemical studies. alpha B-crystallin mRNA and protein expression were uniform in the developing atria and ventricles without regional differences or gradients. alpha B-crystallin expression was absent in the endocardial cushion, pulmonary trunk, aorta, and endothelium. Examination of muscle precursors revealed expression throughout the dorsoventral aspect of the myotomes and in developing skeletal muscle. Our findings suggest that alpha B-crystallin may serve pivotal roles as a structural protein and a molecular chaperone in myofiber stabilization of metabolically active tissues during early embryogenesis. Thus, early alpha B-crystallin expression in myogenic lineages supports the hypothesis that the putative functions of alpha B-crystallin are coupled to the activation of genetic programs responsible for myogenic differentiation and cardiac morphogenesis.