Mural cells of the vascular system include vascular smooth muscle cells (SMCs) and pericytes whose role is to stabilize and/or provide contractility to blood vessels. One of the earliest markers of mural cell development in vertebrates is α smooth muscle actin (acta2; αsma), which is expressed by pericytes and SMCs. In vivo models of vascular mural cell development in zebrafish are currently lacking, therefore we developed two transgenic zebrafish lines driving expression of GFP or mCherry in acta2-expressing cells. These transgenic fish were used to trace the live development of mural cells in embryonic and larval transgenic zebrafish. acta2:EGFP transgenic animals show expression that largely mirrors native acta2 expression, with early pan-muscle expression starting at 24 hpf in the heart muscle, followed by skeletal and visceral muscle. At 3.5 dpf, expression in the bulbus arteriosus and ventral aorta marks the first expression in vascular smooth muscle. Over the next 10 days of development, the number of acta2:EGFP positive cells and the number of types of blood vessels associated with mural cells increases. Interestingly, the mural cells are not motile and remain in the same position once they express the acta2:EGFP transgene. Taken together, our data suggests that zebrafish mural cells develop relatively late, and have little mobility once they associate with vessels.

Smooth muscle is important for the contractility and elasticity of visceral organs. The zebrafish is an excellent model for understanding embryonic development, yet due to a lack of appropriate markers, visceral smooth muscle development remains poorly characterized. Here, we develop markers and trace the development of gut and swim bladder smooth muscle in embryonic and juvenile fish. The first smooth muscle marker we detect in the vicinity of the gut is the myoblast marker nonmuscle myosin heavy chain-b at 50 hours postfertilization (hpf), followed by the early smooth muscle markers SM22␣-b, and ␣-smooth muscle actin at 56 and 60 hpf, respectively. Markers of more differentiated smooth muscle, smoothelin-b and cpi-17, appear by 3 days postfertilization (dpf). Tropomyosin, a relatively late marker, is first expressed at 4 dpf. We find that smooth muscle marker expression in the swim bladder follows the same sequence of marker expression as the gut, but markers have a temporal delay reflecting the later formation of swim bladder smooth muscle. Developmental Dynamics 236:1623-1632, 2007.