This study tested the hypothesis that halothane anesthesia would cause decreased acetylcholine (ACh) release within the medial pontine reticular formation (mPRF). ACh was collected by microdialysis and measured by high pressure liquid chromatography during wakefulness and during halothane-induced anesthesia. The electroencephalogram (EEG) showed that spindles were a reliable indicator of anesthetic depth. There was a statistically significant disease in ACh release during halothane anesthesia compared with ACh release during wakefulness. Spindles always disappeared during noxious stimulation and during emergence from anesthesia when pontine ACh levels began to increase. These results are consistent with previous data concerning brain stem cholinergic influences on thalamocortical spindle generation, and suggest that similar mechanisms generate cortical spindles during natural sleep and halothane anesthesia.