Objective: To determine the effect of cardiac resynchronization therapy (CRT) on sympathetic nervous function in heart failure (HF). Background: Neurohormonal dysregulation and cardiac autonomic dysfunction are associated with HF and contribute to HF progression and its poor prognosis. We hypothesized that mechanical resynchronization improves cardiac sympathetic function in HF. Methods: Sixteen consecutive patients receiving CRT for advanced cardiomyopathy and 10 controls were included in this prospective study. NYHA class, 6-minute walk distance, echocardiographic parameters, plasma norepinephrine (NE) were assessed at baseline, 3-month and 6-month follow-up. Cardiac sympathetic function was determined by 123iodine metaiodobenzylguanidine (123I-MIBG) scintigraphy and 24-hour ambulatory electrocardiography. Results: Along with improvement in NYHA class (3.1 ± 0.3 to 2.1 ± 0.4, P < 0.001) and LVEF (23 ± 6% to 33 ± 12%, P < 0.001 delayed heart/mediastinum (H/M) 123 I-MIBG ratio increased significantly (1.8 ± 0.7 to 2.1 ± 0.6, P = 0.04) while the H/M 123I-MIBG washout rate decreased significantly (54 ± 25% to 34± 24%, P = 0.01) from baseline to 6-month follow-up. The heart rate variability (HRV) measured in SD of normal-to-normal intervals also increased significantly from baseline (82 ± 30 ms) to follow-up (111 ± 32 ms, P = 0.04). The improvement in NYHA after CRT was significantly associated with baseline 123I-MIBG H/M washout rate (r = 0.65, P = 0.03). The improvement in LVESV index was associated with baseline 123I-MIBG delayed H/M ratio (r = −0.67, P = 0.02) and H/M washout rate (r = 0.65, P = 0.03). Conclusion: After CRT, improvements in cardiac symptoms and LV function were accompanied by rebalanced cardiac autonomic control as measured by 123I-MIBG and HRV.