Gregory R. Adams

Skeletal muscle gene expression in space-flown rats

Skeletal muscles are vulnerable to marked atrophy under microgravity. This phenomenon is due to the transcriptional alteration of skeletal muscle cells to weightlessness. To further investigate this issue at a subcellular level, we examined the expression of ~26,000 gastrocnemius muscle genes in space‐flown rats by DNA microarray analysis. Comparison of the changes in gene expression among spaceflight, tail‐suspended, and denervated rats revealed that such changes were unique after spaceflight and not just an extension of simulated weightlessness. The microarray data showed two spaceflight‐specific gene expression patterns: 1) imbalanced expression of mitochondrial genes with disturbed expression of cytoskeletal molecules, including putative mitochondria‐anchoring proteins, A‐kinase anchoring protein, and cytoplasmic dynein, and 2) up‐regulated expression of ubiquitin ligase genes, MuRF‐1, Cbl‐b, and Siah‐1A, which are rate‐limiting enzymes of muscle protein degradation. Distorted expression of cytoskeletal genes during spaceflight resulted in dislocation of the mitochondria in the cell. Several oxidative stress‐inducible genes were highly expressed in the muscle of spaceflight rats. We postulate that mitochondrial dislocation during spaceflight has deleterious effects on muscle fibers, leading to atrophy in the form of insufficient energy provision for construction and leakage of reactive oxygen species from the mitochondria.

Effects of creatine loading and training on running performance and biochemical properties of rat skeletal muscle

Several reports have shown that the use of oral creatine (Cr) supplementation can increase performance during brief high intensity exercise in humans. The purpose of this study was to examine the separate and combined effects of Cr supplementation and high intensity run training on the performance capacity and biochemical properties of rodent skeletal muscle. Running performance was assessed following acute (10-d) and chronic (4-wk) Cr supplementation. Results indicate that Cr supplementation alone has ergogenic effects and the combination of run training plus Cr results in a more pronounced enhancement of performance than either intervention alone. The benefits of Cr supplementation were seen most clearly during repetitive bouts of high intensity interval running. Cr concentrations increased in both the slow soleus and fast plantaris muscles (P < 0.05) in response to Cr supplementation. Increased creatine concentrations appeared to be reflected in increased phosphorylated creatine (PCr). Citrate synthase (CS) activity was increased in both the soleus and plantaris muscles following training (P < 0.05). CS activity of the untrained soleus but not the plantaris responded to the dietary stimulus. There were no significant changes in either creatine phosphokinase activity or myosin heavy chain isoform distribution following training or supplementation. These results indicate that the gains in high intensity running performance seen following Cr loading are a combined result of increased aerobic (CS) and anaerobic (Cr and PCr) energy buffering capacity of the muscle.

Skeletal muscle growth in young rats is inhibited by chronic exposure to IL-6 but preserved by concurrent voluntary endurance exercise

Childhood diseases are often accompanied by chronic inflammation, which is thought to negatively impact growth. Interleukin-6 (IL-6) is typically cited as an indicator of inflammation and is linked to impaired growth. This study was designed to isolate and identify potential effects of chronic IL-6 exposure on skeletal muscle growth during development. A second aim was to determine if endurance exercise, thought to antagonize chronic inflammation, would interact with any effects of IL-6. The muscles of one leg of rapidly growing rats were exposed to IL-6 or vehicle for 14 days. Subgroups of IL-6-infused rats were provided access to running wheels. Local IL-6 infusion resulted in approximately 13% muscle growth deficit (myofibrillar protein levels). Exercise (>4,000 m/day) prevented this deficit. IL-6 infusion increased mRNA for suppressor of cytokine signaling-3 (SOCS3) and tumor necrosis factor-alpha (TNF-alpha), and this was not prevented by exercise. IL-6 infusion increased the mRNAs for atrogin, insulin-like growth factor-I (IGF-I), and IGF binding protein-4 (IGFBP4), and these effects were mitigated by exercise. Exercise stimulated an increase in total RNA ( approximately 19%) only in the IL-6-infused muscle, suggesting that a compensatory increase in translational capacity was required to maintain muscle growth. This study indicates that IL-6 exposure during periods of rapid growth in young animals can retard growth possibly via interactions with key growth factors. Relatively high volumes of endurance-type exercise do not exacerbate the negative effects of IL-6 and in fact were found to be beneficial in protecting muscle growth.

Characterization and Regulation of Mechanical Loading-Induced Compensatory Muscle Hypertrophy

In mammalian systems, skeletal muscle exists in a dynamic state that monitors and regulates the physiological investment in muscle size to meet the current level of functional demand. This review attempts to consolidate current knowledge concerning development of the compensatory hypertrophy that occurs in response to a sustained increase in the mechanical loading of skeletal muscle. Topics covered include: defining and measuring compensatory hypertrophy, experimental models, loading stimulus parameters, acute responses to increased loading, hyperplasia, myofiber-type adaptations, the involvement of satellite cells, mRNA translational control, mechanotransduction, and endocrinology. The authors conclude with their impressions of current knowledge gaps in the field that are ripe for future study.

Voluntary exercise during chronic renal failure in rats

PURPOSE Chronic renal failure (CRF) patients often experience a significant degradation in quality of life that is associated with decreased physical fitness. Previous animal studies have used forced running or swimming as modalities to investigate the interactions between exercise and CRF. These modalities generally include stress responses unrelated to the exercise itself. The purpose of the current work was to determine whether, and to what extent, rats experiencing the onset of CRF would participate in voluntary wheel running exercise. An additional objective was to examine physiological parameters related to skeletal muscle and cardiovascular adaptation in the context of CRF and exercise. METHODS Groups of rats were assigned to sham-operated or 5/6 nephrectomy groups, and further divided into running or nonrunning subgroups. Blood, heart, and muscle tissues were collected 30 d after the exercise groups were returned to running wheel-equipped cages. RESULTS The results demonstrated that rats experiencing the early stages of CRF will voluntarily exercise to the same extent as sham-operated animals (e.g., sham, 7.2+/-0.8 vs CRF, 6.8+/-0.7 km.d). CRF resulted in increased systolic blood pressure that was not normalized by exercise. CRF induced a decrease in hemoglobin concentration that was prevented by exercise. Voluntary running resulted in an apparently nonpathological left ventricular hypertrophy in both the sham-operated and CRF rats. In locomotor skeletal muscles, CRF resulted in a 31% decrease in citrate synthase activity that was completely blunted by voluntary running activity. CONCLUSION Rats experiencing the onset of CRF will run voluntarily. This exercise appears to provide some potentially palliative effects on the skeletal muscle and cardiovascular responses to CRF.

Exercise and leukocyte interchange among central circulation, lung, spleen, and muscle.

Circulating leukocytes increase rapidly with exercise then quickly decrease when the exercise ends. We tested whether exercise acutely led to bidirectional interchange of leukocytes between the circulation and the lung, spleen, and active skeletal muscle. To accomplish this it was necessary to label a large number of immune cells (granulocytes, monocytes, and lymphocytes) in a way that resulted in minimal perturbation of cell function. Rats were injected intravenously with a single bolus of carboxyfluorescein diacetate succinamidyl ester (CFSE) dye which is rapidly and irreversibly taken up by circulating cells. The time course of the disappearance of labeled cells and their reappearance in the circulation following exercise was determined via flow cytometry. The majority of circulating leukocytes were labeled at 4h. post-injection and this proportion slowly declined out to 120 h. At both 24 and 120 h, running resulted in an increase in the proportion of labeled leukocytes in the circulation. Analysis of the skeletal muscle, spleen and lung indicated that labeled leukocytes had accumulated in those tissues and were mobilized to the circulation in response to exercise. This indicates that there is an ongoing exchange of leukocytes between the circulation and tissues and that exercise can stimulate their redistribution. Exchange was slower with muscle than with spleen and lung, but in all cases, influenced by exercise. Exercise bouts redistribute leukocytes between the circulation and the lung, spleen and muscle. The modulatory effects of exercise on the immune system may be regulated in part by the systemic redistribution of immune cells.

Insulin-Like Growth Factor I Signaling in Skeletal Muscle and the Potential for Cytokine Interactions

Recent research has demonstrated that intracellular signaling components associated with several proinflammatory cytokines have the potential to interact with signaling pathways that regulate anabolic processes in skeletal muscle. This presentation and the ensuing brief review are intended to present a selection of the potential interactions between these two critical processes.

Effect of exercise training on aortic tone in chronic renal insufficiency.

BACKGROUND Chronic renal insufficiency (CRI) is associated with a high incidence of hypertension (HTN), endothelial dysfunction, atherosclerosis and cardiovascular disease. Sedentary life style increases, whereas regular exercise reduces the risk of cardiovascular disease. This study was designed to test the effect of regular exercise on vasodilatory and vasoconstrictive responses of the thoracic aorta in rats with renal mass reduction. METHODS One week after 5/6 nephrectomy (CRI) or sham operation (control), rats were housed in either regular cages or cages equipped with running wheels for 4 weeks. Thereafter, thoracic aorta was harvested and contractile response to potassium and phenylephrine (PhE), and relaxation response to acetylcholine (ACh) and sodium nitroprusside (SNP) were determined. RESULTS Compared with the control animals, sedentary CRI animals exhibited significant azotemia, proteinuria, HTN, oxidative stress, and increased sensitivity to potassium and PhE, and reduced sensitivity to ACh and SNP. Exercise training for 4 weeks reduced oxidative stress, reversed CRI-induced heightened sensitivity of the aorta to PhE and potassium, and restored its sensitivity to ACh (but not SNP) without affecting arterial pressure or renal function. CONCLUSIONS CRI results in heightened sensitivity to potassium- and alpha-1 adrenergic-mediated contractility and depressed sensitivity to endothelium-dependent relaxation in the aorta. Regular exercise improves these abnormalities without affecting arterial pressure or renal function. These observations suggest that exercise training can improve vascular function in animals, and perhaps humans, with chronic kidney disease.

Growth inhibition and compensation in response to neonatal hypoxia in rats.

Background:Hypoxia (Hx) is an important disease mechanism in prematurity, childhood asthma, and obesity. In children, Hx results in chronic inflammation.Methods:We investigated the effects of Hx (12% O2) during postnatal days 2–20 in rats. Control groups were normoxic control (Nc), and normoxic growth restricted (Gr) (14-pup litters).Results:The Hx-exposed and Gr rats had similar decreases in growth. Hx increased plasma tumor necrosis factor-α (TNF-α) and interleukin 6 (IL-6) levels and decreased insulin-like growth factor 1 (IGF-I) and vascular endothelial growth factor (VEGF) levels. Hx resulted in hypertrophy of the right ventricle (RV) but disproportionate decrements in limb skeletal muscle (SM) growth. miR-206 was depressed in the hypertrophied RV of Hx rats but was increased in growth-retarded SM. Hx resulted in decreased RV messenger RNA (mRNA) level for myostatin but had no effect on SM myostatin. The mRNA for Hx-sensitive factors such as hypoxia inducible factor-1α (HIF-1α) was depressed in the RV of Hx rats, suggesting negative feedback.Conclusion:The results indicate that Hx induces a proinflammatory state that depresses growth-regulating mechanisms and that tissues critical for survival, such as the heart, can escape from this general regulatory program to sustain life. This study identifies accessible biomarkers for evaluating the impact of interventions designed to mitigate the long-term deleterious consequences of Hx that all too often occur in babies born prematurely.

A rat model of exercise-induced asthma: a nonspecific response to a specific immunogen.

Exercise-induced bronchoconstriction (EIB) is common; however, key aspects of its pathogenesis are still unclear. We investigated the feasibility of adapting an established animal model of asthma to investigate the earliest stages of EIB. The hypothesis was that a single exposure to a normally innocuous, and brief, exercise challenge could trigger EIB symptoms in rats previously sensitized to ovalbumin (OVA) but otherwise unchallenged. Brown-Norway rats were sensitized by intraperitoneal injection of OVA at 0 and 2 wk. At week 3, animals were exposed to either aerosolized OVA (SS) or exercise (EXS). A trained, blinded, clinical observer graded EIB by respiratory sounds. Plasma and lung cytokine levels were analyzed. No control rats with or without exercise (EX, CON) showed evidence of EIB. Eighty percent of the SS group demonstrated abnormal breath sounds upon exposure to aerosolized OVA. Approximately 30% of EXS rats sensitized to OVA but exposed only to exercise had abnormal breath sounds. Lung tissue levels of TNF-α, IL-1α, growth-related oncogene/keratinocyte/chemoattractant, and IFN-γ were significantly higher (P < 0.001) in the SS group, relative to all other groups. Changes in most of these cytokines were not notable in the EXS rats, suggesting a different mechanism of EIB. Remarkably, IFN-γ, but not the other cytokines measured, was significantly elevated following brief exercise in both sensitized and unsensitized rats. Exercise led to detectable breathing sound abnormalities in sensitized rats, but less severe than those observed following classical OVA challenge. Precisely how this immune crossover occurs is not known, but this model may be useful in elucidating essential mechanisms of EIB.