P2 purinoceptor-mediated depolarization of rat supraoptic neurosecretory cells in vitro.

1. Intracellular recordings were obtained from supraoptic magnocellular neurosecretory cells (MNCs) in superfused explants of rat hypothalamus. Application of ATP and UTP, but not adenosine, produced TTX-insensitive depolarizations accompanied by increases of input conductance. 2. The P2X agonists alpha,beta-methylene ATP, beta,gamma-methylene ATP and 2-methylthio ATP mimicked the effects of ATP in > 77% of the cells tested. Depolarizing responses to ATP were reversibly inhibited by PPADS (pyridoxal-phosphate-6-azophenyl-2',4'- disulphonic acid; IC50 approximately 0.5 microM), a selective P2X antagonist. 3. The reversal potential of responses to ATP (-37 mV) was not strongly affected by intracellular Cl- injection or by removal of Cl- from the external solution. The reversal potential of responses to the most potent P2X agonist, alpha,beta-methylene ATP, was -29 mV. These values suggest the involvement of non-selective cationic channels, a finding which is consistent with the ionotropic cationic channel structure of cloned P2X purinoceptors. 4. The reversal potential of UTP-mediated responses (-33 mV) was also consistent with the involvement of non-selective cationic channels. Since cloned P2U receptors display homology with G-protein-coupled receptors, cationic channels modulated by UTP are probably different from those mediating P2X responses.