<p>Fatigue, a complex and multifaceted symptom, profoundly influences quality of life, particularly among individuals suffering from chronic medical conditions or neurological disorders. This symptom not only exacerbates existing conditions but also hinders daily functioning, thereby perpetuating a vicious cycle of worsening symptoms and reduced physical activity. Given the pivotal role of the motor cortex (M1) in coordinating and executing voluntary movements, understanding how the cortex regulates fatigue is crucial. Despite its importance, the neural mechanisms underlying fatigue remain inadequately explored. In this study, we employed electrophysiological recordings in the M1 region of mice to investigate how excitation–inhibition dynamics and neural oscillations are regulated during exercise-induced fatigue. We observed that fatigue led to decreased voluntary physical activity and cognitive performance, manifesting as reduced running wheel distance, mean speed, exercise intensity, and exploratory behaviour. At the neural level, we detected increased firing frequencies for M1 neurons, including both pyramidal neurons and interneurons, along with heightened beta-band oscillatory activity and stronger coupling between beta-band oscillations and interneurons. These findings enhance our understanding of the mechanisms underlying fatigue, offering insights into behavioural, excitability, and oscillatory changes. The results of this study could pave the way for the development of novel intervention strategies to combat fatigue.</p>