BACKGROUND. Non-invasive hemodynamic measurements require placing animals in restrainers. To minimize restraint-induced hemodynamic artifacts several adaptation protocols have been proposed. However, so far there is no comparative study between various adaptation protocols using continuous hemodynamic measurements in rodents. METHODS. We investigated hemodynamic effects of three habituation protocols to repeated restraint stress with telemetry in rats. Male, 16-week old, Sprague-Dawley rats were implanted with telemetric transmitters under general anaesthesia with IP ketamine 100 mg/kg bw and IP xylazine 10 mg/kg bw. Continuous recordings of heart rate (HR) and mean arterial blood pressure (MABP) were made before and during restraint protocols. Stress adaptation protocols included placement of rats in plastic restrainers used for tail-cuff measurements in the middle of the dark phase. Restraint lasted for 60 min, and was repeated every second day. Rats were subjected either to 3-day, 2-week or 4-week stress protocol. RESULTS. Restraint stress significantly increased MABP and HR. The smallest hemodynamic response was found between 40 and 60 min after placement of rats in restrainers. All tested habituation protocols failed to prevent restraint-induced increase in MABP and HR. Repeated restraint affected diurnal MABP and HR variation, but it had no significant effect on average 24-hour MABP and HR. CONCLUSIONS. For the first time various adaptation protocols to restraint stress in rodents were compared with the use of continuous hemodynamic recordings. No complete hemodynamic adaptation to restraint regardless the number of restraint exposures was found. Repeated restraint altered diurnal blood pressure variation, however did not affect average 24-hour blood pressure.