The present study aimed to assess the function of muscular and nervous systems in high-maneuvering jet fighters with the chosen method of clinical neurophysiology, which methodologically has not yet been presented in detail. Fifteen pilots with the experience of more than 1700 flying hours at 7G overloading on average and fifteen healthy subjects for the comparison of health status declared to participate in this study. The dermatomal perception from C4 to S1 was examined using von Frey’s filaments tactile method. Surface electromyography (sEMG) recordings examined the activity of proximal and distal muscles in the upper and lower extremities, the transmission of motor nerve impulses peripherally was diagnosed by electroneurography (ENG), the efferent transmission from C5–C7 and L4–L5 spinal centers to muscles was entirely verified with recordings of motor-evoked potentials induced oververtebrally with the magnetic field (MEP). The pilots estimated more lumbosacral than cervical pain at about 2 on the 10-point visual analog scale (VAS). Sensory perception studies did not reveal abnormal symptoms in the C2–S1 dermatomes innervation. Clinical neurophysiology studies indicated, in general, the lack of pathology during sEMG tests in comparison to healthy subjects or even better muscle motor unit contractile properties in pilots, both in the upper and lower extremities. In pilots, the parameters of ENG and MEP examinations show a statistically significant sensitivity for detecting the slight changes and their consequences in the transmission of neural impulses within L4–L5 ventral root fibers. The research results enable specifying the algorithm of future preventing rehabilitative treatment in high-maneuvering jet fighters with an average flight experience of 2000 h and working conditions at 7G on average. This study, for the first time, describes the application of a set of diagnostic neurophysiological methods with the particular importance of MEPs in the clinical evaluation of the jet fighters’ health status.