An actin barrier to resealing

K. Miyake, P. L. McNeil, K. Suzuki, R. Tsunoda, N. Sugai

Research output: Contribution to journalArticle

78 Scopus citations

Abstract

Plasma membrane disruption is a common form of cell injury in many normal biological environments, including many mammalian tissues. Survival depends on the initiation of a rapid resealing response that is mounted only in the presence of physiological levels of extracellular Ca 2+. Vesicle-vesicle and vesicle-plasma membrane fusion events occurring in cortical cytoplasm surrounding the defect are thought to be a crucial element of the resealing mechanism. However, in mammalian cells, the vesicles used in this fusion reaction (endosomes/lysosomes) are not present in a 'pre-docked' configuration and so must be brought into physical contact with one another and with the plasma membrane. We propose that a requisite prelude to fusion is the disassembly in local cell cortex of the physical barrier constituted by filamentous actin. Consistent with this hypothesis, we found that rat gastric epithelial (RGM1) cell cortical staining with phalloidin was apparently reduced at presumptive disruption sites. Moreover, flow cytofluorometric analysis of wounded RGM1 populations revealed a small, but significant, Ca 2+-dependent reduction in whole cell phalloidin staining. The functional significance of this disruption-induced depolymerization response was confirmed in several independent tests. Introduction into RGM1 cells of the filamentous actindepolymerizing agent, DNase1, enhanced resealing, although cytochalasin treatment, by itself, had no effect. By contrast, when the filamentous actin cytoskeleton was stabilized experimentally, using phalloidin or jasplakinolide, resealing was strongly inhibited. Cells in wounded cultures displayed an enhanced cortical array of filamentous actin, and resealing by such cells was enhanced strongly by both cytochalasin and DNase 1, demonstrating the specific reversibility of a biologically mediated, polymerization-induced inhibition of resealing. We conclude that localized filamentous actin disassembly removes a cortical barrier standing in the way of membrane-membrane contacts leading to resealing-requisite homotypic and exocytotic fusion events.

Original languageEnglish (US)
Pages (from-to)3487-3494
Number of pages8
JournalJournal of Cell Science
Volume114
Issue number19
StatePublished - Nov 12 2001

    Fingerprint

Keywords

  • Actin
  • Disruption
  • Exocytosis
  • Plasma membrane
  • Resealing

ASJC Scopus subject areas

  • Cell Biology

Cite this

Miyake, K., McNeil, P. L., Suzuki, K., Tsunoda, R., & Sugai, N. (2001). An actin barrier to resealing. Journal of Cell Science, 114(19), 3487-3494.