ATP depletion

A novel method to study junctional properties in epithelial tissues. I. Rearrangement of the actin cytoskeleton

Robert Bacallao, Alan Garfinkel, Steven Monke, Guido Zampighi, Lazaro J. Mandel

Research output: Contribution to journalArticle

131 Citations (Scopus)

Abstract

The effect of cellular injury caused by depletion of intracellular ATP stores was studied in the Madin-Darby canine kidney (MDCK) and JTC cell lines. In prior studies, it was shown that ATP depletion uncouples the gate and fence functions of the tight junction. This paper extends these observations by studying the changes in the actin cytoskeleton and tight junction using electron microscopy and confocal fluorescence microscopy in combination with computer-aided three-dimensional reconstruction. Marked regional differences in the sensitivity to the effects of ATP depletion were observed in the actin cytoskeleton. Actin depolymerization appears to first affect the cortical actin network running along the apical basal axis of the cell. The next actin network that is disrupted is the stress fibers found at the basal surface of the cell. Finally, the actin ring at the level of the zonulae occludens and adherens is compromised. The breakup of the actin ring correlates with ultrastructural changes in tight junction strands and the loss of the tight junction's role as a molecular fence. During the process of actin network dissolution, polymerized actin aggregates form in the cytoplasm. The changes in the junctional complexes and the potential to reverse the ATP depletion suggest that this may be a useful method to study junctional complex formation and its relationship to the actin cytoskeletal network.

Original languageEnglish (US)
Pages (from-to)3301-3313
Number of pages13
JournalJournal of Cell Science
Volume107
Issue number12
StatePublished - Dec 1994
Externally publishedYes

Fingerprint

Actin Cytoskeleton
Actins
Epithelium
Adenosine Triphosphate
Tight Junctions
Adherens Junctions
Stress Fibers
Madin Darby Canine Kidney Cells
Fluorescence Microscopy
Confocal Microscopy
Electron Microscopy
Cytoplasm
Cell Line
Wounds and Injuries

Keywords

  • Actin cytoskeleton
  • ATP depletion
  • Tight junction

ASJC Scopus subject areas

  • Cell Biology

Cite this

ATP depletion : A novel method to study junctional properties in epithelial tissues. I. Rearrangement of the actin cytoskeleton. / Bacallao, Robert; Garfinkel, Alan; Monke, Steven; Zampighi, Guido; Mandel, Lazaro J.

In: Journal of Cell Science, Vol. 107, No. 12, 12.1994, p. 3301-3313.

Research output: Contribution to journalArticle

Bacallao, Robert ; Garfinkel, Alan ; Monke, Steven ; Zampighi, Guido ; Mandel, Lazaro J. / ATP depletion : A novel method to study junctional properties in epithelial tissues. I. Rearrangement of the actin cytoskeleton. In: Journal of Cell Science. 1994 ; Vol. 107, No. 12. pp. 3301-3313.
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