Heart rate variability (HRV) and heart rate (HR) responses following a 60 degree head-up tilt were measured in 60 infants at 1 and 3 mo of age to investigate the effects on these of age, sleep state, sleep position, and mother's smoking status. HRV was determined from Poincaré plots of 500 sequential RR intervals to measure overall variability derived from the SDRR of this plot, and instantaneous variability derived from the SDdeltaRR. HR responses to the tilt were measured as changes in RR interval length from rest to immediately following the tilt and again once a stable pattern was reached. SDRR and SDdeltaRR increased 20 and 40%, respectively, with age (p < 0.0001), SDRR was higher in active sleep (AS) than quiet sleep (QS, +72%, p < 0.0001) but both measures of variability (SDRR and SDdeltaRR) were lower in the prone position compared with supine (-18%, p < 0.0001). However, several findings were dependent on the basal RR interval, thus the age effect disappeared once RR interval was taken into account, sleep state remained an important factor and the lower variability when prone now became a difference of -3% (p = 0.034). The tilt generally provoked a reflex tachycardia followed by a bradycardia and settling to a stable HR level below, at, or above baseline within 30 s. The more unusual responses were no HR change, sustained tachycardia or sustained bradycardia (15% of total). These were more likely to occur in younger infants (p = 0.008) and in AS (p < 0.0001). No changes were seen in any of the cardiac indices related to maternal smoking status. The findings confirm several reports indicating that prone sleeping damps some physiologic responses. The data emphasize the need to consider basal heart rate, and sleep position as well as sleep state in autonomic function testing during infant sleep.