Interrelated but Not Time-Aligned Response in Myogenic Regulatory Factors Demethylation and mRNA Expression after Divergent Exercise Bouts.

INTRODUCTION: DNA methylation regulates exercise-induced changes in the skeletal muscle transcriptome. However, the specificity and the time course responses in the myogenic regulatory factors DNA methylation and mRNA expression after divergent exercise modes are unknown. PURPOSE: This study aimed to compare the time course changes in DNA methylation and mRNA expression for selected myogenic regulatory factors ( MYOD1 , MYF5 , and MYF6 ) immediately after, 4 h after, and 8 h after a single bout of resistance exercise (RE), high-intensity interval exercise (HIIE), and concurrent exercise (CE). METHODS: Nine healthy but untrained males (age, 23.9 ± 2.8 yr; body mass, 70.1 ± 14.9 kg; peak oxygen uptake [V̇O 2peak ], 41.4 ± 5.2 mL·kg -1 ·min -1 ; mean ± SD) performed a counterbalanced, randomized order of RE (4 × 8-12 repetition maximum), HIIE (12 × 1 min sprints at V̇O 2peak running velocity), and CE (RE followed by HIIE). Skeletal muscle biopsies (vastus lateralis) were taken before (REST) immediately (0 h), 4 h, and 8 h after each exercise bout. RESULTS: Compared with REST, MYOD1 , MYF5 , and MYF6 , mean methylation across all CpGs analyzed was reduced after 4 and 8 h in response to all exercise protocols ( P < 0.05). Reduced levels of MYOD1 methylation were observed after HIIE and CE compared with RE ( P < 0.05). Compared with REST, all exercise bouts increased mRNA expression over time ( MYOD1 at 4 and 8 h, and MYF6 at 4 h; P < 0.05). MYF5 mRNA expression was lower after 4 h compared with 0 h and higher at 8 h compared with 4 h ( P < 0.05). CONCLUSIONS: We observed an interrelated but not time-aligned response between the exercise-induced changes in myogenic regulatory factors demethylation and mRNA expression after divergent exercise modes. Despite divergent contractile stimuli, changes in DNA methylation and mRNA expression in skeletal muscle were largely confined to the late (4-8 h) recovery period and similar between the different exercise challenges.

Differential expression patterns of myogenic regulatory factors in the postnatal longissimus dorsi muscle of Jeju Native Pig and Berkshire breeds along with their co-expression with Pax7

BACKGROUND: Myogenic regulatory factors (MRFs) such as MyoD, Myf6 and Myf5 play a vital role in the growth and development of muscles. Jeju Native Pig (JNP) is the top ranker in Korea amongst the indigenous livestock reared for meat purpose. Few studies covering transcript abundance of the MRFs and related to their co-expression with Pax7 in JNP have been conducted. Despite having better quality pork, JNP does not have a comparative growth rate with respect to western breeds. Therefore, the present study was designed with the objective to study the relative transcript levels of MRFs in the postnatal myogenesis of longissimus dorsi muscles in JNP and Berkshire breeds. RESULTS: Relative transcript levels were analyzed by qRT-PCR and blot expression analysis through Western blotting. Immunocytochemistry was performed to analyze their expressions at cellular levels. ToppCluster aided in the analysis of gene ontology of biological processes. The quantitative transcript levels of MyoD and Pax7 were significantly (P < 0.05) higher in Berkshire than in JNP. Myotube formation was observed under the co-expression of MyoD and Pax7. ToppCluster helped in the understanding of the linking of biological processes of the MRFs with the different signaling pathways. MyBPH had significantly (P < 0.05) high transcript levels during the chosen age groups in JNP than Berkshire. CONCLUSIONS: The current study can be helpful in understanding the genetic basis for myogenesis in postnatal stage. Moreover, it can act as stepping stone for the identification of marker genes related to body growth and meat quality in JNP.

Satellite cells deficiency and defective regeneration in dynamin 2-related centronuclear myopathy.

Dynamin 2 (DNM2) is a ubiquitously expressed protein involved in many functions related to trafficking and remodeling of membranes and cytoskeleton dynamics. Mutations in the DNM2 gene cause the autosomal dominant centronuclear myopathy (AD-CNM), characterized mainly by muscle weakness and central nuclei. Several defects have been identified in the KI-Dnm2R465W/+ mouse model of the disease to explain the muscle phenotype, including reduction of the satellite cell pool in muscle, but the functional consequences of this depletion have not been characterized until now. Satellite cells (SC) are the main source for muscle growth and regeneration of mature tissue. Here, we investigated muscle regeneration in the KI-Dnm2R465W/+ mouse model for AD-CNM. We found a reduced number of Pax7-positive SCs, which were also less activated after induced muscle injury. The muscles of the KI-Dnm2R465W/+ mouse regenerated more slowly and less efficiently than wild-type ones, formed fewer new myofibers, and did not recover its normal mass 15 days after injury. Altogether, our data provide evidence that the muscle regeneration is impaired in the KI-Dnm2R465W/+ mouse and contribute with one more layer to the comprehension of the disease, by identifying a new pathomechanism linked to DNM2 mutations which may be involved in the muscle-specific impact occurring in AD-CNM.

Toxoplasma gondii Impairs Myogenesis in vitro, With Changes in Myogenic Regulatory Factors, Altered Host Cell Proliferation and Secretory Profile.

Toxoplasma gondii is the causative agent of toxoplasmosis, a parasitic disease with a wide global prevalence. The parasite forms cysts in skeletal muscle cells and neurons, although no evident association with inflammatory infiltrates has been typically found. We studied the impact of T. gondii infection on the myogenic program of mouse skeletal muscle cells (SkMC). The C2C12 murine myoblast cell line was infected with T. gondii tachyzoites (ME49 strain) for 24 h followed by myogenic differentiation induction. T. gondii infection caused a general decrease in myotube differentiation, fusion and maturation, along with decreased expression of myosin heavy chain. The expression of Myogenic Regulatory Factors Myf5, MyoD, Mrf4 and myogenin was modulated by the infection. Infected cultures presented increased proliferation rates, as assessed by Ki67 immunostaining, whereas neither host cell lysis nor apoptosis were significantly augmented in infected dishes. Cytokine Bead Array indicated that IL-6 and MCP-1 were highly increased in the medium from infected cultures, whereas TGF-ß1 was consistently decreased. Inhibition of the IL-6 receptor or supplementation with recombinant TGF-ß failed to reverse the deleterious effects caused by the infection. However, conditioned medium from infected cultures inhibited myogenesis in C2C12 cells. Activation of the Wnt/ß-catenin pathway was impaired in T. gondii-infected cultures. Our data indicate that T. gondii leads SkMCs to a pro-inflammatory phenotype, leaving cells unresponsive to ß-catenin activation, and inhibition of the myogenic differentiation program. Such deregulation may suggest muscle atrophy and molecular mechanisms similar to those involved in myositis observed in human patients.

Activation of protein synthesis, regeneration, and MAPK signaling pathways following repeated bouts of eccentric cycling.

The aim of this study was to examine the activation of skeletal muscle signaling pathways related to protein synthesis and the gene expression of regeneration/degradation markers following repeated bouts of eccentric cycling. Nine untrained men (25.4 ± 1.9 yr) performed two 30-min eccentric cycling bouts (ECC1, ECC2) at 85% of maximal concentric workload, separated by 2 wk. Muscle biopsies were taken from the vastus lateralis before and 2 h after each bout. Indirect markers of muscle damage were assessed before and 24-48 h after exercise. Changes in the Akt/mammalian target of rapamycin (mTOR)/rbosomal protein S6 kinase 1 (S6K1)/ribosomal protein S6 (rpS6) and MAPK signaling pathways were measured by Western blot and changes in mRNA expression of IL-6 and IL-1ß, and myogenic regulatory factors (MRFs) were measured by real-time PCR. ECC1 induced greater increases in indirect markers of muscle damage compared with ECC2. Phosphorylation of S6K1 and rpS6 increased after both exercise bouts (P < 0.05), whereas phosphorylation of mTOR increased after ECC2 only (P = 0.03). Atrogin-1 mRNA expression decreased after ECC1 and ECC2 (P < 0.05) without changes in muscle RING-finger protein-1 mRNA. Basal mRNA levels of myoblast determination protein-1 (MyoD), MRF4, and myogenin were higher 2 wk after ECC1 (P < 0.05). MRF4 mRNA increased after ECC1 and ECC2 (P < 0.05), whereas MyoD mRNA expression increased only after ECC1 (P = 0.03). Phosphorylation of JNK and p38 MAPK increased after both exercise bouts (P < 0.05), similar to IL-6 and IL-1ß mRNA expression. All together, these results suggest that differential regulation of the mTOR pathway and MRF expression could mediate the repeated bout effect observed between an initial and secondary bout of eccentric exercise.

Human Regulatory Protein Ki-1/57 Is a Target of SUMOylation and Affects PML Nuclear Body Formation.

Ki-1/57 is a nuclear and cytoplasmic regulatory protein first identified in malignant cells from Hodgkin's lymphoma. It is involved in gene expression regulation on both transcriptional and mRNA metabolism levels. Ki-1/57 belongs to the family of intrinsically unstructured proteins and undergoes phosphorylation by PKC and methylation by PRMT1. Previous characterization of its protein interaction profile by yeast two-hybrid screening showed that Ki-1/57 interacts with proteins of the SUMOylation machinery, the SUMO E2 conjugating enzyme UBC9 and the SUMO E3 ligase PIAS3, which suggested that Ki-1/57 could be involved with this process. Here we identified seven potential SUMO target sites (lysine residues) on Ki-1/57 sequence and observed that Ki-1/57 is modified by SUMO proteins in vitro and in vivo. We showed that SUMOylation of Ki-1/57 occurred on lysines 213, 276, and 336. In transfected cells expressing FLAG-Ki-1/57 wild-type, its paralog FLAG-CGI-55 wild-type, or their non-SUMOylated triple mutants, the number of PML-nuclear bodies (PML-NBs) is reduced compared with the control cells not expressing the constructs. More interestingly, after treating cells with arsenic trioxide (As2O3), the number of PML-NBs is no longer reduced when the non-SUMOylated triple mutant Ki-1/57 is expressed, suggesting that the SUMOylation of Ki-1/57 has a role in the control of As2O3-induced PML-NB formation. A proteome-wide analysis of Ki-1/57 partners in the presence of either SUMO-1 or SUMO-2 suggests that the involvement of Ki-1/57 with the regulation of gene expression is independent of the presence of either SUMO-1 or SUMO-2; however, the presence of SUMO-1 strongly influences the interaction of Ki-1/57 with proteins associated with cellular metabolism, maintenance, and cell cycle.

Differentiated evolutionary relationships among chordates from comparative alignments of multiple sequences of MyoD and MyoG myogenic regulatory factors.

MyoD and MyoG are transcription factors that have essential roles in myogenic lineage determination and muscle differentiation. The purpose of this study was to compare multiple amino acid sequences of myogenic regulatory proteins to infer evolutionary relationships among chordates. Protein sequences from Mus musculus (P10085 and P12979), human Homo sapiens (P15172 and P15173), bovine Bos taurus (Q7YS82 and Q7YS81), wild pig Sus scrofa (P49811 and P49812), quail Coturnix coturnix (P21572 and P34060), chicken Gallus gallus (P16075 and P17920), rat Rattus norvegicus (Q02346 and P20428), domestic water buffalo Bubalus bubalis (D2SP11 and A7L034), and sheep Ovis aries (Q90477 and D3YKV7) were searched from a non-redundant protein sequence database UniProtKB/Swiss-Prot, and subsequently analyzed using the Mega6.0 software. MyoD evolutionary analyses revealed the presence of three main clusters with all mammals branched in one cluster, members of the order Rodentia (mouse and rat) in a second branch linked to the first, and birds of the order Galliformes (chicken and quail) remaining isolated in a third. MyoG evolutionary analyses aligned sequences in two main clusters, all mammalian specimens grouped in different sub-branches, and birds clustered in a second branch. These analyses suggest that the evolution of MyoD and MyoG was driven by different pathways.

Temporal study following burn injury in young rats is associated with skeletal muscle atrophy, inflammation and altered myogenic regulatory factors.

INTRODUCTION: Burn injury (BI) greater than 40% has been associated with protein catabolism and it is characterized by a hypermetabolic response followed for muscle loss. OBJECTIVE: The purpose of this study was to investigate the temporal effects of extensive experimental BI in the skeletal muscle distant from lesion, through morphological analysis, expression of genes related to muscle atrophy, inflammation and the myogenic regulatory factors. MATERIALS AND METHODS: A total of 60 young male wistar rats were distributed into two groups, control (C) and subjected to scald burn injury (SBI). The animals were euthanized 1, 4 and 14 days post-sham or 45% of the total body surface BI. The medial head of gastrocnemii muscles were submitted to histopathological, morphometric (muscle fibers area and density), MyoD and myogenin immunoexpression, and gene expression for TNF-α, iNOS and E3 ubiquitin ligases (MuRF1 and MAFbx). RESULTS: Histopathological findings were consistent with increased amount of connective tissue and inflammatory process. Muscle fiber area of SBI groups was smaller than C and no differences were found in fiber muscle density. TNF-α was higher in SBI groups, one and 14 days post-injury; iNOS expression was higher on the first and fourth day post-injury. MuRF-1 was higher on the day four and MAFbx on the day 14. CONCLUSION: In conclusion, BI causes inflammation, atrophy and myogenesis stimulation in muscle as a result of systemic host response.

Morphology and expression of genes related to skeletal muscle growth in juveniles of pirarucu (Arapaima gigas, Arapaimatidae, Teleostei) / Morfologia e expressão de genes relacionados ao crescimento do músculo esquelético em juvenis de pirarucu (Arapaima gigas, Arapaimatidae, Teleostei)

Skeletal muscle growth in the pirarucu (Arapaima gigas) is highly interesting to fish farmers because it provides information about how the mechanism in muscle mass increase, characteristic of the pecies, is regulated. Pirarucu has specific muscle growth that highlights the speciess significance and commercial value. Current research evaluates the morphology and the growth-related gene expression in the red and white skeletal muscles of the pirarucu. Muscle samples were collected from the lateral anterior region and frozen in liquid nitrogen. Histological sections were performed and stained by HE for morphological analysis. Red and white muscle samples were used to determine MyoD, myogenin, and myostatin genes expression by Real-time Polymerase Chain Reaction. Although MyoD and myogenin were not statistically different in the two types of muscles, myostatin was significantly higher in the white rather than in the red muscle. Results show the muscle growth characteristics of the species and may be helpful for improving aquaculture management programs.(AU)

A caracterização do crescimento muscular no pirarucu (Arapaima gigas) é de elevado interesse para os piscicultores, pois fornece informações de extrema importância sobre como esse mecanismo é regulado e permite o rápido aumento na massa muscular característico da espécie. O pirarucu possui um crescimento muscular típico, o que destaca sua importância e seu valor comercial. O objetivo do presente trabalho foi avaliar a morfologia e a expressão de genes relacionados ao crescimento da musculatura esquelética vermelha e branca do pirarucu. As amostras de músculo vermelho e branco foram obtidas da região lateral anterior superficial e profunda, respectivamente, e estas foram congeladas em nitrogênio líquido. Para análise morfológica, cortes histológicos obtidos em criostato foram submetidos à coloração com HE. A expressão dos genes MyoD, miogenina e miostatina foi feita por PCR em tempo real após transcrição reversa (RT-qPCR). Em relação à expressão de MyoD e miogenina, não houve diferença estatística na comparação entre os músculos; por outro lado, a expressão da miostatina foi significativamente maior no músculo branco, em comparação com o músculo vermelho. Estes resultados refletem as características de crescimento muscular do pirarucu e podem ser úteis na tentativa de melhorar as condições de criação e a sobrevivência da espécie.(AU)

Morphology and expression of genes related to skeletal muscle growth in juveniles of pirarucu (Arapaima gigas, Arapaimatidae, Teleostei) / Morfologia e expressão de genes relacionados ao crescimento do músculo esquelético em juvenis de pirarucu (Arapaima gigas, Arapaimatidae, Teleostei)

Skeletal muscle growth in the pirarucu (Arapaima gigas) is highly interesting to fish farmers because it provides information about how the mechanism in muscle mass increase, characteristic of the pecies, is regulated. Pirarucu has specific muscle growth that highlights the species’s significance and commercial value. Current research evaluates the morphology and the growth-related gene expression in the red and white skeletal muscles of the pirarucu. Muscle samples were collected from the lateral anterior region and frozen in liquid nitrogen. Histological sections were performed and stained by HE for morphological analysis. Red and white muscle samples were used to determine MyoD, myogenin, and myostatin genes expression by Real-time Polymerase Chain Reaction. Although MyoD and myogenin were not statistically different in the two types of muscles, myostatin was significantly higher in the white rather than in the red muscle. Results show the muscle growth characteristics of the species and may be helpful for improving aquaculture management programs.

A caracterização do crescimento muscular no pirarucu (Arapaima gigas) é de elevado interesse para os piscicultores, pois fornece informações de extrema importância sobre como esse mecanismo é regulado e permite o rápido aumento na massa muscular característico da espécie. O pirarucu possui um crescimento muscular típico, o que destaca sua importância e seu valor comercial. O objetivo do presente trabalho foi avaliar a morfologia e a expressão de genes relacionados ao crescimento da musculatura esquelética vermelha e branca do pirarucu. As amostras de músculo vermelho e branco foram obtidas da região lateral anterior superficial e profunda, respectivamente, e estas foram congeladas em nitrogênio líquido. Para análise morfológica, cortes histológicos obtidos em criostato foram submetidos à coloração com HE. A expressão dos genes MyoD, miogenina e miostatina foi feita por PCR em tempo real após transcrição reversa (RT-qPCR). Em relação à expressão de MyoD e miogenina, não houve diferença estatística na comparação entre os músculos; por outro lado, a expressão da miostatina foi significativamente maior no músculo branco, em comparação com o músculo vermelho. Estes resultados refletem as características de crescimento muscular do pirarucu e podem ser úteis na tentativa de melhorar as condições de criação e a sobrevivência da espécie.

Polymorphism of MyoD1 and Myf6 genes and associations with carcass and meat quality traits in beef cattle.

Myogenic determination factor 1 (MyoD1) and myogenic factor 6 (Myf6) genes belong to the myogenic differentiation (MyoD) gene family, which play key roles in growth and muscle development. The study aimed to investigate the effects of variants in cattle MyoD1 and Myf6 on carcass and meat traits. We screened single nucleotide polymorphisms (SNPs) of both genes in 8 cattle populations, including Simmental, Angus, Hereford, Charolais, Limousin, Qinchuan, Luxi, and Jinnan by sequencing. The G782A locus was identified in exon 1 of MyoD1 (MyoD1-BglI) as well as the T186C locus in exon 1 of Myf6 (Myf6-ApaLI). For the two SNPs, the A allele was significantly more frequent than the B allele in the populations tested. The χ(2) test showed that the MyoD1-BglI locus conformed to Hardy-Weinberg equilibrium in the 8 populations, as did the Myf6-ApaLI locus, with the exception of the Simmental population (P > 0.05). Association analysis revealed that the MyoD1-BglI locus was significantly associated with loin muscle area (LMA) (P < 0.05), and the Myf6-ApaLI locus was significantly associated with carcass length (CL) (P < 0.05). Animals with BB and AB genotypes for the MyoD1-BglI locus had larger LMAs compared to animals with AA genotype. Individuals with BB genotype had longer CLs compared to those with AA and AB genotypes. We conclude that the two SNPs might provide useful genetic markers, opening up new possibilities for cattle breeding and improvements in gene-assisted selection.

Developmental characteristics of pectoralis muscle in Pekin duck embryos.

To confirm the entire developmental process and transition point of embryonic Pekin duck pectoral muscle, and to investigate the association between pectoral muscle development and their regulating genes, anatomical and morphological analyses of embryonic Pekin duck skeletal muscles were performed, and the expression patterns of its regulating genes were investigated. The anatomical analysis revealed that body weight increased with age, while increases in pectoral muscle weight nearly ceased after the embryo was 20 days of hatching (E20). The developmental morphological characteristics of Pekin duck pectoral muscle at the embryonic stage showed that E20 was the transition point (from proliferation to fusion) of Pekin duck pectoral muscle. The expression patterns of MRF4, MyoG, and MSTN indicated that E19 or E20 was the fastest point of pectoral muscle development and the crucial transition for Pekin duck pectoral muscle development during the embryonic stage. Together, these findings imply that E20 is the crucial transition point (from proliferation to fusion) of Pekin duck pectoral muscle and that there is no muscle fiber hypertrophy after E20. Results of this study provide further understanding of the developmental process and transition point of Pekin duck pectoral muscle during the embryo stage.

Effect of nandrolone decanoate on skeletal muscle repair.

This study analyzed the effect of nandrolone decanoate (ND) on muscle repair and the expression of myogenic regulatory factors following cryoinjury in rat skeletal muscle. Adult male Wistar rats were randomly divided into 4 groups: control group, sham group, cryoinjured group treated with ND and non-injured group treated with ND. Treatment consisted of subcutaneous injections of ND (5 mg/kg) twice a week. After sacrifice, the tibialis anterior muscle was removed for the isolation of total RNA and analysis of myogenic regulatory factors using real-time PCR as well as morphological analysis using the hematoxylin-eosin assay. There was a significant increase in MyoD mRNA after 7 days and in myogenin mRNA after 21 days in the cryoinjured ND group in comparison to other groups in the same period. The morphological analysis revealed no edema or myonecrosis after 7 days as well as no edema or inflammatory infiltrate after 14 days in the cryoinjured ND group. In conclusion the anabolic steroid nandrolone decanoate can modulate the muscle repair process in rats following cryoinjury by influencing the expression of regulatory myogenic factors and phases of muscle repair.

Heart failure-induced skeletal myopathy in spontaneously hypertensive rats.

BACKGROUND: Although skeletal muscle atrophy and changes in myosin heavy chain (MyHC) isoforms have often been observed during heart failure, their pathophysiological mechanisms are not completely defined. In this study we tested the hypothesis that skeletal muscle phenotype changes are related to myogenic regulatory factors and myostatin/follistatin expression in spontaneously hypertensive rats (SHR) with heart failure. METHODS: After developing tachypnea, SHR were subjected to transthoracic echocardiogram. Pathological evidence of heart failure was assessed during euthanasia. Age-matched Wistar-Kyoto (WKY) rats were used as controls. Soleus muscle morphometry was analyzed in histological sections, and MyHC isoforms evaluated by electrophoresis. Protein levels were assessed by Western blotting. STATISTICAL ANALYSIS: Student'st test and Pearson correlation. RESULTS: All SHR presented right ventricular hypertrophy and seven had pleuropericardial effusion. Echocardiographic evaluation showed dilation in the left chambers and left ventricular hypertrophy with systolic and diastolic dysfunction in SHR. Soleus weight and fiber cross sectional areas were lower (WKY 3615 ± 412; SHR 2035 ± 224 µm(2); P<0.001), and collagen fractional volume was higher in SHR. The relative amount of type I MyHC isoform was increased in SHR. Myogenin, myostatin, and follistatin expression was lower and MRF4 levels higher in SHR. Myogenin and follistatin expression positively correlated with fiber cross sectional areas and MRF4 levels positively correlated with I MyHC isoform. CONCLUSION: Reduced myogenin and follistatin expression seems to participate in muscle atrophy while increased MRF4 protein levels can modulate myosin heavy chain isoform shift in skeletal muscle of spontaneously hypertensive rats with heart failure.

Muscle-specific growth hormone receptor (GHR) overexpression induces hyperplasia but not hypertrophy in transgenic zebrafish.

Even though growth hormone (GH) transgenesis has demonstrated potential for improved growth of commercially important species, the hormone excess may result in undesired collateral effects. In this context, the aim of this work was to develop a new model of transgenic zebrafish (Danio rerio) characterized by a muscle-specific overexpression of the GH receptor (GHR) gene, evaluating the effect of transgenesis on growth, muscle structure and expression of growth-related genes. In on line of transgenic zebrafish overexpressing GHR in skeletal muscle, no significant difference in total weight in comparison to non-transgenics was observed. This can be explained by a significant reduction in expression of somatotrophic axis-related genes, in special insulin-like growth factor I (IGF-I). In the same sense, a significant increase in expression of the suppressors of cytokine signaling 1 and 3 (SOCS) was encountered in transgenics. Surprisingly, expression of genes coding for the main myogenic regulatory factors (MRFs) was higher in transgenic than non-transgenic zebrafish. Genes coding for muscle proteins did not follow the MRFs profile, showing a significant decrease in their expression. These results were corroborated by the histological analysis, where a hyperplasic muscle growth was observed in transgenics. In conclusion, our results demonstrated that GHR overexpression does not induce hypertrophic muscle growth in transgenic zebrafish probably because of SOCS impairment of the GHR/IGF-I pathway, culminating in IGF-I and muscle proteins decrease. Therefore, it seems that hypertrophy and hyperplasia follow two different routes for entire muscle growth, both of them triggered by GHR activation, but regulated by different mechanisms.

Calcium-regulated transcriptional pathways in the normal and pathologic heart.

The cytosolic calcium concentration ([Ca(2+)](c)) is key for the regulation of many cellular processes, such cell signaling and proliferation, metabolism, and muscle contraction. In cardiomyocytes, Ca(2+) is an important regulator in many cellular functions such electrophysiological processes, excitation-contraction coupling, regulation of contractile proteins activity, energy metabolism, cell death, and transcriptional regulation by the activation of Ca(2+)-dependent transcriptional pathways. In cardiomyocytes, the two main Ca(2+) -dependent pathways are the Ca(2+)/calmodulin-calcineurin-NFAT and the Ca(2+) /calmodulin-dependent kinases-MEF2. Both pathways are involved in the transcriptional control of many cardiac genes. Cardiac hypertrophy (CH) and heart failure (HF) are characterized by alterations in calcium handling such a low sarcoplasmic reticulum Ca(2+) content, decreased rate of Ca(2+) removal from the sarcoplasm, increased diastolic [Ca(2+)](c), and decreased systolic [Ca(2+)](c), all of them contributing to diminished contractibility and force generation in failing heart. At gene expression level, there are also many changes such decreased levels of SERCA2a and activation of a fetal gene expression program in cardiomyocytes. A variety of Ca(2+)-dependent signaling pathways have been implicated in CH and HF, but whether these pathways are interrelated and whether there is specificity among them are still unclear and under investigation. The focus of this review is to make an analysis of the current knowledge about the role of Ca(2+) signaling pathways in the regulation of cardiac gene expression making special emphasis in novel strategies to correct Ca(2+) handling alterations by means of SERCA2a gene therapy.

Evidence for the association of the human regulatory protein Ki-1/57 with the translational machinery.

Ki-1/57 is a cytoplasmic and nuclear protein of 57 kDa first identified in malignant cells from Hodgkin's lymphoma. Based on yeast-two hybrid protein interaction we found out that Ki-1/57 interacts with adaptor protein RACK1 (receptor of activated kinase 1), CIRP (cold-inducible RNA-binding protein), RPL38 (ribosomal protein L38) and FXR1 (fragile X mental retardation-related protein 1). Since these proteins are involved in the regulation of translation we suspected that Ki-1/57 may have a role in it. We show by immunoprecipitation the association of Ki-1/57 with FMRP. Confocal microscopy revealed that Ki-1/57 colocalizes with FMRP/FXR1/2 to stress granules. Furthermore Ki-1/57 cosediments with free ribosomal particles and enhances translation, when tethered to a reporter mRNA, suggesting that Ki-1/57 may be involved in translational regulation.

Adaptações morfofuncionais e respostas moleculares do músculo esquelético de ratos submetidos ao treinamento resistido / Morphofunctional adaptations an molecular responses of skeletal muscle of rats submitted to resistance training

Embora fortes evidências demonstrem que os fatores de regulação miogênica (MRFs) e o fator de crescimento semelhante à insulina (IGF-I) apresentem um importante papel na resposta hipertrófica após treinamento resistido (TR) agudo, permanece desconhecido se a resposta dos MRFs e IGF-I também ocorre durante a adaptação ao TR a longo-prazo. Portanto, o objetivo deste estudo foi testar a hipótese de que a resposta hipertrófica e modulação das fibras do músculo esquelético após TR a longo-prazo poderia estar associada ao aumento na expressão gênica dos MRFs e IGF-I. Ratos Wistar (80 dias de idade, 250-300 g) foram divididos em quatro grupos: Controle 8 semana (C8, n = 8), Treinado 8 semanas (T8, n = 8), Controle 12 semanas (C12, n = 8) e Treinado 12 semanas (T12, n = 8). Os grupos T8 e T12 foram submetidos a um programa de TR progressivo (3 dias/semana) durante 8 e 12 semanas, respectivamente. O protocolo de treinamento consistiu de quatro séries de 10-12 repetições, com um período de descanso de 40 segundos entre cada série, realizado a 65-75% de uma repetição máxima (1RM). Ao término do experimento, os animais foram sacrificados e o músculo plantar coletado para as análises morfológica e molecular. O TR durante 8 e 12 semanas não promoveu qualquer alteração (p > 0,05) significante no ganho de peso corporal e consumo alimentar dos grupos T8 e T12 em relação aos grupos C8 e C12, respectivamente...

Although strong evidence show that the myogenic regulatory factors (MRFs) and insulin-like growth factor (IGF-I) have important roles in the hypertrophy response after acute resistance training, it is still unclear if response of MRFs and IGF-I also occurs during the adaptation to prolonged periods of resistance training (RT). Therefore, the purpose of this study was to test the hypothesis that fiber-types transition and hypertrophy during long-term RT could be associated with increased MRFs and IGF-I mRNA expression in the skeletal muscle. Male Wistar rats (80 days old, 250-300 g) were divided into four groups: 8 weeks control (C8, n = 8), 8-weeks trained (T8, n = 8), 12-weeks control (C12, n = 8), 12-weeks trained (T12, n = 8). T8 and T12 groups were submitted to a progressive RT program (3 day/week) for 8 and 12 weeks, respectively. The training protocol consisted of four sets of 10–12 repetitions, with a 40 s rest period between each set, performed at 65–75% of one repetition maximum (1RM). At the end of the experiment, animals were sacrificed and the plantaris muscle collected for morphological and molecular analysis. The RT did not change (p > 0.05) in body weight gain and food intake in the T8 and T12 compared to the C8 and C12 groups, respectively...

Chronic heart failure-induced skeletal muscle atrophy, necrosis, and changes in myogenic regulatory factors.

BACKGROUND: Although intrinsic skeletal muscle abnormalities can influence exercise intolerance during heart failure (HF), the factors responsible for muscle changes have not been elucidated. In this study we evaluated the expression of myogenic regulatory factors (MRF), myosin heavy chain (MyHC) isoforms, and fiber trophism in the soleus muscle of rats with myocardial infarction-induced heart failure. METHOD/RESULTS: Six months after surgery, 2 groups of rats were studied: sham, and infarcted rats with HF (MI/HF+, MI size: 41.1±6.3% of total left ventricular area). In the infarcted group, microscopic evaluation revealed scattered foci of fiber necrosis in combination with inflammatory cells, phagocytosis, and increased fibrous tissue. The frequency of necrotic fibers was significantly higher in the MI/HF+ group than in the sham. The MI/HF+ group had atrophy of type I, IC/IIC, and IIA fibers compared to the sham group (P<0.05). MyoD gene expression was higher in the MI/HF+ group (sham: 1.00±0.49; MI/HF+: 2.53±0.71 arbitrary units; P<0.001). Myogenin and MRF4 gene expression was similar in both groups. Myogenin protein levels were reduced in the MI/HF+ group (sham: 1.00±0.21; MI/HF+: 0.74±0.21 arbitrary units; P=0.026). MyoD and MRF4 protein levels, as well as the MyHC distribution, were not different between groups. The MI/HF+ group had higher TNF-α and IL-6 serum concentrations than the sham group. CONCLUSIONS: Heart failure-induced skeletal muscle atrophy is combined with fiber necrosis, increased MyoD gene expression and decreased myogenin protein levels.