Differentiation of the myocardial rudiment of mouse embryos: an ultrastructural study including freeze-fracture replication.

The differentiation of the myocardial rudiment was examined in mouse embryos, isolated between the afternoons of the eighth and ninth days of gestation, by means of transmission electron microscopy of ultrathin sections and of freeze-fracture replicas; some of the material used for sectioning was labelled with ruthenium red. Formation of the presumptive pericardial cavity commences during the late presomite stage (afternoon of the eighth day) and the myocardial rudiment originates in situ as a thickening of the splanchnic pericardial lining. Initially, the myocardium comprises an epithelium or plate directly exposed to the pericardial lumen and overlying a separate layer of endocardial elements. As the heart tube bulges into the pericardial coelom, it becomes surrounded by a sleeve of myocardium which thickens and stratifies during the ninth day and subsequently (on the tenth day) acquires an epicardial covering of flattened cells. The myocardium commences pulsations at or about the 3-4 somite stage (morning of the ninth day) by which time the myoblasts already contain striated myofibrillae and specialised cell junctions. From its earliest appearance, the myocardial plate contains tight junctions and desmosomes between the lateral borders of the apical parts of the myoblasts. Gap junctions soon appear in the same regions and they increase in number and extent, as the myoblasts elongate and divide, thus establishing contact in various planes; they are supplemented by the formation of fasciae adherentes and of more desmosomes. On the other hand, tight junctions decline in extent and are eventually confined to the epicardium. Other features such as caveolae and T-tubules are not established by the end of the ninth day and coupling arrangements of the sarcoplasmic reticulum are only in the rudimentary stages of formation.