An intercellular regenerative calcium wave in porcine coronary artery endothelial cells in primary culture

Andrea Alberto Domenighetti, Jean Louis Bény*, Fabienne Chabaud, Maud Frieden

*Corresponding author for this work

Research output: Contribution to journalArticle

49 Scopus citations

Abstract

1. A regenerative calcium wave is an increase in cytosolic free calcium concentration ([Ca2+](i)) which extends beyond the stimulated cells without decrement of amplitude, kinetics of [Ca2+](i) increase and speed of propagation. 2. The aim of the present study was to test the hypothesis that such a wave could be evoked by bradykinin stimulation and by scraping cultured endothelial cells from porcine coronary arteries. 3. Calcium imaging was performed using the calcium-sensitive dye fura-2. A wound or a delivery of bradykinin to two to three cells on growing clusters of ~ 300 cells caused an increase in [Ca2+](i) which was propagated throughout the cluster in a regenerative manner over distances up to 400 μm. This wave spread through gap junctions since it was inhibited by the cell uncoupler palmitoleic acid. 4. The same experiments performed in confluent cultures caused a rise in [Ca2+](i) which failed to propagate in a regenerative way. The wave propagation probably failed because the confluent cells were less dye coupled than the growing cells. This was confirmed by immunohistology which detected a dramatic decrease in the number of connexin 40 gap junctions in the confluent cultures. 5. The regenerative propagation of the wave was blocked by inhibitors of calcium-induced calcium release (CICR) and phospholipase C (PLC), and by suppression of extracellular calcium, but not by clamping the membrane potential with high-potassium solution. 6. We conclude that regenerative intercellular calcium waves exist in cultured islets but not in confluent cultures of endothelial cells. An increase in [Ca2+](i) is not sufficient to trigger a regenerative propagation. The PLC pathway, CICR and extracellular calcium are all necessary for a fully regenerated propagation.

Original languageEnglish (US)
Pages (from-to)103-116
Number of pages14
JournalJournal of Physiology
Volume513
Issue number1
DOIs
StatePublished - Nov 15 1998

ASJC Scopus subject areas

  • Physiology

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