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In vitro pharmacologic separation of corneal endothelial migration and spreading responses.

Abstract
Repair of corneal endothelial wounds involves two forms of cell translocation: (1) "migration," in which individual cells at the wound edge break contacts with neighboring cells and move as individuals into the wound defect, and (2) "spreading," in which cells within the confluent monolayer adjacent to the wound move as a group into the wound area. The authors combined morphometric analysis of Giemsa-stained cultures, phase-contrast video microscopy, and Rh-phalloidin staining of actin filaments to study the effects of epidermal growth factor (EGF) and indomethacin on the migratory and spreading responses to wounding using an in vitro wound-closure model which mimics the amitotic state and general behavior of human corneal endothelium. They found that EGF stimulated the migration of individual cells from the wound edge, induced cellular elongation, and promoted a diffuse distribution of actin filaments. Indomethacin promoted spreading of the confluent monolayer into the wound defect, induced enlargement and flattening of cells, and promoted the formation of long, thick actin stress fibers. These results provide evidence that the migration and spreading responses of corneal endothelial cells to wounding can be pharmacologically separated. The findings suggest that migration of individual cells during wound repair may result from an endogenous form of EGF-like stimulation and that the elongated shape associated with this form of translocation results, at least in part, from an EGF-like alteration in actin-filament organization. Spreading of the confluent monolayer to cover the wound defect may result from a decrease in cyclic adenosine monophosphate induced by a transient reduction in prostaglandin E2 synthesis. This form of translocation may result, in part, from enlargement and flattening of corneal endothelial cells secondary to an enhancement of actin stress-fiber formation.
AuthorsN C Joyce, B Meklir, A H Neufeld
JournalInvestigative ophthalmology & visual science (Invest Ophthalmol Vis Sci) Vol. 31 Issue 9 Pg. 1816-26 (Sep 1990) ISSN: 0146-0404 [Print] United States
PMID2211027 (Publication Type: Comparative Study, Journal Article, Research Support, U.S. Gov't, P.H.S.)
Chemical References
  • Actins
  • Epidermal Growth Factor
  • Indomethacin
Topics
  • Actins (metabolism)
  • Analog-Digital Conversion
  • Animals
  • Cell Division (drug effects)
  • Cell Movement (drug effects)
  • Cells, Cultured
  • Endothelium, Corneal (cytology, drug effects, metabolism)
  • Epidermal Growth Factor (pharmacology)
  • Indomethacin (pharmacology)
  • Male
  • Microscopy, Phase-Contrast
  • Rabbits
  • Signal Transduction
  • Video Recording
  • Wound Healing (drug effects)

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