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J. Biol. Chem., Vol. 275, Issue 27, 20268-20273, July 7, 2000
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From the Laboratory of Molecular and Cellular Neurobiology,
Department of Cell Biology and Pathology, Medical School, Hospital of
Bellvitge, University of Barcelona, Campus of Bellvitge, Feixa Llarga
s/n, L'Hospitalet de Llobregat, E-08907 Barcelona, Spain
The presence of ATP within cells is well
established. However, ATP also operates as an intercellular signal via
specific purinoceptors. Furthermore, nonsecretory cells can release ATP
under certain experimental conditions. To measure ATP release and
membrane currents from a single cell simultaneously, we used
Xenopus oocytes. We simultaneously recorded membrane
currents and luminescence. Here, we show that ATP release can be
triggered in Xenopus oocytes by hyperpolarizing pulses. ATP
release (3.2 ± 0.3 pmol/oocyte) generated a slow inward current
(2.3 ± 0.1 µA). During hyperpolarizing pulses, the permeability
for ATP4- was more than 4000 times higher than that for
Cl-. The sensitivity to GdCl3 (0.2 mM) of hyperpolarization-induced ionic current, ATP release
and E-ATPase activity suggests their dependence on stretch-activated
ion channels. The pharmacological profile of the current inhibition
coincides with the inhibition of ecto-ATPase activity. This enzyme is
highly conserved among species, and in humans, it has been cloned and
characterized as CD39. The translation, in Xenopus oocytes,
of human CD39 mRNA encoding enhances the ATP-supported current,
indicating that CD39 is directly or indirectly responsible for the
electrodiffusion of ATP.
ATP Crossing the Cell Plasma Membrane Generates an Ionic Current
in Xenopus Oocytes*
§,,
*
This work was supported by Dirección General de
Enseñanza Superior e Investigación Científica from
the Spanish Government, the Comissió Interdepertamental de
Recerca i Innovació Tecnològica from the local Government
of the Generalitat de Catalunya, and Fundació La Marató de
TV3 Spain.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
These authors contributed equally to this work.
§
Recipient of a grant from Fundació August Pi i Sunyer.
¶
To whom correspondence should be addressed. Tel.:
34-93-402-4279 or 34-93-403-5809; Fax: 34-93-403-5810; E-mail:
solsona@bellvitge. bvg.ub.es.
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