Virtual Reality (VR) is a computer simulation of a three-dimensional environment, often displayed by means of a headset with integrated screens. While VR is known to consistently stimulate the autonomic nervous system, little is known regarding a possible effect on corticospinal excitability, or regarding a potential link between autonomous and voluntary nervous system modulations during VR. Fifteen healthy young participants were enrolled in a single-session experiment. They were seated in a comfortable chair and equipped with a VR headset that displayed a simulated roller coaster ride. Galvanic Skin Response (GSR) and cardiovascular markers (heart rate, heart rate variability) were monitored throughout the experiment. Corticospinal excitability was quantified by measuring the amplitude of the motor evoked potential (MEP), elicited by transcranial magnetic stimulation on the cortical representation of the right First Dorsal Interosseous (FDI) muscle. Results showed modulation in skin conductance, according to the phase of the roller coaster. Corticospinal excitability was increased during downhill and decreased during uphill phases, as compared to flat sections. The evolution of MEP/Mmax was concomitant and correlated to the evolution of GSR. No effect was observed on any of the cardiovascular markers. The present study showed that VR can be an efficient stimulus to modulate corticospinal excitability, even in the absence of a motor simulated situation. These data suggest a potential link between voluntary and autonomic nervous system regulation during VR of stressful situations. This study highlights the attractiveness of VR as an efficient stressor for both autonomous and motor systems, in sport training and in rehabilitation.