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Am J Physiol Cell Physiol (March 27, 2002). doi:10.1152/ajpcell.00002.2002
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Articles in PresS, published online ahead of print March 27, 2002
Am J Physiol Cell Physiol, 10.1152/ajpcell.00002.2002
Submitted on January 4, 2002
Accepted on March 22, 2002

Generation of a Cell Line with Smooth Muscle Phenotype from Hypertrophied Urinary Bladder

Yongmu Zheng1, Wilfried T. Weber2, Shuqin Wang2, Alan J. Wein1, Stephen A. Zderic3, Samuel Chacko4, and Michael E. DiSanto1*

1 Urology, University of Pennsylvania, Philadelphia, PA, USA
2 Pathobiology, University of Pennsylvania, Philadelphia, PA, USA
3 Urology, Children's Hospital of Philadelphia, Philadelphia, PA, USA
4 Pathobiology, University of Pennsylvania, Philadelphia, PA, USA; Urology, University of Pennsylvania, Philadelphia, PA, USA

* To whom correspondence should be addressed. E-mail: mdisanto{at}mail.med.upenn.edu.

We have established a cell line from hypertrophied rabbit urinary bladder smooth muscle (SM) that stably expresses SM myosin (SMM). These cells, termed BSM, are spindle-shaped and form swirls, similar to the "hills and valleys" described for cultured aortic SM cells. SDS-PAGE and Western blotting revealed that BSM expresses the N-terminal SMM heavy chain isoform SM-B, the C-terminal SM1 and SM2 isoforms, and SM {alpha}-actin. In addition, they express cGMP-dependent protein kinase G, made by contractile SM cells in vitro, but not by non-contractile cells synthesizing extracellular matrix. Immunofluorescence studies indicate a homogeneous population of cells expressing {alpha}-actin and SMM, including the SM-B isoform, and karyotyping demonstrates a stable 4N chromosomal pattern. These cells also express calcium-dependent myosin light chain kinase and phosphatase activity and contract in response to the muscarinic agonist bethanechol. To our knowledge, BSM is the first visceral SM cell line that expresses the SM-B isoform and might serve as a useful model to study the transcriptional regulation of tissue-specific SMM isoforms in differentiation and pathological SM.




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