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Endurance exercise differentially stimulates heart and axial muscle development in zebrafish (Danio rerio).

Mechanical load is an important factor in the differentiation of cells and tissues. To investigate the effects of increased mechanical load on development of muscle and bone, zebrafish were subjected to endurance swim training for 6 h/day for 10 wk starting at 14 days after fertilization. During the first 3 wk of training, trained fish showed transiently increased growth compared with untrained (control) fish. Increased expression of proliferating cell nuclear antigen suggests that this growth is realized in part through increased cell proliferation. Red and white axial muscle fiber diameter was not affected. Total cross-sectional area of red fibers, however, was increased. An improvement in aerobic muscle performance was supported by an increase in myoglobin expression. At the end of 10 wk of training, heart and axial muscle showed increased expression of the muscle growth factor myogenin and proliferating cell nuclear antigen, but there were major differences between cardiac and axial muscle. In axial muscle, expression of the "slow" types of myosin and troponin C was increased, together with expression of erythropoietin and myoglobin, which enhance oxygen transport, indicating a shift toward a slow aerobic phenotype. In contrast, the heart muscle shifts to a faster phenotype but does not become more aerobic. This suggests that endurance training differentially affects heart and axial muscle.

Animals↗

rst and its paralogue kirre act redundantly during embryonic muscle development in Drosophila.

The polynucleate myotubes of vertebrates and invertebrates form by fusion of myoblasts. We report the involvement of the Drosophila melanogaster Roughest (Rst) protein as a new membrane-spanning component in this process. Rst is strongly expressed in mesodermal tissues during embryogenesis, but rst null mutants display only subtle embryonic phenotypes. Evidence is presented that this is due to functional redundancy between Rst and its paralogue Kirre. Both are highly related single-pass transmembrane proteins with five extracellular immunoglobulin domains and three conserved motifs in the intracellular domain. The expression patterns of kirre and rst overlap during embryonic development in muscle founder cells. Simultaneous deletion of both genes causes an almost complete failure of fusion between muscle founder cells and fusion-competent myoblasts. This defect can be rescued by one copy of either gene. Moreover, Rst, like Kirre is a myoblast attractant.

Amino Acid Sequence↗

Expression of dystrophin-associated glycoproteins during human fetal muscle development: a preliminary immunocytochemical study.

An immunocytochemical study was performed on quadriceps muscle from eight fetuses ranging from 12 weeks of gestation to term, using antibodies against the dystrophin-associated proteins, in order to evaluate the developmental expression of these proteins. For comparison, antibodies against dystrophin and utrophin were also used. The expression of the 59 kDa dystrophin-associated protein was simultaneous with that of dystrophin, which is also a subsarcolemmal protein. The extracellular glycoprotein of 156 kDa (alpha-dystroglycan) and the transmembrane glycoprotein of 43 kDa (beta-dystroglycan) appeared to be expressed later. The transmembrane glycoproteins of 50 kDa (adhalin) and 35 kDa were fully expressed at an even later stage of fetal muscle development. This study suggests that the subsarcolemmal proteins may have an essential role in the assembly of the transmembrane and extracellular components of the dystrophin-glycoprotein complex during fetal muscle development. The knowledge obtained from observing the developmental expression of these proteins may contribute to the understanding of the molecular mechanism of their different involvement in muscle disorders.

Cytoskeletal Proteins↗

Jaw muscle development as evidence for embryonic repatterning in direct-developing frogs.

The Puerto Rican direct-developing frog Eleutherodactylus coqui (Leptodactylidae) displays a novel mode of jaw muscle development for anuran amphibians. Unlike metamorphosing species, several larval-specific features never form in E. coqui; embryonic muscle primordia initially assume an abbreviated, mid-metamorphic configuration that is soon remodelled to form the adult morphology before hatching. Also lacking are both the distinct population of larval myofibres and the conspicuous, larval-to-adult myofibre turnover that are characteristic of muscle development in metamorphosing species. These modifications are part of a comprehensive alteration in embryonic cranial patterning that has accompanied life history evolution in this highly speciose lineage. Embryonic 'repatterning' in Eleutherodactylus may reflect underlying developmental mechanisms that mediate the integrated evolution of complex structures. Such mechanisms may also facilitate, in organisms with a primitively complex life cycle, the evolutionary dissociation of embryonic, larval, and adult features.

Animals↗

Nerve-muscle interactions during flight muscle development in Drosophila.

During Drosophila pupal metamorphosis, the motoneurons and muscles differentiate synchronously, providing an opportunity for extensive intercellular regulation during synapse formation. We examined the existence of such interactions by developmentally delaying or permanently eliminating synaptic partners during the formation of indirect flight muscles. When we experimentally delayed muscle development, we found that although adult-specific primary motoneuron branching still occurred, the higher order (synaptic) branching was suspended until the delayed muscle fibers reached a favourable developmental state. In reciprocal experiments we found that denervation caused a decrease in the myoblast pool. Furthermore, the formation of certain muscle fibers (dorsoventral muscles) was specifically blocked. Exceptions were the adult muscles that use larval muscle fibers as myoblast fusion targets (dorsal longitudinal muscles). However, when these muscles were experimentally compelled to develop without their larval precursors, they showed an absolute dependence on the motoneurons for their formation. These data show that the size of the myoblast pool and early events in fiber formation depend on the presence of the nerve, and that, conversely, peripheral arbor development and synaptogenesis is closely synchronized with the developmental state of the muscle.

Animals↗

Expression of fibroblast growth factor 2 and its receptor during skeletal muscle development from turkeys with different growth rates.

Fibroblast growth factor 2 (FGF2) is a key regulator of muscle cell proliferation and differentiation. To address how FGF2 and fibroblast growth factor receptor 1 (FGFR1) gene expression influences skeletal muscle development and growth, pectoralis major muscle was isolated at embryonic days (ED) 14, 16, 18, 20, 22, and 24, and at 1-, 8-, 12-, and 16-week posthatch from a turkey line (F) selected only for increased 16-week body weight and its genetic control line (RBC2). The mRNA levels of FGF2 and FGFR1 were measured by semi-quantitative reverse transcription polymerase chain reaction. Compared to the RBC2 line males, the F line males had higher FGF2 mRNA levels at ED 14 and 16, and higher FGFR1 mRNA levels at ED 18, but down-regulated FGF2 and FGFR1 gene expression at ED 22. Although no FGF2 mRNA was detected in posthatch muscle tissue, the F line turkeys had more FGFR1 gene expression at 8-, 12-, and 16-week posthatch than the RBC2 line turkeys. During myogenic satellite cell proliferation, the F line cells had higher FGF2 and FGFR1 mRNA levels than the RBC2 line cells. The satellite cell responsiveness to FGF2 treatment was evaluated by the ability of the cells to proliferate. The male satellite cells were more responsive to FGF2 stimulation than the female cells in both lines. These results suggest that the F line turkeys have increased FGF2 signaling that may affect muscle cell proliferation and differentiation, which may also lead to an enhancement in muscle development and growth rate.

Animals↗

Muscle flexibility as a risk factor for developing muscle injuries in male professional soccer players. A prospective study.

BACKGROUND: Muscular tightness is frequently postulated as an intrinsic risk factor for the development of a muscle injury. However, very little prospective data exist to prove this. HYPOTHESIS: Increased muscle tightness identifies a soccer player at risk for a subsequent musculoskeletal lesion. STUDY DESIGN: Prospective cohort study. METHODS: We examined 146 male professional soccer players before the 1999-2000 Belgian soccer competition. None of the players had a history of muscle injury in the lower extremities in the previous 2 years. The flexibility of the hamstring, quadriceps, adductor, and calf muscles of these players was measured goniometrically before the start of the season. All of the examined players were monitored throughout the season to register subsequent injuries. RESULTS: Players with a hamstring (N = 31) or quadriceps (N = 13) muscle injury were found to have significantly lower flexibility in these muscles before their injury compared with the uninjured group. No significant differences in muscle flexibility were found between players who sustained an adductor muscle injury (N = 13) or a calf muscle injury (N = 10) and the uninjured group. CONCLUSIONS: These results indicate that soccer players with an increased tightness of the hamstring or quadriceps muscles have a statistically higher risk for a subsequent musculoskeletal lesion. CLINICAL SIGNIFICANCE: Preseason hamstring and quadriceps muscle flexibility testing can identify male soccer players at risk of developing hamstring and quadriceps muscle injuries.

Athletic Injuries↗

Neonatal and adult myosin heavy chains form homodimers during avian skeletal muscle development.

Myosin isoforms contribute to the heterogeneity and adaptability of skeletal muscle fibers. Besides the well-characterized slow and fast muscle myosins, there are those isoforms that appear transiently during the course of muscle development. At a stage of development when two different myosins are coexpressed, the possibility arises for the existence of heterodimers, molecules containing two different heavy chains, or homodimers, molecules with two identical heavy chains. The question of whether neonatal and adult myosin isoforms can associate to form a stable heterodimer was addressed by using stage-specific monoclonal antibodies in conjunction with immunological and electron microscopic techniques. We find that independent of the ratio of adult to neonatal myosin, depending on the age of the animal, the myosin heavy chains form predominantly homodimeric molecules. The small amount of hybrid species present suggests that either the rod portion of the two heavy chain isoforms differs too much in sequence to form a stable alpha-helical coiled coil, or that the biosynthesis of the heavy chains precludes the formation of heterodimeric molecules.

Animals↗

Genetic approaches to understanding muscle development.

The analysis of both naturally occurring and experimentally induced mutants has greatly advanced our understanding of muscle development. Molecular biological techniques have led to the isolation of genes associated with inherited human diseases that affect muscle tissues. Analysis of the encoded proteins in conjunction with the mutant phenotypes can provide powerful insights into the function of the protein in normal muscle development. Systematic searches for muscle mutations have been made in experimental systems, most notably the fruit fly Drosophila melanogaster and the nematode Caenorhabditis elegans. In addition, known muscle protein genes from other organisms have been used to isolate homologs from genetically manipulatable organisms, allowing mutant analysis and the study of protein function in vivo. Mutations in transcription factor genes that affect mesoderm development have been isolated and genetic lesions affecting myofibril assembly have been identified. Genetic experiments inducing mutations and rescuing them by transgenic methods have uncovered functions of myofibrillar protein isoforms. Some isoforms perform muscle-specific functions, whereas others appear to be replaceable by alternative isoforms. Mutant analysis has also uncovered a relationship between proteins at the cell membrane and the assembly and alignment of the myofibrillar apparatus. We discuss examples of each of these genetic approaches as well as the developmental and evolutionary implications of the results.

Animals↗

The human and mouse fibroblast growth factor 6 (FGF6) genes and their products: possible implication in muscle development.

FGF6 is structurally very similar to the other members of the FGF gene family, and particularly to the FGF4 gene, which was instrumental in its isolation. Its longest open reading frame encodes a 208 amino acid residues long protein, both in man and in the mouse. It is expressed as a 4.8 kb transcript in skeletal muscle. In developing muscle, expression starts at the myotomal stage and culminates in differentiated fetal muscle masses. In culture, FGF6 protein is mitogenic and has a transforming capacity for fibroblasts. It represses the terminal differentiation of myoblasts. Action of FGF6 could be mediated by the FGFR4 receptor, which binds FGF6 and whose gene is also expressed in developing skeletal muscle.

Animals↗

Differential regulation of epaxial and hypaxial muscle development by paraxis.

In vertebrates, skeletal muscle is derived from progenitor cell populations located in the epithelial dermomyotome compartment of the each somite. These cells become committed to the myogenic lineage upon delamination from the dorsomedial and dorsolateral lips of the dermomyotome and entry into the myotome or dispersal into the periphery. Paraxis is a developmentally regulated transcription factor that is required to direct and maintain the epithelial characteristic of the dermomyotome. Therefore, we hypothesized that Paraxis acts as an important regulator of early events in myogenesis. Expression of the muscle-specific myogenin-lacZ transgene was used to examine the formation of the myotome in the paraxis-/- background. Two distinct types of defects were observed that mirrored the different origins of myoblasts in the myotome. In the medial myotome, where the expression of the myogenic factor Myf5 is required for commitment of myoblasts, the migration pattern of committed myoblasts was altered in the absence of Paraxis. In contrast, in the lateral myotome and migratory somitic cells, which require the expression of MyoD, expression of the myogenin-lacZ transgene was delayed by several days. This delay correlated with an absence of MyoD expression in these regions, indicating that Paraxis is required for commitment of cells from the dorsolateral dermomyotome to the myogenic lineage. In paraxis-/-/myf5-/- neonates, dramatic losses were observed in the epaxial and hypaxial trunk muscles that are proximal to the vertebrae in the compound mutant, but not those at the ventral midline or the non-segmented muscles of the limb and tongue. In this genetic background, myoblasts derived from the medial (epaxial) myotome are not present to compensate for deficiencies of the lateral (hypaxial) myotome. Our data demonstrate that Paraxis is an important regulator of a subset of the myogenic progenitor cells from the dorsolateral dermomyotome that are fated to form the non-migratory hypaxial muscles.

Animals↗

Effects of rumen fluid transplantation on longissimus dorsi muscle development in Xizang sheep: An association analysis based on transcriptomic and serum metabolomic profiles.

This study aimed to investigate the effects of rumen fluid transplantation (RFT) on the growth and development of the longissimus dorsi muscle in female Xizang sheep. After RFT, muscle lightness differed significantly between the two groups, with the LDC group showing significantly higher lightness than the LDT group. In contrast, no significant differences were observed between groups in other muscle phenotypic traits, including drip loss, pH, cooking loss, shear force, redness, and yellowness. Antioxidant-related indices (SOD, GSH-PX, MDA, CAT, and T-AOC) also showed no significant differences between groups. Histological analysis revealed that muscle fiber length, width, and density were significantly greater in the experimental group than in the control group. Transcriptomic analysis identified 515 differentially expressed genes (DEGs), of which 419 were downregulated. KEGG analysis indicated that genes involved in muscle development-related pathways, such as cell adhesion and the PI3K-Akt signaling pathway, were predominantly downregulated. Key serum metabolites (L-kynurenine, IPA, allantoin, and propionylcarnitine) showed highly significant positive correlations with muscle fiber growth indices. In contrast, metabolites such as l-carnitine, acetylcarnitine, and citrulline were negatively correlated with muscle fiber growth, but positively correlated with the expression of muscle structure-related genes (COL11A1 and EFNA5) and with meat lightness. Overall, this study provides new insights into the potential molecular basis by which RFT influences muscle growth and development. However, the mechanisms by which RFT affects muscle development and meat quality-related traits remain unclear and warrant further investigation.

Animals↗

Scanning electron microscopy of prenatal muscle development in the mouse.

Scanning electron microscopy was used to study prenatal muscle development in mouse fetuses ranging from 12 days to 18 days gestation. Some transmission electron micrographs were also used for comparison. At 12 days no myofibres were evident although at 13 days long fibres surrounded by many mononucleated cells could be seen. At 14 days bundles of primary myofibres were observed with new myofibres forming in crevices between adjacent fibres. At 16 days the primary myofibres had separated but smaller secondary myofibres could be seen forming along their surfaces. The myofibres were very compact at 18 days and often appeared fused but this was due to the basal lamina ensheathing clusters of primary myofibres with their secondaries. The scanning electron micrographs appeared to illustrate the theories of muscle development which have arisen out of the various studies employing sectioning techniques.

Animals↗

Nerve-muscle interactions regulate motor terminal growth and myoblast distribution during muscle development.

Interactions between motoneurons and muscles influence many aspects of neuromuscular development in all animals. These interactions can be readily investigated during adult muscle development in holometabolous insects. In this study, the development of the dorsolongitudinal flight muscle (DLM) and its innervation is investigated in the moth, Manduca sexta, to address the specificity of neuromuscular interactions. The DLM develops from an anlage containing both regressed larval template fibers and imaginal myoblasts. In the adult, each fiber bundle (DLM1-5) is innervated by a single motoneuron (MN1-MN5), with the dorsal-most fiber bundle (DLM5) innervated by a mesothoracic motoneuron (MN5). The DLM failed to develop following complete denervation because myoblasts failed to accumulate in the DLM anlage. After lesioning MN1-4, MN5 retained its specificity for the DLM5 region of the anlage and failed to rescue DLM1-4. Thus specific innervation of the DLM fiber bundles does not depend on interactions among motoneurons. Myoblast accumulation, but not myonuclear proliferation, increased around the MN5 terminals, producing a hypertrophied adult DLM5. Therefore, motoneurons compete for uncommitted myoblasts. MN5 terminals subsequently grew more rapidly over the hypertrophied DLM5 anlage, indicating that motoneuron terminal expansion is regulated by the size of the target muscle anlage.

Animals↗

Transient expression of collagen type XIV during muscle development and its reappearance after denervation and degeneration.

In the formation of muscle pattern, the architectural arrangement is believed to be controlled by the local connective tissue cells. In this study we examined the immunohistological localization of Type XIV collagen recognized by a monoclonal antibody, MAb DBM, in embryonic chick hind limbs from stage (St.) 27 to 2 weeks post hatching. DBM staining was transiently observed in the epimysium from St. 30, in the perimysium of the dorsal region from St. 37, and in the entire perimysium from St. 39. After hatching, DBM staining was notably diminished in both epimysium and perimysium. In contrast, DBM staining and in situ hybridization signals for Type XIV collagen mRNA increased in the muscle connective tissues after denervation and around the regenerating muscle fibers. Therefore, Type XIV collagen expression appears to coincide with muscle activity and muscle regenerating conditions, and Type XIV collagen is considered to play roles in muscle development and regeneration.

Amino Acid Sequence↗

Regulation of fetal muscle development by thyroxine.

We have demonstrated that fetal hypophysectomy (hypox) induces deficiencies in skin and adipose tissue development and that chronic thyroxine (T4) treatment either normalizes or enhances skin and adipose tissue development in hypox fetuses. Since hypox also induces deficiencies in muscle development we report herein results of a histochemical and biochemical analysis of muscle development conducted in studies of T4-treated hypox fetuses. Pig fetuses were hypophysectomized on day 70 of gestation and treated with T4 during two periods, i.e. 70-90 days and 90-105 days gestational age, by implanting slow-release T4 (15 mg) pellets in the lateral musculature of the hind limb. Blood, muscle and other tissue samples were obtained upon removal of fetuses at days 90 and 105 of gestation. Weights of hypox, control and T4-treated hypox fetuses were similar at the end of the early period (70-90 days) whereas T4 significantly depressed body weight (p < 0.05) during the later period (90-105 days). Muscle weights, protein, DNA and RNA concentrations and muscle fiber size were not influenced by hypox or T4 treatment. Hypox-induced deficiencies in muscle capillary-to-fiber ratios, percentage of dry matter and general morphology were normalized by T4 (p < 0.05) during both treatment periods. Lipid deposition (staining) and histochemical reactivity for an oxidative marker (NADH-TR) were enhanced (p < 0.05) by T4 during both treatment periods as revealed by computer-assisted image analysis. Histochemistry for myofibrillar (acid) ATPase showed that the hypox-impaired type II to type I fiber type conversion was not influenced by T4 treatment. Muscle developmental traits enhanced or normalized by T4 were directly or indirectly related to lipid and/or oxidative metabolism. Effects of T4 on muscle and other tissues indicate, therefore, that thyroid hormones may be key regulators of oxidative and/or lipid metabolism in the neonatal animal.

Animals↗

The effect of testosterone on skeletal muscle development and protein synthesis in rabbits.

The effect of testosterone administration on muscle development and protein synthesis was studied with intact male rabbits. Rabbits were randomized into three experimental groups; day 0 control (G-1), day 15 control (G-2) and day 15 testosterone (G-3). Testosterone (delta4-Androsten-17beta-ol-3-one in silastic tubing) was surgically implanted into the G-3 rabbits. On day (G-1) and day 15 (G-2 and 3) of the experiment all rabbits were injected IP with 50 mu Ci-3H-leucine. Rabbits were sacrificed at intervals after the 3H-leucine injection. Right and left semitendinosus muscles, pituitary and adrenal glands, testicles and blood samples were obtained for analysis. Testosterone improved (P less than .01) gain and feed efficiency, depressed testicle weight (P less than .01), increased serum testosterone and insulin (P less than 0.01) but did not affect adrenal and pituitary weights. Muscle moisture, fat and total protein, myofibrillar, sarcoplasmic and stromal protein fractions and RNA and DNA content were not significantly affected by testosterone although G-3 semitendinosus muscle had the highest total and myofibrillar protein, RNA, and DNA content. The testosterone treated rabbits (G-3) had higher (P less than 0.05) 3H-leucine incorporation into myofibrillar protein and lower 3H-leucine incorporation into sarcoplasmic protein than the day 0 and day 15 controls.

Adrenal Glands↗

A BAC transgenic analysis of the Mrf4/Myf5 locus reveals interdigitated elements that control activation and maintenance of gene expression during muscle development.

The muscle-specific transcription factors Myf5 and Mrf4 are two of the four myogenic regulatory factors involved in the transcriptional cascade responsible for skeletal myogenesis in the vertebrate embryo. Myf5 is the first of these four genes to be expressed in the mouse. We have previously described discrete enhancers that drive Myf5 expression in epaxial and hypaxial somites, branchial arches and central nervous system, and argued that additional elements are required for proper expression (Summerbell, D., Ashby, P. R., Coutelle, O., Cox, D., Yee, S. P. and Rigby, P. W. J. (2000) Development 127, 3745-3757). We have now investigated the transcriptional regulation of both Myf5 and Mrf4 using bacterial artificial chromosome transgenesis. We show that a clone containing Myf5 and 140 kb of upstream sequences is sufficient to recapitulate the known expression patterns of both genes. Our results confirm and reinforce the conclusion of our earlier studies, that Myf5 expression is regulated differently in each of a considerable number of populations of muscle progenitors, and they begin to illuminate the evolutionary origins of this complex regulation. We further show that separate elements are involved in the activation and maintenance of expression in the various precursor populations, reflecting the diversity of the signals that control myogenesis. Mrf4 expression requires at least four elements, one of which may be shared with Myf5, providing a possible explanation for the linkage of these genes throughout vertebrate phylogeny. Further complexity is revealed by the demonstration that elements which control Mrf4 and Myf5 are embedded in an unrelated neighbouring gene.

Animals↗