PURPOSE: This study investigates the acute and longitudinal effects of resistance training on occupational muscle activity in office workers with chronic pain. METHODS: 30 female office workers with chronic neck and shoulder pain participated for 10 weeks in high-intensity elastic resistance training for 2 minutes per day (n = 15) or in control receiving weekly email-based information on general health (n = 15). Electromyography (EMG) from the splenius and upper trapezius was recorded during a normal workday. RESULTS: Adherence to training and control interventions were 86% and 89%, respectively. Compared with control, training increased isometric muscle strength 6% (P < 0.05) and decreased neck/shoulder pain intensity by 40% (P < 0.01). The frequency of periods with complete motor unit relaxation (EMG gaps) decreased acutely in the hours after training. By contrast, at 10-week follow-up, training increased average duration of EMG gaps by 71%, EMG gap frequency by 296% and percentage time below 0.5%, and 1.0% EMGmax by 578% and 242%, respectively, during the workday in m. splenius. CONCLUSION: While resistance training acutely generates a more tense muscle activity pattern, the longitudinal changes are beneficial in terms of longer and more frequent periods of complete muscular relaxation and reduced pain.