A role for phospholipase C activity but not ryanodine receptors in the initiation and propagation of intercellular calcium waves

M. Hansen, Scott A Boitano, E. R. Dirksen, M. J. Sanderson

Research output: Contribution to journalArticle

69 Citations (Scopus)

Abstract

Mechanical stimulation of a single cell in an airway epithelial culture initiates an increase in intracellular Ca2+ concentration ([Ca2+](i)) that propagates from cell to cell as an intercellular Ca2+ wave. These Ca2+ waves appear to require an increase in intracellular inositol 1,4,5-trisphosphate (IP3) concentration ([IP3](i)) in the stimulated cell and are propagated between cells by the diffusion of IP3 through gap junctions. To test the hypothesis that the activation of phospholipase C (PLC) contributes to the elevation of [IP3](i) and initiation of an intercellular Ca2+ wave, changes in [Ca2+](i) induced by mechanical stimulation were measured by digital fluorescence microscopy in the presence of the PLC inhibitor, aminosteroid U73122. Following exposure to U73122 mechanical stimulation elevated [Ca2+](i) of the stimulated cell, but did not initiate the propagation of an intercellular Ca2+ wave. By contrast, in the presence of U73343, a similar aminosteroid that does not inactivate PLC, mechanical stimulation increased the [Ca2+](i) of the stimulated cell and initiated an intercellular Ca2+ wave. U73122 also blocked the elevation of [Ca2+](i) of airway epithelial cells in response to ATP, a P2-receptor agonist that activates PLC to elevate [IP3](i) and [Ca2+](i). In addition, the propagation of intercellular Ca2+ waves was not affected by the ryanodine-receptor agonists, caffeine or ryanodine. The hypotheses that: (1) an elevation of [IP3](i) is required to initiate intercellular Ca2+ waves; (2) mechanical stimulation activates PLC; and (3) Ca2+ wave propagation in airway epithelial cells involves Ca2+ release from intracellular stores primarily via IP3 receptors are supported by these results.

Original languageEnglish (US)
Pages (from-to)2583-2590
Number of pages8
JournalJournal of Cell Science
Volume108
Issue number7
StatePublished - 1995
Externally publishedYes

Fingerprint

Ryanodine Receptor Calcium Release Channel
Calcium Signaling
Type C Phospholipases
Epithelial Cells
Inositol 1,4,5-Trisphosphate Receptors
Ryanodine
Inositol 1,4,5-Trisphosphate
Gap Junctions
Caffeine
Fluorescence Microscopy
Adenosine Triphosphate
1-(6-((3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione

Keywords

  • Airway epithelium
  • Aminosteroid U73122
  • Caffeine
  • Inositol trisphosphate
  • Intercellular communication
  • Mechanical stimulation

ASJC Scopus subject areas

  • Cell Biology

Cite this

A role for phospholipase C activity but not ryanodine receptors in the initiation and propagation of intercellular calcium waves. / Hansen, M.; Boitano, Scott A; Dirksen, E. R.; Sanderson, M. J.

In: Journal of Cell Science, Vol. 108, No. 7, 1995, p. 2583-2590.

Research output: Contribution to journalArticle

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N2 - Mechanical stimulation of a single cell in an airway epithelial culture initiates an increase in intracellular Ca2+ concentration ([Ca2+](i)) that propagates from cell to cell as an intercellular Ca2+ wave. These Ca2+ waves appear to require an increase in intracellular inositol 1,4,5-trisphosphate (IP3) concentration ([IP3](i)) in the stimulated cell and are propagated between cells by the diffusion of IP3 through gap junctions. To test the hypothesis that the activation of phospholipase C (PLC) contributes to the elevation of [IP3](i) and initiation of an intercellular Ca2+ wave, changes in [Ca2+](i) induced by mechanical stimulation were measured by digital fluorescence microscopy in the presence of the PLC inhibitor, aminosteroid U73122. Following exposure to U73122 mechanical stimulation elevated [Ca2+](i) of the stimulated cell, but did not initiate the propagation of an intercellular Ca2+ wave. By contrast, in the presence of U73343, a similar aminosteroid that does not inactivate PLC, mechanical stimulation increased the [Ca2+](i) of the stimulated cell and initiated an intercellular Ca2+ wave. U73122 also blocked the elevation of [Ca2+](i) of airway epithelial cells in response to ATP, a P2-receptor agonist that activates PLC to elevate [IP3](i) and [Ca2+](i). In addition, the propagation of intercellular Ca2+ waves was not affected by the ryanodine-receptor agonists, caffeine or ryanodine. The hypotheses that: (1) an elevation of [IP3](i) is required to initiate intercellular Ca2+ waves; (2) mechanical stimulation activates PLC; and (3) Ca2+ wave propagation in airway epithelial cells involves Ca2+ release from intracellular stores primarily via IP3 receptors are supported by these results.

AB - Mechanical stimulation of a single cell in an airway epithelial culture initiates an increase in intracellular Ca2+ concentration ([Ca2+](i)) that propagates from cell to cell as an intercellular Ca2+ wave. These Ca2+ waves appear to require an increase in intracellular inositol 1,4,5-trisphosphate (IP3) concentration ([IP3](i)) in the stimulated cell and are propagated between cells by the diffusion of IP3 through gap junctions. To test the hypothesis that the activation of phospholipase C (PLC) contributes to the elevation of [IP3](i) and initiation of an intercellular Ca2+ wave, changes in [Ca2+](i) induced by mechanical stimulation were measured by digital fluorescence microscopy in the presence of the PLC inhibitor, aminosteroid U73122. Following exposure to U73122 mechanical stimulation elevated [Ca2+](i) of the stimulated cell, but did not initiate the propagation of an intercellular Ca2+ wave. By contrast, in the presence of U73343, a similar aminosteroid that does not inactivate PLC, mechanical stimulation increased the [Ca2+](i) of the stimulated cell and initiated an intercellular Ca2+ wave. U73122 also blocked the elevation of [Ca2+](i) of airway epithelial cells in response to ATP, a P2-receptor agonist that activates PLC to elevate [IP3](i) and [Ca2+](i). In addition, the propagation of intercellular Ca2+ waves was not affected by the ryanodine-receptor agonists, caffeine or ryanodine. The hypotheses that: (1) an elevation of [IP3](i) is required to initiate intercellular Ca2+ waves; (2) mechanical stimulation activates PLC; and (3) Ca2+ wave propagation in airway epithelial cells involves Ca2+ release from intracellular stores primarily via IP3 receptors are supported by these results.

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