Role of endothelin-1/endothelin-A receptor-mediated signaling pathway in the aortic arch patterning in mice

Abstract

The intercellular signaling mediated by endothelins and their G protein- coupled receptors has recently been shown to be essential for the normal embryonic development of subsets of neural crest cell derivatives. Endothelin-1 (ET-1) is proteolytically generated from its inactive precursor by endothelin-converting enzyme-1 (ECE-1) and acts on the endothelin-A (ET(A)) receptor. Genetic disruption of this ET-1/ECE-1/ET(A) pathway results in defects in branchial arch-derived craniofacial tissues, as well as defects in cardiac outflow and great vessel structures, which are derived from cephalic (cardiac) neural crest. In this study, in situ hybridization of ET(A)(-/-) and ECE-1(-/-) embryos with a cardiac neural crest marker, cellular retinoic acid-binding protein-1, shows that the migration of neural crest cells from the neural tube to cardiac outflow tract is not affected in these embryos. Immunostaining of an endothelial marker, platelet endothelial cell adhesion molecule CD-31, shows that the initial formation of the branchial arch arteries is not disturbed in ET(A)(-/-) or ECE-1(-/-) embryos. To visualize the subsequent patterning of arch vessels in detail, we generated ET(A)(-/-) or ECE-1(-/-) embryos that expressed an SM22α-lacZ marker transgene in arterial smooth muscle cells. Whole-mount X-gal staining of these mutant embryos reveals that the abnormal regression and persistence of specific arch arteries results in disturbance of asymmetrical remodeling of the arch arteries. These defects include abnormal regression of arch arteries 4 and 6, enlargement of arch artery 3, and abnormal persistence of the bilateral ductus caroticus and right dorsal aorta. These abnormalities eventually lead to various types of great vessel malformations highly similar to those seen in neural crest-ablated chick embryos and human congenital cardiac defects. This study demonstrates that ET-1/ET(A)-mediated signaling plays an essential role in a complex process of aortic arch patterning by affecting the postmigratory cardiac neural crest cell development.