BackgroundNociception in rats is frequently measured in terms of latency of withdrawal reaction to radiant heat (thermal nociceptive threshold). The aim of this study was to determine how much housing acclimatization and ambient temperature affect the results of thermal pain threshold testing. MethodsAll experiments used adult male Wistar rats. Thermal pain thresholds were tested using the radiant heat withdrawal reaction at three different body sites: forepaws, hind paws and tail. Skin temperature was measured using an Infrared thermometer and ambient temperature was set at 18, 20, 24 or 26 degrees C. ResultsThe results demonstrate that (1) thermal pain threshold was inversely related to both ambient and skin temperature; (2) housing acclimatization and repeated testing had no effect on nociceptive thresholds at any of the three body sites; (3) a resting, cranio-caudal distribution, of nociceptive sensitivity was observed; (4) hind paws and tail were more sensitive to changes of skin and ambient temperature than forepaws. ConclusionThese findings show the importance of recording laboratory conditions in experiments and their influence on results.