Sympathetic neural responses to mental stress are well documented but controversial, while sympathetic neural responses to emotional stress are unknown. The purpose of this study was to investigate neural and cardiovascular responses to emotional stress evoked by negative pictures and reexamine the relationship between muscle sympathetic nerve activity (MSNA) and perceived stress. Mean arterial pressure (MAP), heart rate (HR), MSNA, and perceived stress levels were recorded in 18 men during three randomized trials: 1) neutral pictures, 2) negative pictures, and 3) mental stress. MAP and HR increased during mental stress (14±2 mmHg and 15±2 beats/min; P<0.001), but did not change during negative or neutral pictures. MSNA did not change during neutral pictures (1±1 burst/min; n=16), negative pictures (0±1 burst/min; n=16), or mental stress (1±2 burst/min; n=13). Perceived stress levels were higher during mental stress (3±0 a.u.) compared to negative pictures (2±0 a.u.; P<0.001). Perceived stress levels were not correlated to changes in MSNA during negative pictures (r=0.10; P=0.84) or mental stress (r=0.36; P=0.23). In conclusion, our results demonstrate robust increases in MAP and HR during mental stress, but not during emotional stress evoked by negative pictures. Although the influence of mental stress on MSNA remains unresolved, our findings challenge the concept that perceived stress levels modulate MSNA during mental stress.