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-cells communicate via intermittent release of ATP
1 Department of Medical Cell Biology, University of Uppsala, Uppsala, Sweden
* To whom correspondence should be addressed. E-mail: Eva.Grapengiesser{at}medcellbiol.uu.se.
The role of external ATP for intercellular communication was studied in glucose-stimulated
pancreatic 
-cells isolated from ob/ob-mice. Digital image analyses with
fura-2 revealed spontaneous transients of cytoplasmic Ca2+ appearing in synchrony
in the absence of cell contacts. After removing slow oscillations with methoxy-verapamil,
addition of ATP (0.1 - 100 µM) resulted in prompt firing of a transient,
followed by suppression of the generation and synchronization of spontaneously
occurring transients. It was possible to trigger transients during the suppressive
phase by raising the concentration of ATP. The dual action of ATP was mimicked by
ADP or 2-methylthio-ATP but not by AMP or UTP. The number of spontaneous
transients and their synchronization were reduced in the presence of the
dephosphorylating agent apyrase. Additional evidence that intermittent release of
ATP participates in the generation of spontaneous Ca2+ transients was obtained from
the suppression observed using antagonists of the purinoceptors - suramin (0.3 - 30
µM), pyridoxalphosphate-6-azophenyl-2, 4-disulphonic acid (PPADS; 10 - 30 µM)
and 2-deoxy-N-methyladenosine (MRS 2179; 0.3 - 30 µM) - or counteracting -cell
release of ATP by inhibiting exocytosis with 100 nM adrenaline, 100 nM somatostatin
or lowering the temperature below 30 C. The data indicate that ATP has time-dependent
actions (prompt stimulation followed by inhibition) on the generation
of Ca2+ transients mediated by P2Y receptors. It is proposed that -cells both receive a
neural ATP signal with co-ordinating effects on their Ca2+ oscillations and propagate
this message to adjacent cells via intermittent release of ATP combined with gap
junction coupling.
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