The mechanisms of cell membrane resealing in rabbit corneal epithelial cells

Curr Eye Res. 2005 Jul;30(7):543-54. doi: 10.1080/02713680590968574.

Abstract

Purpose: To examine membrane repair mechanisms in rabbit corneal epithelial (RCE) cells.

Methods: Microneedle puncture and fluorescent dye loss were used to wound membranes and assay resealing, respectively. Different repair mechanisms were detected pharmacologically and with antisense oligonucleotides.

Results: The RCE cells rapidly reseal plasma membranes by calcium-dependent exocytotic mechanisms that exhibit both facilitated and potentiated responses to multiple wounding. The facilitated response was inhibited by specific inhibitors of protein kinase C (PKC) and brefeldin A, and the potentiated response was blocked by inhibitors of cAMP-dependent protein kinase (PKA). Reduction of myosin IIA inhibited the facilitated response, and reduction of IIB inhibited the initial resealing.

Conclusions: RCE cells rapidly repair plasma membrane disruptions. At a second wound at the same site, PKC stimulated vesicle formation from the Golgi apparatus, resulting in more rapid membrane resealing for a facilitated response. The RCE cell also contains a PKA-dependent global potentiation of membrane resealing.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Blotting, Western
  • Calcium / pharmacology
  • Cell Line
  • Cell Membrane / drug effects
  • Cell Membrane / physiology*
  • Cyclic AMP-Dependent Protein Kinases / antagonists & inhibitors
  • Cyclic AMP-Dependent Protein Kinases / metabolism
  • Enzyme Inhibitors / pharmacology
  • Epithelium, Corneal / cytology*
  • Epithelium, Corneal / drug effects
  • Exocytosis / physiology
  • Fluorescent Dyes
  • Nonmuscle Myosin Type IIA / metabolism
  • Nonmuscle Myosin Type IIB / metabolism
  • Oligonucleotides, Antisense / pharmacology
  • Protein Kinase C / antagonists & inhibitors
  • Protein Kinase C / metabolism
  • Pyridinium Compounds
  • Quaternary Ammonium Compounds
  • Rabbits
  • Wound Healing / physiology*

Substances

  • Enzyme Inhibitors
  • FM1 43
  • Fluorescent Dyes
  • Oligonucleotides, Antisense
  • Pyridinium Compounds
  • Quaternary Ammonium Compounds
  • Cyclic AMP-Dependent Protein Kinases
  • Protein Kinase C
  • Nonmuscle Myosin Type IIA
  • Nonmuscle Myosin Type IIB
  • Calcium