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Am J Physiol Renal Physiol 285: F230-F240, 2003. First published April 22, 2003; doi:10.1152/ajprenal.00024.2003
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Membrane repolarization is delayed in proximal tubules after ischemia-reperfusion: possible role of microtubule-organizing centers

Flavia A. Wald, Yolanda Figueroa, Andrea S. Oriolo, and Pedro J. I. Salas

Department of Cell Biology and Anatomy, University of Miami School of Medicine, Miami, Florida 33136

Submitted 17 January 2003 ; accepted in final form 17 April 2003

We have previously shown that microtubule-organizing centers (MTOCs) attach to the apical network of intermediate filaments (IFs) in epithelial cells in culture and in epithelia in vivo. Because that attachment is important for the architecture of microtubules (MTs) in epithelia, we analyzed whether chemical anoxia in LLC-PK1 and CACO-2 cells or unilateral ischemia-reperfusion in rat kidney (performed under fluorane anesthesia) had an effect on the binding and distribution of MTOCs. In culture, we found that chemical anoxia induces MTOC detachment from IFs by morphological and biochemical criteria. In reperfused rat proximal tubules, noncentrosomal MTOCs were fully detached from the cytoskeleton and scattered throughout the cytoplasm at 3 days after reperfusion, when brush borders were mostly reassembled. At that time, MTs were also fully reassembled but, as expected, lacked their normal apicobasal orientation. Two apical membrane markers expressed in S2 and S3 segments were depolarized at the same stage. At 8 days after reperfusion, membrane polarity, MTOCs, and MTs were back to normal. Na+-K+-ATPase was also found redistributed, not to the apical domain but rather to an intracellular compartment, as described by others (Alejandro VS, Nelson W, Huie P, Sibley RK, Dafoe D, Kuo P, Scandling JD Jr., and Myers BD. Kidney Int 48: 1308–1315, 1995). The prolonged depolarization of the apical membrane may have implications in the pathophysiology of acute renal failure.

ischemic acute renal failure; apical polarity; intermediate filaments



Address for reprint requests and other correspondence: P. J. I. Salas, Dept. of Cell Biology and Anatomy, R-124 -RMSB 4090, Univ. of Miami School of Medicine, 1600 NW 10th Ave., Miami, FL 33136 (E-mail: psalas{at}miami.edu).




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