Cell surface glycoconjugates and the extracellular matrix of the developing mouse embryo epicardium

Cell surface glycoconjugates and the extracellular matrix (ECM) of the proepicardium and the developing epicardium were studied in early mouse embryos by light and electron microscopy with histochaemical and immunocytochaemical techniques. The extracardially located proepicardium consists of polarized mesothelial cells forming the proepicardial vesicles. These vesicles contain a fine proteoglycan network and an acellular ECM rich in hyaluronic acid. Membrane-bound glycoconjugates are shown with cuprolinic blue, alcian blue and ruthenium red on the apical (outer) cell surface, while fibronectin and laminin are present on the basal (luminal) cell surface. These membrane and matrix components of the proepicardium might be involved in specific attachment of proepicardial cells to the bare heart tube and might facilitate the initial migration of epicardial cells over the myocardial surface. In the cell coat of the cardiomyocytes of the bare heart tube the fibronectin and laminin are concentrated in patches. The formation of the epicardial covering is a rapid process, requiring only about 2 days (9-11 days) to ensheath the entire heart tube from the inflow to the outflow segment. The subepicardial matrix between the newly formed epicardial covering and myocardial layer is acellular at first, but contains a condensing proteoglycan network, membrane and matrix fibronectin, type IV collagen and laminin on the myocardial cell surface. The formation and the distribution of the subepicardial ECM show regional characteristics. The accumulating ECM forms wide subepicardial spaces and protuberances in the atrioventricular and interventricular sulci. The sulci of the heart seem to provide the optimum microenvironment for haematopoiesis and vasculogenesis. Haematopoietic islands and coronary vessel forerunners appear and concentrate in the regularly spaced surface protuberances. The vasculogenesis proceeds from the inflow to the outflow segment of the heart. The first blood capillaries appear in the sinoatrial sulcus of the 10-day embryo. By 11-13 days the subepicardial blood vessels form an interconnected network and establish the coronary artery orifices.