Hyaluronan and atrioventricular canal morphogenesis

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The acellular cardiac jelly matrix that separates the myocardium and endocardium in the primitive heart forms endocardial cushion swellings in two regions of the heart, the atrioventricular (AV) canal and the outflow tract. As development proceeds, a population of the endothelial cells lining the cushions undergoes an epithelial to mesenchymal transformation under the influence of myocardial-derived stimuli. Resultant migratory mesenchymal cells invade the cushion matrix to become cardiac valve precursors. Molecular genetic studies in the mouse demonstrate that both myaluronan (HA) and versican are required for formation on the cardiac cushions. Animals lacking these matrix molecules versican are required for formation of the cardiac cushions. Animals lacking these matrix molecules the compelling evidence for the role of the extracellular matrix in endocardial cushion formation, it is unclear if matrix molecules actively participate in the formation of valve structures or in the matrix provides only a supportive environment for cellular migration and differentiation. We hypothesize that HA plays two roles in AV canal morphogenesis: a) By binding to other matrix components (versican, and type VI collagen) and thus forming a structural component of the cardiac jelly and b) By signaling via a cell surface receptor-mediated mechanism upstream of Ras. Experiments are designed to examine the function and regulation of HA in each of these proposed capacities. The specific aims are: 1. Characterize the developmental expression patterns for the principal source of HA during heart development, hyaluronan synthase-2 (Has2). In addition, define the expression of molecular markers of cushion morphogenesis in mouse and chick endocardial cushions using an in vitro collagen gel transformation assay to establish the applicability of this assay for the mammalian AV canal. 2. Determine the functional role of HA during in vitro endocardial cushion morphogenesis. 3. Characterize the cis regulatory elements of the Has2 gene that are responsible for expression in the cardiac cushions. Taken together, these studies will provide a greater understanding of the biologic role of the extracellular matrix in the developing heart and provide unique reagents and techniques for future analysis of AV canal morphogenesis.