Humans with traumatic spinal myelopathy exhibit intralesional conduction block and autonomic failure as pathophysiologic sequelae of their injury. Analysis of heart rate variability (HRV) provides a means of assessing changes in the function of the autonomic nervous system (ANS) and the cardiac sequelae of injury. Thirteen patients with long-standing spinal cord injury (SCI) and 13 able-bodied controls were studied. Each patient received a single 10-mg dose of an immediate release (IR) formulation of 4-aminopyridine (4-AP). Twenty-four hour heart rate (HR) and HRV data were acquired using a Holter ambulatory electrocardiographic (ECG) monitor. Analysis of acquired data was carried out using a minicomputer programmed to separate ECG R-R intervals into frequency patterns that appear as peaks dispersed along a frequency range of 0.0 to 1.0 Hz. Twenty-four hour baseline, pretreatment low-frequency (LF) HRV power was diminished in all patients with SCI compared with able-bodied-controls and was significantly decreased in tetraplegic patients (P = 0.03). This difference in LF HRV power disappeared during the 24 hours immediately after administration of 4-AP, and mean LF HRV power in tetraplegic patients became indistinguishable from LF HRV power in controls. 4-Aminopyridine appears to influence ANS function and LF HRV in humans with long-standing SCI.