INVESTIGADORES
ESPELT Maria Victoria
congresos y reuniones científicas
Título:
Kinetic of extracellular ATP accumulation and volume regulation of hypotonically challenged hepatoma cells
Autor/es:
SANCHEZ ALBERTI G; ALVAREZ C; ESPELT M.V; SCHWARZBAUM P.J
Lugar:
Los Cocos, Córdoba
Reunión:
Congreso; First South American Spring Symposium in Signal Transduction and Molecular Medicine (SISTAM2010); 2010
Resumen:
Kinetic of extracellular ATP accumulation from
hypotonically challenged hepatoma cells.
Sanchez Alberti G., Alvarez C., Espelt M.V.,
Schwarzbaum P.J. IQUIFIB. Facultad de Farmacia y Bioquímica, Universidad de Buenos
Aires. Junin 956, 1113 Buenos Aires, Argentina. Contact e-mail: pjs@qb.ffyb.uba.ar
In most animal cells, release of cytosolic ATP into
the extracellular space occurs in response to osmotic gradients, with either
cAMP and/or Ca2+i signaling such release. In this study,
we evaluated the main processes governing the kinetic of extracellular ATP
(ATPe) accumulation for Huh-7 hepatoma cells exposed to hypotonicity. Exposure
of cells to a 180 mOsm hypotonic medium led to an exponential increase of
[ATP]e to a maximum (33.4 ± 11.33), followed by a slower non linear decrease (t1/2=11
min). ATPe accumulation was inhibited 75% by carbenoxolone, an inhibitor of
Pannexin 1, whereas pre-treatment of cells with the purinergic receptor P2Y13
agonist 2-Methylthio adenosine 5diphosphate (which inhibits adenylate cyclase)
led to a 29% inhibition. Cells display ectoATPase activity ranging from 0.5×10-3
nmoles×(sec×106cells)-1 (at 60.5 nM) to 4.9×10-3
nmoles×(sec×106cells)-1 (at 970 nM). Since under these
conditions cell mortality is negligible, the kinetic of ATPe is mainly controlled
by ectoATPase activity (which consumes ATPe) and non lytic ATP efflux (which
generates ATPe), with this latter efflux being partially inhibited by P2Y13
activation and partially mediated by Pannexin 1.