[Purpose] During exercise, skeletal muscle motor units are recruited based on afferent sensory input following peripheral metabolic by-product accumulation. The purpose of this study was to investigate whether lactate plays a role in conveying fatigue-related information to the brain. [Subjects] Eleven healthy adults participated in this study. [Methods] Subjects performed handgrip exercises at 10%, 30%, and 50% maximal voluntary contraction for 120 s. They were monitored for brachial artery blood pressure, respiratory quotient, muscle fatigue (integrated electromyogram, median power frequency), blood lactate levels, muscle blood flow, and brain activity. [Results] The handgrip exercise protocol caused significant muscle fatigue based on 28% and 37% reductions in median power frequency detected at 30% and 50% maximal voluntary contraction, respectively. Subjects exhibited intensity-dependent increases in blood pressure, respiratory quotient, muscle blood flow, and circulating lactate concentrations. Furthermore, brain activity increased at 30% and 50% maximal voluntary contraction. Multiple regression analysis identified muscle blood flow at 30% maximal voluntary contraction and lactate at 50% maximal voluntary contraction with standardized partial regression coefficients of −0.64 and 0.75, respectively. [Conclusion] These data suggest that blood lactate concentration and muscle blood flow, which reflect muscle metabolism, may convey load intensity information to the brain during muscle fatigue.