Although women have been performing increasingly more manual labor in the workplace in the past 2 decades, their physiological responses and gender-based differences in muscle microvascularity during occupational activities have not yet been extensively documented. This study assessed gender differences and tissue heterogeneity in peripheral circulatory responses from 2 muscle groups during pushing and pulling exercise until volitional exhaustion. In healthy men and women, near-infrared spectroscopy was used to determine peripheral responses, oxygenation, and blood volume simultaneously from the right biceps brachii and lumbar erector spinae. Pulmonary oxygen uptake was assessed using a metabolic measurement cart. Although the 11 men who participated in the study demonstrated greater pulmonary oxygen uptake and power output at volitional exhaustion, their peak peripheral responses for both muscles were similar to those of the 11 women participating. In both sexes, oxygenations trends decreased in both muscles with an increase in workload. However, whereas blood volume increased in the biceps, it decreased in the lumbar muscle in both sexes. At 20% to 60% levels of peak pulmonary oxygen uptake, the percent change in peripheral bicep responses was greater for men than for women (P < 0.05). In contrast, women demonstrated greater change in lumbar muscle oxygenation compared with men at 40% to 60% of peak pulmonary oxygen uptake (P < 0.05). Similar peripheral responses for biceps and lumbar muscles at the point of volitional exhaustion suggest that gender differences in pulmonary oxygen uptake are independent of oxygen extraction or delivery across the muscle groups monitored. However, at submaximal levels of exercise, the peripheral changes in each muscle were gender dependent. Although biceps and lumbar muscles are 2 discrete muscle groups, based on the heterogeneity found in the blood volume trends it is likely that oxygen supply and demand are regulated by muscle location and muscle fiber characteristics. Overall, gender-based assessment of occupational activities should incorporate both pulmonary and peripheral circulatory responses to understand each sex's performance effectiveness.