Background-Although there have been few studies in which the hemodynamic effects of right ventricular (RV) and left ventricular (LV) pacing were compared with those of biventricular (BV) pacing, the autonomic changes during these different pacing modes remain unknown. We hypothesized that BV pacing results in improved hemodynamics and a decrease in sympathetic nerve activity (SNA) compared with single-site pacing. Methods and Results-Thirteen men with a mean ejection fraction of 0.28Ϯ0.7 were enrolled in the study. Arterial blood pressure (BP), central venous pressure (CVP), and SNA were recorded during 3 minutes of right atrial (RA)-RV, RA-LV, and RA-BV pacing at a rate 10 beats faster than sinus rhythm. BP was greater during LV (151Ϯ7/ 85Ϯ3 mm Hg) and BV (151Ϯ6/85Ϯ3 mm Hg) pacing than during RV pacing (146Ϯ7/82Ϯ3 mm Hg) (PϽ0.05). There were no differences in CVP among all pacing modes (Pϭ0.27). SNA was significantly less (PϽ0.02) during both LV (606Ϯ35 U) and BV (582Ϯ41 U) pacing compared with RV pacing (685Ϯ32 U) (PϽ0.02). Although not statistically significant (Pϭ0.08 to 0.14), there was a trend for patients with a narrow QRS to have a lower mean BP and higher SNA during LV pacing than during BV pacing (rϭ0.42 to 0.49). Conclusions-LV-based pacing results in improved hemodynamics and a decrease in SNA compared with RV pacing in patients with LV dysfunction regardless of the QRS duration.