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Glial-cell line derived neurotrophic factor (GDNF) is a neurotrophic factor produced by skeletal muscle. GDNF has been shown to be potent survival factor for motor neurons. We have previously shown that GDNF content in skeletal muscle is altered following exercise and GDNF levels 24 hours after the last bout of exercise are significantly different than those measured immediately after exercise. We sought to determine how long after exercise do GDNF protein levels remain altered. We removed extensor digitorum longus (EDL) and Soleus (SOL), 24 hr, 72 hr and 1 week following the last bout of involuntary exercise. Muscles were processed for GDNF protein content using enzyme linked immunosorbant assay. Contralateral EDL and SOL were removed at the same time points for immunohistochemical analysis. The results showed that 72 hr following exercise there was a significant increase of GDNF protein content in SOL and a significant decrease in EDL (p≤ 0.05). The results suggest exercise causes a relatively long-lived effect on GDNF protein content and that expression of GDNF protein in type I skeletal muscle maybe regulated differently than in type II skeletal muscle. This work was supported by NIH Grant 1R15AG022908-01A2, MSU-KCMS, and Western Michigan University.