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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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