Extracellular ATP stimulates three different receptor-signal transduction systems in FRTL-5 thyroid cells. Activation of phospholipase C, and inhibition and activation of adenylate cyclase.

In FRTL-5 thyroid cells, extracellular ATP, a P2-agonist, not only stimulates phospholipase C but also inhibits forskolin- or thyrotropin (TSH)-induced stimulation of adenylate cyclase in a pertussis toxin-sensitive manner [Okajima, Sato, Nazarea, Sho, & Kondo (1989) J. Biol. Chem. 264, 13029-13037]. We have now found that, in pertussis toxin-treated cells, ATP can directly stimulate adenylate cyclase. Although adenylate cyclase modulation occurs through ATP metabolites such as AMP and adenosine, we show that extracellular ATP itself also regulates cyclic AMP production, based on the following: (1) the actions of ATP were imitated by hydrolysis-resistant ATP analogues, (2) the elimination of adenosine by adenosine deaminase decreased the effect of ATP only partially, at least at concentrations greater than 10 microM-ATP, and (3) the amount of AMP produced from ATP was too low to account for the ATP effects. To identify the respective receptors for the three different actions of ATP, we established an antagonist profile. Suramin, which has been reported to be a P2-receptor antagonist, inhibited ATP-induced phospholipase C activation in a competitive fashion, but did not affect ATP-induced adenylate cyclase modulation. On the other hand, 8-cyclopentyl-1,3-diphenylxanthine competitively antagonized both the stimulatory and inhibitory ATP actions on cyclic AMP levels, but did not influence the activation of phospholipase C by ATP. The order of potency for various xanthine derivatives was clearly different with respect to their antagonistic effects on the stimulation and inhibition of adenylate cyclase induced by ATP. We conclude that ATP activates three receptors, each of which is coupled to a different signal transduction system in FRTL-5 cells, i.e. phospholipase C activation, and adenylate cyclase activation and inhibition.