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.
Mishima
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N
Department of Cellular Biology and Anatomy, Medical College of Georgia, Augusta, 30912-2000, USA.
Mikawa
T
T
Kirby
M L
ML
D000199:Actins / Q000096:biosynthesis* / Q000235:genetics; D000818:Animals; D016375:Antisense Elements (Genetics); D001011:Aorta / Q000166:cytology; D001483:Base Sequence; D048429:Cell Size / Q000235:genetics / Q000502:physiology*; D002478:Cells, Cultured; D002642:Chick Embryo; D015536:Down-Regulation; D018014:Gene Transfer Techniques; D008969:Molecular Sequence Data; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000378:metabolism*; D009432:Neural Crest / Q000166:cytology / Q000378:metabolism*; D016337:RNA, Catalytic / Q000235:genetics; D012190:Retroviridae / Q000235:genetics
D000199:Actins; D016375:Antisense Elements (Genetics); D016337:RNA, Catalytic