Dietary nitrate preserves mitochondrial bioenergetics and mitochondrial protein synthesis rates during short-term immobilization in mice.

Skeletal muscle disuse reduces muscle protein synthesis rates and induces atrophy, events associated with decreased mitochondrial respiration and increased reactive oxygen species (ROS). Since dietary nitrate can improve mitochondrial bioenergetics, we examined whether nitrate supplementation attenuates disuse-induced impairments in mitochondrial function and muscle protein synthesis rates. Female C57Bl/6N mice were subject to single-limb casting (3 or 7 days) and consumed drinking water with or without 1 mM sodium nitrate. Compared to the contralateral control limb, 3 days of immobilization lowered myofibrillar fractional synthesis rates (FSR, p<0.0001), resulting in muscle atrophy. While FSR and mitophagy-related proteins were higher in subsarcolemmal (SS) compared to intermyofibrillar (IMF) mitochondria, immobilization for 3 days decreased FSR in both SS (p = 0.009) and IMF (p = 0.031) mitochondria. Additionally, 3 days of immobilization reduced maximal mitochondrial respiration and protein content, and increased maximal mitochondrial ROS emission without altering mitophagy-related proteins in muscle homogenate or isolated mitochondria (SS, IMF). While nitrate consumption did not attenuate the decline in muscle mass or myofibrillar FSR, intriguingly, nitrate completely prevented immobilization-induced reductions in SS and IMF mitochondrial FSR. In addition, nitrate prevented alterations in mitochondrial content and bioenergetics following both 3 and 7 days of immobilization. However, in contrast to 3 days of immobilization, nitrate did not prevent the decline in SS and IMF mitochondrial FSR following 7 days. Therefore, while nitrate supplementation was not sufficient to prevent muscle atrophy, nitrate may represent a promising therapeutic strategy to maintain mitochondrial bioenergetics and transiently preserve mitochondrial protein synthesis rates during short-term muscle disuse. KEY POINTS: Alterations in mitochondrial bioenergetics (decreased respiration and increased reactive oxygen species) are thought to contribute to muscle atrophy and reduced protein synthesis rates during muscle disuse. Since dietary nitrate can improve mitochondrial bioenergetics, we examined if nitrate supplementation could attenuate immobilization-induced skeletal muscle impairments in female mice. Dietary nitrate prevented short-term (3 day) immobilization-induced declines in mitochondrial protein synthesis rates, reductions in markers of mitochondrial content, and alterations in mitochondrial bioenergetics. Despite these benefits, and the preservation of mitochondrial content and bioenergetics during more prolonged (7 day) immobilization, nitrate consumption did not preserve skeletal muscle mass or myofibrillar protein synthesis rates. Overall, while dietary nitrate did not prevent atrophy, nitrate supplementation represents a promising nutritional approach to preserve mitochondrial function during muscle disuse. Abstract figure legend In female mice consuming standard drinking water (H 2 O), 3 and 7 days of single-limb immobilization decreased mitochondrial (mito) protein fractional synthesis rate (FSR), myofibrillar (myofib) protein FSR, and mitochondrial respiration, and increased mitochondrial reactive oxygen species (ROS). In contrast, sodium nitrate (NO 3 ) prevented the immobilization-induced alterations in mitochondrial bioenergetics (respiration, ROS) at both timepoints (3- and 7-day). In addition, mitochondrial protein FSR was transiently (3 day) preserved in the immobilized limb of nitrate-consuming mice. Combined, while dietary nitrate was not sufficient to prevent muscle atrophy, nitrate preserved mitochondrial bioenergetics and mitochondrial protein synthesis rates during short-term muscle disuse in mice. This article is protected by copyright. All rights reserved.