The effects of exercise and a physiological increase in plasma insulin concentration on muscle lipoprotein lipase activity (mLPLA), leg exchange of glucose, and serum lipoprotein levels were investigated in healthy young men. During euglycemic hyperinsulinemia (n = 7) at 44 mU.liter-1, m-LPLA in non-exercised muscle decreased from 30 +/- 7.4 mU.g-1 wet weight (w.w.) (mean +/- SE) to 19 +/- 3.3 (P less than 0.05). Furthermore, the decrease in m-LPLA correlated closely (r = 0.97, P less than 0.05) with the increase in leg glucose uptake. Moreover, basal m-LPLA correlated with the insulin-induced increase in leg glucose uptake (r = 0.93, P less than 0.05). In the control group (n = 6) in which saline was infused in place of insulin and glucose, m-LPLA in nonexercised muscle did not change with time. No change in m-LPLA was observed immediately after one-legged knee extension exercise, but 4 h after exercise m-LPLA was higher (P less than 0.05) in the exercised thigh (47 +/- 17.8 mU.g-1 w.w.) compared with the contralateral nonexercised thigh (29 +/- 6.3 mU.g-1 w.w.). This difference was not found 8 h after exercise. The triacylglycerol content of serum lipoproteins decreased during insulin infusion. It is concluded that in contrast to the effect on adipose tissue, physiological concentrations of insulin decrease m-LPLA in proportion to the effect of insulin on muscle glucose uptake, while muscle contractions cause a local, delayed, and transient increase in m-LPLA. Further-more, basal m-LPLA is an indicator of muscle insulin sensitivity.