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Am J Physiol Cell Physiol 284: C389-C403, 2003. First published October 3, 2002; doi:10.1152/ajpcell.00052.2002
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Vol. 284, Issue 2, C389-C403, February 2003

Fluid shear stress remodels expression and function of junctional proteins in cultured bone cells

Mia M. Thi1,2, Takashi Kojima2, Stephen C. Cowin1, Sheldon Weinbaum1, and David C. Spray1,2

1 New York Center for Biomedical Engineering, City College of the City University of New York, New York, 10031; and 2 Department of Neuroscience, Albert Einstein College of Medicine, Yeshiva University, Bronx, New York 10461

We tested the hypothesis that fluid shear stress (tau ) modifies the expression, function, and distribution of junctional proteins [connexin (Cx)43, Cx45, and zona occludens (ZO)-1] in cultured bone cells. Cell lines with osteoblastic (MC3T3-E1 cells) and osteocytic (MLO-Y4 cells) phenotypes were exposed to tau -values of 5 or 20 dyn/cm2 for 1-3 h. Immunostaining indicated that at 5 dyn/cm2, the distribution of Cx43, Cx45, and ZO-1 was moderately disrupted at cell membranes; at 20 dyn/cm2, disruption was more severe. Intercellular coupling was significantly decreased at both shear stress levels. Western blots showed the downregulation of membrane-bound Cx43 and ZO-1 and the upregulation of cytosolic Cx43 and Cx45 at different levels of shear stress. Similarly, Northern blots revealed that expression of Cx43, Cx45, and ZO-1 was selectively up- and downregulated in response to different shear stress levels. These results indicate that in cultured bone cells, fluid shear stress disrupts junctional communication, rearranges junctional proteins, and determines de novo synthesis of specific connexins to an extent that depends on the magnitude of the shear stress. Such disconnection from the bone cell network may provide part of the signal whereby the disconnected cells or the remaining network initiate focal bone remodeling.

gap junctions; connexin; zona occludens-1; mechanotransduction; bone remodeling


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