Effects of Multiple Bouts of Long-duration Hindlimb Unloading and Recovery on Rat Plantaris Muscle

Exposure to microgravity results in a rapid reduction of muscle mass. However, few studies exist designed to examine the effects of multiple long-term exposures to microgravity with alternating recovery periods on skeletal muscle. To determine what happens to the recovery of skeletal muscle when faced with subsequent unloading and recovery periods. Male Sprague-Dawley (6 mo) were assigned to the following groups as shown in figure 1 below: 28d hindlimb unloading (1HU), 28d HU session followed by a 56d recovery bout of normal cage ambulation at 1g (1HU+REC), 2 cycles of 28d HU with a 56d recovery period between unloadings (2HU), 2 cycles of 28d HU as in the 2HU group, but followed by an additional 56d recovery at 1g (2HU+REC), and an age- and housing-matched control group (CON). On the final day of the experimental period, plantaris muscles were excised and weighed. The 1HU+REC (0.548 ± 0.012), 2HU+REC (0.562 ± 0.015), and CON (0.550 ± 0.013) showed no statistical difference (p\u3e0.05) between each other. The 1 HU (0.442 ± 0.020) and 2 HU (0.431 ± 0.011) groups were significantly less (p\u3c0.001) than recovery and aged control animals but were not significantly different from each other. The results show that the plantaris muscle presented reduction of muscle mass with initial and subsequent exposures to microgravity. However, with the recovery period, animals were able to regain lost muscle mass, similar to age-matched controls. These findings would be relevant for astronauts participating in multiple long-duration missions throughout their career

Effects of Voluntary Resistance Exercise Training During Recovery From Hindlimb Unloading on Rat Gastrocnemius Muscle

As research continues to examine the deleterious impact of long-duration spaceflight on human muscle mass and function, there remain gaps in our knowledge of muscle physiology, especially in examining how muscle’s ability to recover or rehabilitate from unloading may alter the results of multiple exposures to microgravity followed by 1g recovery. The purpose of this study was to analyze the effects of resistance exercise training of gastrocnemius muscle mass and anabolism during the initial recovery period immediately following a bout of unloading, as well as to examine the role that exercise may have on a subsequent period of weightlessness. This was achieved in rodent models of simulated spaceflight (0g), recovery (1g), and resistance training (\u3e1g) using male Sprague-Dawley (6 mo) rats randomly assigned to the following groups: 28d hindlimb unloading (HU), 28d HU followed by a 56d recovery period of normal cage ambulation at 1g (1HU+REC), 2 cycles of 28d HU with a 56d recovery period between unloading (2HU), 2HU followed by an additional 56d recovery at 1g (2HU+REC), or an age- and housing-matched control group (CON). In addition, following the initial 28d HU period, two groups of animals were given 7d recovery at 1g followed by a 7wk (3 sessions/wk) moderate-intensity, moderate-volume voluntary resistance exercise program (EX) in which the animals were trained to perform a squat-like motion with full extension of the lower limb and resistance was applied incrementally by weighted pouches over the scapula to ~65% bodyweight. At the conclusion of the experiments, gastrocnemius muscles were carefully excised, weighed, and evaluated for cumulative (24h) rates of protein synthesis (FSR). Values of both muscle mass and FSR were lower than control during periods of unloading (p\u3c0.05), but with recovery, control values were reached for mass and surpassed for FSR. Interestingly, there was no significant difference between the mass of 2HU and 2HU+EX (p\u3e0.05), and both were diminished in comparison to control animals, suggesting that benefits of exercise during periods of ambulatory reloading after disuse/microgravity may not be additive. In conclusion, our data suggest that given adequate recovery, microgravity-induced losses of muscle mass can be fully restored to control values, and this adaptational response persists even with multiple exposures. These findings may have important implications not only for career astronauts, but also for individuals who have been subjected to casting of a limb or a period of bed rest following severe injury or illness

DEPTOR Expression Correlates with Muscle Protein Synthesis

Mammalian target of rapamycin (mTOR) has long been declared a focal point of muscle protein synthesis. mTORC1 (an mTOR complex consisting of mTOR, raptor, PRAS40, and mLST8) has been associated with regulation of protein translation in muscle, altering expression and activity levels of key downstream targets S6K1 and eIF-4E-BP1. mTORC1 has been shown to be affected by various stimuli, including nutritional status, growth factors, and mechanical loading. But in past incidents we have found disconnects in muscle protein synthesis and mTOR signaling, stimulating discussions that mTOR content and activation alone may not be able to fully account for muscle protein synthesis. Gaining popularity as a target for anti-cancer therapies, we became interested in DEPTOR, an endogenous inhibitor of mTORC1. Pharmacological inhibition of DEPTOR in cell culture and mouse studies has displayed increases of anabolic signaling in response to atrophic circumstances. We present two unique catabolic conditions in which we explore DEPTOR expression and muscle protein synthesis and demonstrate the first known data proposing that DEPTOR expression is not only influenced by physiological stimuli, including mechanical loading and insulin sensitivity, but that DEPTOR expression strongly correlates with 24-hr cumulative muscle protein synthesis rates. In one study, male Sprague Dawley rats were subjected to various conditions of musculoskeletal unloading, reloading, and overload, in which hindlimb unloading (HU) was utilized to mimic chronic disuse atrophy (28-d), followed by ambulatory reloading (56-d post HU) with and without the addition of resistance exercise prescribed to assist in recovery (3 sessions/wk for 7-wks; progressive increases in added resistance up to ~60% BW). DEPTOR expression was assessed via Immunoblotting. 24-hr cumulative muscle protein synthesis (FSR) was measured via stable isotope labeling and quantified by gas chromatogram/mass spectrometry. DEPTOR demonstrated a strong negative correlation with FSR in the gastrocnemius (r = - 0.93261; p \u3c0.01). In our second study, male obese Zucker rats were divided into their lean and obese phenotypes, as well as placed into sedentary and resistance exercised groups. DEPTOR and FSR were assessed as described above following operant conditioning and four progressive exercise sessions over 9-d. Gastrocnemius DEPTOR/FSR was again significant (r = - 0.75723; p\u3c0.01). Collectively, these results are the first to associate physiologic changes in DEPTOR expression with alterations of FSR, which may have important implications towards the design of therapeutic targets for the control of muscle mass or in evaluating muscle anabolism

Partial Support Ventilation and Mitochondrial-Targeted Antioxidants Protect against Ventilator-Induced Decreases in Diaphragm Muscle Protein Synthesis

Mechanical ventilation (MV) is a life-saving intervention in patients in respiratory failure. Unfortunately, prolonged MV results in the rapid development of diaphragm atrophy and weakness. MV-induced diaphragmatic weakness is significant because inspiratory muscle dysfunction is a risk factor for problematic weaning from MV. Therefore, developing a clinical intervention to prevent MV-induced diaphragm atrophy is important. In this regard, MV-induced diaphragmatic atrophy occurs due to both increased proteolysis and decreased protein synthesis. While efforts to impede MV-induced increased proteolysis in the diaphragm are well-documented, only one study has investigated methods of preserving diaphragmatic protein synthesis during prolonged MV. Therefore, we evaluated the efficacy of two therapeutic interventions that, conceptually, have the potential to sustain protein synthesis in the rat diaphragm during prolonged MV. Specifically, these experiments were designed to: 1) determine if partial-support MV will protect against the decrease in diaphragmatic protein synthesis that occurs during prolonged full-support MV; and 2) establish if treatment with a mitochondrial-targeted antioxidant will maintain diaphragm protein synthesis during full-support MV. Compared to spontaneously breathing animals, full support MV resulted in a significant decline in diaphragmatic protein synthesis during 12 hours of MV. In contrast, diaphragm protein synthesis rates were maintained during partial support MV at levels comparable to spontaneous breathing animals. Further, treatment of animals with a mitochondrial-targeted antioxidant prevented oxidative stress during full support MV and maintained diaphragm protein synthesis at the level of spontaneous breathing animals. We conclude that treatment with mitochondrial-targeted antioxidants or the use of partial-support MV are potential strategies to preserve diaphragm protein synthesis during prolonged MV

Twenty-One Days of Lunar Environment Alters Muscle Fiber Areas in Mouse Gastrocnemius

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Regulation of mitochondrial quality following repeated bouts of hindlimb unloading

Muscle disuse impairs muscle quality and is associated with increased mortality. Little is known regarding additive effects of multiple bouts of disuse, which is a common occurrence in patients experiencing multiple surgeries. Mitochondrial quality is vital to muscle health and quality; however, to date mitochondrial quality control has not been investigated following multiple bouts of disuse. Therefore, the purpose of this study was to investigate mitochondrial quality controllers during multiple bouts of disuse by hindlimb unloading. Male rats (n ∼ 8/group) were assigned to the following groups: hindlimb unloading for 28 days, hindlimb unloading with 56 days of reloading, 2 bouts of hindlimb unloading separated by a recovery phase of 56 days of reloading, 2 bouts of hindlimb unloading and recovery after each disuse, or control animals with no unloading. At designated time points, tissues were collected for messenger RNA and protein analysis of mitochondrial quality. Measures of mitochondrial biogenesis, such as proliferator-activated receptor gamma coactivator 1 alpha, decreased 30%–40% with unloading with no differences noted between unloading conditions. Measures of mitochondrial translation were 40%–50% lower in unloading conditions, with no differences noted between bouts of unloading. Measures of mitophagy were 40%–50% lower with reloading, with no differences noted between reloading conditions. In conclusion, disuse causes alterations in measures of mitochondrial quality; however, multiple bouts of disuse does not appear to have additive effects. Novelty Disuse atrophy causes multiple alterations to mitochondrial quality control. With sufficient recovery most detriments to mitochondrial quality control are fixed. In general, multiple bouts of disuse do not produce additive effects.The accepted manuscript in pdf format is listed with the files at the bottom of this page. The presentation of the authors' names and (or) special characters in the title of the manuscript may differ slightly between what is listed on this page and what is listed in the pdf file of the accepted manuscript; that in the pdf file of the accepted manuscript is what was submitted by the author

Ratio of p-mTOR to total mTOR protein in the diaphragm following 12 hours spontaneous breathing or mechanical ventilation.

<p>Representative western blots are shown below the graph. Values are expressed as mean ± SE change from percent control. Symbols: * significantly different (p<0.05) from all other groups.</p