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

Jing Chen, Mark Wagner

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

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

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Renal Physiology
Volume280
Issue number4 49-4
StatePublished - Apr 2001
Externally publishedYes

Fingerprint

Blister
Cytoskeleton
Kidney
Membranes
Myosins
Plakins
Interference Microscopy
Microvilli
Dextrans
Permeability
Lasers
Salts
Rabbits
ezrin

Keywords

  • Adenosine 5′-triphosphate depletion
  • Ezrin
  • Ischemia
  • Microvilli
  • Myosin-1

ASJC Scopus subject areas

  • Physiology
  • Physiology (medical)

Cite this

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

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