Objective: Cardiac arrhythmias are the most common cause of death in hemodialysis. Autonomic dysfunction plays a central role in this arrhythmogenic background. Previous studies on hemodialysis-related changes in heart-rate-variability(HRV) give contradictory results. This study investigated HRV indices both at rest and in response to physical and mental stimulation maneuvers at multiple time-points around and during the hemodialysis procedure. Design and method: Autonomic function was assessed by linear and non-linear HRV indices at pre-dialysis, during-dialysis(3 equal time-periods), post-dialysis, and on the non-dialysis day in 36 hemodialysis patients. Continuous measurement of beat-by-beat heart-rate was recorded with Finometer-PRO(The Netherlands) at rest and after orthostatic, sit-to-stand, handgrip and mental-arithmetic test. Results: The RMSSD, SD1, and SD2 indices significantly increased during dialysis(early-HD, mid-HD, late-HD periods) compared with the pre-dialysis levels(p<0.05) and returned to baseline post-dialysis(RMSSD: 54.39±83.73 vs 137.98±109.53* vs 119.85±97.34* vs 144.47±88.74* vs 85.82±121.43msec, *p<0.05 vs pre-dialysis and post-dialysis). No differences were detected in the above indices between the pre-dialysis and non-dialysis day. However, post-dialysis, the HRV responses to orthostatic and sit-to-stand tests were more exaggerated than in the pre-dialysis measurements (p<0.05). The HRV responses both at resting and physical tests in the non-dialysis day were similar to the pre-dialysis levels. HRV indices in mental-arithmetic test during hemodialysis were much higher than at the non-dialysis day(RMSSD: 77.05 (180.41) vs 19.75 (105.47)msec; p = 0.031) were detected. Conclusions: Hemodialysis causes marked changes in autonomic function. Resting HRV indices return to baseline post-dialysis, but HRV responses to physical stress remain exaggerated and return to baseline on the non-dialysis day. Detecting patients with significant autonomic dysfunction may help towards reduction of arrhythmia risk through individualized approaches.