The physiological role of brain Ang II and acetylcholine in mediating water deprivation-induced drinking was assessed in male Sprague-Dawley rats. Specific receptor antagonists were intracerebroventricularly (i.c.v.) administered in 48-h water-deprived rats. When water was given 20 min after i.c.v. injection, PD 123319 almost totally blocked the drinking response. However, losartan and CGP 42112A produced an approx. 20% inhibition of water intake. Central blockade of AT1 receptor with KR 31080 and cholinergic receptor with atropine attenuated water intake more than 50% which was significantly greater than inhibition produced by losartan and CGP 42112A. Atropine given alone or mixed with losartan and CGP-42112A produced a similar magnitude of inhibition of water intake. When water was given 90 min after i.c.v. injection, losartan or CGP-42112A produced a significantly greater inhibition of water intake than when water was given 20 min after injection. The present results suggest that both the central angiotensinergic and cholinergic system play an important role in the physiological drinking response after water deprivation. Both brain AT1 and AT2 receptors are involved in dehydration-induced drinking, but relative contribution of the receptors remains to be clarified.