The purpose of this study was to determine the influence of local contractile activity on lipoprotein lipase (LPL) regulation in skeletal muscle. Short-term voluntary run training increased LPL mRNA concentration and LPL immunoreactive mass about threefold in white skeletal muscles of the rat hindlimb (all P < 0.01). Training also increased total and heparin-releasable LPL enzyme activity in white hindlimb muscles and in postheparin plasma (P < 0.05). Training did not enhance LPL regulation in a white muscle that was not recruited during running (masseter). LPL levels were already high in red skeletal muscles of control rats, and training did not result in a further rise. In resting rats, local electrical stimulation of a motor nerve to a predominantly white muscle caused a significant rise in LPL mRNA, immunoreactive mass, and enzyme activity relative to the contralateral control muscle of the same animals (all P < 0.01). Finally, LPL expression was several times greater in a red muscle (soleus) of rats with normal postural activity than rats with immobilized hindlimbs (P < 0.01). In summary, these studies support the hypothesis that local contractile activity is required for increasing LPL expression during exercise training and for maintaining a high level of LPL expression in postural muscles.