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Actin concentration and monomer-polymer ratio in developing chicken skeletal muscle.

The actin concentration and monomer-polymer ratio in developing chicken skeletal muscle were determined by means of a DNase I inhibition assay. The concentration of G-actin in embryonic muscle was much higher than the critical concentration for polymerization of purified actin. As muscle development progressed, the amount of total actin remarkably increased, whereas the concentration of G-actin markedly decreased, and finally in adults reached the critical concentration for polymerization of purified actin. When the monomeric actin in the soluble fraction of embryonic muscle was purified, the critical concentration for polymerization of the embryonic actin decreased to the same value as that of adult skeletal muscle actin. On the other hand, there was no difference between the crude and purified actin in the type of actin. They consisted of alpha-, beta-, and gamma-actins; their amounts were in the order, beta greater than gamma greater than alpha. Furthermore, polymerization of the monomeric actin in the soluble fraction of embryonic muscle was induced by the addition of myosin or HMM. The large amount of monomeric actin in the embryonic skeletal muscle may be due to the presence of some factor(s) which inhibits actin polymerization and also to an insufficiency of myosin.

Actins↗

Whole-body skeletal muscle mass: development and validation of total-body potassium prediction models.

BACKGROUND: A substantial proportion of total body potassium (TBK) in humans is found in skeletal muscle (SM), thus affording a means of predicting total-body SM from whole-body counter-measured (40)K. There are now > 30 whole-body counters worldwide that have large cross-sectional and longitudinal TBK databases. OBJECTIVE: We explored 2 SM prediction approaches, one based on the assumption that the ratio of TBK to SM is stable in healthy adults and the other on a multiple regression TBK-SM prediction equation. DESIGN: Healthy subjects aged >or= 20 y were recruited for body-composition evaluation. TBK and SM were measured by whole-body (40)K counting and multislice magnetic resonance imaging, respectively. A conceptual model with empirically derived data was developed to link TBK and adipose tissue-free SM as the ratio of TBK to SM. RESULTS: A total of 300 subjects (139 men and 161 women) of various ethnicities with a mean (+/- SD) body mass index (in kg/m(2)) of 25.1 +/- 5.4 met the study entry criteria. The mean conceptual model-derived TBK-SM ratio was 122 mmol/kg, which was comparable to the measurement-derived TBK-SM ratios in men and women (119.9 +/- 6.7 and 118.7 +/- 8.4 mmol/kg, respectively), although the ratio tended to be lower in subjects aged >or= 70 y. A strong linear correlation was observed between TBK and SM (r = 0.98, P < 0.001), with sex, race, and age as small but significant prediction model covariates. CONCLUSIONS: Two different types of prediction models were developed that provide validated approaches for estimating SM mass from (40)K measurements by whole-body counting. These methods afford an opportunity to predict SM mass from TBK data collected in healthy adults.

Absorptiometry, Photon↗

Lactate genesis by rat liver and muscle during development.

Lactate has been shown to be an important fuel for brain metabolism during early postnatal development (1). In an attempt to identify the source(s) of lactate in the postnatal rat, we have studied the in vitro catabolism of glucose, galactose, fructose, alanine, glycerol, and octanoate in liver and muscle minces prepared from suckling rat pups. Whereas galactose, fructose, and octanoate were found to be lactagenic (lactate generating) in liver, glucose was the sole lactate precursor in muscle. Galactose was most effective as a hepatic lactate source at 3 d of age. Thereafter, the production of lactate from galactose decreased to reach control levels by 15 d of age. In contrast, fructose or octanoate were lactagenic throughout development. Lactate formation from galactose was completely halted by iodoacetate, inhibited by high galactose concentrations, and suppressed by fasting. The absence of oxygen increased lactate production from either fructose or octanoate, but it did not affect lactagenesis from galactose. Muscle minces produced lactate from glucose in an age-dependent manner similar to the development pattern of lactate formation from galactose by liver. Because lactose-derived galactose is readily available during suckling, it is suggested that galactose-based hepatic lactagenesis serves a unique role in maintaining the supply of lactate during early postnatal development. This hepatic capability may augment glucose-based muscle lactate synthesis at a time when lactate is a major brain fuel.

Aging↗

Quantitative expression analysis of genes affecting muscle growth during development of rainbow trout(Oncorhynchus mykiss).

The molecular characterization of the hyperplasia and hypertrophy that characterize postembryonic muscle development in rainbow trout is of great interest to aquaculturists because of the commercial value of the species. Determination of temporal expression levels of the genes that control muscle development is an important step in molecular analysis. Real-time quantitative reverse transcriptase polymerase chain reaction was used to characterize expression in the muscle of 3 MRF, 2 MEF, and 2 myostatin genes during 9 stages of trout development. Expression of genes that promote muscle growth (MRF and MEF) peaked in swim-up fry, and in some cases again in 25-g, 140-g, and spawning fish. Myostatin genes, which restrict muscle growth, were expressed at very low levels early in development, but their expression levels were elevated in 140-g and spawning fish. Expression levels and the known function of each tested gene were used to infer the extent of hyperplasia, hypertrophy, and restriction of muscle growth during each stage. Both hyperplasia and hypertrophy appeared to peak in swim-up fry and spawning females, and hyperplasia also appeared to peak in 25-g fish. These results should provide valuable information for developmental biologists and those interested in understanding muscle growth in fish.

Analysis of Variance↗

Development of rat muscle during short- and long-term hindlimb suspension.

Histochemical and contractile properties of developing rat soleus (Sol) and plantaris (P) muscles were studied after hindlimb suspension to determine the effects of reduced activity levels on muscle development. Suspension (S) began at age 18 days and lasted for 14, 28, and 206 days, and results were compared with age-matched controls. Body weights were normal until 14 days and Sol growth was inhibited more than P, weighing 38 and 47% of controls at 46 and 224 days compared with 68 and 59% in P. The Sol did not develop into a slow-twitch (ST) muscle as evidenced by faster times to peak tension and half-relaxation times, faster times to develop 50% of maximum tetanic tension (Po) and a mean of 33% fewer ST fibers. Twitch tension and Po were lower in S-Sol and S-P, but force/cross-sectional area was unchanged. Fiber areas were smaller, but no structural changes characteristic of disuse atrophy were found. Fiber type populations were unchanged in P, and contractile properties were only minimally affected, demonstrating the greater importance of activity for ST muscles during development.

Adenosine Triphosphatases↗

Identification of a subpopulation of merozoites of Sarcocystis singaporensis that invades and partially develops inside muscle cells in vitro.

The affinity of merozoites of Sarcocystis singaporensis obtained from the lungs of acutely infected rats to muscle cells and other cell lines grown in vitro was examined. Two distinct types of mature schizonts developed in the lungs 11-13 days p.i. with sporocysts: those containing PAS- merozoites (type 1) which mainly reacted with antibodies prepared against sporozoites, and others containing PAS+ merozoites (type 2) which were antigenically close to bradyzoites. When inoculated onto cell cultures, type 1-merozoites induced schizogonic development in brain capillary endothelial cells of the rat. In contrast, type 2-merozoites invaded L6 myoblasts. In long-term cultures (50 days) of L6 cells, zoites transformed to a 8-15 microns long uninucleate stage which, tentatively, could be unizoite sarcocysts. Although the observed dichotomy in merozoite development is unprecedented in this form, evidence from previous work suggests that these observations are relevant to other Sarcocystis species. The presented cell culture system could be a first step towards successful growth of sarcocysts in vitro.

Animals↗

Direct regulation of the muscle-identity gene apterous by a Hox protein in the somatic mesoderm.

Hox genes control segment identity in the mesoderm as well as in other tissues. Most evidence indicates that Hox genes act cell-autonomously in muscle development, although this remains a controversial issue. We show that apterous expression in the somatic mesoderm is under direct Hox control. We have identified a small enhancer element of apterous (apME680) that regulates reporter gene expression in the LT1-4 muscle progenitors. We show that the product of the Hox gene Antennapedia is present in the somatic mesoderm of the second and third thoracic segments. Through complementary alterations in the Antennapedia protein and in its binding sites on apME680, we show that Antennapedia positively regulates apterous in a direct manner, demonstrating unambiguously its cell-autonomous role in muscle development. Finally, we determine that LT1-4 muscles contain more nuclei in the thorax than in the abdomen and we propose that one of the segmental differences under Hox control is the number of myoblasts allocated to the formation of specific muscles in different segments.

Animals↗

Interaction of the disintegrin and cysteine-rich domains of ADAM12 with integrin alpha7beta1.

We describe a novel interaction between the disintegrin and cysteine-rich (DC) domains of ADAM12 and the integrin alpha7beta1. Integrin alpha7beta1 extracted from human embryonic kidney 293 cells transfected with alpha7 cDNA was retained on an affinity column containing immobilized DC domain of ADAM12. 293 cells stably transfected with alpha7 cDNA adhered to DC-coated wells, and this adhesion was partially inhibited by 6A11 integrin alpha7 function-blocking antibody. The X1 and the X2 extracellular splice variants of integrin alpha7 supported equally well adhesion to the DC protein. Integrin alpha7beta1-mediated cell adhesion to DC had different requirements for Mn2+ than adhesion to laminin. Furthermore, integrin alpha7beta1-mediated cell adhesion to laminin, but not to DC, resulted in efficient cell spreading and phosphorylation of focal adhesion kinase (FAK) at Tyr397. We also show that adhesion of L6 myoblasts to DC is mediated in part by the endogenous integrin alpha7beta1 expressed in these cells. Since integrin alpha7 plays an important role in muscle cell growth, stability, and survival, and since ADAM12 has been implicated in muscle development and regeneration, we postulate that the interaction between ADAM12 and integrin alpha7beta1 may be relevant to muscle development, function, and disease. We also conclude that laminin and the DC domain of ADAM12 represent two functional ligands for integrin alpha7beta1, and adhesion to each of these two ligands via integrin alpha7beta1 triggers different cellular responses.

ADAM Proteins↗

Transgenic mouse models of muscle aging.

In the last decade transgenic animals have been become a powerful and exciting research model to study the molecular mechanisms underlying the cellular and physiological processes such as cell growth, differentiation, apoptosis, and the regulation of specific gene expression. In the context of skeletal muscle development, transgenic mice and gene-targeting approaches have led to the definition of specific roles for Muscle Regulatory Factors (MRFs) during embryogenesis, although less is known about the molecular mechanism underlying skeletal muscle aging. Recent studies using specific models of transgenic mice have added new insights into the muscle aging process, providing a baseline for designing appropriate strategies to attenuate or to reverse the cumulative effects of aging. In this review we discuss some of the transgenic models currently available to address the molecular mechanisms of skeletal muscle senescence. Given the complexity of the aging process, this review should be regarded as a presentation of works in progress rather than a comprehensive description of muscle aging.

Aging↗

Slow troponin T mRNA in striated muscles is expressed in both cell type and developmental stage specific manner.

We have cloned cDNA sequences of both rat and mouse slow troponin T gene. These sequences share a high level of homology with each other and with the human slow troponin T gene although we were unable to detect an alternatively spliced exon present at 3' end of human slow troponin T cDNA in either mouse or rat cDNAs. Northern blot analysis detected a high level expression of slow troponin T in adult mouse Soleus with a lower level expression in mixed postnatal skeletal muscles. Unlike late fetal and postnatal skeletal muscles in which slow troponin T expression is restricted to slow muscle fibre rich regions only, in situ hybridisation analysis detected this isoform to be highly expressed in somitic myotome and all muscle masses at 10-14 days of gestation after which its expression was rapidly downregulated. The unexpected expression of slow troponin T mRNA in fetal heart was apparent by both northern blotting and in situ hybridisation analyses. Slow troponin T mRNA in fetal heart was first detected at 10 day in utero reaching maximum levels of expression at 12-15 days gestation. The slow troponin T in the heart was mainly expressed in the ventral ventricles until day 15 after which low level expression was also observed in both atria. Slow troponin T mRNA in both atrium and ventricle was mainly expressed in outer wall of the myocardium although it was also expressed in interventricular septum. This study therefore shows that in addition to being a cell type specific marker during later fetal and postnatal skeletal muscle development, slow troponin T represented one of the major developmental isoforms expressed in embryonic and fetal skeletal muscle as well as in the cardiac muscle.

Age Factors↗

Localization of regenectin in regenerates of American cockroach (Periplaneta americana) legs.

The localization of regenectin, a sucrose-binding C-type lectin, in the regeneration of the cockroach leg was investigated by immunoblotting and immunofluorescence studies. Regenectin was found to appear transiently around developing muscle cells in regenerating legs in the late stage of regeneration. With maturation of the muscles, it disappeared and was not detectable in completely regenerated legs. These findings suggest that regenectin is a cementing substance connecting developing muscle cells. Regenectin was not detected in embryos or nymphal legs at various developmental stages, suggesting that it might not be involved in normal development of embryos and legs.

Animals↗

Cross-coupling between voltage-dependent Ca2+ channels and ryanodine receptors in developing ascidian muscle blastomeres.

1. Ascidian blastomeres of muscle lineage express voltage-dependent calcium channels (VDCCs) despite isolation and cleavage arrest. Taking advantage of these large developing cells, developmental changes in functional relations between VDCC currents and intracellular Ca2+ stores were studied. 2. Inactivation of ascidian VDCCs is Ca2+ dependent, as demonstrated by two pieces of evidence: (1) a bell-shaped relationship between prepulse voltage and amplitude during the test pulse in Ca2+, but not in Ba2+, and (2) the decay kinetics of Ca2+ currents (ICa) obtained as the size of tail currents. 3. During replacement in the external solution of Ca2+ with Ba2+, the inward current appeared biphasic: it showed rapid decay followed by recovery and slow decay. This current profile was most evident in the mixed bath solution (2 % Ca2+ and 98 % Ba2+, abbreviated to '2Ca/98Ba'). 4. The biphasic profile of I2Ca/98Ba was significantly attenuated in caffeine and in ryanodine, indicating that Ca2+ release is involved in shaping the current kinetics of VDCCs. After washing out the caffeine, the biphasic pattern was reproducibly restored by depolarizing the membrane in calcium-rich solution, which is expected to refill the internal Ca2+ stores. 5. The inhibitors of endoplasmic reticulum (ER) Ca2+-ATPase (SERCAs) cyclopiazonic acid (CPA) and thapsigargin facilitated elimination of the biphasic profile with repetitive depolarization. 6. At a stage earlier than 36 h after fertilization, the biphasic profile of I2Ca/98Ba was not observed. However, caffeine induced a remarkable decrease in the amplitude of I2Ca/98Ba and this suppression was blocked by microinjection of the Ca2+ chelator BAPTA, showing the presence of caffeine-sensitive Ca2+ stores at this stage. 7. Electron microscopic observation shows that sarcoplasmic membranes (SR) arrange closer to the sarcolemma with maturation, suggesting that the formation of the ultrastructural machinery underlies development of the cross-coupling between VDCCs and Ca2+ stores.

Animals↗

Early innervation of abdominal swimmeret muscles in developing lobsters.

The swimmerets in the abdomen of the lobster Homarus americanus are paired external appendages whose back and forth propulsive movements are brought about largely by a group of power and return stroke muscles located in the lateral abdominal cavity. We find functional innervation of these muscles by several excitatory axons and a single inhibitor in embryonic and stage 1 larval lobsters before the external appendages are even formed. This early innervation is via a few nerve bundles in which branches of the motor axons are intertwined in a complex manner. As the swimmerets develop to maturity in later larval and juvenile stages, the innervation consisting usually of several excitor and a single inhibitor synaptic terminals becomes localized to individual muscles. Patterned synaptic activity in these muscles was not seen in the embryonic and larval stages but has been shown in early juvenile stages, when it coincides with the onset of rhythmic movement of the swimmerets. Consequently, such early innervation of the swimmeret muscles may be influential in establishing the central circuitry for the generation of patterned activity, a possibility that was discounted in a previous study (Proc. Natl. Acad. Sci. USA, 70:954-958).

Abdomen↗

Force development in muscle strips of the disused and nondistended rat urinary bladder.

In rats the flow of urine was diverted from the bladder over a period of 7-10 days. Active length-tension curves of muscle strips of such disused bladders did not differ from those of controls, provided lengths were related to optimum length for force development. When, however, lengths were related to length in situ the active length-tension curves were shifted to the left. The muscle strips of the disused bladders showed a higher degree of stiffness than those of controls. The present findings are related to previous ones made on disused but distended bladders showing changes in the opposite direction. Taken together they demonstrate the importance of filling and periodic emptying for the preservation of normal length-tension relations in the bladder.

Animals↗

Breed differences in the histochemical properties of the M. pubo-ischio-femoralis pars medialis myofibre of domestic cocks.

1. Histochemical properties of M. pubo-ischio-femoralis pars medialis (PIF muscle) were compared in 7 breeds of cocks. This muscle was largely composed of Type I fibres and their transitional form (Type I tr). Type IIA fibres were observed in the cranial part. 2. The weight and cross-sectional area of the PIF muscle increased with increasing body weight. However, the relative muscle development to body weight differed among the 7 breeds. 3. A quarter of Type I fibres were of the transitional variety in bantam fowls. Conversely, few, if any, Type I tr fibres were observed in the large breeds where the muscle was poorly developed. 4. As the histochemical properties of Type I fibres made an effective response to the different body weights and the relative PIF muscle development among breeds, it was concluded that PIF muscle performed an important function in supporting the body weight and maintaining posture.

Adenosine Triphosphatases↗

Mechanical work and fatigue: their roles in the development of muscle work capacity.

To compare the effects of mechanical work versus fatigue on the improvement of work capacity of muscle, young, healthy men were selected to life a 45-pound weight with their quadriceps for 30 sessions, followed by 5 sessions of testing. The study consisted of two phases. Subjects in the first phase did equal amounts of mechanical work with both quadriceps, but had different amounts of fatigue from side to side because the rate of work was different. Subjects in the second phase fatigued both quadriceps, but one quadriceps was fatigued with a rest period between contraction cycles, and one was fatigued without a rest period, thus allowing the side with the rest cycle to do more mechanical work. To test the relative effects of training, a transfer-of-training design was used in both phases. The results suggest that fatigue, defined operationally as the inability or unwillingness of the subject to continue the prescribed task under given reinforcement conditions, plays a larger role in the development of work capacity of muscle than the amount of mechanical work per se.

Adolescent↗

Differential effects of neonatal denervation on intrafusal muscle fibers in the rat.

The response of developing muscle spindles to denervation was studied by sectioning the nerve to the medial gastrocnemius muscle of rats at birth. The denervated spindles were examined daily throughout the first postnatal week for changes in ultrastructure and expression of several isoforms of myosin heavy chain (MHC). Each of the three different types of intrafusal muscle fiber exhibited a different response to denervation. Within 5 days after the nerve section nuclear bag2 fibers degenerated completely; nuclear bag1 fibers persisted, but ceased to express the 'spindle-specific' slow-tonic MHC isoform and thereby could not be differentiated from extrafusal fibers; nuclear chain fibers did not form. The capsules of spindles disassembled, hence spindles or their remnants could no longer be identified 1 week after denervation. Neonatal deefferentation has little effect on these features of developing spindles, so removal of afferent innervation is presumably the factor that induces the loss of spindles in denervated muscles. Degeneration of the bag2 fiber, but not bag1 or extrafusal fibers, reflects a greater dependence of the bag2 fiber than the bag1 fiber on afferent innervation for maintenance of its structural integrity. This difference in response of the two types of immature bag fiber to denervation might reflect an origin of the bag2 fibers from a lineage of myogenic cells distinct from that giving rise to bag1 or extrafusal fibers, or a difference in the length of contact with afferents between the two types of bag fiber prior to nerve section.

Animals↗

Patterning muscles using organizers: larval muscle templates and adult myoblasts actively interact to pattern the dorsal longitudinal flight muscles of Drosophila.

Pattern formation in muscle development is often mediated by special cells called muscle organizers. During metamorphosis in Drosophila, a set of larval muscles function as organizers and provide scaffolding for the development of the dorsal longitudinal flight muscles. These organizers undergo defined morphological changes and dramatically split into templates as adult fibers differentiate during pupation. We have investigated the cellular mechanisms involved in the use of larval fibers as templates. Using molecular markers that label myoblasts and the larval muscles themselves, we show that splitting of the larval muscles is concomitant with invasion by imaginal myoblasts and the onset of differentiation. We show that the Erect wing protein, an early marker of muscle differentiation, is not only expressed in myoblasts just before and after fusion, but also in remnant larval nuclei during muscle differentiation. We also show that interaction between imaginal myoblasts and larval muscles is necessary for transformation of the larval fibers. In the absence of imaginal myoblasts, the earliest steps in metamorphosis, such as the escape of larval muscles from histolysis and changes in their innervation, are normal. However, subsequent events, such as the splitting of these muscles, fail to progress. Finally, we show that in a mutant combination, null for Erect wing function in the mesoderm, the splitting of the larval muscles is aborted. These studies provide a genetic and molecular handle for the understanding of mechanisms underlying the use of muscle organizers in muscle patterning. Since the use of such organizers is a common theme in myogenesis in several organisms, it is likely that many of the processes that we describe are conserved.

Actins↗