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Articles in PresS, published online ahead of print November 20, 2001
Am J Physiol Cell Physiol, 10.1152/ajpcell.00361.2001
Submitted on July 30, 2001
Accepted on November 7, 2001
1 Surgery, Physiology, and Pathology, University of Maryland School of Medicine, Baltimore, MD, USA
2 Medicine, University of California at San Diego, San Diego, CA, USA
* To whom correspondence should be addressed. E-mail: jwang{at}smail.umaryland.edu.
Early mucosal restitution is a primary repair modality in the gastrointestinal tract and occurs by epithelial cell migration to reseal the superficial wounds after injury. We have recently demonstrated that differentiated intestinal epithelial cells induced by forced expression of the Cdx2 gene migrate over the wounded edge much faster than undifferentiated parental cells in an in vitro model, but the exact mechanism involved is still unclear. The current study tests the hypothesis that these differentiated intestinal epithelial cells exhibit increased migration after wounding by altering voltage-gated K+ (Kv) channel expression and cytosolic free Ca2+ concentration ([Ca2+]cyt ). Studies were conducted in parental IEC-6 and stable Cdx2-transfected IEC-6 cells (IEC-Cdx2L1). IEC-Cdx2L1 cells with highly differentiated phenotype expressed higher basal levels of Kv1.1 and Kv1.5 mRNAs and proteins than those observed in undifferentiated parental IEC-6 cells. Neither IEC-Cdx2L1 cells nor parental IEC-6 cells expressed voltage-dependent Ca2+ channels (VDCC). The increased expression of Kv channels in differentiated IEC-Cdx2L1 cells was associated with an increase in whole cell K+ currents (IK(V)), membrane hyperpolarization, and rise of resting [Ca2+]cyt . The migration rates in differentiated IEC-Cdx2L1 cells were ~4 times of parental IEC-6 cells. Inhibition of Kv channel expression by depletion of cellular polyamines reduced [Ca2+]cyt , resulted in the reorganization of myosin II, along with a marked reduction in stress fibers, and inhibited cell migration. In contrast, elevation of [Ca2+]cyt by the Ca2+ ionophore, ionomycin, promoted formation of myosin II stress fibers, and increased cell migration. These results suggest that increased migration of differentiated intestinal epithelial cells after wounding is mediated, at least partially, by increasing Kv channel activity and membrane hyperpolarization, thus enhancing the Ca2+ driving force for Ca2+ influx during restitution.
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