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Resistance exercises eliciting eccentric overload (EO) are considered to strongly promote muscular hypertrophy and broad neuromuscular adaptations but typically require specialized equipment. The aims of these experiments were to assess whether EO is achieved during common high-speed stretch-shortening cycle (SSC) exercises (rebound bench press throw [RBPT] and squat jump [SJ]), and to test the effect of the external load on the EO achieved. Twenty-nine under 18 handball players and fifteen physically active males (24.9 ± 3.2 years) took part in the experiments. Testing consisted of a single set of 6 repetitions with light (25%-30% 1-RM), moderate (50% 1-RM), and heavy (70%-75% 1-RM) loads. Eccentric and concentric force near the zero-velocity point (50-200 ms) as well as eccentric-concentric force ratio (EO; %) were calculated. In RBPT, higher EO values were found at 50% 1-RM than 70% 1-RM in the time interval 50 ms before and after the zero-velocity point. Higher EO values were also found at 50% 1-RM than both 30% 1-RM and 70% 1-RM 100 ms before and after the zero-velocity point. For the SJ, higher EO values were found at 50% 1-RM and 75% 1-RM than 25% 1-RM 100 ms before and after the zero-velocity point. Higher EO values were found at 50% 1-RM than 25% 1-RM 200 ms before and after the zero-velocity point. However, the higher EO values in the SJ were found far from the zero-velocity point. High-speed SSC resistance training provides similar EO to other methods and thus should promote muscle hypertrophy and other neuromuscular adaptations.

This systematic review and meta-analysis determined resistance training (RT) load effects on various muscle hypertrophy, strength, and neuromuscular performance task [e.g., countermovement jump (CMJ)] outcomes. Relevent studies comparing higher-load [60% 1-repetition maximum (RM) or15-RM] and lower-load (≤60% 1-RM or ≥ 15-RM) RT were identified, with 45 studies (from 4713 total) included in the meta-analysis. Higher- and lower-load RT induced similar muscle hypertrophy at the whole-body (lean/fat-free mass; [ES (95% CI) = 0.05 (-0.20 to 0.29)

Background Most papers examining the lateral abdominal muscles (LAMs) and low back pain (LBP) are cross-sectional, with groups of participants being divided into a control and an LBP group. We hypothesized that morphological measurements of the LAMs in adolescent soccer players may predict future LBP incidence. The aim of this study was to examine the associations between the morphology of LAMs and LBP incidence rate among adolescent soccer players. Methods Ninety-seven adolescent male soccer players with no LBP at baseline were recruited into the prospective cohort study. The thickness of the LAMs was measured at baseline by ultrasound imaging in a supine rest position. Nine cases of LBP occurred during the follow-up 6 month observation. Results An obliquus internus (OI) asymmetry was related to increasing LBP risk (odds ratio = 19.99; 95%CI: 2.4–167.9). Spearman correlation also showed a linear relationship between OI asymmetry value and duration of LBP ( R = 0.75, p = 0.02). An OI side-to-side difference greater than 1.25 mm suggests possible LBP incidence in the 6-month observation among adolescent soccer players. Conclusion The morphological changes of the OI may be related to LBP’s incidence in adolescent soccer players. The presence of OI asymmetry increases the odds of LBP by at least 2.4times. Hypertrophy of the OI on one side of the body may contribute to trunk muscle imbalance.

The optimal pattern of sedentarism displacement and mechanisms underlying its health effects are poorly understood. Therefore, the aim of this study was to quantify muscle-tendon adaptation in response to two different sedentarism displacement interventions and relate any adaptations to functional outcomes. Thirty-four older women (73±5yrs) underwent skeletal muscle-tendon size and functional assessments. Participants were randomly allocated to: Sedentary behaviour fragmentation (SBF), Light intensity physical activity (LIPA), or Control groups. Measures were taken at weeks 0 and 8. Gait speed significantly increased (p=0.003), in both experimental groups (SBF: 0.06 ± 0.08m/s, 6±10%, LIPA: 0.06 ± 0.07m/s, 6±6%), but not control (-0.02 ± 0.12m/s, -2±9%). Accordingly, the relative change in Vastus Lateralis muscle volume, accounted for 30% (p=0.027), and 45% (p=0.0006) of the explained variance in the relative change in gait speed, for SBF and LIPA respectively. Gastrocnemius Medialis fascicle length changes were positively associated with gait speed changes, following LIPA exclusively (R2= 0.50, p=0.009). This is the first study to show SBF and LIPA are adequate loading in older women, with related muscle adaptation and clinically relevant gait speed improvements. Such adaptations appear similar irrespective of whether sedentarism displacement is prescribed in a single bout (LIPA) or in frequent micro-bouts (SBF).

Resistance training is a commonly used strategy for improving both athletic performance and general health. While the contribution of resistance training intensity and volume to muscle strength and hypertrophy have been extensively investigated, training frequency only recently received sufficient attention, especially in older adults. A meta-regression was conducted to compare muscle strength and hypertrophic adaptations to resistance training programmes performed with different training frequencies in adults over 60 years of age. The systematic literature search identified 14 articles for meta-regression. For each outcome, an effect size (ES) was calculated as the pre-test-post-test change, divided by the pooled pre-test standard deviation (SD). Random-effects meta-regressions for multilevel data structures, using study as the clustering variable, were performed using package metafor in R. Maximal strength shows a significant effect of frequency (p = 0.001), with an increase in effect size of 0.14 for every day increase in frequency (CI: 0.08, 0.21). For muscle hypertrophy, no significant effect of frequency was found (p = 0.67). Considering that muscle hypertrophy was not affected, while maximum strength was only slightly improved with additional training days, it seems unlikely that more than two weekly resistance training sessions would provide any further benefits for older adults.

Menopause transition may impair muscle function, decreasing exercise tolerance. The torque-duration relationship (hyperbolic curve) forms a practical framework within which exercise tolerance may be explored. In this regard, resistance training (RT) increases the curvature constant of this relationship (W'). Muscle hypertrophy and strength gains have been suggested as possible mediators of RT-induced improvement in W', however, it is unclear what the main mediator is. Higher-volume RT (HV-RT), beyond that recommended by RT-guidelines (i.e. three sets per exercise), may promote greater hypertrophy, but not higher strength gains. Hence, this study aimed to investigate whether greater hypertrophy in HV-RT maximises W' gain when compared to LVRT in postmenopausal women (PW). Fifty-eight PW were randomised to the control group (CTRL), HV-RT (six sets per exercise) or LV-RT (three sets per exercise). They underwent a 12-week RT program and were assessed for W', thigh lean body mass (TLBM) and maximal isometric voluntary contraction (MIVC). The TLBM gain was higher (

Macrophages are one of the top players when considering immune cells involved with tissue homeostasis. Recently, increasing evidence has demonstrated that macrophages could also present two major subsets during tissue healing: proliferative macrophages (M1-like), which are responsible for increasing myogenic cell proliferation, and restorative macrophages (M2-like), which are involved in the end of the mature muscle myogenesis. The participation and characterization of these macrophage subsets are critical during myogenesis to understand the inflammatory role of macrophages during muscle recovery and to create supportive strategies that can improve mass muscle maintenance. Indeed, most of our knowledge about macrophage subsets comes from skeletal muscle damage protocols, and we still do not know how these subsets can contribute to skeletal muscle adaptation. Thus, this narrative review aims to collect and discuss studies demonstrating the involvement of different macrophage subsets during the skeletal muscle damage/regeneration process, showcasing an essential role of these macrophage subsets during muscle adaptation induced by acute and chronic exercise programs.

Introduction: Creatine supplementation has been widely used to improve athletic performance. Furthermore, recent discoveries indicate that this supplement has an important therapeutic effect in many diseases characterized by atrophy, muscle weakness and metabolic diseases (muscle, bone, lung and brain). Aim: to analyze through a literature review the effects of chronic use of creatine on kidney function. Methodology: this is an argumentative literature review, based on quantitative descriptive-exploratory technique. The researched samples came from national and international journals, published between the years 2011 and 2021. The journals were acquired from the Scielo and PubMed electronic databases. Results: Several studies, mostly executed on men performing resistance training three times a week, were supplemented with creatine monohydrate at different dosis for 7 days and compared with matched controls supplemented with dextrosol. Several tests were performed, in some of them, blood and urine samples were collected before and 30 days after supplementation in which numerous biochemical parameters and renal function were evaluated. Creatine monohydrate supplementation did not produce adverse events and, as expected, promoted an increase in performance and body weight. Conclusions: the present paper did not find evidence in the literature to support that creatine may represent a risk to the health of healthy men. However, cases in the literature suggest that creatine can harm kidney function with indiscriminate use, so as not to bring health risks, it is recommended that healthy individuals who regularly use this supplement do not exceed 5g/day. Introducción: la suplementación con creatina se ha utilizado ampliamente para mejorar el rendimiento deportivo. Además, descubrimientos recientes indican que este suplemento tiene un importante efecto terapéutico en muchas enfermedades caracterizadas por atrofia, debilidad muscular y enfermedades metabólicas (musculares, óseas, pulmonares y cerebrales). Objetivo: analizar a través de una revisión de la literatura los efectos del uso crónico de creatina sobre la función renal. Metodología: se trata de una revisión de la literatura argumentativa, basada en la técnica cuantitativa descriptiva-exploratoria. Las muestras investigadas provienen de revistas nacionales e internacionales, publicadas entre los años 2011 y 2021. Las revistas fueron adquiridas de las bases de datos electrónicas Scielo y PubMed. Resultados: varios estudios, en su mayoría realizados en hombres que realizaban entrenamiento de resistencia tres veces por semana, se suplementaron con monohidrato de creatina a diferentes concentraciones durante 7 días y se compararon con controles emparejados suplementados con dextrosol. Se realizaron varias pruebas, en algunas de ellas se recolectaron muestras de sangre y orina antes y 30 días después de la suplementación en las que se evaluaron numerosos parámetros bioquímicos y función renal. La suplementación con monohidrato de creatina no causó eventos adversos y, como se esperaba, promovió un aumento en el rendimiento y el peso corporal. Conclusiones: el presente estudio no encontró evidencia en la literatura que sustente que la creatina pueda representar un riesgo para la salud de hombres sanos. Sin embargo, los casos en la literatura sugieren que la creatina puede dañar la función renal con un uso indiscriminado, por lo que para no traer riesgos para la salud, se recomienda que las personas sanas que usen regularmente este suplemento no superen los 5g / día. Introdução: a suplementação da creatina tem sido amplamente utilizada para melhorar o desempenho atlético. Além disso, descobertas recentes indicam que este suplemento tem um importante efeito terapêutico em muitas doenças caracterizadas por atrofia, fraqueza muscular e doenças metabólicas (músculos, ossos, pulmão e cérebro). Objetivo: analisar por meio de uma revisão da literatura os efeitos do uso crônico da creatina sobre a função renal. Metodologia: trata-se de uma revisão argumentativa da literatura, de base técnica quantitativa descritivo-exploratória. As amostras pesquisadas foram advindas de periódicos, nacionais e internacionais, publicados entre os anos de 2011 e 2021. Os periódicos foram adquiridos dos bancos de dados eletrônicos Scielo e PubMed. Resultados: Diversos estudos, a maioria realizado em homenss realizando treinamento resistido três vezes por semana foram suplementados com monohidrato de creatina em diferentes concentrações, por 7 dias e comparados com controles pareados suplementados com dextrosol. Vários testes foram realizados, em alguns deles, amostras de sangue e urina foram coletadas antes e 30 dias após a suplementação no qual inúmeros parâmetros bioquímicos e função renal foram avaliados. A suplementação de creatina monohidratada não causou eventos adversos e, como esperado, promoveu aumento do desempenho e do peso corporal. Conclusões: o presente estudo não encontrou na literatura, evidências que sustentem que a creatina pode representar um risco para a saúde de homens saudáveis. No entanto, casos na literatura sugerem que a creatina pode prejudicar a função renal com o uso indiscriminado, para não trazer riscos à saúde, recomenda-se que indivíduos saudáveis que fazem uso regular desse suplemento não ultrapassem 5g / dia.

This paper aimed to compare the effect of drop-set (DS) and rest-pause (RP) systems versus traditional resistance training (TRT) with equalized total training volume on maximum dynamic strength (1RM) and thigh muscle thickness (MT). Twenty-eight resistance-trained males were randomly assigned to either RP (n = 10), DS (n = 9) or TRT (n = 9) protocols performed twice a week for 8 weeks. 1RM and MT of the proximal, middle and distal portions of the lateral thigh were assessed at baseline and post-intervention. A significant time × group interaction was observed for 1RM (P = 0.001) in the barbell back squat after 8-weeks. Post hoc comparisons revealed that RP promoted higher 1RM than TRT (P = 0.001); no statistical differences in strength were observed between the other conditions. A significant main effect of time was revealed for MT at the proximal (P = 0.0001) and middle (P = 0.0001) aspects of the lateral thigh for all training groups; however, the distal portion did not show a time effect (P = 0.190). There were no between-group interactions for MT. Our findings suggest that RP promotes slightly superior strength-related improvements compared with TRT, but hypertrophic adaptations are similar between conditions. Novelty: Rest-pause elicited a slightly superior benefit for strength adaptations compared with traditional resistance training. Resistance training systems do not promote superior hypertrophic adaptations when total training volume is equalized. Muscle thickness in distal portion of thigh is similar to baseline. Although modest, effect sizes tended to favor rest-pause.

The skeletal muscle hypertrophic response to resistance exercise training (RT) is highly variable across individuals. The molecular underpinnings of this heterogeneity are unclear. This study investigated transcriptional networks linked to RT-induced muscle hypertrophy, classified as 1) predictive of hypertrophy, 2) responsive to RT independent of muscle hypertrophy, or 3) plastic with hypertrophy. Older adults ( n = 31, 18 F/13 M, 70 ± 4 yr) underwent 14-wk RT (3 days/wk, alternating high-low-high intensity). Muscle hypertrophy was assessed by pre- to post-RT change in mid-thigh muscle cross-sectional area (CSA) [computed tomography (CT), primary outcome] and thigh lean mass [dual-energy X-ray absorptiometry (DXA), secondary outcome]. Transcriptome-wide poly-A RNA-seq was performed on vastus lateralis tissue collected pre- ( n = 31) and post-RT ( n = 22). Prediction networks (using only baseline RNA-seq) were identified by weighted gene correlation network analysis (WGCNA). To identify Plasticity networks, WGCNA change indices for paired samples were calculated and correlated to changes in muscle size outcomes. Pathway-level information extractor (PLIER) was applied to identify Response networks and link genes to biological annotation. Prediction networks ( n = 6) confirmed transcripts previously connected to resistance/aerobic training adaptations in the MetaMEx database while revealing novel member genes that should fuel future research to understand the influence of baseline muscle gene expression on hypertrophy. Response networks ( n = 6) indicated RT-induced increase in aerobic metabolism and reduced expression of genes associated with spliceosome biology and type-I myofibers. A single exploratory Plasticity network was identified. Findings support that interindividual differences in baseline gene expression may contribute more than RT-induced changes in gene networks to muscle hypertrophic response heterogeneity. Code/Data: https://github.com/kallavin/MASTERS_manuscript/tree/master .

Made available in DSpace on 2021-06-25T10:15:01Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-03-01 The purpose was to compare the effects of protein (whey protein) and carbohydrate supplementation and protein alone both combined with resistance training on muscle strength, muscle mass and total training volume progression in untrained young men. Resistance training was performed using the leg press and knee extension until concentric failure (8-12 repetition maximum), three times a week for eight weeks. Muscle strength and muscle cross-sectional area were assessed before and after training. Total training volume progression was calculated considering the first and eighth week. Seventeen men completed the study (protein and carbohydrate, n=9, age 23.44 ± 4.56 years, weight: 62.13±6.17 kg, height: 1.75±0.02 m, body mass index: 20.29±2.08 kg/m 2; protein, n=8, age 24.63±2.39 years, weight: 69.01±5.57 kg, height: 1.77±0.07 m; body mass index: 21.64±1.05 kg/m 2. Both protocols showed similar increases in muscle strength (effect size: protein and carbohydrate=1.28; protein=0.97; p0.05). Protein and carbohydrate supplementation combined with resistance training does not induce greater gains in muscle strength, hypertrophy and total training volume compared to resistance training combined with protein alone in untrained individuals. Department of Food and Nutrition School of Pharmaceutical Science Sao Paulo State University (UNESP) MUSCULAB Laboratory of Neuromuscular Adaptations to Resistance Training Department of Physical Education Federal University of Sao Carlos University of Sao Paulo School of Physical Education and Sport of Ribeirao Preto Department of Food and Nutrition School of Pharmaceutical Science Sao Paulo State University (UNESP)

Fucoidan is a sulfated polysaccharide found in a range of brown algae species. Growing evidence supports the long‐term supplementation of fucoidan as an ergogenic aid to improve skeletal muscle performance. The aim of this study was to investigate the effect of fucoidan on the skeletal muscle of mice. Male BL/6 mice (N = 8–10) were administered a novel fucoidan blend (FUC, 400 mg/kg/day) or vehicle (CON) for 4 weeks. Treatment and control experimental groups were further separated into exercise (CON+EX, FUC+EX) or no‐exercise (CON, FUC) groups, where exercised groups performed 30 min of treadmill training three times per week. At the completion of the 4‐week treatment period, there was a significant increase in cross‐sectional area (CSA) of muscle fibers in fucoidan‐treated extensor digitorum longus (EDL) and soleus fibers, which was accompanied by a significant increase in tibialis anterior (TA) muscle force production in fucoidan‐treated groups. There were no significant changes in grip strength or treadmill time to fatigue, nor was there an effect of fucoidan or exercise on mass of TA, EDL, or soleus muscles. In gastrocnemius muscles, there was no change in mRNA expression of mitochondrial biogenesis markers PGC‐1α and Nrf‐2 in any experimental groups; however, there was a significant effect of fucoidan supplementation on myosin heavy chain (MHC)‐2x, but not MHC‐2a, mRNA expression. Overall, fucoidan increased muscle size and strength after 4 weeks of supplementation in both exercised and no‐exercised mice suggesting an important influence of fucoidan on skeletal muscle physiology., Fucoidan is a complex polysaccharide found in brown seaweed. While fucoidan has many bioactive properties, here we show that in mice, 4 weeks of supplementation with a novel blend of fucoidan species increased both size and strength of skeletal muscle.

This study aimed to analyze the effect of resistance training (RT) performed until volitional failure with low, moderate, and high loads on muscle hypertrophy and muscle strength in healthy adults and to assess the possible participant-, design-, and training-related covariates that may affect the adaptations.Using Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines, MEDLINE, CINAHL, EMBASE, SPORTDiscus, and Web of Science databases were searched. Including only studies that performed sets to volitional failure, the effects of low- (15 repetitions maximum (RM)), moderate- (9-15 RM), and high-load (≤8 RM) RTs were examined in healthy adults. Network meta-analysis was undertaken to calculate the standardized mean difference (SMD) between RT loads in overall and subgroup analyses involving studies deemed of high quality. Associations between participant-, design-, and training-related covariates with SMD were assessed by univariate and multivariate network meta-regression analyses.Twenty-eight studies involving 747 healthy adults were included. Although no differences in muscle hypertrophy between RT loads were found in overall (P = 0.113-0.469) or subgroup analysis (P = 0.871-0.995), greater effects were observed in untrained participants (P = 0.033) and participants with some training background who undertook more RT sessions (P = 0.031-0.045). Muscle strength improvement was superior for both high-load and moderate-load compared with low-load RT in overall and subgroup analysis (SMD, 0.60-0.63 and 0.34-0.35, respectively; P0.001-0.003), with a nonsignificant but superior effect for high compared with moderate load (SMD, 0.26-0.28, P = 0.068).Although muscle hypertrophy improvements seem to be load independent, increases in muscle strength are superior in high-load RT programs. Untrained participants exhibit greater muscle hypertrophy, whereas undertaking more RT sessions provides superior gains in those with previous training experience.

Cannabidiol (CBD) has proven clinical benefits in the treatment of seizures, inflammation, and pain. The recent legalization of CBD in many countries has caused increased interest in the drug as an over-the-counter treatment for athletes looking to improve recovery. However, no data on the effects of CBD on the adaptive response to exercise in muscle are available. To address this gap, we eccentrically loaded the tibialis anterior muscle of 14 rats, injected them with a vehicle (n = 7) or 100 mg/kg CBD (n = 7), and measured markers of injury, inflammation, anabolic signaling, and autophagy 18 hr later. Pro-inflammatory signaling through nuclear factor kappa B (NF-kB) (Ser536) increased with loading in both groups; however, the effect was significantly greater (36%) in the vehicle group (p p = .66). The ribosomal protein S6 phosphorylation (240/244) increased with stimulation (p p = .09). The ubiquitin-binding protein p62 levels were not modulated by stimulation (p = .6), but they were 46% greater in the CBD compared with the vehicle group (p = .01). Although liver weight did not differ between the groups (p = .99) and levels of proteins associated with stress were similar, we did observe serious side effects in one animal. In conclusion, an acute dose of CBD decreased pro-inflammatory signaling in the tibialis anterior without blunting the anabolic response to exercise in rats. Future research should determine whether these effects translate to improved recovery without altering adaptation in humans.

Muscle differentiation is a multifaceted and tightly controlled process required for the formation of skeletal muscle fibers. Satellite cells are the direct cellular contributors to muscle repair in injuries or disorders. Here, we show that autotaxin (Atx) expression and activity is required for satellite cell differentiation. Conditional ablation of Atx or its pharmacological inhibition impairs muscle repair. Mechanistically, we identify LPAR1 as the key receptor in Atx-LPA signaling. Myogenic gene array and pathway analysis identified that Atx-LPA signaling activates ribosomal protein S6 kinase (S6K), an mTOR-dependent master regulator of muscle cell growth via LPAR1. Furthermore, Atx transgenic mice show muscle hypertrophic effects and accelerated regeneration. Intramuscular injections of Atx/LPA show muscle hypertrophy. In addition, the regulatory effects of Atx on differentiation are conserved in human myoblasts. This study identifies Atx as a critical master regulator in murine and human muscles, identifying a promising extracellular ligand in muscle formation, regeneration, and hypertrophy.

Evidence from clinical trials and observational studies suggests that both progressive resistance exercise training (PRT) and metformin delay a variety of age-related morbidities. Previously, we completed a clinical trial testing the effects of 14 weeks of PRT + metformin (metPRT) compared to PRT with placebo (plaPRT) on muscle hypertrophy in older adults. We found that metformin blunted PRT-induced muscle hypertrophic response. To understand potential mechanisms underlying the inhibitory effect of metformin on PRT, we analyzed the muscle transcriptome in 23 metPRT and 24 plaPRT participants. PRT significantly increased expression of genes involved in extracellular matrix remodeling pathways, and downregulated RNA processing pathways in both groups, however, metformin attenuated the number of differentially expressed genes within these pathways compared to plaPRT. Pathway analysis showed that genes unique to metPRT modulated aging-relevant pathways, such as cellular senescence and autophagy. Differentially expressed genes from baseline biopsies in older adults compared to resting muscle from young volunteers were reduced following PRT in plaPRT and were further reduced in metPRT. We suggest that although metformin may blunt pathways induced by PRT to promote muscle hypertrophy, adjunctive metformin during PRT may have beneficial effects on aging-associated pathways in muscle from older adults.

MiR-33a is found as a regulator of cell proliferation in many cancer cells. However, it remains unknown if and how miR-33a plays a role in myoblast proliferation. To investigate the effect of miR-33a on myoblast proliferation, miR-33a mimic or inhibitor was co-administered with or without insulin-like growth factor 1 (IGF1) to simulation myoblasts. Our study showed that up-regulation of miR-33a impaired myoblast proliferation, while down-regulation of miR-33a enhanced myoblast proliferation. Mechanistically, we examined that miR-33a can inhibit the transcription of IGF1, follistatin (FST) and cyclin D1 (CCND1) by targeting their 3′UTR region in both HEK293T cells and duck myoblasts. Moreover, up-regulation of miR-33a decreased and its down-regulation increased the mRNA expression of PI3K, Akt, mTOR and S6K. Importantly, the decreased PI3K, Akt, mTOR and S6K expression by miR-33a mimics was abrogated by co-administered with IGF1. Altogether, our results demonstrated that miR-33a may directly target IGF1, FST and CCND1 to inhibit myoblast proliferation via PI3K/Akt/mTOR signaling pathway. In conclusion, miR-33a is a potential negative regulator of myoblast proliferation and by modulating its expression could promote the early development of skeletal muscle.

This study aimed to analyse changes in sprint performance, muscle volumes (MVs) and sprint mechanical parameters (SMPs) in national-level sprinters performing a 5-month indoor sprint-based training...

SUMMARY Mechanical signals, such as those evoked by maximal-intensity contractions (MICs), can induce an increase in muscle mass. Rapamycin-sensitive signaling events are widely implicated in the regulation of this process; however, recent studies indicate that rapamycin-insensitive signaling events are also involved. Thus, to identify these events, we generate a map of the MIC-regulated and rapamycin-sensitive phosphoproteome. In total, we quantify more than 10,000 unique phosphorylation sites and find that more than 2,000 of these sites are significantly affected by MICs, but remarkably, only 38 of the MIC-regulated events are mediated through a rapamycin-sensitive mechanism. Further interrogation of the rapamycin-insensitive phosphorylation events identifies the S473 residue on Tripartite Motif-Containing 28 (TRIM28) as one of the most robust MIC-regulated phosphorylation sites, and extensive follow-up studies suggest that TRIM28 significantly contributes to the homeostatic regulation of muscle size and function as well as the hypertrophy that occurs in response to increased mechanical loading., Graphical Abstract, In brief By comprehensively mapping the rapamycin-sensitive and contraction regulated phosphoproteome, Steinert et al. identify TRIM28 as a non-canonical regulator of muscle size and function. This study provides a resource for further delineating the signaling components through which mechanical stimuli regulate mTOR and muscle growth.

Orientador: Maeli Dal Pai Silva Tese ( doutorado) - Universidade Estadual de Campinas, Instituto de Biologia Resumo: O pirarucu (Arapaima gigas) caracteriza-se como uma espécie promissora para a Aqüicultura, devido principalmente às suas características de rápido crescimento e rusticidade. Sua criação em regime intensivo tem obtido enorme sucesso, podendo alcançar até 10 quilos de peso corporal em apenas um ano de criação. O pirarucu é considerado hoje uma das mais importantes espécies de peixes da bacia Amazônica e, por esta razão, é primordial que se investigue os mecanismos celulares e moleculares que controlam o rápido crescimento muscular, contribuindo com novas estratégias de criação e com a manutenção da espécie. O crescimento do músculo estriado esquelético nos peixes pode ocorrer por dois mecanismos: hipertrofia e/ou hiperplasia das fibras, a partir de células satélites ou mioblastos. Esse processo é controlado por Fatores Reguladores Miogênicos (MRFs) e pelo fator de crescimento Miostatina. O objetivo do presente estudo foi avaliar as características morfológicas e o crescimento muscular hipertrófico e hiperplásico, bem como analisar a expressão gênica e protéica da MyoD, da Miogenina e da Miostatina na musculatura esquelética do pirarucu (A. gigas), em diferentes fases de crescimento. Os animais utilizados no presente estudo foram provenientes de duas pisciculturas: na primeira, foram obtidos os "alevinos" (pós-larvas; 48 g); na segunda, os animais em diferentes estágios de crescimento, divididos em quatro grupos de acordo com o peso corporal. Grupo A (50 gramas, n=7), grupo B (420 gramas, n=7), grupo C (5,5 quilogramas, n=7) e grupo D (9,1 quilogramas, n=7). As amostras musculares foram coletadas, congeladas e submetidas à coloração HE, para avaliação do padrão morfológico das fibras, e à reação para a enzima NADH-TR, para avaliar o metabolismo oxidativo das fibras. Para avaliar o padrão de crescimento hiperplásico e hipertrófico da musculatura branca, foi calculado o menor diâmetro de uma população de fibras por animal, e estas foram distribuídas em classes, na dependência do seu diâmetro. A análise da expressão gênica de MyoD, miogenina e miostatina foi feita por Reação em Cadeia da Polimerase após Transcrição Reversa (RTqPCR); para análise da expressão protéica, foi utilizado o Western Blot. A distribuição das fibras em classes de diâmetro exibiu o seguinte padrão: o grupo A mostrou a maior parte das fibras na classe 20, o grupo B, na classe 50, o grupo C, nas classes 50 e 80, e o grupo D, na classe 80. Isso indica uma alta taxa de hiperplasia das fibras nos grupos menores (A e B) e alta hipertrofia das fibras nos grupos maiores (C e D). Para a análise da expressão gênica de MyoD e miogenina no músculo vermelho e branco dos "alevinos", não houve diferença estatística; para a miostatina, houve expressão diferencial, com os maiores níveis encontrados no músculo branco em comparação com o músculo vermelho. Na avaliação da expressão de MyoD e miogenina, tanto a expressão gênica como a expressão protéica não mostraram diferença significativa. Por outro lado, a expressão gênica da miostatina foi menor no grupo A e maior nos demais, e a expressão da proteína miostatina foi maior no grupo A, diminuindo nos demais grupos. Estes resultados refletem as características de crescimento muscular da espécie e sugerem que a expressão dos MRFs e da miostatina são responsáveis pelo balanço entre a hiperplasia e a hipertrofia das fibras, contribuindo para o rápido crescimento da espécie e a manutenção das características do filé Abstract: Pirarucu (Arapaima gigas) is a promising fish species for Aquaculture programs mainly by the fast growing feature and rusticity. Their rearing under intensive conditions generated much successful results, as they reach up to 10 kilograms in just one year. Considered one of the most important fish species from Amazon basin, it is of primary interest to investigate the cellular and molecular mechanisms that control the fast muscle growth in pirarucu, providing information for new rearing strategies and species conservation. In most fish, skeletal muscle growth occurs by two mechanisms: hypertrophy and hyperplasia, from satellite cells or myoblasts. These process are under control by Myogenic Regulatory Factors (MRFs) and by the growth factor Myostatin. The animals were obtained from two pisciculture, where we got the alevin pirarucu (n=7; 48 grams weight), and the specimens at different growth stages, divided into groups according body weight. Group A (50 grams, n=7), group B (420 grams, n=7), group C (5,5 kilograms, n=7) and group D (9,1 kilograms, n=7). Muscle samples were collected, frozen and stained with HE for morphological analysis, and submitted to NADH-TR enzyme reaction for oxidative methabolism analysis. To evaluate hyperplasic and hypertrophic muscle growth, it was measured the smallest diameter from a set of fibers, which were grouped into diameter classes. Gene expression analysis of MyoD, Myogenin and Myostatin were performed by Quantitative Polimerase Chain Reaction after Reverse Transcription (RT-qPCR); protein content analysis was by Western Blot technique. Muscle fibers distribution in classes showed the following pattern: group A showed most fibers in class 20, group B, in class 50, group C, in classes 50 and 80, and group D, in class 80. This is an indicative of high fiber hiperplasia rate in groups A and B, and high hypertrophy in groups C and D. There was no statistical difference in MyoD and Myogenin genes expression in red and white muscles of pirarucu; however, Myostatin expression showed high levels in white muscle compared to red muscle. Evaluating MyoD and Myogenin expression in white muscle of pirarucu at different growth stages, both gene and protein levels were similar. Myostatin gene expression was low in group A and high in the other groups; on the other hand, Myostatin protein was high in group A and low in the other groups. These results reflect the muscle growth characteristics in pirarucu and suggest that the MRFs and Myostatin expression are controlling the balance between hyperplasic and hypertrophic mechanisms, promoting the fast rate feature of pirarucu and their fillet quality Doutorado Biologia Celular Doutor em Biologia Celular e Estrutural

Orientador: Maeli Dal Pai Silva Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia Resumo: Nos peixes, o conhecimento dos fatores que controlam o crescimento muscular e a análise das proteínas miofibrilares, é importante para entender a dinâmica do crescimento, a plasticidade e as adaptações musculares, principalmente, em espécies com grande valor comercial como a tilápia do Nilo (Oreochromis niloticus). No presente estudo, utilizou-se a tilápia do Nilo em quatro estágios: alevinos de 35 dias (0.65g ± 0.08); juvenis de 60 dias (13.67g ± 1.35); adultos de 90 dias (73.18g ± 4.70) e adultos de 190 dias (349.76g ± 34.62). Em cada estágio, fragmentos musculares foram coletados e submetidos às seguintes análises: morfométrica, para caracterizar o crescimento muscular hiperplásico e hipertrófico no músculo branco; imunohistoquímica, para analisar a expressão dos fatores de regulação miogênica MyoD e miogenina e a expressão da proteína PCNA no músculo branco; histoquímica da ATPase miofibrilar (mATPase) e à eletroforese em gel de poliacrilamida ¿ duodecil sulfato de sódio (SDS-PAGE) para observar as características da mATPase e da cadeia pesada da miosina nos músculos branco e vermelho, respectivamente. Os resultados indicaram que a expressão de MyoD e miogenina foi similar em alevinos, juvenis e adultos de 90 dias, porém, em adultos de 190 dias a expressão de miogenina foi maior do que a de MyoD. A expressão do PCNA, em cada estágio, foi mais acentuada do que MyoD e miogenina com picos no estágio de alevinos e adultos de 90 dias. A expressão de MyoD e miogenina nos estágios de alevinos, juvenis e adultos de 90 dias, mostrou que a hiperplasia e a hipertrofia ocorreram como resultado da proliferação e da diferenciação dos mioblastos. O aumento da expressão de miogenina em adultos de 190 dias, indicou que a diferenciação celular e a hipertrofia foi mais significativa nesse estágio. A análise da mATPase indicou, além da presença de fibras musculares vermelhas e brancas, fibras híbridas tanto no músculo vermelho como no músculo branco, ao longo do crescimento muscular da tilápia. A partir de alevinos, o músculo vermelho da região superficial mostrou a presença de cadeia pesada da miosina slow e o músculo branco, que forma a maior parte da massa muscular, cadeia pesada da miosina fast. Essas isoformas apresentaram massa molecular semelhante à cadeia pesada da miosina do tipo I do músculo sóleo de rato. No músculo branco, a partir dos alevinos, foi observada outra isoforma de miosina de massa molecular superior à cadeia pesada da miosina do tipo I do músculo sóleo de rato. No músculo vermelho a partir dos adultos, observou-se outra isoforma de miosina de massa molecular semelhante à cadeia pesada da miosina do tipo II do músculo sóleo de rato. A expressão das isoformas de cadeias pesadas da miosina no músculo estriado esquelético da tilápia do Nilo durante o crescimento, pode estar relacionada com a plasticidade fenotípica que ocorre durante o crescimento muscular e reflete na capacidade desses peixes de se adaptar às variações ambientais, importantes para a sobrevivência Abstract: In fish, the knowledge of factors that control the muscle growth and the myofibrillar proteins analyze is important to understand the dynamic of growth, the plasticity and the muscle adaptations, mainly, in species with high commercial valuable, as the Nile tilapia (Oreochromis niloticus). In the present study, Nile tilapia into four age stages were used: 35 day alevins (0.65g ± 0.08); 60 day juveniles (13.67g ± 1.35); 90 day adults (73.18g ± 4.70) and 190 day adults (349.76g ± 34.62). In each stage, muscle fragments were collected and submitted to the following analyzes: morphometric, to characterize the hyperplastic and hypertrophyc growth in the white muscle; immunohistochemical, to analize the myogenic regulatory factors MyoD and myogenin expression, and the PCNA protein expression in white muscle; histochemical of the myofibrillar ATPase (mATPase) and electrophoresis by sodium duodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) in the red and white muscle to observed mATPase and myosin heavy chain characteristics, respectively. The results indicate that MyoD and myogenin expression was similar in alevins, juveniles and 90 day adults, however, in 190 day adults the myogenin was higher than the MyoD expression. The PCNA expression, in each stage, was higher than MyoD and myogenin with peaks in alevins and 90 day adults. The MyoD expression in alevins, juveniles and 90 day adults, showed that the hyperplasia and hypertrophy occurred due to the results of myoblasts proliferation and differentiation. The increased of myogenin expression in 190 day adults indicated that cellular differentiation and the hypertrophy was more expressive in this stage. The mATPase showed, beyond red and white muscle fibers, hybrid fibers in both red and white muscle during growth. From alevins, the red muscle showed slow myosin heavy chain (MHCs) and the white muscle, fast myosin heavy chain (MHCf). These isoforms had a molecular mass similar to the type I myosin heavy chain (MHCI) of soleus rat muscle. In the white muscle, from alevins was observed other myosin isoform with molecular mass superior to the MHCI of soleus rat muscle. In the red muscle, in adults, was observed other myosin isoform with molecular mass similar to the type II myosin heavy chain (MHC II) of soleus rat muscle. The expression of myosin isoforms in the skeletal muscle of Nile tilapia during growth, can be related to the phenotypic plasticity that occur during muscle growth and reflects this fish capacity to adapt to changes in environmental conditions which are important for its survival Doutorado Histologia Doutor em Biologia Celular e Estrutural