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Articles in PresS, published online ahead of print October 2, 2002
Am J Physiol Cell Physiol, 10.1152/ajpcell.00052.2002
Submitted on February 1, 2002
Accepted on September 30, 2002
1 Center for Biomedical Engineering, The City University of New York, CUNY Graduate Center, New York, NY, USA; Department of Neuroscience, Albert Einstein College of Medicine, Yeshiva University, Bronx, NY, USA
2 Department of Neuroscience, Albert Einstein College of Medicine, Yeshiva University, Bronx, NY, USA
3 Center for Biomedical Engineering, The City University of New York, CUNY Graduate Center, New York, NY, USA
4 Department of Neuroscience, Albert Einstein College of Medicine, Yeshiva University, Bronx, NY, USA; Center for Biomedical Engineering, The City University of New York, CUNY Graduate Center, New York, NY, USA
* To whom correspondence should be addressed. E-mail: spray{at}aecom.yu.edu.
We have tested the hypothesis that fluid shear stress modifies expression, function and distribution of junctional proteins (Cx43, Cx45, and ZO-1) in cultured bone cells. Cell lines with osteoblastic (MC3T3-E1) and osteocytic (MLO-Y4) phenotypes were exposed to shear stress of 5 or 20 dyn/cm2 for 1-3 h. Immunostaining indicated that at 5 dyn/cm2 distribution of Cx43, Cx45 and ZO-1 were 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 down-regulation of membrane bound Cx43 and ZO-1 and up-regulation of cytosolic Cx43 and Cx45 at different levels of shear stress. Similarly, northern blots revealed that expressions of Cx43, Cx45, and ZO-1 were selectively up- and down-regulated 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.
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