Introduction and Background The objective of this study is to identify neurophysiological markers generated in laryngeal muscles by navigated transcranial magnetic stimulation (nTMS) of motor speech related cortical areas, opercular part of Broca's region and primary motor region (M1) for laryngeal muscles in healthy subjects. The specific objectives were: a) to determine the neurophysiological parameters of stimulation and registration of short latency response (SLR) in the laryngeal muscle after nTMS of primary motor region (M1) for the laryngeal muscles of the dominant hemisphere, b) to determine the neurophysiological parameters of stimulation and registration of the long latency response (LLR) in the laryngeal muscle after nTMS of opercular part of Broca’s region of the dominant hemisphere, c) to induce a pseudo speech arrest during stimulation of M1 for laryngeal muscle whose stimulation has previously generated SLR in CTHY muscle, and d) to induce speech arrest during stimulation of opercular part of Broca’s region whose stimulation has previously generated the LLR in CTHY muscle. Methodology and Results a) In all 19 healthy subjects short latency response (SLR) was recorded from laryngeal muscle by stimulation of M1 for laryngeal muscles with two applied methodologies (single stimulus and train of stimuli). In 11 patients we used the nTMS methodology with single stimulus during facilitation task (vocalization). The results of parameters of stimulation of SLR in laryngeal muscle: stimulation intensity 45.64 ± 17.68% of the maximum stimulator output and the intensity of the electric filed (EF) 23:58 ± 99.91 V / m, and the results of the parameters of registration of SLR in laryngeal muscle: latency of SLR 11.90 ± 1.27 ms and amplitude of SLR 0:20 ± 0.17 mV. In the other 8 subjects and 2 subjects who had applied the previous methodology, we used a new methodology of train of stimuli during visual object naming task (VON). Results showed no significant differences between two used methodologies. b) Moving the site of stimulation in front of the localization of M1 for laryngeal muscles toward Broca’s region of inferior frontal gyrus, LLR was recorded in laryngeal muscle in all 10 healthy subjects in whom we used a new methodology of train of stimuli during VON. The parameters of stimulation for eliciting LLR in laryngeal muscle were: stimulation intensity of 36.34 ± 4.94% of maximum stimulator output, the intensity of the electric field (EF) of 10:58 ± 57.68 V / m. The latency of LLR in laryngeal muscle was 58.50 ± 5.90 ms. In addition to the latency of LLR, we have measured the time in which LLR was induced in relation to the beginning of visually presented object. The results showed that there was no statistically significant difference in latency of LLR between the time difference in which LLR was induced and naming latency of VPO measured with the first appearance of the microphone onset. c) In all 10 subjects in whom M1 for laryngeal muscles was stimulated generating SLR in laryngeal muscle, the same location was again stimulated with a train of stimuli inducing the pseudo speech arrest during counting task and naming of VPO. Pseudo speech arrest was characterized by contractions of facial and lip muscles. d) In 10 subjects in whom opercular part of Boca’s region was stimulated generating LLR in laryngeal muscle, the same location was again stimulated with a train of stimuli inducing speech arrest in one subject during counting task and naming of VPO. In three subjects qualitative language disturbances were induced. Conclusion For the first time neurophysiologic markers of motor speech related cortical areas are identified by names: a) neurophysiologic marker for M1 for laryngeal muscle is SLR recorded in laryngeal muscle, and b) neurophysiologic marker for the opercular part of the Boca’s region is LLR recorded in laryngeal muscle. Significance The methodology was developed for identification of neurophysiologic markers of motor speech related cortical areas by names in healthy volunteers. This methodology is a direct contribution to the methodology of intraoperative direct and transcranial electrical stimulation for eliciting responses from laryngeal muscles. It is expected that this methodology will be used in preoperative mapping of motor speech related cortical areas to facilitate surgical planning, and mapping of motor speech related cortical areas during neurosurgical operations, in order to preserve these areas from injuries and to prevent postoperative speech deficits. The results of this study will contribute to our understanding of the underlying cognitive mechanisms responsible for the processing and production of speech.