Excitatory effect of ATP on acutely dissociated ventromedial hypothalamic neurons of the rat

The ATP-induced increase in the cytosolic Ca(2+) concentration ([Ca]i) and current in acutely dissociated ventromedial hypothalamic rats neurons were investigated using fura-2 microfluorometry and the nystatin-perforated patch recording method, respectively. The ATP-induced [Ca]i increase was mimicked by dimethyl-thio-ATP and ATPgammaS, and was inhibited by P2 purinoreceptor antagonists. The ATP-induced [Ca]i increase was markedly reduced by removal of external Na(+) or Ca(2+), and by addition of various Ca(2+) channel antagonists. ATP induced a transient inward current exhibiting a strong inward rectification at membrane potentials more positive than -20 mV. The ATP-induced current at a holding potential of -70 mV was concentration-dependent with a half-maximum effective concentration of 26 microM. Increasing the external Ca(2+) concentration to 10 mM shifted the dose-response relationship to the right. ATP induced only a small current and a small increase in [Ca]i, even at 10 mM Ca(2+), when external Na(+) was removed, suggesting the relatively low permeability to Ca(2+) of purinoceptor channels. These results suggest that ATP activates non-selective cation channels by acting on P2X purinoceptors on dissociated ventromedial hypothalamic neurons, which in turn increases [Ca]i by increasing Ca(2+) influx through voltage-dependent Ca(2+) channels.