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PubMed2001

Differential effects of diminished oestrogen and androgen levels on development of skeletal muscle fibres in hypogonadal mice.

Androgen and oestrogen hormones influence skeletal muscle size and the characteristics of skeletal muscle fibre types. These effects have typically been assessed by producing acute shortages (castration/ovariectomy) or by hormone supplementation. Little evidence exists, however, on how sex hormone shortages affect muscle development from early stages through to adulthood. Using the hypogonadal mouse model (hpg) we examined the effects of diminished androgen and oestrogen upon muscle size and fibre type composition in murine gastrocnemius and soleus muscles. Hypogonadal male soleus muscle was significantly smaller than normal males, and approximated the normal and hypogonadal females weight and fibre type characteristics. The hypogonadal male gastrocnemius muscle, however, was significantly small in comparison with normal and hypogonadal female gastrocnemius muscles, with the type IIB fibre diameters decreased most markedly. The hypogonadal female soleus muscle approximated the normal female phenotype, but the gastrocnemius muscle was larger than the normal female, approximating the size of the normal male gastrocnemius muscle. Here too, the type IIB fibres showed the most alteration, with greatly increased fibre diameters. Appropriate amounts of androgens were necessary for gender-specific patterns of growth in male muscles, whilst similar amounts of oestrogen were necessary for female gastrocnemius muscle growth, but not for female soleus muscle. Hypogonadism in this murine model generally retards muscle development in males, but has no apparent influence or enhances muscle development in females. Type IIB fibres are most dependent upon sex hormones for appropriate development, but this relationship is muscle-specific.

Adenosine Triphosphatases↗
PubMed2004

Maternal UNC-45 is involved in cytokinesis and colocalizes with non-muscle myosin in the early Caenorhabditis elegans embryo.

The Caenorhabditis elegans UNC-45 protein contains tetratricopeptide repeats and a domain with similarity to fungal proteins, and it differentially colocalizes with myosin heavy chain B in the body wall muscles of adult worms. Although it is essential for normal myosin filament assembly in body wall muscle development, strong mutants show a previously unexplained maternal effect. We show here that the UNC-45 protein is maternally contributed and is present in all cells of the early embryo whereas zygotic UNC-45 expression is only detected in the developing muscle cells. Embryos produced from adults with reduced germline expression of UNC-45 exhibit cytokinesis defects suggesting that UNC-45 has a novel role in the early embryo in addition to muscle development. Yeast two-hybrid screens show that UNC-45 can directly interact with NMY-2, a non-muscle type II myosin, and UNC-45 and NMY-2 colocalize at cell boundaries in early embryos. Localization of UNC-45 at these boundaries is dependent upon the presence of NMY-2. Our results suggest that UNC-45 interacts with more than one type of myosin and functions in the embryo to regulate cytoplasmic myosin assembly and/or stability during cytokinesis.

Actins↗
PubMed1996

Innervation regulates myosin heavy chain isoform expression in developing skeletal muscle fibers.

The influence of innervation on primary and secondary myogenesis and its relation to fiber type diversity were investigated in two specific wing muscles of quail embryo, the posterior (PLD) and anterior latissimus dorsi (ALD). In the adult, these muscles are composed almost exclusively of pure populations of fast and slow fibers, respectively. When slow ALD and fast PLD muscles developed in ovo in an aneurogenic environment induced after neural tube ablation, the cardiac ventricular myosin heavy chain (MHC) isoform was not expressed. The adult slow MHC isoform, SM2, appeared by embryonic day 7 (ED 7) in normal innervated slow ALD but was not expressed in denervated muscle. Analysis of in vitro differentiation of myoblasts from fast PLD and slow ALD muscles isolated from ED 7 control and neuralectomized quail embryos showed no fundamental differences in the pattern of MHC isoform expression. Newly differentiated fibers accumulated cardiac ventricular, embryonic fast, slow SM1 and SM3 MHC isoforms. Nevertheless, the expression of slow SM2 isoform in myotubes formed from slow ALD myoblasts only occurred when myoblasts were cultured in the presence of embryonic spinal cord. Our studies demonstrate that the neural tube influences primary as well as secondary myotube differentiation in avian forelimb and facilitates the expression of different MHC, particularly slow SM2 MHC gene expression in slow myoblasts.

Acetylcholinesterase↗
PubMed1981

The in situ staining of muscles during development with the cholinesterase technique.

Developing muscles from forelegs of 11- to 18-day-old mouse embryos were stained in situ for cholinesterase with the copper-ferrocyanide technique. The skin of the legs represents a diffusion barrier for the incubation medium. Therefore, in older embryos the skin was mechanically removed after trypsin digestion. In younger embryos the skin remained on the forelegs after trypsin treatment. With this technique it is possible to follow the establishment of the muscular pattern in the legs.

Animals↗
PubMed1994

Regulation of the rat muscle glycogen phosphorylase-encoding gene during muscle cell development.

The muscle isozyme of glycogen phosphorylase (MGP) catalyzes the hydrolysis hydrolysis of intracellular glycogen in mammalian tissues and is produced in skeletal muscle, brain and heart. The MGP gene is developmentally and neutrally regulated in skeletal muscle, but little is known about the gene's transcriptional regulation. We have isolated and characterized the 5' flanking region of rat MGP. Truncated portions of the MGP 5' flanking region were coupled to the bacterial cat reporter gene and used in transient transfection assays in the mouse muscle C2C12 cell line. The region between -211 and +62 contained the smallest regulatory domain capable of demonstrating developmentally regulated myogenic expression in C2C12 cells. This was in contrast with findings from another investigation that transfected this cell line with human MGP [Lockyer and McCracken, J. Biol. Chem. 266 (1991) 20262-20269]. A 172-nucleotide (nt) region between -839 and -666 functioned as a potent enhancer in C2C12 cells when coupled to its cognate promoter, but not when coupled to a simian virus 40 promoter. This rat MGP enhancer region is 78% identical to a comparable region of the human MGP 5' flanking region, but contains only one putative regulatory element that has been previously identified in other muscle genes. These data suggest that rat MGP transcription in C2C12 muscle cells is modulated by a potent enhancer that utilizes novel regulatory elements.

Amino Acid Sequence↗
PubMed1985

Gating kinetics of nonjunctional acetylcholine receptor channels in developing Xenopus muscle.

The development of nonjunctional acetylcholine (ACh) receptor gating kinetics was studied in myotomal muscle of Xenopus laevis by spectral analysis of ACh-induced noise. The results are consistent with the idea that there is a developmental shift in relative numbers of receptors with fast and slow gating kinetics. For about the first 20 hr following the onset of ACh sensitivity, the majority of channel openings contributing to the ACh noise were slow, having an apparent mean open time of about 3 msec. Shortly before the time of hatching, there began to appear a significant contribution to the noise from channels having a mean open time of less than 1 msec. The relative amount of fast channel activity increased during the 2 days following hatching, and by the age of maturity of the myotomal synapse, most of the ACh noise arose from fast channel activity. A comparison of nonjunctional kinetics with synaptic currents at maturity suggests that the control of channel gating time is similar at junctional and nonjunctional regions.

Animals↗
PubMed1979

Synthesis of the calcium transport ATPase of sarcoplasmic reticulum and other muscle proteins during development of muscles cells in vivo and in vitro.

The effect of medium Ca2+ concentration upon the concentration and the rate of synthesis of muscle proteins was investigated in chicken pectoralis muscle cultures. There is an easily identifiable class of muscle protein which includes the Ca2+-ATPase of sarcoplasmic reticulum, myosin, troponin C, ATP : creatine phosphotransferase, muscle specific actin, tropomysin 1 and 2, and muscle hemagglutinin, which show a large increase in concentration during normal development. The increased synthesis of these proteins was inhibited, without inhibition of cell proliferation, in culture media of relatively low Ca2+ concentration, 0.05--0.3 mM, where fusion was prevented. Similar medium Ca2+ concentration was required for the expression of all these proteins, suggesting their coordinate regulation. The proteins are denoted as 'calcium-modulated proteins'. The increased Ca2+ transport activity of sarcoplasmic reticulum in cultured chicken pectoralis muscle cells during development at 1.8 mM medium calcium concentration represents de novo synthesis of the Ca2+ transport ATPase, as shown by immunoprecipitation, active site labeling and direct identification of the Ca2+ transport ATPase on two-dimensional gel electropherograms of whole muscle homogenates. The concentration and the turnover rate of the majority of the muscle proteins is not affected significantly by medium Ca2+ concentration between 0.06 and 1.8 mM. It is proposed that increase in cytoplasmic free Ca2+ concentration during fusion plays a central role in the regulation of the synthesis of calcium-modulated proteins.

Acetylcholinesterase↗
PubMed1997

Neural regulation of the formation of skeletal muscle phosphorylase kinase holoenzyme in adult and developing rat muscle.

Neural influences on the co-ordination of expression of the multiple subunits of skeletal muscle phosphorylase kinase and their assembly to form the holoenzyme complex, alpha4beta4gamma4delta4, have been examined during denervation and re-innervation of adult skeletal muscle and during neonatal muscle development. Denervation of the tibialis anterior and extensor digitorum longus muscles of the rat hindlimb was associated with a rapid decline in the mRNA for the gamma subunit, and an abrupt decrease in gamma-subunit protein. The levels of the alpha- and beta-subunit proteins in the denervated muscles also declined rapidly, their time course of reduction being similar to that for the gamma-subunit protein, but they did not decrease to the same extent. In contrast with the rapid decline in gamma-subunit mRNA upon denervation, alpha- and beta-subunit mRNAs stayed at control innervated levels for approx. 8-10 days, but then decreased rapidly. Their decline coincided very closely with the onset of re-innervation. Re-innervation of the denervated muscles, which occurs rapidly and uniformly after the sciatic nerve crush injury, produced an eventual slow and prolonged recovery of the mRNA for all three subunits and parallel increases in each of the subunit proteins. A similar co-ordinated increase of both subunit mRNA and subunit proteins of the phosphorylase kinase holoenzyme was observed during neonatal muscle development, during the period when the muscles were attaining their adult pattern of motor activity. The phosphorylase kinase holoenzyme remains in a non-activated form during all of these physiological changes, as is compatible with the presence of the full complement of the regulatory subunits. These data are consistent with a model whereby the transcriptional and translational expression of phosphorylase kinase gamma subunit occurs only with concomitant expression of the alpha and beta subunits. This would ensure that free and unregulated, activated gamma subunit alone, which would give rise to unregulated glycogenolysis, is not produced. The data also suggest that control of phosphorylase kinase subunit expression and the formation of the holoenzyme in skeletal muscle is provided by the motor nerve, probably through imposed levels or patterns of muscle activity.

Aging↗
PubMed1994

Development of the locomotory muscle of the chaetognath Sagitta. 1. Quantitative and qualitative aspects of the body and muscle tissue development within the phylum.

The Chaetognath primary muscle constitutes the main tissue of the body-wall. It is made up of four main longitudinal bands. Muscle fibres are separated from the nervous system. During muscle growth, four myogenic zones produce two types of fibres A and B, forming units. These units ordered in an epithelial-like tissue, show various kind of intercellular junctions. For a given animal, the overall number of A and B fibres units is practically the same in each band. The four myogenic areas are the centre for a two-phase production of A and B fibres which appear at a regular rate in each quadrant, by the simultaneous production of three sets of A fibres and two sets of B fibres. The former differentiate at once, the latter some time later, and always at the same moment in their lives. In this way the number of sets of A and B fibres increases during the animal's life. This hyperplasia is accompanied by the hypertrophy of the cells which make up each group. For any given animal, the ventral quadrants are thicker than the dorsal quadrants. Chaetognath locomotory muscle is a good model on which to study the development of a skeletal muscle. Its development is closely related to the hydroskeleton growth, to the animal's shape and various other constructional factors which play a central role in this very isolated phylum.

Animals↗
PubMed1999

The MADS domain containing transcription factor cMef2a is expressed in heart and skeletal muscle during embryonic chick development.

Muscle enhancer factor 2 (MEF2) proteins are important transcription factors for muscle-specific gene activation. Four family members are known in mammals, referred to as MEF2A, MEF2B, MEF2C, and MEF2D. Here we report the isolation and expression pattern of the chick Mef2a gene (cMef2a). cMef2a expression starts in precardiac mesoderm of HH stage 8 embryos. During further embryonic development expression continues in the heart tube and later in atrium and ventricle. A second cMef2a expression domain appears in somites of stage 13 embryos. Somitic cMef2a expression is limited to the myotome and is not found in newly formed somites until the muscle-specific transcription factors MyoD and myogenin are present. This suggests that activation of the cMef2a gene in skeletal muscle is dependent on these basic helix-loop-helix transcription factors. cMef2a expression in heart and skeletal muscle continues into adulthood when it is also seen in intestinal mesenchyme and in brain.

Amino Acid Sequence↗
PubMed1995

The response of adult and developing rat plantaris muscle to overload.

The effect of overload on the rat plantaris muscle was studied in animals of different ages. Overload was induced by removal of gastrocnemius and soleus muscles. As expected, when the operation was carried out in adults, the plantaris muscle became heavier and stronger. These changes occured within 30 days after the operation. In animals in which the operation was carried out 1-12 days after birth and the muscle examined 6-20 weeks later, different results were obtained. In the group operated at 1-9 days of age, the muscles developed a lower maximal twitch and tetanic tension than the contralateral plantaris muscle. There was no difference in the time to peak or muscle weight between the overloaded and the contralateral muscles. Similar changes were observed in animals where the overload was induced at 11 or 12 days of age except for the weight which was significantly higher than that of the control plantaris muscles. The number of slow fibers increased in animals where overload was induced 11-12 days postnatally or in adults, but not when muscles were overloaded at 9 days of age. The possible reasons for the different response of adult and neonatal muscles to overload are discussed.

Animals↗
PubMed1998

Differential regulation of potassium currents by FGF-1 and FGF-2 in embryonic Xenopus laevis myocytes.

1. Fibroblast growth factors (FGFs) are involved in the regulation of many aspects of muscle development. This study investigated their role in regulating voltage-dependent K+ currents in differentiating Xenopus laevis myocytes. Both FGF-1 and FGF-2 are expressed by developing muscle cells, so their actions were compared. Experiments were performed on cultured myocytes isolated from stage 15 embryos. 2. Long-term exposure of the embryonic myocytes to FGF-1 downregulated inward rectifier K+ current (IK(IR)) density as well as both sustained and inactivating voltage-dependent outward K+ currents (IK,S and IK,I, respectively) and their densities. In contrast, FGF-2 upregulated these currents, although, because of an increase in capacitance caused by FGF-2, current density did not change with this factor. 3. The regulation of IK(IR) by FGF-1 was prevented by the cytoplasmic tyrosine kinase inhibitor herbimycin A, but that of IK,S and IK,I was unaffected, indicating that FGF-1 achieves its regulatory effects on electrical development via separate signalling pathways. The receptor tyrosine kinase inhibitor genistein in isolation suppressed K+ currents, but this may have occurred through a channel-blocking mechanism. 4. In many cells, IK, S was found to be composed of two components with differing voltage dependencies of activation. The FGFs brought about an alteration in the amount of total IK,S by equal effects on each component. Conversely, herbimycin A increased the proportion of low voltage-activated current without affecting total current amplitude. Therefore, we suggest that a single species of channel whose voltage dependence is shifted by tyrosine phosphorylation generates IK,S. 5. In summary, FGF-1 and FGF-2 exert opposite effects on voltage-dependent K+ currents in embryonic myocytes and, furthermore, FGF-1 achieves its effects on different K+ currents via separate second messenger pathways.

Animals↗
PubMed1993

Isolation and characterization of a regulated form of actin depolymerizing factor.

Actin depolymerizing factor (ADF) is an 18.5-kD protein with pH-dependent reciprocal F-actin binding and severing/depolymerizing activities. We previously showed developing muscle down-regulates ADF (J. R. Bamburg and D. Bray. 1987. J. Cell Biol. 105: 2817-2825). To further study this process, we examined ADF expression in chick myocytes cultured in vitro. Surprisingly, ADF immunoreactivity increases during the first 7-10 d in culture. This increase is due to the presence of a new ADF species with higher relative molecular weight which reacts identically to brain ADF with antisera raised against either brain ADF or recombinant ADF. We have purified both ADF isoforms from myocytes and have shown by peptide mapping and partial sequence analysis that the new isoform is structurally related to ADF. Immunoprecipitation of both isoforms from extracts of cells prelabeled with [32P]orthophosphate showed that the new isoform is radiolabeled, predominantly on a serine residue, and hence is called pADF. pADF can be converted into a form which comigrates with ADF on 1-D and 2-D gels by treatment with alkaline phosphatase. pADF has been quantified in a number of cells and tissues where it is present from approximately 18% to 150% of the amount of unphosphorylated ADF. pADF, unlike ADF, does not bind to G-actin, or affect the rate or extent of actin assembly. Four ubiquitous protein kinases failed to phosphorylate ADF in vitro suggesting that ADF phosphorylation in vivo is catalyzed by a more specific kinase. We conclude that the ability to regulate ADF activity is important to muscle development since myocytes have both pre- and posttranslational mechanisms for regulating ADF activity. The latter mechanism is apparently a general one for cell regulation of ADF activity.

Actin Depolymerizing Factors↗
PubMed2002

The structure and functional significance of variations in the connective tissue within muscle.

The amount of intramuscular connective tissue (IMCT) and its morphological distribution is highly variable between muscles of differing function. The functional roles of this component of muscle have been poorly understood, but a picture is gradually emerging of the central role this component has in growth, transmission of mechanical signals to muscle cells and co-ordination of forces between fibres within a muscle. The aim of this review is to highlight recent advances that begin to show the functional significance of some of the variability in IMCT. IMCT has a number of clearly defined roles. It patterns muscle development and innervation, and mechanically integrates the tissue. In developing muscles, proliferation and growth of muscle cells is stimulated and guided by cell-matrix interactions. Recent work has shown that the topography of collagen fibres is an important signal. The timing and rates of expression of connective tissue proteins also show differences between muscles. Discussion of mechanical roles for IMCT has traditionally been limited to the passive elastic response of muscle. However, it is now clear that IMCT provides a matrix to integrate the contractile function of the whole tissue. Mechanical forces are co-ordinated and passed between adjacent muscle cells via cell-matrix interactions and the endomysial connective tissue that links the cells together. An emerging concept is that division of a muscle into fascicles by the perimysial connective tissue is related to the need to accommodate shear strains as muscles change shape during contraction and extension.

Animals↗
PubMed1988

The diagnosis of temporomandibular joint disease: two-compartment arthrography and MR.

The reliability and accuracy of two-compartment temporomandibular joint (TMJ) arthrography was compared with MR imaging on the basis of an analysis of surgical findings obtained from joints that had been studied preoperatively with arthrography or MR or, in some cases, both procedures. Seven hundred forty-three consecutive TMJ arthrograms were successfully obtained in a total of 443 patients by using a single 27-gauge needle and a two-compartment technique in each joint. There was a 100% correlation with surgical findings in 218 radiologically abnormal joints operated on within 90 days of arthrography with respect to the presence or degree of meniscus displacement and normal or abnormal disk morphology and function. In 604 patients 1052 TMJs were studied with high-field-strength surface-coil MR. Surgical findings were available for correlation in 170 of the joints studied. Forty-three joints were studied with both two-compartment arthrography and MR. Eight operated joints had been imaged successfully with both two-compartment arthrography and MR. Both methods of evaluation provided highly reliable and accurate information regarding meniscus position and shape. Arthrography was superior to MR in detecting capsular adhesions and the presence or absence of perforation of the disk or meniscus attachments. Simple meniscectomy (with or without insertion of a temporary Silastic TMJ implant) was the most frequently performed surgical procedure in the series, followed by meniscus repositioning procedures. Joint effusions, failed TMJ implants, and avascular necrosis were demonstrated best with MR. Soft-tissue lesions, including intrinsic degeneration of the meniscus, anomalous muscle development, muscle atrophy, tendinitis, and injuries such as contusions and hematomas, were demonstrated only with MR. Partial-flip-angle GRASS (gradient-recalled acquisition in the steady state) techniques permit both fast scanning and study of functional joint dynamic. Joint fluid may appear as high signal intensity on GRASS images because of T2*-weighting. We recommend MR as the procedure of choice for diagnosis of uncomplicated internal derangements of the TMJ. Two-compartment arthrography with videofluoroscopy is an important ancillary procedure that should be performed whenever capsular adhesions or perforations are suspected and not demonstrated with MR and whenever MR is inconclusive.

Arthrography↗