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Am J Physiol Renal Physiol 280: F619-F627, 2001;
0363-6127/01 $5.00
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Vol. 280, Issue 4, F619-F627, April 2001

Altered membrane-cytoskeleton linkage and membrane blebbing in energy-depleted renal proximal tubular cells

Jing Chen1 and Mark C. Wagner2

1 Department of Life Sciences, Indiana State University, Terre Haute 47809; and 2 Division of Nephrology, Department of Medicine, Indiana University School of Medicine, Indianapolis, Indiana 46202

The effects of energy depletion on two membrane-cytoskeletal linker proteins (ezrin and myosin-1beta ) and membrane bleb formation were studied in isolated rabbit proximal tubule cells. Measurements of cytoskeletal-membrane interactions by using the laser optic trap method revealed a stronger association of control tubule membrane with the apical cytoskeleton compared with the basal cytoskeleton. Energy depletion weakened the apical membrane-cytoskeleton interactions to a greater degree. Biochemical studies demonstrated that energy depletion altered both ezrin and myosin-1beta . The salt-insensitive ezrin fraction dissociated from the cytoskeleton; myosin-1beta redistributed from the peripheral cytoskeleton to a perinuclear/nuclear complex. These changes in ezrin and myosin-1beta and the weakening of the membrane-cytoskeleton interactions correlated with the release of brush-border membrane blebs observed by differential interference contrast microscopy. Permeability of membrane blebs was also evaluated during energy depletion and indicated an increased permeabilization of basal blebs to 3-kDa dextrans. These results support the hypothesis that alterations in membrane-cytoskeleton linkers facilitate the formation and detachment of blebs by weakening membrane-cytoskeleton interactions.

ezrin; myosin-1; microvilli; adenosine 5'-triphosphate depletion; ischemia


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