Local modulation of intracellular calcium levels near a single-cell wound in human endothelial monolayers

Paul D. Drumheller, Jeffrey A. Hubbell

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

An endothelial cell monolayer with a single mechanically lysed cell was used as a model to examine the extent, kinetics, and nature of local calcium mobilization in the neighborhood of a wound. Individual endothelial cells from confluent monolayers were mechanically lysed with a minutien needle coupled to a micromanipulator while producing no observable mechanical trauma to the neighboring cells. Changes in calcium levels in individual cells surrounding the wound site were monitored by epifluorescence microphotometry with the calcium-sensitive fluorophore indo-1. Individual cells adjacent to the wound site showed a substantial increase in their intracellular calcium levels, almost as high as the calcium levels attained by ionophore controls. The magnitude of intracellular calcium mobilization in confluent monolayers decreased with distance from the wound site, and those cells located at a radius greater than seven cells from the wound site showed no change in their calcium levels. Thus, lysis of a single cell resulted in calcium mobilization in approximately 200 neighboring cells. The time necessary for intracellular calcium to reach maximum levels also increased with distance from the wound site. Calcium mobilization was partly intracellular and was inhibited by disrupting cell-cell coupling or by increasing gap junction resistance by heptanol. This mobilization was greatly attenuated in subconfluent endothelial monolayers, and it was not observed in fibroblasts or smooth muscle cells; furthermore, the effect was defective in monolayers intentionally contaminated with smooth muscle cells. This study examines the extent and possible mechanisms of local endothelial activation near a microscopic endothelial wound.

Original languageEnglish
Pages (from-to)1258-1265
Number of pages8
JournalArteriosclerosis, Thrombosis, and Vascular Biology
Volume11
Issue number5
StatePublished - Dec 1 1991
Externally publishedYes

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Calcium
Wounds and Injuries
Smooth Muscle Myocytes
Endothelial Cells
Heptanol
Gap Junctions
Ionophores
Needles
Fibroblasts

Keywords

  • Calcium
  • Endothelial cells
  • Indo-1
  • Injury

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine

Cite this

Local modulation of intracellular calcium levels near a single-cell wound in human endothelial monolayers. / Drumheller, Paul D.; Hubbell, Jeffrey A.

In: Arteriosclerosis, Thrombosis, and Vascular Biology, Vol. 11, No. 5, 01.12.1991, p. 1258-1265.

Research output: Contribution to journalArticle

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