A hallmark of chronic heart failure (CHF) is an increased sympathetic tone resulting in autonomic imbalance. Renal denervation (DNx) in CHF patients has resulted in symptomatic improvement, but the protective mechanisms remain unclear. We hypothesized in CHF, unilateral renal DNx would improve cardiac autonomic balance. The present study utilized conscious, chronically instrumented New Zealand White rabbits undergoing renal DNx prior to pacing-induced CHF. Four treatment groups were used: non-pace non-DNx (Sham-INV), non pace DNx (sham-DNx), pace non-DNx (CHF-INV) or pace DNx (CHF-DNx). We examined several markers indicative of autonomic balance. Baroreflex sensitivity and time domain heart rate variability (HRV) were both decreased in the CHF-INV group compared to sham-INV and restored to sham levels by renal DNx. Power spectral analysis indicated an increase in LF/HF ratio in the CHF-INV as compared to the sham-INV, which was normalized to sham levels by DNx. In order to assess if this was due to a withdrawal of sympathetic tone or an increase in parasympathetic tone, the heart rate response was measured after an IV bolus of metoprolol or atropine. Bradycardia induced by IV metoprolol (indicative of cardiac sympathetic tone) was exacerbated in CHF-INV rabbits compared to sham-INV but was normalized in CHF-DNx. Conversely, the tachycardia in response to IV atropine (indicative of cardiac vagal tone) was not improved in CHF-DNx vs CHF-INV animals. Renal DNx also prevented the increase in circulating plasma NE seen in CHF-INV rabbits. These results suggest renal DNx improves cardiac autonomic balance in CHF by a reduction sympathetic tone.