Soleus biopsies were obtained from four male astronauts 45 days before and within 2 h after a 17 day spaceflight. For all astronauts, single chemically skinned post‐flight fibres expressing only type I myosin heavy chain (MHC) developed less average peak Ca2+ activated force (Po) during fixed‐end contractions (0.78 ± 0.02 vs. 0.99 ± 0.03 mN) and shortened at a greater mean velocity during unloaded contractions (Vo) (0.83 ± 0.02 vs. 0.64 ± 0.02 fibre lengths s−1) than pre‐flight type I fibres. The flight‐induced decline in absolute Po was attributed to reductions in fibre diameter and/or Po per fibre cross‐sectional area. Fibres from the astronaut who experienced the greatest relative loss of peak force also displayed a reduction in Ca2+ sensitivity. The elevated Vo of the post‐flight slow type I fibres could not be explained by alterations in myosin heavy or light chain composition. One alternative possibility is that the elevated Vo resulted from an increased myofilament lattice spacing. This hypothesis was supported by electron micrographic analysis demonstrating a reduction in thin filament density post‐flight. Post‐flight fibres shortened at 30 % higher velocities than pre‐flight fibres at external loads associated with peak power output. This increase in shortening velocity either reduced (2 astronauts) or prevented (2 astronauts) a post‐flight loss in fibre absolute peak power (μN (fibre length) s−1). The changes in soleus fibre diameter and function following spaceflight were similar to those observed after 17 days of bed rest. Although in‐flight exercise countermeasures probably reduced the effects of microgravity, the results support the idea that ground‐based bed rest can serve as a model of human spaceflight. In conclusion, 17 days of spaceflight decreased force and increased shortening velocity of single Ca2+‐activated muscle cells expressing type I MHC. The increase in shortening velocity greatly reduced the impact that impaired force production had on absolute peak power.

We examined intramuscular endomysial collagen, cross-linking, and advanced glycation end products, as well as the general and contractile protein concentration of 20 young (25 +/- 3 yr) and 22 old (78 +/- 6 yr, range: 70-93 yr) sedentary men and women to better understand the underlying basis of changes in skeletal muscle mass and function that occur with aging. The old individuals had an impaired ability (increased time) (P < 0.05) to climb stairs (80%), rise from a chair (56%), and walk (44%), as well as lower (P < 0.05) quadriceps muscle volume (-29%), muscle strength (-35%), muscle power (-48%), and strength (-17%) and power (-33%) normalized to muscle size. Vastus lateralis muscle biopsies revealed that intramuscular endomysial collagen (young: 9.6 +/- 1.1, old: 10.2 +/- 1.2 microg/mg muscle wet wt) and collagen cross-linking (hydroxylysylpyridinoline) (young: 395 +/- 65, old: 351 +/- 45 mmol hydroxylysylpyridinoline/mol collagen) were unchanged (P > 0.05) with aging. The advanced glycation end product, pentosidine, was increased (P < 0.05) by approximately 200% (young: 5.2 +/- 1.3, old: 15.9 +/- 4.5 mmol pentosidine/mol collagen) with aging. While myofibrillar protein concentration was lower (-5%, P < 0.05), the concentration of the main contractile proteins myosin and actin were unchanged (P > 0.05) with aging. These data suggest that the synthesis and degradation of proteins responsible for the generation (myosin and actin) and transfer (collagen and pyridinoline cross-links) of muscle force are tightly regulated in aging muscle. Glycation-related cross-linking of intramuscular connective tissue may contribute to altered muscle force transmission and muscle function with healthy aging.

This investigation examined the effects of acute resistance exercise (RE), progressive resistance training (PRT), and age on the human skeletal muscle Transcriptome. Two cohorts of young and old adults [study A: 24 yr, 84 yr (n = 28); study B: 25 yr, 78 yr (n = 36)] were studied. Vastus lateralis biopsies were obtained pre- and 4 h post-RE in conjunction with the 1st and 36th (last) training session as part of a 12-wk PRT program in study A, whereas biopsies were obtained in the basal untrained state in study B. Additionally, the muscle fiber type specific (MHC I and MHC IIa) Transcriptome response to RE was examined in a subset of young and old women from study A. Transcriptome profiling was performed using HG U133 Plus 2.0 Arrays. The main findings were 1) there were 661 genes affected by RE during the 1st and 36th training bout that correlated with gains in muscle size and strength with PRT (termed the Transcriptome signature of resistance exercise adaptations); 2) the RE gene response was most pronounced in fast-twitch (MHC IIa) muscle fibers and provided additional insight into the skeletal muscle biology affected by RE; 3) skeletal muscle of young adults is more responsive to RE at the gene level compared with old adults and age also affected basal level skeletal muscle gene expression. These skeletal muscle Transcriptome findings provide further insight into the molecular basis of sarcopenia and the impact of resistance exercise at the mixed muscle and fiber type specific level.

We examined the effect of two commonly consumed over-the-counter analgesics, ibuprofen and acetaminophen, on muscle protein synthesis and soreness after high-intensity eccentric resistance exercise. Twenty-four males (25 +/- 3 yr, 180 +/- 6 cm, 81 +/- 6 kg, and 17 +/- 8% body fat) were assigned to one of three groups that received either the maximal over-the-counter dose of ibuprofen (IBU; 1,200 mg/day), acetaminophen (ACET; 4,000 mg/day), or a placebo (PLA) after 10-14 sets of 10 eccentric repetitions at 120% of concentric one-repetition maximum with the knee extensors. Postexercise (24 h) skeletal muscle fractional synthesis rate (FSR) was increased 76 +/- 19% (P < 0.05) in PLA (0.058 +/- 0.012%/h) and was unchanged (P > 0.05) in IBU (35 +/- 21%; 0.021 +/- 0.014%/h) and ACET (22 +/- 23%; 0.010 +/- 0.019%/h). Neither drug had any influence on whole body protein breakdown, as measured by rate of phenylalanine appearance, on serum creatine kinase, or on rating of perceived muscle soreness compared with PLA. These results suggest that over-the-counter doses of both ibuprofen and acetaminophen suppress the protein synthesis response in skeletal muscle after eccentric resistance exercise. Thus these two analgesics may work through a common mechanism to influence protein metabolism in skeletal muscle.

Muscle biopsies were obtained from the vastus lateralis before and after 84 days of bed-rest from six control (BR) and six resistance-exercised (BRE) men to examine slow-and fast-twitch muscle fibre contractile function. BR did not exercise during bed-rest and had a 17 and 40% decrease in whole muscle size and function, respectively. The BRE group performed four sets of seven maximal concentric and eccentric supine squats 2-3 days per week (every third day) that maintained whole muscle strength and size. Slow (MHC I) and fast (MHC IIa) muscle fibres were studied at 15• C for diameter, peak force (P o ), contractile velocity (V o ) and force-power parameters. SDS-PAGE was performed on each single fibre after the functional experiments to determine MHC isoform composition. MHC I and IIa BR fibres were, respectively, 15 and 8% smaller, 46 and 25% weaker (P o ), 21 and 6% slower (V o ), and 54 and 24% less powerful after bed-rest (P < 0.05). BR MHC I and IIa P o and power normalized to cell size were lower (P < 0.05). BRE MHC I fibres showed no change in size or V o after bed-rest; however, P o was 19% lower (P < 0.05), resulting in 20 and 30% declines (P < 0.05) in normalized P o and power, respectively. BRE MHC IIa fibres showed no change in size, P o and power after bed-rest, while V o was elevated 13% (P < 0.05). BRE MHC IIa normalized P o and power were 10 and 15% lower (P < 0.05), respectively. MHC isoform composition shifted away from MHC I fibres, resulting in an increase (P < 0.05) in MHC I/IIa (BR and BRE) and MHC IIa/IIx (BR only) fibres. These data show that the contractile function of the MHC I fibres was more affected by bed-rest and less influenced by the resistance exercise protocol than the MHC IIa fibres. Considering the large differences in power of human MHC I and IIa muscle fibres (5-to 6-fold), the maintenance of whole muscle function with the resistance exercise programme is probably explained by (1) the maintenance of MHC IIa power and (2) the shift from slow to fast (MHC I → MHC I/IIa) in single fibre MHC isoform composition.

Skeletal muscle is a heterogeneous tissue comprised of muscle fiber and mononuclear cell types that, in addition to movement, influences immunity, metabolism and cognition. We investigated the gene expression patterns of skeletal muscle cells using RNA-seq of subtype-pooled single human muscle fibers and single cell RNA-seq of mononuclear cells from human vastus lateralis, mouse quadriceps, and mouse diaphragm. We identified 11 human skeletal muscle mononuclear cell types, including two fibro-adipogenic progenitor (FAP) cell subtypes. The human FBN1+ FAP cell subtype is novel and a corresponding FBN1+ FAP cell type was also found in single cell RNA-seq analysis in mouse. Transcriptome exercise studies using bulk tissue analysis do not resolve changes in individual cell-type proportion or gene expression. The cell-type gene signatures provide the means to use computational methods to identify cell-type level changes in bulk studies. As an example, we analyzed public transcriptome data from an exercise training study and revealed significant changes in specific mononuclear cell-type proportions related to age, sex, acute exercise and training. Our single-cell expression map of skeletal muscle cell types will further the understanding of the diverse effects of exercise and the pathophysiology of muscle disease.Skeletal muscle is a complex heterogeneous tissue consisting of multinucleated muscle fibers, immune cells, endothelial cells, muscle stem cells (satellite cells), non-myogenic mesenchymal progenitors (e.g., fibro-adipogenic progenitors, or FAPs), and other mononuclear cells 1 . To improve the understanding of skeletal muscle cell types and their transcriptional signatures, we studied human and mouse skeletal muscle mononuclear cells by single-cell RNA-sequencing and single human muscle fiber subtypes by RNA-seq.The majority of skeletal muscle is composed of the multinucleated fibers that facilitate movement. These muscle fibers include several fiber types of differing metabolic and functional properties 2-4 . While slow-twitch (or Type I) muscle fibers possess high oxidative capacity, fast-twitch (or Type II) muscle fibers have a high glycolytic capacity and are capable of supplying more power than Type I fibers 2-4 . Fiber-type composition differs across individuals and can change by as much as 10-30% during exercise training regimens 5-7 . Furthermore, the transcriptomic response to physical activity is different in each fiber-type as each fiber-type responds differently to different modes of exercise 8,9 . Crucially, muscle fibers secrete myokines, which both act locally within muscle tissue as well as influence other organs and tissues via hormone-like signaling 10 . Myokines may be responsible for the immune-, metabolism-, and cognition-related benefits of physical activity, as well as the chronic diseases that are caused by lack of physical activity (insulin resistance, cardiovascular disease, etc.) 10 .Besides multinucleated fibers, skeletal muscle contains many mononuclear cells, such as immune cells,...

Trappe TA, Carroll CC, Dickinson JM, LeMoine JK, Haus JM, Sullivan BE, Lee JD, Jemiolo B, Weinheimer EM, Hollon CJ. Influence of acetaminophen and ibuprofen on skeletal muscle adaptations to resistance exercise in older adults. Am J Physiol Regul Integr Comp Physiol 300: R655-R662, 2011. First published December 15, 2010; doi:10.1152/ajpregu.00611.2010.-Evidence suggests that consumption of over-the-counter cyclooxygenase (COX) inhibitors may interfere with the positive effects that resistance exercise training has on reversing sarcopenia in older adults. This study examined the influence of acetaminophen or ibuprofen consumption on muscle mass and strength during 12 wk of knee extensor progressive resistance exercise training in older adults. Thirty-six individuals were randomly assigned to one of three groups and consumed the COX-inhibiting drugs in double-blind placebo-controlled fashion: placebo (67 Ϯ 2 yr; n ϭ 12), acetaminophen (64 Ϯ 1 yr; n ϭ 11; 4 g/day), and ibuprofen (64 Ϯ 1 yr; n ϭ 13; 1.2 g/day). Compliance with the resistance training program (100%) and drug consumption (via digital video observation, 94%), and resistance training intensity were similar (P Ͼ 0.05) for all three groups. Drug consumption unexpectedly increased muscle volume (acetaminophen: 109 Ϯ 14 cm 3 , 12.5%; ibuprofen: 84 Ϯ 10 cm 3 , 10.9%) and muscle strength (acetaminophen: 19 Ϯ 2 kg; ibuprofen: 19 Ϯ 2 kg) to a greater extent (P Ͻ 0.05) than placebo (muscle volume: 69 Ϯ 12 cm 3 , 8.6%; muscle strength: 15 Ϯ 2 kg), when controlling for initial muscle size and strength. Follow-up analysis of muscle biopsies taken from the vastus lateralis before and after training showed muscle protein content, muscle water content, and myosin heavy chain distribution were not influenced (P Ͼ 0.05) by drug consumption. Similarly, muscle content of the two known enzymes potentially targeted by the drugs, COX-1 and -2, was not influenced (P Ͼ 0.05) by drug consumption, although resistance training did result in a drug-independent increase in COX-1 (32 Ϯ 8%; P Ͻ 0.05). Drug consumption did not influence the size of the nonresistance-trained hamstring muscles (P Ͼ 0.05). Over-thecounter doses of acetaminophen or ibuprofen, when consumed in combination with resistance training, do not inhibit and appear to enhance muscle hypertrophy and strength gains in older adults. The present findings coupled with previous short-term exercise studies provide convincing evidence that the COX pathway(s) are involved in the regulation of muscle protein turnover and muscle mass in humans.cyclooxygenase; prostaglandin; sarcopenia SARCOPENIA, DEFINED AS THE loss of skeletal muscle mass, is a debilitating clinical condition associated with old age (13,40). This age-related loss of muscle mass is associated with a loss of muscle strength and function, a reduced functional independence, and a financial burden to the US healthcare system estimated at $20 billion per year (19,23,24,51). One of the most proven treatments for sarcopenia is resistance exercise training, which in...

Harber MP, Konopka AR, Douglass MD, Minchev K, Kaminsky LA, Trappe TA, Trappe S. Aerobic exercise training improves whole muscle and single myofiber size and function in older women. Am J Physiol Regul Integr Comp Physiol 297: R1452-R1459, 2009. First published August 19, 2009 doi:10.1152/ajpregu.00354.2009.-To comprehensively assess the influence of aerobic training on muscle size and function, we examined seven older women (71 Ϯ 2 yr) before and after 12 wk of cycle ergometer training. The training program increased (P Ͻ 0.05) aerobic capacity by 30 Ϯ 6%. Quadriceps muscle volume, determined by magnetic resonance imaging (MRI), was 12 Ϯ 2% greater (P Ͻ 0.05) after training and knee extensor power increased 55 Ϯ 7% (P Ͻ 0.05). Muscle biopsies were obtained from the vastus lateralis to determine size and contractile properties of individual slow (MHC I) and fast (MHC IIa) myofibers, myosin light chain (MLC) composition, and muscle protein concentration. Aerobic training increased (P Ͻ 0.05) MHC I fiber size 16 Ϯ 5%, while MHC IIa fiber size was unchanged. MHC I peak power was elevated 21 Ϯ 8% (P Ͻ 0.05) after training, while MHC IIa peak power was unaltered. Peak force (Po) was unchanged in both fiber types, while normalized force (Po/cross-sectional area) was 10% lower (P Ͻ 0.05) for both MHC I and MHC IIa fibers after training. The decrease in normalized force was likely related to a reduction (P Ͻ 0.05) in myofibrillar protein concentration after training. In the absence of an increase in Po, the increase in MHC I peak power was mediated through an increased (P Ͻ 0.05) maximum contraction velocity (Vo) of MHC I fibers only. The relative proportion of MLC1s (Pre: 0.62 Ϯ 0.01; Post: 0.58 Ϯ 0.01) was lower (P Ͻ 0.05) in MHC I myofibers after training, while no differences were present for MLC 2s and MLC 3f isoforms. These data indicate that aerobic exercise training improves muscle function through remodeling the contractile properties at the myofiber level, in addition to pronounced muscle hypertrophy. Progressive aerobic exercise training should be considered a viable exercise modality to combat sarcopenia in the elderly population.