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PMID: 10764752 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

ATP crossing the cell plasma membrane generates an ionic current in xenopus oocytes.

The Journal of biological chemistry ·Vol. 275 ·No. 27 ·2000-07-07 ·Pages 20268-73

Bodas E, Aleu J, Pujol G, Martin-Satué M, Marsal J, Solsona C

Abstract

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 microA). During hyperpolarizing pulses, the permeability for ATP(4-) was more than 4000 times higher than that for Cl(-). The sensitivity to GdCl(3) (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.

MeSH Terms
Adenosine Triphosphatases/genetics,metabolism Adenosine Triphosphate/metabolism,pharmacology Animals Antigens, CD/genetics,metabolism Apyrase Cell Membrane/metabolism Cell Membrane Permeability Cyanides/pharmacology Humans Ion Channels/metabolism Oocytes/enzymology,metabolism Patch-Clamp Techniques Protein Biosynthesis Xenopus
Chemicals
Antigens, CD Cyanides Ion Channels Adenosine Triphosphate Adenosine Triphosphatases ectoATPase Apyrase CD39 antigen
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Bodas E
Laboratory of Molecular 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.
Aleu J
Pujol G
Martin-Satué M
Marsal J
Solsona C
Article Info
Journal
The Journal of biological chemistry
Abbr.
J Biol Chem
ISSN
0021-9258
Published
2000-07-07
Pages
20268-73
Language
English
Region
United States
NLM ID
2985121R
Subset
IM
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