P2Y-purinoceptor regulation of surfactant secretion from rat isolated alveolar type II cells is associated with mobilization of intracellular calcium

Abstract

1 The effect of methylene, thio, and imido substituted analogues of adenosine 5'-triphosphate (ATP) on surfactant phospholipid secretion and calcium mobilization in rat isolated alveolar Type II cells was studied. 2 ATP was the most potent secretagogue of adenine nucleotides studied. The rank order of agonist potency for [3H]-phosphatidylcholine secretion was ATP greater than adenosine 5'-O-(3-thiotriphosphate) (gamma S-ATP) greater than beta, gamma-imido adenosine 5'-triphosphate (AMPPNP) greater than beta, gamma-methylene adenosine 5'-triphosphate (beta, gamma-CH2-ATP) greater than alpha, beta-methylene adenosine 5'-triphosphate (alpha, beta-CH2-ATP). The respective EC50S were 10(-6) M, 2 X 10(-6) M, 2 X 10(-5) M, and greater than 2.5 X 10(-4) M. 3 Exogenous ATP also induced a rapid mobilization of intracellular calcium monitored by changes in Fura 2 fluorescence. The rank order of agonist potency for calcium mobilization was similar to the rank order of agonist potency for surfactant secretion: ATP = gamma S-ATP greater than AMPPNP greater than alpha, beta-CH2-ATP. 4 There was no effect of EGTA on ATP-induced calcium mobilization, consistent with the hypothesis that exogenous ATP induces release of calcium from intracellular stores. 5 These data are consistent with a P2Y-purinoceptor regulating surfactant secretion from isolated Type II cells via mobilization of intracellular calcium, since: (a) non-hydrolyzed analogues of ATP are potent secretagogues, (b) beta, gamma-CH2-ATP was a more potent secretagogue than alpha, beta-CH2-ATP and (c) the rank orders of agonist potency for calcium mobilization and phospholipid secretion were the same.