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Membrane crystals of Ca2+-ATPase in sarcoplasmic reticulum of developing muscle.

The vanadate-induced crystallization of Ca2+-ATPase was analyzed on sarcoplasmic reticulum vesicles isolated between 10 and 28 days of development from pectoralis muscles of chicken. After exposure to Na3VO4 in a Ca2+-free medium, Ca2+-ATPase crystals begin to appear on portions of the surface of a few vesicles, isolated at 18 days of development. Thereafter, the number of vesicles containing Ca2+-ATPase crystals rapidly increases and after 1 week of postnatal development (28 days), it reaches the adult level of about 30% of the vesicle population. These observations are discussed with reference to the mechanism of Ca2+-ATPase crystallization and the regulation of sarcoplasmic reticulum biosynthesis.

Animals↗

Skeletal muscle development in the mouse embryo.

In this review we discuss the recent findings concerning the mechanisms that restrict somitic cells to the skeletal muscle fate, the myogenic regulatory factors controlling skeletal muscle differentiation and specification of myogenic cell lineages, the nature of inductive signals and the role of secreted proteins in embryonic patterning of the myotome. More specifically, we review data which strongly support the hypothesis that Myf-5 plays a unique role in development of epaxial muscle, that MyoD plays a unique role in development of hypaxial muscles derived from migratory myogenic precursor cells, and that both genes are responsible for development of intercostal and abdominal muscles (hypaxial muscles that develop from the dermatomal epithelia). In addition, while discussing upstream and post-translational regulation of myogenic regulatory factors (MRFs), we suggest that correct formation of the myotome requires a complex cooperation of DNA binding proteins and cofactors, as well as inhibitory function of non-muscle cells of the forming somite, whose proteins would sequester and suppress the transcription of MRFs. Moreover, in the third part of our review, we discuss embryonic structures, secreted proteins and myogenic induction. However, although different signaling molecules with activity in the process of somite patterning have been identified, not many of them are found to be necessary during in vivo embryonic development. To understand their functions, generation of multiple mutants or conditional/tissue-specific mutants will be necessary.

Animals↗

Soluble age-related factors from skeletal muscle which influence muscle development.

Successful regeneration of damaged striated muscle in adult mice is dependent on the regeneration of newly differentiated myofibers from proliferating satellite cells and inhibition of scar tissue formation by fibroblasts. As with most tissues, the ability of skeletal muscle to regenerate decreases in older animals. In this study, we have analysed soluble extracts from intact and regenerating skeletal muscle from mice of different ages for their ability to affect avian myogenesis in tissue culture. We were interested in determining whether an age-dependent difference could be detected with this tissue culture bioassay system. Total cell proliferation in the cultures, measured by [3H]thymidine incorporation was increased equally by muscle extracts from both young and older mice but the resulting cell populations differed in proportion of cell types. The ratio of myoblasts to fibroblasts was significantly greater in cultures exposed to extracts from younger mouse muscle as compared with cultures exposed to extracts from older animals. This age-related activity was found to reside in a low molecular weight (MW) (greater than 12 kD) component of the extract. This fraction had dissimilar effects on myoblasts and fibroblasts. Relative to saline controls, myoblast proliferation was increased and fibroblast proliferation decreased. The low MW fraction from younger mouse muscle extracts stimulated myogenic cell proliferation and myotube formation to a greater extent than the similar fraction prepared from older mouse muscle. Conversely, younger mouse muscle fractions had significantly greater inhibitory activity against fibroblast proliferation than did older mouse muscle fractions.

Age Factors↗

Control of facial muscle development by MyoR and capsulin.

Members of the MyoD family of basic helix-loop-helix (bHLH) transcription factors control the formation of all skeletal muscles in vertebrates, but little is known of the molecules or mechanisms that confer unique identities to different types of skeletal muscles. MyoR and capsulin are related bHLH transcription factors expressed in specific facial muscle precursors. We show that specific facial muscles are missing in mice lacking both MyoR and capsulin, reflecting the absence of MyoD family gene expression and ablation of the corresponding myogenic lineages. These findings identify MyoR and capsulin as unique transcription factors for the development of specific head muscles.

Animals↗

Rapid lateral diffusion of extrajunctional acetylcholine receptors in the developing muscle membrane of Xenopus tadpole.

We have studied the lateral diffusion of acetylcholine (ACh) receptors in the extrajunctional region of developing myotomal muscle cell membrane of Xenopus tadpoles by a technique of local inactivation. The myotomal muscle cell surfaces of Xenopus tadpoles were exposed to external solution by gently removing the skin of the tail. The density of ACh receptors was monitored by membrane depolarizations in response to iontophoretically applied pulses of ACh. A pulse of alpha-bungarotoxin was pressure ejected onto the exposed fiber surface, resulting in a rapid local inactivation of the ACh receptors. With time, the functional ACh receptors diffused into the region of inactivation, producing a recovery of ACh response. That the observed recovery of ACh sensitivity is due to diffusion of ACh receptors from the unexposed undersurface of the fiber to the inactivated region was evidenced by the following: (1) no recovery was observed following prolonged toxin application; (2) pretreatment of the muscle cells with concanavalin A, which cross-links and immobilizes ACh receptors, prevented recovery; (3) mapping of ACh response along the muscle cell axis showed that the recovery cannot be accounted for by diffusion along the longitudinal axis of the fiber; and (4) the diffusion coefficients observed after scaling the recovery rate with fiber radius fell within a small range (1.5 to 4.0 X 10(-9) cm2/sec), consistent with diffusion of ACh receptors around the fiber circumference. This finding of rapid lateral diffusion within developing tadpole myotomal muscle membrane supports the notion that the localization of ACh receptors induced by innervation could be achieved by a "diffusion-trap" mechanism where the nerve contact region serves as a trap for rapidly diffusing receptors in the membrane.

Acetylcholine↗

Six myosin heavy chain isoforms are expressed during chick breast muscle development.

Two major embryonic myosin heavy chains are expressed in embryonic chick breast muscle until the first week after hatching. Of these, one is already detected in the 8-day-old embryo. The other appears on day 12. Two putative slow embryonic isoforms represent minor components transiently expressed between days 8 and 12. A neonatal heavy chain is expressed at low concentrations on day 8 and increases with development. It is the only isoform two weeks after hatching, and is ultimately replaced by the fast myosin heavy chain in the adult muscle.

Animals↗

Effects of nicotine administration in developing muscle fibers of rats offspring.

The authors studied the effects of nicotine in developing skeletal muscles of rats. Pregnant Wistar rats received an enteral dose of nicotine (2.5, 5.0 or 10mg/kg, respectively, in groups G2, G5, and G10) from the 10th to the 18th or 20th days of pregnancy. Myotube atrophy was observed mainly in 20-day-old fetuses of G10. Twenty-day-old control fetus and of G2 and G5 had a high sarcoplasmic expression of desmin (weaker in G10). Vimentin expression was similar in 18-day-old fetuses of the control, G2, and G5 groups, but it was weaker in 20-day-old fetuses of the G2 and G5 groups. This would indicate an acceleration of the maturation pattern of vimentin expression in these intoxicated fetus. In conclusion, high doses of nicotine induce myotube atrophy and decrease of the expression of intermediate filaments, whereas relatively low doses of nicotine (G2 and G5) induce an early decrease of vimentin expression with no myofiber atrophy.

Animals↗

Diaphragm muscle development in bottlenose dolphins (Tursiops truncatus).

Being born directly into the aquatic environment creates unique challenges for the breathing muscles of neonatal cetaceans. Not only must these muscles be active at the instant of birth to ventilate the lungs, but their activities must also be coordinated with those of the locomotor muscles such that breathing takes place only at the water's surface. At least one major locomotory muscle of bottlenose dolphins (Tursiops truncatus) has been demonstrated to be well developed and, therefore, able to power the neonatal dolphin's early movements (Dearolf et al. [2000] J Morphol 244:203-215). Thus, because of the demands for coordinated behavior with the locomotor muscles, it is hypothesized that the breathing muscles of bottlenose dolphins, represented in this study by the diaphragm, will also demonstrate adult morphology at birth. However, histochemical and biochemical analyses demonstrate that neonatal dolphins have immature diaphragms, with only 52% of the adult slow fiber-type profile (neonates: 34% slow-twitch fibers; adults: 66% slow-twitch fibers). The developmental state of the dolphin diaphragm is compared to those of other neonatal mammals, using a muscle development index (% slow-twitch fibers in neonatal muscle / % slow-twitch fibers in adult muscle). Fiber-type profiles reported in the literature are used to calculate index values for the diaphragms of altricial rats, rabbits, and cats, intermediate baboons and humans, and precocial sheep and horses. The dolphin is not unique in having an immature diaphragm at birth; however, there is a positive relationship between the developmental state of the diaphragm and the overall developmental state of the neonate. The presence of type IIc ("undifferentiated") fibers in the diaphragms of altricial developers (e.g., rats, rabbits, and cats) is correlated with the slow contraction speeds recorded from their diaphragms. The diaphragms of neonatal horses and dolphins express little to no type IIc fibers and, thus, may have the ability to contract at the speeds required for their increased ventilation rates. These results lead to the modification of the criterion for evaluating the developmental state of a muscle at birth. Thus, the developmental state of a neonatal muscle should be based on both its value of Dearolf et al.'s (2000) developmental index, as well as the percentage of type IIc fibers found in that muscle.

Animals↗

Muscle development in Caenorhabditis elegans: mutants exhibiting retarded sarcomere construction.

We have studied the structural changes within the body-wall muscle cells of Caenorhabditis elegans during postmitotic development. In wildtype, the number of sarcomeres progressively increases, and each sarcomere appears to grow in length and depth continuously during this period. In mature wild-type cells, the anterior-most body-wall muscle cells have 6--7 sarcomeres; the rest have 9--10 sarcomeres per cell. Twelve mutants in the unc-52 II gene exhibit markedly retarded sarcomere construction and progressive paralysis. Several unc-52 mutants, such as the severely paralyzed SU200, produced only 2--3 sarcomeres per body-wall muscle cell, while the other mildly paralyzed unc-52 mutants, such as SU250, build 3--4 sarcomeres per muscle cell. Other structures such as the pharynx and even the noncontractile organelles of the body-wall muscle cells do not appear to be structurally or functionally altered. The unc-52 body-wall sarcomeres become moderately disorganized as they are outstripped by cell growth; sufficient order is preserved, however, so that the majority of thick and thin filaments still interdigitate. The myosin heavy chains of SU200 body-wall muscle fail to accumulate normally, while the pharyngeal myosin heavy chains do not appear to be specifically affected. This biochemical result correlates well with the specificity of morphological changes in the mutant. A model is discussed in which the biochemical and morphological deficits are explained by a simple regulatory mechanism.

Animals↗

Aggregation of myonuclei and the spread of slow-tonic myosin immunoreactivity in developing muscle spindles.

The pattern of regional expression of a slow-tonic myosin heavy chain (MHC) isoform was studied in developing rat soleus intrafusal muscle fibers. Binding of the slow-tonic antibody (ATO) began at the equator of prenatal intrafusal fibers where sensory nerve endings are located, and spread into the polar regions of nuclear bag2 and bag1 fibers but not nuclear chain fibers during ontogeny. The onset of the ATO reactivity coincided with the appearance of equatorial clusters of myonuclei (nuclear bag formations) in bag1 and bag2 fibers. Moreover, the intensity of the ATO reaction was strongest in the region of equatorial myonuclei and decreased with increasing distance from the equator of bag1 and bag2 fibers at all stages of prenatal and postnatal development. The polar expansion of ATO reactivity continued throughout the postnatal development of bag1 fibers, but ceased shortly after birth in bag2 fiber coincident with innervation by motor axons. Thus, afferents that innervate the equator might induce the slow-tonic MHC isoform in bag2 and bag1 fibers by regulating the myosin gene expression by equatorial myonuclei, and efferents or twitch contractile activity might inhibit the spread of the slow-tonic MHC isoform into the poles of bag2 but not bag1 fibers. Absence of ATO binding in chain fibers suggests that chain myotubes may not be as susceptible to the effect of afferents as are myotubes that develop into bag2 and bag1 fibers. The different patterns of slow-tonic MHC expression in the three types of intrafusal fiber may therefore result from the interaction of three elements: sensory neurons, motor neurons, and intrafusal myotubes.

Aging↗

Myofibril and sarcoplasmic reticulum changes during muscle development: activity vs inactivity.

1. The purpose of this study was to determine whether biochemical changes of skeletal muscle that occur as a result of exercise in young rats persist into adulthood. 2. Littermates (10 days old) were assigned to a 3, 6 and 12 week control or training group. In addition, a rest-exercise group (R-E) and exercise-rest (E-R) group were included. 3. The rest-exercise and exercise-rest rats were maintained for the 12 weeks with the first 6 weeks being either rest or exercise and the condition reversed during the last 6 weeks of the experiment. 4. Myofibril ATPase activity of rat plantaris increased from the 10d to 12 week animals (P less than 0.05). As anticipated, training resulted in a lowered activity at 6 and 12 weeks compared to controls. 5. The Ca2+ uptake and Ca2+-ATPase activity of the sarcoplasmic reticulum followed a similar pattern. 6. With regard to the exercise-rest rats, the myofibril and SR ATPase activities at 12 weeks were comparable to the 12 weeks control rats. 7. The rest-exercise group approximated the 12 week training group with regard to myofibril and SR ATPase activities (P less than 0.05). 8. The results suggest that the training adaptations that occur during development of skeletal muscle return to normal, when training ceases in the adult rat. 9. Furthermore, animals that started to train prior to puberty do not have a greater capacity to adapt than animals which initiated training during adulthood.

Aging↗

Muscle development genes: their relevance in neuromuscular disorders.

Myogenesis is a complex cascade of events that involves the specification and differentiation of muscle precursor cells or myoblasts, their fusion to form primary and secondary myotubes and subsequent maturation into muscle fibres. In addition, the development of axial muscle requires the migration of muscle precursor cells. These events are under strict genetic control. The contribution of individual genes to this process has been highlighted both by the phenotype of mice with targeted inactivation of individual myogenic regulatory factors and by rare human disorders in which the involvement of these genes has been demonstrated. The inactivation of known myogenic regulatory genes is associated with abnormal regulation of skeletal muscle differentiation and has an effect on regeneration but does not cause progressive muscle weakness or wasting. This review summarises recent developments in this field and will be of particular relevance to those interested in neuromuscular disorders. We also examine the possibility that some rare human conditions associated with abnormal muscle formation may be due to genetic defects in one of the myogenic regulatory genes.

Animals↗

Spatial and temporal changes in myosin heavy chain gene expression in skeletal muscle development.

Seven myosin heavy chains (MyHC) are expressed in mammalian skeletal muscle in spatially and temporally regulated patterns. The timing, distribution, and quantitation of MyHC expression during development and early postnatal life of the mouse are reported here. The three adult fast MyHC RNAs (IIa, IIb, and IId/x) are expressed in the mouse embryo and each mRNA has a distinct temporal and spatial distribution. In situ hybridization analysis demonstrates expression of IIb mRNA by 14.5 dpc, which proceeds developmentally in a rostral to caudal pattern. IId/x and IIa mRNAs are detectable 2 days later. Ribonuclease protection assays demonstrate that the three adult fast genes are expressed at approximately equal levels relative to each other in the embryo but at quite low levels relative to the two developmental isoforms, embryonic and perinatal. Just after birth major changes in the relative proportions of different MyHC RNAs and protein occur. In all cases, RNA expression and protein expression appear coincident. The changes in MyHC RNA and protein expression are distinct in different muscles and are restricted in some cases to particular regions of the muscle and do not always reflect their distribution in the adult.

Animals↗

Semitendinosus muscle development in fetally decapitated pigs.

The effect of decapitation on semitendinosus muscle growth and development was studied in the fetal pig. Pig fetuses were decapitated at 45 d of gestation and histochemical and biochemical analyses conducted at 110 d of gestation. Decapitation prevented or delayed the conversion of secondary fibers from Type II to Type I histochemistry as is characteristic of the normal semitendinosus. Morphological analyses indicated either a prolonged stage of secondary myofiber formation or an inhibition of secondary myofiber hypertrophy in muscle from decapitated fetuses. Biochemical and histochemical analyses indicated that muscle from decapitated fetuses was biochemically and chemically immature compared with that from normal fetuses.

Adenosine Triphosphatases↗

Growth of axons into developing muscles of the chick forelimb is preceded by cells that stain with Schwann cell antibodies.

A study has been made of the development of limb and muscle nerves in relation to the first appearance of Schwann cells in the flexor digitorum profundus (fdp) and flexor carpi ulnaris (fcu) muscles of the avian forelimb. Schwann cells were identified by immunofluorescent techniques with antibodies to the glycoprotein HNK-1. Myotubes and nerves were identified by using antibodies to myosin and to neurofilament, respectively. At stage 24/25 the brachialis longus inferior (Bli n) and superior (Bls n) nerve trunks within proximal regions of the forelimb were surrounded by Schwann cells. These cells extended in a column for a distance of approximately 100 microns beyond the growing ends of nerves. At stage 26 both interosseus nerve (in n) and the medial-ulnar nerve (m-u n) had formed from the Bli n; each of these branches was surrounded by Schwann cells, which again extended approximately 100 microns beyond the growing ends of the nerves. By stage 26/27 the fdp and fcu muscles were clearly delineated by groups of myotubes. No nerves were detected within these groups; however, Schwann cells were observed between the myotubes. At stage 27 axons had left the in n and m-u n and grown into the fdp and fcu muscles, respectively. These axons were surrounded by Schwann cells. The present observations show that Schwann cells are located ahead of the main limb and muscle nerves as they grow into the fdp and fcu muscles of the limb. It is possible that these Schwann cells play a role in guiding nerves to their correct muscles in the developing chick forelimb.

Animals↗

A zebrafish unc-45-related gene expressed during muscle development.

We report the isolation and expression pattern of zebrafish unc45r, a gene related to Caenorhabditis elegans unc-45. UNC-45 is a muscle-specific protein thought to interact with myosin and promote the assembly of muscle thick filaments during C. elegans development. Zebrafish Unc45r shares sequence features with C. elegans UNC-45, including three tetratricopeptide repeats and a CRO1/She4p homology domain. unc45r is expressed in mesoderm adjacent to the dorsal midline during late gastrula stages and is coexpressed with muscle specific genes in somitic mesoderm during development of trunk skeletal muscle. unc45r is also expressed in cranial skeletal muscle as well as in cardiac and smooth muscle. The isolation of a muscle-specific unc-45 related gene from zebrafish suggests a common mechanism for muscle filament assembly between vertebrates and invertebrates.

Amino Acid Sequence↗