Background: Increasing the activity of the gamma acetylcholine receptor subunit and semaphorin a3 by affecting the transmission of nerve messages to the muscle decreases skeletal muscle function. Evidence shows that regular physical activity can promote muscle function by affecting the expression of these two genes. This study investigated the effect of increasing resistance training on the expression of acetylcholine receptor gamma (CHRNG) and semaphorin-a3 (Sema3A) subunit genes in the gastrocnemius muscle of male rats. Methods: In an experimental study, 12 six-week-old male rats with an average weight of 195-220 grams, were randomly divided into 2 groups (6 in each group), resistance training (RT) and control (Control). The resistance training group performed incremental resistance training 5 days a week for 4 weeks. Twenty-four hours after the last training and recovery session, the sacrificial and biceps muscles of the subjects were extracted to determine the expression of CHRNG and Sema3A genes in real-time. Results: The expression of CHRNG (P=0.044) and Semaphorine-a3 (P=0.040) genes decreased significantly in the resistance training group compared with the control group. Conclusion: this study showed that incremental resistance training can improve neuromuscular function by reducing the expression of CHRNG and Sema3A genes at the neuromuscular junction. Based on these findings, increasing resistance exercises are recommended to improve muscle performance.