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Coordination of intercellular Ca(2+) signaling in endothelial cell tubes of mouse resistance arteries.


ABSTRACT: To test the hypothesis that Ca(2+) responses to GPCR activation are coordinated between neighboring ECs of resistance arteries.EC tubes were freshly isolated from superior epigastric arteries of C57BL/6 mice. Intercellular coupling was tested using microinjection of propidium iodide. Following loading with fluo-4 dye, intracellular Ca(2+) responses to ACh were imaged with confocal microscopy.Cell-to-cell transfer of propidium iodide confirmed functional GJCs. A 1 ?m ACh stimulus evoked Ca(2+) responses (9.8 ± 0.8/min, F/F(0) = 3.11 ± 0.2) which pseudo-line-scan analysis revealed as composed of Ca(2+) waves and spatially restricted Ca(2+) release events. A 100 nm ACh stimulus induced Ca(2+) responses of lower frequency (4.5 ± 0.7/min) and amplitude (F/F(0) = 1.95 ± 0.11) composed primarily of spatially restricted events. The time interval between Ca(2+) waves in adjacent cells (0.79 ± 0.12 s) was shorter (p < 0.05) than that between nonadjacent cells (1.56 ± 0.25 s). Spatially restricted Ca(2+) release events had similar frequencies and latencies between adjacent and nonadjacent cells. Inhibiting intracellular Ca(2+) release with 2-APB, Xestospongin C or thapsigargin eliminated Ca(2+) responses.With moderate GPCR stimulation, localized Ca(2+) release events predominate among cells. Greater GPCR stimulation evokes coordinated intercellular Ca(2+) waves via the ER. Calcium signaling during GPCR activation is complex among cells, varying with stimulus intensity and proximity to actively signaling cells.

SUBMITTER: Socha MJ 

PROVIDER: S-EPMC3502682 | biostudies-literature | 2012 Nov

REPOSITORIES: biostudies-literature

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Coordination of intercellular Ca(2+) signaling in endothelial cell tubes of mouse resistance arteries.

Socha Matthew J MJ   Domeier Timothy L TL   Behringer Erik J EJ   Segal Steven S SS  

Microcirculation (New York, N.Y. : 1994) 20121101 8


<h4>Objective</h4>To test the hypothesis that Ca(2+) responses to GPCR activation are coordinated between neighboring ECs of resistance arteries.<h4>Methods</h4>EC tubes were freshly isolated from superior epigastric arteries of C57BL/6 mice. Intercellular coupling was tested using microinjection of propidium iodide. Following loading with fluo-4 dye, intracellular Ca(2+) responses to ACh were imaged with confocal microscopy.<h4>Results</h4>Cell-to-cell transfer of propidium iodide confirmed fun  ...[more]

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