Renal sympathetic nerve activity (RSNA) has been measured in anesthetized mice which exhibit poor cardiovascular stability and low blood pressure which confounds proper interpretation of experimental results. We therefore developed a surgical and experimental protocol for measuring RSNA in conscious, unrestrained mice. Male C57Bl6J mice were chronically instrumented with blood pressure telemeters, a jugular venous catheter and a bipolar electrode for recording RSNA. Mice were placed in a home cage and left to recover for 48–72 hrs. Survival rate was 100%; all mice exhibited normal behavior with no sign of distress 24 hrs following surgery. RSNA was successfully recorded in 80% of the mice at 48 and 72 hrs post‐surgery; viable RSNA was reduced to 70 and 50% at 4 and 5 days post‐surgery, respectively. Mean arterial pressure (MAP, 116 ± 2 mmHg, n=10) was consistent with values previously reported for conscious mice. RSNA increased with the normal physical activities of eating and grooming and was validated by pharmacological manipulation of MAP; reduction in MAP to 62 ± 3 mmHg with nitroprusside increased RSNA by 159 ± 41% above baseline (n=5, p<0.05) whereas an increase in MAP to 137 ± 6 mmHg with phenylephrine reduced RSNA to −82 ± 4% compared to baseline (n=5, p<0.05). Thus, we demonstrate an accessible and effective method for direct assessment of RSNA in conscious, unrestrained mice. NIH HL51971, CIHR.