A Time Course for Markers of Protein Synthesis and Degradation with Hindlimb Unloading and the Accompanying Anabolic Resistance to Refeeding
Paul Anthony Roberson, j appl physiol. 14 MAY 2020
INTRODUCTION: Skeletal muscle atrophy is associated with disease, aging, and disuse. Hindlimb unloading (HU) in animals provides an experimental model to study muscle atrophy. A comprehensive time course for how HU affects biomarkers of protein synthesis and degradation acutely and chronically, and the associated resistance to an anabolic stimulus following disuse, remains undocumented.
METHODS: 16-week old C57BL/6 mice underwent 0, 1, 12, 24, 72, 168, or 336h of HU. Following 336h of HU, mice were reloaded for 1, 24, or 72h. Another group of mice underwent 120h of HU, were fasted or refed, and then compared to similar condition control animals (CTL). Protein content and phosphorylation of biomarkers of protein synthesis, degradation, and autophagy were assessed in the soleus muscle.
RESULTS: Gastrocnemius, soleus, and plantaris muscles atrophied within 120h of HU. Protein synthesis trended to decrease following 24h of HU. p70S6K phosphorylation and protein synthesis increased with reloading. Following HU, changes in MAFbx and DEPTOR expression and DEPTOR phosphorylation were consistent with development of a catabolic state. DEPTOR expression recovered following reloading. Animals that underwent 120h of HU exhibited attenuation of refeeding-induced p70S6K phosphorylation compared to CTL counterparts. Following 120h of HU, protein synthesis, 4E-BP1 phosphorylation, and DEPTOR, MAFbx, and Sestrin1 expression indicated a catabolic state. Following 120h of HU, autophagy markers including p62 expression, REDD1 expression, LC3 ratio, and ULK-1 phosphorylation indicated impaired autophagy.
CONCLUSIONS: HU promotes a deleterious balance between protein synthesis and degradation. The time course herein provides scientists information about when the associated biomarkers become affected.