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Hoffmann, R. A wiki for the life sciences where authorship matters. Nature Genetics (2008)

Smooth muscle alpha-actin downregulation in cultured chick aortic smooth muscle and neural crest cells is associated with altered cell shape.

A modified CXL retrovirus was used to clone an antisense smooth muscle alpha-actin ribozyme sequence adjacent to the reporter lacZ sequence. The virus was applied to downregulate alpha-actin expression in cultured smooth muscle cells obtained from chicken aortic arch and cultured neural crest cells. After infection with the ribozyme-containing CXL retrovirus both the smooth muscle and neural crest cells showed beta-galactosidase activity accompanied by a reduction of smooth muscle alpha-actin-positive fibers. Double staining of beta-galactosidase and smooth muscle alpha-actin using immunohistochemistry revealed that single cells infected with the CXL/ribozyme showed little to no smooth muscle alpha-actin protein. The absence of smooth muscle alpha-actin was associated with a distinct change in cellular morphology of the cultured cells, suggesting that expression of smooth muscle alpha-actin in cultured neural crest cells may be associated with cytoskeletal elements rather than vascular smooth muscle phenotype.[1]


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