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

The effects of energy depletion on two membrane-cytoskeletal linker proteins (ezrin and myosin-1 beta) 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-1 beta. The salt-insensitive ezrin fraction dissociated from the cytoskeleton; myosin-1 beta redistributed from the peripheral cytoskeleton to a perinuclear/nuclear complex. These changes in ezrin and myosin-1 beta 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.