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Articles in PresS, published online ahead of print September 5, 2001
Am J Physiol Cell Physiol, 10.1152/ajpcell.00317.2001
Submitted on July 12, 2001
Accepted on September 3, 2001
1 Medicine, University of Montreal, Montreal, Quebec, Canada; Centre hospitalier de l'Universite de Montreal, Montreal, Quebec, Canada
* To whom correspondence should be addressed. E-mail: ryszard.grygorczyk{at}umontreal.ca.
ATP release induced by hypotonic swelling is an ubiquitous phenomenon in eukaryotic cells, but its underlying mechanisms are poorly defined. A mechano-sensitive ATP channel has been implicated because Gd3+, an inhibitor of stretch-activated channels, suppressed ATP efflux monitored by luciferase bioluminescence. We examined the effect of Gd3+ on luciferase bioluminescence and on ATP efflux from hypotonically-swollen cells. We found that luciferase was inhibited by 
10 µM Gd3+, and this may have contributed to the previously-reported inhibition of ATP release. In ATP efflux experiments, luciferase inhibition could be prevented by chelating Gd3+ with EGTA before luminometric ATP determinations. Using this approach, we found that 10 µM to 100 µM Gd3+, i.e. concentrations typically used to block mechano-sensitive channels, actually stimulated hypotonically-induced ATP release from fibroblasts. Inhibition of ATP release required at least 500 µM, 200 µM and 100 µM Gd3+ for fibroblasts, A549 and 16HBE14o- cells, respectively. Such biphasic and cell-specific effects of Gd3+ are most consistent with its action on membrane lipids and membrane-dependent processes such as exocytosis.
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