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Proteomic Analysis of Bladder Smooth Muscle Cell Response to Cyclic Hydrostatic Pressure

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Abstract

We and other laboratories previously demonstrated that multiple signaling pathways are at work in SMC responses to applied mechanical stimuli, indicating the need for a high-throughput approach to study the complex mechanotransduction mechanism. In the present study, the feasibility of proteomic techniques to examine the mechanotransduction of bladder smooth muscle cells (SMC) was tested for the first time. Specifically, two-dimensional (2D) gel electrophoresis and image analyses were employed to separate and identify protein profiles in control and in cells exposed to cyclic pressure (0.1 Hz, 40 cmH2O) for 1 h. The results provide evidence that 20 (out of 155) spots exhibited changes (up- or down-regulation) in intensity between control and cyclic pressure groups. The mass spectrometry analysis provided sequences for 118 tryptic peptides from these 20 spots, and peptide mapping provided a positive match of peptide sequences to 24 distinct proteins. Moreover, the results of Western analysis independently confirmed up-regulation of nuclear matrix protein, lamin A/C in SMC exposed to cyclic pressure. Since the protein samples were predominantly of the cytosolic fraction, we concluded that the observed up-regulation was due to the translocation of either membrane-bound or nuclear localized proteins into the cytosol. Based on these observations, the present study demonstrated that the proteomic approach can be a powerful analytical tool for mechanobiology research that allows analysis of a large number of variables, thereby supporting development of new hypotheses to guide future investigation.

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Acknowledgments

The authors wish to thank Dr. Bruce Gao, Department of Bioengineering, Clemson University, for providing the rats used as the source of bladder cells. The funding for this research was provided by Paralyzed Veterans of America (2289-02) and by Clemson University.

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Authors and Affiliations

  1. Department of Bioengineering, Clemson University, 501 Rhodes Engineering Research Center, Clemson, SC, 29634-0905, USA

    Jiro Nagatomi & Margaret Gray

  2. Department of Genetics and Biochemistry, Clemson University, Clemson, SC, 29634, USA

    Yonnie Wu

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  1. Jiro Nagatomi
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Correspondence to Jiro Nagatomi.

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Nagatomi, J., Wu, Y. & Gray, M. Proteomic Analysis of Bladder Smooth Muscle Cell Response to Cyclic Hydrostatic Pressure. Cel. Mol. Bioeng. 2, 166–173 (2009). https://doi.org/10.1007/s12195-009-0043-0

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  • DOI: https://doi.org/10.1007/s12195-009-0043-0

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