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Am J Physiol Cell Physiol 282: C219-C226, 2002. First published September 5, 2001; doi:10.1152/ajpcell.00317.2001
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Vol. 282, Issue 1, C219-C226, January 2002

SPECIAL COMMUNICATION
Cell swelling-induced ATP release and gadolinium-sensitive channels

Francis Boudreault and Ryszard Grygorczyk

Centre hospitalier de l'Université de Montréal and Department of Medicine, Université de Montréal, Montréal, Québec, Canada H2W 1T8

ATP release induced by hypotonic swelling is an ubiquitous phenomenon in eukaryotic cells, but its underlying mechanisms are poorly defined. A mechanosensitive (MS) ATP channel has been implicated because gadolinium (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 10-100 µM Gd3+, i.e., concentrations typically used to block MS channels, actually stimulated hypotonically induced ATP release from fibroblasts. Inhibition of ATP release required at least 500, 200, or 100 µM Gd3+ for fibroblasts, A549 cells, 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.

mechanosensitive adenosine 5'-triphosphate release; luciferase bioluminescence


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