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Am J Physiol Cell Physiol 284: C156-C167, 2003. First published September 4, 2002; doi:10.1152/ajpcell.00233.2002
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Vol. 284, Issue 1, C156-C167, January 2003

Developmentally regulated expression of calponin isoforms and the effect of h2-calponin on cell proliferation

M. Moazzem Hossain1, Daw-Yang Hwang1, Qi-Quan Huang1, Yasuharu Sasaki2, and Jian-Ping Jin1

1 Department of Physiology and Biophysics, Case Western Reserve University School of Medicine, Cleveland, Ohio 44106-4970; and 2 Department of Pharmacology, Kitasato University, 5-9-1, Sirogane, Minatoku 108-8641, Japan

h2-Calponin is found in both smooth muscle and nonmuscle cells, and its function remains to be established. Western blots with specific monoclonal antibodies detected significant expression of h2-calponin in the growing embryonic stomach and urinary bladder and the early pregnant uterus. Although the expression of h1-calponin is upregulated in the stomach and bladder during postnatal development, the expression of h2-calponin is decreased to low levels in quiescent smooth muscle cells. To investigate a hypothesis that h2-calponin regulates the function of the actin cytoskeleton during cytokinesis, a smooth muscle-originated cell line (SM3) lacking calponin was transfected to express either sense or antisense h2-calponin cDNA and the effects on the rates of cell proliferation were examined. Both stable and transient sense cDNA-transfected cells had a significantly decreased proliferation rate compared with the antisense cDNA-transfected or nontransfected cells. Immunofluorescence microscopy showed that the force-expressed h2-calponin was associated with actin-tropomyosin microfilaments. The number of binuclear cells was significantly greater in the sense cDNA-transfected culture, in which h2-calponin was concentrated in a nuclear ring structure formed by actin filaments. The results suggest that h2-calponin may regulate cytokinesis by inhibiting the activity of the actin cytoskeleton.

smooth muscle development; cytokinesis; tropomyosin; actin cytoskeleton; monoclonal antibody; transfective expression


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