We investigated the effect of resistance training and fish protein intake on the motor unit firing pattern and motor function in elderly. Fifty healthy elderly males and females (69.2 ± 4.7 years) underwent 6 weeks of intervention. We applied the leg-press exercise as resistance training and fish protein including Alaska pollack protein (APP) as nutritional supplementation. Subjects were divided into four groups: fish protein intake without resistance training (APP-CN, n = 13), placebo intake without resistance training (PLA-CN, n = 12), fish protein intake with resistance training (APP-RT, n = 12), and placebo intake with resistance training (PLA-RT, n = 13). Motor unit firing rates were calculated from multi-channel surface electromyography by the Convolution Kernel. For the chair-stand test, while significant increases were observed at 6 weeks compared with 0 week in all groups (p < 0.05), significant increases from 0 to 3 weeks and 6 weeks were observed in APP-RT (18.2 ± 1.9 at 0 week to 19.8 ± 2.2 at 3 weeks and 21.2 ± 1.9 at 6 weeks) (p < 0.05). Increase and/or decrease in the motor unit firing rate were mainly noted within motor units with a low-recruitment threshold in APP-RT and PLA-RT at 3 and 6 weeks (12.3 pps at 0 week to 13.6 pps at 3 weeks and 12.1 pps at 6 weeks for APP-RT and 12.9 pps at 0 week to 13.9 pps at 3 weeks and 14.1 pps at 6 weeks for PLA-RT at 50% of MVC) (p < 0.05), but not in APP-CN or PLA-CN (p > 0.05). Time courses of changes in the results of the chair-stand test and motor unit firing rate were different between APP-RT and PLA-RT. These findings suggest that, in the elderly, the effect of resistance training on the motor unit firing rate is observed in motor units with a low-recruitment threshold, and additional fish protein intake modifies these adaptations in motor unit firing patterns and the motor function following resistance training.