Although inhalation of nitrous oxide (N2O) causes hypothermia in rats, there is a paucity of information as to whether tolerance develops to this effect. The purpose of this study was to determine whether tolerance to N2O hypothermia develops within a single administration as well as over repeated administrations. Temperature was measured telemetrically by implanting intraperitoneal thermal sensors/transmitters in male Long-Evans rats. Experimental rats received an initial 2-h exposure to 60% N2O and became hypothermic relative to controls breathing placebo gas. Only a few rats demonstrated evidence of acute tolerance over the 120 min. Over the next 10 days, the experimental rats received five additional 30-min exposures to 60% N2O and five 30-min exposures to placebo while the control rats received only placebo gas exposures. Chronic tolerance developed to N2O hypothermia over these repeated administrations. A test for Pavlovian drug conditioning found no evidence that conditioned temperature effects contributed to chronic tolerance development. In a second experiment, naive rats were given a 380-min exposure to 60% N2O and a 380-min exposure to placebo gas in a counterbalanced order. Acute tolerance did develop to N2O hypothermia, with the recovery of temperature beginning after a mean of 141 min of gas administration. Hence, both acute and chronic tolerance develop to N2O's hypothermic effects in rats.