Genotype Score for Iron Status Is Associated with Muscle Fiber Composition in Women

Human muscle fiber composition is heterogeneous and mainly determined by genetic factors. A previous study reported that experimentally induced iron deficiency in rats increases the proportion of fast-twitch muscle fibers. Iron status has been reported to be affected by genetic factors. As the TMPRSS6 rs855791 T/C and HFE rs1799945 C/G polymorphisms are strongly associated with iron status in humans, we hypothesized that the genotype score (GS) based on these polymorphisms could be associated with the muscle fiber composition in humans. Herein, we examined 214 Japanese individuals, comprising of 107 men and 107 women, for possible associations of the GS for iron status with the proportion of myosin heavy chain (MHC) isoforms (I, IIa, and IIx) as markers of muscle fiber composition. No statistically significant correlations were found between the GS for iron status and the proportion of MHC isoforms in all participants. When the participants were stratified based on sex, women showed positive and negative correlations of the GS with MHC-IIa (age-adjusted p = 0.020) and MHC-IIx (age-adjusted p = 0.011), respectively. In contrast, no correlation was found in men. In women, a 1-point increase in the GS was associated with 2.42% higher MHC-IIa level and 2.72% lower MHC-IIx level. Our results suggest that the GS based on the TMPRSS6 rs855791 T/C and HFE rs1799945 C/G polymorphisms for iron status is associated with muscle fiber composition in women.

Skeletal Muscle Adaptations and Performance Outcomes Following a Step and Exponential Taper in Strength Athletes

Before major athletic events, a taper is often prescribed to facilitate recovery and enhance performance. However, it is unknown which taper model is most effective for peaking maximal strength and positively augmenting skeletal muscle. Thus, the purpose of this study was to compare performance outcomes and skeletal muscle adaptations following a step vs. an exponential taper in strength athletes. Sixteen powerlifters (24.0 ± 4.0 years, 174.4 ± 8.2 cm, 89.8 ± 21.4 kg) participated in a 6-week training program aimed at peaking maximal strength on back squat [initial 1-repetition-maximum (1RM): 174.7 ± 33.4 kg], bench press (118.5 ± 29.9 kg), and deadlift (189.9 ± 41.2 kg). Powerlifters were matched based on relative maximal strength, and randomly assigned to either (a) 1-week overreach and 1-week step taper or (b) 1-week overreach and 3-week exponential taper. Athletes were tested pre- and post-training on measures of body composition, jumping performance, isometric squat, and 1RM. Whole muscle size was assessed at the proximal, middle, and distal vastus lateralis using ultrasonography and microbiopsies at the middle vastus lateralis site. Muscle samples (n = 15) were analyzed for fiber size, fiber type [myosin-heavy chain (MHC)-I, -IIA, -IIX, hybrid-I/IIA] using whole muscle immunohistochemistry and single fiber dot blots, gene expression, and microRNA abundance. There were significant main time effects for 1RM squat (p < 0.001), bench press (p < 0.001), and deadlift, (p = 0.024), powerlifting total (p < 0.001), Wilks Score (p < 0.001), squat jump peak-power scaled to body mass (p = 0.001), body mass (p = 0.005), fat mass (p = 0.002), and fat mass index (p = 0.002). There were significant main time effects for medial whole muscle cross-sectional area (mCSA) (p = 0.006) and averaged sites (p < 0.001). There was also a significant interaction for MHC-IIA fiber cross-sectional area (fCSA) (p = 0.014) with post hoc comparisons revealing increases following the step-taper only (p = 0.002). There were significant main time effects for single-fiber MHC-I% (p = 0.015) and MHC-IIA% (p = 0.033), as well as for MyoD (p = 0.002), MyoG (p = 0.037), and miR-499a (p = 0.033). Overall, increases in whole mCSA, fCSA, MHC-IIA fCSA, and MHC transitions appeared to favor the step taper group. An overreach followed by a step taper appears to produce a myocellular environment that enhances skeletal muscle adaptations, whereas an exponential taper may favor neuromuscular performance.

Effect of Eccentric and Concentric Contraction Mode on Myogenic Regulatory Factors Expression in Human Vastus Lateralis Muscle

Background: Skeletal muscle contractions are caused to release myokines by muscle fiber. This study investigated the myogenic regulatory factors, as MHC I, IIA, IIX, Myo-D, MRF4, Murf, Atrogin-1, Decorin, Myonection, and IL-15 mRNA expression in the response of eccentric vs. concentric contraction. Methods: Eighteen healthy men were randomly divided into two eccentric and concentric groups, each of 9 persons. Isokinetic contraction protocols included maximal single-leg eccentric or concentric knee extension tasks at 60°/s with the dominant leg. Contractions consisted of a maximum of 12 sets of 10 reps, and the rest time between each set was 30 seconds. The baseline biopsy was performed four weeks before the study, and post-test biopsies were taken immediately after exercise protocols from Vastus Lateralis muscle. The gene expression levels evaluated using Real-Time PCR methods. Results: A significant difference in MyoD, MRF4, Myonection, and Decorin mRNA, were observed following eccentric or concentric contractions (P≤0.05). The MHC I, MHC IIA, IL-15 mRNA has been changed significantly compared to the pre-exercise in the concentric group (P≤0.05). While only MHC IIX and Atrogin-1 mRNA changed significantly in the eccentric group (P≤0.05). Additionally, the results showed a significant difference in MyoD, MRF4, IL-15, and Decorin were observed at the follow-up values between eccentric or concentric groups (P≤0.05). Conclusion: Our findings highlight the growing importance of elucidating the different responses of muscle growth factors associated with a myogenic activity such as MHC IIA, Decorin, IL-15, Myonectin, Decorin, MuRF1, and MHC IIX mRNA in following to various types of exercise.

Contractility of permeabilized rat vastus intermedius muscle fibres following high-fat, high-sucrose diet consumption

Obesity is a worldwide health concern associated with impaired physical function. It is not clear if contractile protein dysfunction contributes to the impairment of muscle function observed with obesity. The purpose of this study was to examine if diet-induced obesity affects contractile function of chemically permeabilized vastus intermedius fibres of male Sprague Dawley rats expressing fast myosin heavy chain (MHC) IIa or slow MHC I. Rats consumed either a high-fat, high-sucrose (HFHS) diet or a standard (CHOW) diet beginning as either weanlings (7-week duration: WEAN7 cohort, or 14-week duration: WEAN14 cohort) or young adults (12-week duration: ADULT12 cohort, 24-week duration: ADULT24 cohort). HFHS-fed rats had higher (P<0.05) whole-body adiposity (derived from dual-energy X-ray absorptiometry) than CHOW-fed rats in all cohorts. Relative to CHOW diet groups, the HFHS diet was associated with impaired force production in a) MHC I fibres in the ADULT24 cohort, and b) MHC IIa fibres in the ADULT12 and ADULT24 cohorts combined. However, the HFHS diet did not significantly affect the Ca2+-sensitivity of force production, unloaded shortening velocity, or ratio of active force to active stiffness in any cohort. We conclude that diet-induced obesity can impair force output of permeabilized muscle fibres of adult rats. Novelty Bullets: • We assessed contractile function of permeabilized skeletal muscle fibres in a rat model of diet-induced obesity. • The high-fat, high-sucrose diet was associated with impaired force output of fibres expressing MHC I or MHC IIa in some cohorts of rats. • Other measures of contractile function were not significantly affected by diet.

Evaluation of vitamin A status on myogenic gene expression and muscle fiber characteristics

A randomized complete block design experiment with 30 yearling crossbred steers (average BW = 436.3 ± 39.8 kg) fed a steam-flaked corn-based diet was used to evaluate the effects dietary vitamin A (Rovimix A 1000; DSM Nutritional Products Ltd., Sisseln, SUI) supplementation on myogenic gene expression and skeletal muscle fiber characteristics during the finishing phase. Steers were blocked by BW (n = 5 blocks; 6 steers/block), randomly assigned to pens (n = 2 steers/pen), and one of the following treatments: no added vitamin A (0 IU; 0.0 IU/kg of dietary dry matter intake of additional vitamin A), vitamin A supplemented at the estimated requirement (2,200 IU; 2,200 IU/kg of dietary dry matter (DM) of additional vitamin A), and vitamin A supplemented at 5× the estimated requirement (11,000 IU; 11,000 IU/kg of dietary DM of additional vitamin A). After all treatments underwent a 91-d vitamin A depletion period, additional vitamin A was top-dressed at feeding via a ground corn carrier. Blood, longissimus muscle, and liver biopsy samples were obtained on days 0, 28, 56, 84, and 112. Biopsy samples were used for immunohistochemical and mRNA analysis. Sera and liver samples were used to monitor circulating vitamin A and true vitamin A status of the cattle. Expression for myosin heavy chain (MHC)-I diminished and rebounded (P = 0.04) over time. The intermediate fiber type, MHC-IIA, had a similar pattern of expression (P = 0.01) to that of MHC-I. On day 84, C/EBPβ expression was also the greatest (P = 0.03). The pattern of PPARγ (P &lt; 0.01) and PPARδ (P &lt; 0.01) expression seemed to mimic that of MHC-I expression, increasing from days 84 to 112. Distribution of MHC-IIA demonstrated a change over time (P = 0.02). Muscle fiber cross-sectional area increased by day (P &lt; 0.01) for each MHC with the notable increase between days 0 and 56. Total nuclei density decreased (P = 0.02) over time. Cells positive for only Myf5 increased (P &lt; 0.01) in density early in the feeding period, then declined, indicating that satellite cells were fusing into fibers. The dual-positive (PAX7+Myf5) nuclei also peaked (P &lt; 0.01) around day 56 then declined. These data indicated that gene expression associated with oxidative proteins may be independent of vitamin A status in yearling cattle.

Influenza Infection has Fiber Type-Specific Effects on Cellular and Molecular Skeletal Muscle Function in Aged Mice

Skeletal muscle myopathies represent a common non-pulmonary manifestation of influenza infection, leading to reduced physical function and hospitalization in older adults. However, underlying mechanisms remain poorly understood. Our study examined the effects of influenza virus A pulmonary infection on contractile function at the cellular (single fiber) and molecular (myosin-actin interactions and myofilament properties) levels in soleus and extensor digitorum longus muscles of aged (20 months) C57BL/6 male mice that were healthy or flu-infected for 7 (7-days post-infection; 7-DPI) or 12 days (12-DPI). Cross-sectional area (CSA) of myosin heavy chain (MHC) IIA and IIB fibers was reduced at 12-DPI relative to 7-DPI and healthy. Maximal isometric force in MHC IIA fibers was also reduced at 12-DPI relative to 7-DPI and healthy, resulting in no change in specific force (maximal isometric force divided by CSA). In contrast, MHC IIB fibers produced greater isometric force and specific force at 7-DPI compared to 12-DPI or healthy. The increased specific force in MHC IIB fibers was likely due to greater myofilament lattice stiffness and/or an increased number or stiffness of strongly bound myosin-actin cross-bridges. At the molecular level, cross-bridge kinetics were slower in MHC IIA fibers with infection, while changes in MHC IIB fibers were largely absent. In both fiber types, greater myofilament lattice stiffness was positively related to specific force. This study provides novel evidence that cellular and molecular contractile function is impacted by influenza infection in a fiber type-specific manner, suggesting potential molecular mechanisms to help explain the impact of flu-induced myopathies.

Sex and fiber type independently influence AMPK, TBC1D1, and TBC1D4 at rest and during recovery from high-intensity exercise in humans

Women and men present different metabolic responses to exercise, yet whether this phenomenon results from differences in fiber type (FT) composition or other sex-specific factors remains unclear. Therefore, our aim was to examine the effects of sex and FT independently on AMP-activated protein kinase (AMPK), acetyl-CoA carboxylase (ACC), Tre-2/BUB2/CDC1 domain family (TBC1D)1, and TBC1D4 in response to acute exercise. Segregated pools of myosin heavy chain (MHC) I and MHC IIa fibers were prepared from vastus lateralis biopsies of young trained men and women at rest and during recovery (0 min, 45 min, 90 min, or 180 min) from high-intensity interval exercise (6 × 1.5 min at 95% maximum oxygen uptake). In resting MHC I vs. IIa fibers, AMPKα2, AMPKγ3, and TBC1D1 were higher and TBC1D4 expression was lower in both sexes, along with higher phospho (p)-TBC1D1Ser660 and lower p-TBC1D4Thr642. Women expressed higher ACC than men in MHC IIa fibers and higher AMPKβ1, AMPKβ2, TBC1D1, and TBC1D4 in both FTs. Immediately after exercise, p-AMPKαThr172 increased only in MHC IIa fibers, whereas p-ACCSer221 increased in both FTs, with no change in p-TBC1D1Ser660 or p-TBC1D4Thr642. During recovery, delayed responses were observed for p-AMPKαThr172 in MHC I (45 min), p-TBC1D4Thr642 in both FTs (45 min), and p-TBC1D1Ser660 (180 min). FT-specific phosphorylation responses to exercise were similar between men and women. Data indicate that sex and FT independently influence expression of AMPK and its substrates. Thus failing to account for sex or FT may reduce accuracy and precision of metabolic protein measurements and conceal key findings. NEW & NOTEWORTHY This investigation is the first to compare muscle fiber type (FT)-specific analysis of proteins between the sexes, providing comprehensive data on AMP-activated protein kinase (AMPK), acetyl-CoA carboxylase (ACC), Tre-2/BUB2/CDC1 domain family (TBC1D)1, and TBC1D4 before and in the hours following high-intensity interval exercise (HIIT). Expression and phosphorylation of specific AMPK isoforms, ACC, TBC1D1, and TBC1D4 were shown to be FT dependent, sex dependent, or both, and TBC1D1 showed an unexpected delay in FT-dependent phosphorylation in the time period following HIIT.

Single-muscle fiber contractile properties in lifelong aerobic exercising women

The purpose of this study was to examine the effects of lifelong aerobic exercise on single-muscle fiber performance in trained women (LLE; n = 7, 72 ± 2 yr) by comparing them to old healthy nonexercisers (OH; n = 10, 75 ± 1 yr) and young exercisers (YE; n = 10, 25 ± 1 yr). On average, LLE had exercised ~5 days/wk for ~7 h/wk over the past 48 ± 2 yr. Each subject had a vastus lateralis muscle biopsy to examine myosin heavy chain (MHC) I and IIa single-muscle fiber size and function (strength, speed, power). MHC I fiber size was similar across all three cohorts (YE = 5,178 ± 157, LLE = 4,983 ± 184, OH = 4,902 ± 159 µm2). MHC IIa fiber size decreased ( P < 0.05) 36% with aging (YE = 4,719 ± 164 vs. OH = 3,031 ± 153 µm2), with LLE showing a similar 31% reduction (3,253 ± 189 µm2). LLE had 17% more powerful ( P < 0.05) MHC I fibers and offset the 18% decline in MHC IIa fiber power observed with aging ( P < 0.05). The LLE contractile power was driven by greater strength (+11%, P = 0.056) in MHC I fibers and elevated contractile speed (+12%, P < 0.05) in MHC IIa fibers. These data indicate that lifelong exercise did not benefit MHC I or IIa muscle fiber size. However, LLE had contractile function adaptations that enhanced MHC I fiber power and preserved MHC IIa fiber power through different contractile mechanisms (strength vs. speed). The single-muscle fiber contractile properties observed with lifelong aerobic exercise are unique and provide new insights into aging skeletal muscle plasticity in women at the myocellular level. NEW & NOTEWORTHY This is the first investigation to examine the effects of lifelong exercise on single-muscle fiber physiology in women. Nearly 50 yr of moderate to vigorous aerobic exercise training resulted in enhanced slow-twitch fiber power primarily by increasing force production, whereas fast-twitch fiber power was preserved primarily by increasing contractile speed. These unique muscle fiber power profiles helped offset the effects of fast-twitch fiber atrophy and highlight the benefits of lifelong aerobic exercise for myocellular health.

Extraordinary Fast-Twitch Fiber Abundance in Elite Weightlifters

ABSTRACTHuman skeletal muscle fibers exist across a continuum of slow → fast-twitch. The amount of each fiber type (FT) influences muscle performance but remains largely unexplored in elite athletes, particularly from strength/power sports. To address this nescience, vastus lateralis (VL) biopsies were performed on World/Olympic (female, n=6, “WCF”) and National-caliber (female, n=9, “NCF”; and male, n=6, “NCM”) American weightlifters. Participant accolades included 3 Olympic Games, 19 World Championships, 25 National records, and >170 National/International medals. Samples were analyzed for myosin heavy chain (MHC) content via SDS-PAGE using two distinct techniques: single fiber (SF) distribution (%) and homogenate (HG) composition. These athletes displayed the highest MHC IIa concentrations ever reported in healthy VL (23±9% I, 5±3% I/IIa, 67±13% IIa, and 6±10% IIa/IIx), with WCF expressing a notable 71±17% (NCF=67±8%, NCM=63±16%). The heavyweights accounted for 91% of the MHC IIa/IIx fibers. When compared to SF, HG overestimated MHC I (23±9 vs. 31±9%) and IIx (0±0 vs. 3±6%) by misclassifying I/IIa fibers as I and IIa/IIx fibers as IIx. These findings suggest athlete caliber (World vs. National), training experience, and body mass determine FT% more than sex and refutes the common pronouncement that women possess more slow and fewer fast-twitch fibers than men. Our results also show the abundance of pure MHC IIa and rarity of IIx in elite strength/power-trained athletes, indicate a potential link between MHC IIa/IIx frequency and body mass, and question the fidelity of HG as a measure of FT% distribution. The extreme fast-twitch abundance partially explains how elite weightlifters generate high forces in rapid time-frames. These data highlight the need for more cellular and molecular muscle research on elite anaerobic athletes.