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Air bubble contact with endothelial cells in vitro induces calcium influx and IP3-dependent release of calcium stores

Sobolewski P, Kandel J, Klinger AL, Eckmann DM. Air bubble contact with endothelial cells in vitro induces calcium influx and IP3-dependent release of calcium stores. Am J Physiol Cell Physiol. 2011 Sep;301(3):C679-86. doi:10.1152/ajpcell.00046.2011. Epub 2011 Jun 1. PubMed PMID: 21633077; PubMed Central PMCID: PMC3273994.

Abstract

Gas embolism is a serious complication of decompression events and clinical procedures, but the mechanism of resulting injury remains unclear. Previous work has demonstrated that contact between air microbubbles and endothelial cells causes a rapid intracellular calcium transient and can lead to cell death. Here we examined the mechanism responsible for the calcium rise. Single air microbubbles (50-150 μm), trapped at the tip of a micropipette, were micromanipulated into contact with individual human umbilical vein endothelial cells (HUVECs) loaded with Fluo-4 (a fluorescent calcium indicator). Changes in intracellular calcium were then recorded via epifluorescence microscopy. First, we confirmed that HUVECs rapidly respond to air bubble contact with a calcium transient. Next, we examined the involvement of extracellular calcium influx by conducting experiments in low calcium buffer, which markedly attenuated the response, or by pretreating cells with stretch-activated channel blockers (gadolinium chloride or ruthenium red), which abolished the response. Finally, we tested the role of intracellular calcium release by pretreating cells with an inositol 1,4,5-trisphosphate (IP3) receptor blocker (xestospongin C) or phospholipase C inhibitor (neomycin sulfate), which eliminated the response in 64% and 67% of cases, respectively. Collectively, our results lead us to conclude that air bubble contact with endothelial cells causes an influx of calcium through a stretch-activated channel, such as a transient receptor potential vanilloid family member, triggering the release of calcium from intracellular stores via the IP3 pathway.

Full Text Link

https://www.ncbi.nlm.nih.gov/pubmed/21633077

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