Reduction In Number Of Fibers Research Articles

Age-Related Alterations at Neuromuscular Junction: Role of Oxidative Stress and Epigenetic Modifications.

With advancing aging, a decline in physical abilities occurs, leading to reduced mobility and loss of independence. Although many factors contribute to the physio-pathological effects of aging, an important event seems to be related to the compromised integrity of the neuromuscular system, which connects the brain and skeletal muscles via motoneurons and the neuromuscular junctions (NMJs). NMJs undergo severe functional, morphological, and molecular alterations during aging and ultimately degenerate. The effect of this decline is an inexorable decrease in skeletal muscle mass and strength, a condition generally known as sarcopenia. Moreover, several studies have highlighted how the age-related alteration of reactive oxygen species (ROS) homeostasis can contribute to changes in the neuromuscular junction morphology and stability, leading to the reduction in fiber number and innervation. Increasing evidence supports the involvement of epigenetic modifications in age-dependent alterations of the NMJ. In particular, DNA methylation, histone modifications, and miRNA-dependent gene expression represent the major epigenetic mechanisms that play a crucial role in NMJ remodeling. It is established that environmental and lifestyle factors, such as physical exercise and nutrition that are susceptible to change during aging, can modulate epigenetic phenomena and attenuate the age-related NMJs changes. This review aims to highlight the recent epigenetic findings related to the NMJ dysregulation during aging and the role of physical activity and nutrition as possible interventions to attenuate or delay the age-related decline in the neuromuscular system.

The imprinted gene Pw1/Peg3 regulates skeletal muscle growth, satellite cell metabolic state, and self-renewal

Pw1/Peg3 is an imprinted gene expressed from the paternally inherited allele. Several imprinted genes, including Pw1/Peg3, have been shown to regulate overall body size and play a role in adult stem cells. Pw1/Peg3 is expressed in muscle stem cells (satellite cells) as well as a progenitor subset of muscle interstitial cells (PICs) in adult skeletal muscle. We therefore examined the impact of loss-of-function of Pw1/Peg3 during skeletal muscle growth and in muscle stem cell behavior. We found that constitutive loss of Pw1/Peg3 function leads to a reduced muscle mass and myofiber number. In newborn mice, the reduction in fiber number is increased in homozygous mutants as compared to the deletion of only the paternal Pw1/Peg3 allele, indicating that the maternal allele is developmentally functional. Constitutive and a satellite cell-specific deletion of Pw1/Peg3, revealed impaired muscle regeneration and a reduced capacity of satellite cells for self-renewal. RNA sequencing analyses revealed a deregulation of genes that control mitochondrial function. Consistent with these observations, Pw1/Peg3 mutant satellite cells displayed increased mitochondrial activity coupled with accelerated proliferation and differentiation. Our data show that Pw1/Peg3 regulates muscle fiber number determination during fetal development in a gene-dosage manner and regulates satellite cell metabolism in the adult.

Adeno-associated Virus Serotype 8 (AAV8) Delivery of Recombinant A20 to Skeletal Muscle Reduces Pathological Activation of Nuclear Factor (NF)-κB in Muscle of mdx Mice

Duchenne muscular dystrophy (DMD) is a genetic muscle disease caused by the absence of a functional dystrophin protein. Lack of dystrophin protein disrupts the dystrophin-glycoprotein complex causing muscle membrane instability and degeneration. One of the secondary manifestations resulting from lack of functional dystrophin in muscle tissue is an increased level of cytokines that recruit inflammatory cells, leading to chronic upregulation of the nuclear factor (NF)-κB. Negative regulators of the classical NF-κB pathway improve muscle health in the mdx mouse model for DMD. We have previously shown in vitro that a negative regulator of the NF-κB pathway, A20, plays a role in muscle regeneration. Here, we show that overexpression of A20 by using a muscle-specific promoter delivered with an adeno-associated virus serotype 8 (AAV8) vector to the mdx mouse decreases activation of the NF-κB pathway in skeletal muscle. Recombinant A20 expression resulted in a reduction in number of fibers with centrally placed nuclei and a reduction in the number of T cells infiltrating muscle transduced with the AAV8-A20 vector. Taken together, we conclude that overexpression of A20 in mdx skeletal muscle provides improved muscle health by reduction of chronic inflammation and muscle degeneration. These results suggest A20 is a potential therapeutic target to ameliorate symptoms of DMD.

Muscles in a mouse model of spinal muscular atrophy show profound defects in neuromuscular development even in the absence of failure in neuromuscular transmission or loss of motor neurons

A mouse model of the devastating human disease “spinal muscular atrophy” (SMA) was used to investigate the severe muscle weakness and spasticity that precede the death of these animals near the end of the 2nd postnatal week. Counts of motor units to the soleus muscle as well as of axons in the soleus muscle nerve showed no loss of motor neurons. Similarly, neither immunostaining of neuromuscular junctions nor the measurement of the tension generated by nerve stimulation gave evidence of any significant impairment in neuromuscular transmission, even when animals were maintained up to 5 days longer via a supplementary diet. However, the muscles were clearly weaker, generating less than half their normal tension. Weakness in 3 muscles examined in the study appears due to a severe but uniform reduction in muscle fiber size. The size reduction results from a failure of muscle fibers to grow during early postnatal development and, in soleus, to a reduction in number of fibers generated. Neuromuscular development is severely delayed in these mutant animals: expression of myosin heavy chain isoforms, the elimination of polyneuronal innervation, the maturation in the shape of the AChR plaque, the arrival of SCs at the junctions and their coverage of the nerve terminal, the development of junctional folds. Thus, if SMA in this particular mouse is a disease of motor neurons, it can act in a manner that does not result in their death or disconnection from their targets but nonetheless alters many aspects of neuromuscular development.

Crossveinless and the TGFβ pathway regulate fiber number in theDrosophilaadult jump muscle

Skeletal muscles are readily characterized by their location within the body and by the number and composition of their constituent muscle fibers. Here, we characterize a mutation that causes a severe reduction in the number of fibers comprising the tergal depressor of the trochanter muscle (TDT, or jump muscle), which functions in the escape response of the Drosophila adult. The wild-type TDT comprises over 20 large muscle fibers and four small fibers. In crossveinless (cv) mutants, the number of large fibers is reduced by 50%, and the number of small fibers is also occasionally reduced. This reduction in fiber number arises from a reduction in the number of founder cells contributing to the TDT at the early pupal stage. Given the role of cv in TGFbeta signaling, we determined whether this pathway directly impacts TDT development. Indeed, gain- and loss-of-function manipulations in the TGFbeta pathway resulted in dramatic increases and decreases, respectively, in TDT fiber number. By identifying the origins of the TDT muscle, from founder cells specified in the mesothoracic leg imaginal disc, we also demonstrate that the TGFbeta pathway directly impacts the specification of founder cells for the jump muscle. Our studies define a new role for the TGFbeta pathway in the control of specific skeletal muscle characteristics.

DIFFUSION TENSOR TRACTOGRAPHY

Benign extracerebral lesions such as meningiomas may cause hemiparesis by compression and deviation without infiltrating the white matter. We used magnetic resonance diffusion tensor imaging and diffusion tensor tractography to investigate the effects of benign extracerebral lesions on the corticospinal tract (CST). Thirteen patients with extracerebral lesions (11 benign meningiomas and 2 benign cysts) underwent magnetic resonance diffusion tensor imaging and diffusion tensor tractography of the CST using fiber assignment by continuous tractography. The CST was reconstructed and assessed by comparing the ipsilateral and unaffected contralateral fibers. The tumor volume, relative fractional anisotropy, fiber deviation, relative fiber number, and relative fiber per voxel were compared between patients without and with temporary presurgical hemiparesis. Seven patients without hemiparesis and five patients with temporary hemiparesis were analyzed; one patient had permanent weakness and was excluded from analysis. There was no significant difference in the tumor volume, relative fractional anisotropy, presence of cerebral edema, or CST deviation between groups. In patients with temporary hemiparesis, the median relative fiber number (mean, 0.35 +/- 0.32) and relative fiber per voxel (mean, 0.49 +/- 0.14) were significantly reduced compared with patients without hemiparesis (0.92 +/- 0.55, P = 0.04; and 0.96 +/- 0.28, P < 0.01, respectively). In patients with benign extracerebral lesions, reduction in fiber number and fiber per voxel, but not fiber deviation, correlated with temporary hemiparesis. Clinical recovery was possible even if the CST fibers detected by diffusion tensor tractography were reduced by benign extracerebral lesions.

The effects of prenatal and postnatal malnutrition on the morphology, differentiation, and metabolism of skeletal striated muscle tissue in rats

To study the contractile properties, metabolism and morphological characteristics of muscles submitted to prenatal and postnatal protein malnutrition. Animals were distributed into two groups: Control, normoprotein diet (CG; n = 15; 5/5/5), and Malnourished, hypoprotein diet (MG; n = 15; 5/5/5), and examined on the 7th, 14th, and 28th days of the experiment. Total body mass, weight, and the contractile properties and morphology of the anterior tibial muscle were assessed. Several 8 microm-thick tissue samples were taken from 7, 14, and 28 day old rats and stained with HE or subjected to NADH-TR or m-ATPase (pH = 4.4) techniques. Body and muscle weight were lower in the malnourished group. On the 7th day of malnutrition, muscle samples exhibited fibers with smaller diameter, higher polymorphism and higher endomysial conjunctive tissue content. Histochemical methods were unable to precisely determine the types of fiber present. On the 14th day, there were smaller muscle fibers, more polymorphism, many of them with central nuclei and moderate endomysial conjunctive tissue content. With reference to contractile properties, the m-ATPase reaction identified both slow and fast fibers. The NADH-TR reaction revealed the following types of fiber: slow oxidative (SO), fast oxidative glycolytic (FOG) and fast glycolytic (FG). On the 28th day smaller, bunched muscle fibers varying shapes. All three types of fiber exhibited unclear recognition limits with respect to contraction and metabolism. Our experimental results suggest that, in addition to the reduction in numbers of fibers, malnutrition retards the differentiation of the morphological, metabolic, and contractile characteristics of skeletal muscle fibers in growing rats.

Myonuclear number and myosin heavy chain expression in rat soleus single muscle fibers after spaceflight.

The effects of 14 days of spaceflight on myonuclear number, fiber size, and myosin heavy chain (MHC) expression in isolated rat soleus muscle fiber segments were studied. Single soleus muscle fibers from rats flown on the Spacelab Life Sciences-2 14-day mission were compared with those from age-matched ground-based control rats by using confocal microscopy and gel electrophoresis. Spaceflight resulted in a significant reduction in the number of fibers expressing type I MHC and an increase in the number of fibers expressing type IIx or IIa MHC. Space-flight also resulted in an increase in the percentage of fibers coexpressing more than one MHC and in the reexpression of the neonatal isoform of MHC in some fibers. Fiber cross-sectional area was significantly reduced in pure type I MHC-expressing fibers and in fibers coexpressing type I+II MHC but not in fibers expressing one or more type II MHC in the flight rats. The number of myonuclei per millimeter was significantly reduced in type I MHC-expressing fibers from the flight rats but was not significantly different in type I+II and type II MHC-coexpressing fibers. Fibers expressing neonatal MHC were similar in size to control fibers but had significantly fewer myonuclei per millimeter than flight fibers not expressing neonatal MHC. In type I MHC-expressing fibers, the reduction in fiber cross-sectional area was greater than the reduction in myonuclear number; thus the average cytoplasmic volume per myonucleus was significantly lower in flight than in control fibers. The reduction in both myonuclear number and fiber size of fibers expressing type I MHC after 14 days of spaceflight supports the hypothesis that changes in the number of myonuclei may be a contributing factor to the reduction in fiber size associated with chronic unloading of the musculature.

Use of the short-term inflammatory response in the mouse peritoneal cavity to assess the biological activity of leached vitreous fibers.

We used a special-purpose glass microfiber sample, Johns-Manville Code 100/475, to study the effects of various acid and alkali treatments on biological activity as assessed by inflammation in the mouse peritoneal cavity, the leaching of Si, and the phase contrast optical microscopy (PCOM) fiber number. We used mild and medium treatments with oxalic acid and Tris buffer and harsh treatment with concentrated HCl and NaOH. Mild oxalic acid and Tris treatment for 2 weeks had no effect on any of the end-points, but prolonging the mild oxalic acid treatment time to 2 months reduced the biological activity and the fiber number. Medium oxalic acid treatment reduced the biological activity and the fiber number and caused a loss of Si. Medium Tris alkali treatment reduced the PCOM-countable fibers and the biological activity but did not cause a substantial loss of Si. Harsh treatment with strong HCl did not affect the fiber number or cause leaching but the biological activity was reduced; strong NaOH reduced the fiber number and biological activity, and caused marked leaching of Si. The medium oxalic acid conditions (pH 1.4) were more acid than those found in lung cells but produced the same effects (reduction in fiber number and biological activity) as the more physiological mild treatment (pH 4.0), when prolonged. This study suggests that medium oxalic acid treatment can be used as a short-term assay to compare loss of Si, reduction in fiber number, and change in biological activity of vitreous fibers.(ABSTRACT TRUNCATED AT 250 WORDS)

Supplementation of a restricted maternal diet with protein or carbohydrate alone prevents a reduction in fetal muscle fibre number in the guinea-pig

A 60% reduction in maternal feed intake is known to cause a reduction of approximately 20% in biceps brachii fibre number in the guinea-pig fetus. This investigation was designed to isolate the dietary component responsible by reducing all dietary components to 60% of the ad lib. level and supplementing the protein, carbohydrate or fat component to the level of the ad lib. intake. Fetal muscles were examined at 50 d gestation to determine numbers of primary and secondary fibres, and at term to determine total fibre number. Fetal and neonatal weights were reduced in all restricted groups (P < 0.05) when compared with ad lib. controls. At term this reduction was significantly less (P < 0.05) in the protein-supplemented group (20%) than in the 60%-restricted and fat-supplemented groups (43%) and the carbohydrate-supplemented group (34%). Biceps brachii fibre numbers were reduced in the 60%-restricted fat-supplemented groups by 14-16%, but fibre numbers were similar in control, protein-supplemented and carbohydrate-supplemented groups. Any reduction in fibre number was in the secondary fibre component of total fibre number. Therefore, biceps brachii fibre numbers were reduced only when maternal diets were deficient in both protein and carbohydrate.

Organization of the proximal, orbital segment of the infraorbital nerve at multiple intervals after axotomy at birth: a quantitative electron microscopic study in rat.

Although much is known of the central consequences of infraorbital nerve (ION) transection at birth, little is known about the effects of this lesion on the organization of the ION itself. To advance our understanding of how deafferentation alters the developing trigeminal neuraxis, 19 newborn rats were subjected to left ION section and perfused 1, 2, 4, 7, 17, or 90 days later. Left IONs were removed in the orbit proximal to the nerve injury site, and axon numbers, types, and fasciculation patterns were assessed with light and electron microscopic methods. Complete axon counts demonstrated that the axotomized ION contained an average (+/- SD) of 13,945 +/- 10,335, 14,112 +/- 3,501, 16,531 +/- 1,904, 9,045 +/- 1,465, 7,018 +/- 4,212, and 8,672 +/- 1,030 axons at the above-listed ages, respectively. These values are well below the 33,059 axons in the normal adult ION (Jacquin et al. [1984] Brain Res. 290:131-135) and the 42,219 axons in the newborn ION (Renehan and Rhoades [1984] Brain Res. 322:369-373). The axotomized ION also contained lower than normal percentages of myelinated axons (26.7% +/- 6.3% on postnatal day 90 vs. 59.7% +/- 6.2% in normal adults). Unmyelinated fibers constituted the vast majority of the remaining fiber types; degenerating fibers never accounted for > 1.6% of all the axons. The number of fascicles making up the axotomized ION overlapped significantly with those found in the normal newborn and adult ION. We conclude that 1) extensive, though variable, axon elimination occurs proximally within one day of the lesion; 2) the 74% reduction in fiber number seen at 90 days is not reliably achieved until postnatal day 7; 3) the higher than normal proportion of unmyelinated axons in the injured ION may underly many of the known effects of neonatal ION injury on the developing whisker-barrel neuraxis; 4) gross changes in ION fasciculation patterns are not prerequisite to injury-induced pattern alterations in the developing trigeminal system.

Why are slow and fast muscles differentially affected during prenatal undernutrition?

The biceps brachii and soleus muscles (fast and slow twitch muscles, respectively) were taken from guinea-pig fetuses, undernourished via maternal dietary intake to 60% of ad lib, and examined for differences from their controls. At birth, muscle fiber number was found to be reduced by 26% in the biceps brachii but was unaltered in the soleus muscle. An examination of the muscles at ages through gestation revealed that any reduction in fiber number was associated with a reduction in the secondary/primary fiber ratio. There was a stage in soleus myogenesis at which fiber number was significantly lower than the control. The deficit was corrected by the continuation of secondary fiber hyperplasia beyond the time of usual cessation. A similar delay was found in the biceps brachii but it did not enable recovery to a full complement of fibers. The difference between the 2 muscles was thought to be related to the relative proportions of primary fibers in the muscles, primary fiber development being apparently unaffected by undernutrition in utero.

Early immobilization of hindleg muscles of dystrophic mice: short-term and long-term effects

Hindleg muscles of normal and dystrophic mice were immobilized unilaterally early during the postnatal period. After 1 week the casts were removed. Of one group of mice hindleg muscles were processed for histopathological and morphometrical evaluation at the end of the immobilization period. Hindlegs of other groups of mice were remobilized for various periods of time before the muscles were examined. In normal mice immobilization of calf muscles that were fixed in a shortened position resulted in atrophy of about 35% compared with untreated muscles. This was accompanied by a reduction in fibre number of about 15%. The antagonists that had been fixed extended, did not show those effects. Immobilization of dystrophic muscles minimized pathology in both agonists and antagonists, although atrophy developed. Upon remobilization the normal muscles resumed postnatal development. They did not deviate from normal, untreated muscles at the age of 3.5 months. Upon remobilization of dystrophic muscles pathology developed, but less severely than during the second and third postnatal week in untreated dystrophic muscles. Significant differences in morphometrical parameters compared with untreated dystrophic muscles were observed during the 3 months remobilization period studied.

Effect of starvation on tissues from the young of four species, with emphasis on the number and diameter of skeletal muscular fibers.

Young rats, rabbits, guinea pigs, and hamsters were decreased in body weight by 39, 45, 34, and 35%, respectively, by a total deprivation of food for 3, 15, 4, and 4.5 days, respectively. The weight of the heart, liver, and kidneys from each of the four species (with the excepetion of the kidneys from the guinea pig) decreased significantly in the starved animals. After starvation in all four species, 0 to 15% of the original weight of the epididymal and perirenal fat pads remained. The effect of total starvation on the weight of skeletal muscles differed for the same muscle in different species and among the three muscles studied within a species. Starvation caused weight losses in the following muscles from the rat, rabbit, guinea pig, and hamster, respectively: soleus, 15, 8, 14, and 30% plantaris, 23, 54, 41 and 24%; biceps brachii, 27, 52, 42, and 29%. The significant loss of weight in the plantaris and biceps brachii muscles from rabbits and guinea pigs were caused by large decreases in the diameter of the fibers, with no change in the number of fibers. Soleus and plantaris muscles from hamsters decreased in weight by a reduction in fiber diameters but no change in the number of fibers; the weight of biceps brachii decreased by a reduction in fiber number only. A reduction in the number of fibers occurred in all muscles from starved rats; the diameter of the fibers was reduced in the plantaris and biceps brachii muscles. No structural damage to the fibers due to starvation was observed under the light microscope in any muscle from the four species.

Numerical analysis of the vestibular nerve in man. A preliminary report.

11 vestibular nerves from individuals of various ages ranging from birth to 85 years have been studied with regard to the numbers of nerve fibers. In this study it has been shown that young children have between 18 773 and 20 212 myelinated vestibular nerve fibers and that there is a clear and statistically significant reduction in number of fibers occurring with increasing age. The reduction averages 40% in the old age group. The study has included a separate investigation of all vestibular branches and the major reduction is found in the ampullary nerve fibers. This part of the observations will be separately published.

Effects of inbreeding on four families of Peppin Merinos

The influence of inbreeding on several fleece and body characters in a strain of Australian Merino sheep is reported. The four families from which data were taken were derived from a single strain of Peppin Merinos. Inbreeding without selection has been carried on since the foundation of the families between 1939 and 1944. Inbreeding is responsible for a reduction in body size, fertility, greasy and clean fleece weight, and wrinkle score, but has relatively little influence on the fleece components (yield, staple length, fibre diameter, and density of fibre population). It follows that the reduction in fleece weight is a direct consequence of a decrease in total fibre number. Since the reduction in fibre number is likely to be a correlated effect of low pre-natal and early post-natal growth rates, it is suggested that the inbreeding has had no genetic effect on the several independent systems involved in adult wool production but has affected only those associated with fitness and vigour. The depression due to inbreeding therefore may not be caused by specific gene fixation but by increased homozygosity per se.