Musicians frequently complain of musculoskeletal pain due to high mechanical demands, with the cervical spine being the most affected. Increased neuromuscular mechanosensitivity due to repetitive mechanical stress over time has been described in neck pain patients. Nevertheless, the association between musculoskeletal pain and neuromuscular mechanosensitivity in musicians is unknown. Therefore, the aim of this study was to analyze the relationship between neuromuscular tissue mechanosensitivity and neck pain in guitarists. Guitarists with chronic neck pain (n = 70) and without pain (n = 70) were enrolled. Pain and disability were measured by the visual analogue scale and the Neck Disability Index, respectively. The pressure pain threshold (PPT) was bilaterally measured for the upper trapezius and median nerve. Finally, the Upper limb neural test one (ULNT1) was bilaterally measured. The analyses included a 2-by-2 mixed analysis of variance, pairwise comparisons with Bonferroni correction, linear regression model, and multiple linear regression. Our data showed that chronic neck pain guitarists have a lower PPT at all locations compared to healthy guitarists. They also showed a bilateral main effect for pain for ULNT1 compared to healthy guitarists. These results were not affected by the mediator variables. Finally, a relationship between upper trapezius PPT and median nerve PPT was found.