Muscle Nuclei Research Articles

Akirin Is Required for Muscle Function and Acts Through the TGF-β Sma/Mab Signaling Pathway in Caenorhabditis elegans Development.

Akirin, a conserved metazoan protein, functions in muscle development in flies and mice. However, this was only tested in the rodent and fly model systems. Akirin was shown to act with chromatin remodeling complexes in transcription and was established as a downstream target of the NFκB pathway. Here we show a role for Caenorhabditis elegans Akirin/AKIR-1 in the muscle and body length regulation through a different pathway. Akirin localizes to somatic tissues throughout the body of C. elegans, including muscle nuclei. In agreement with its role in other model systems, Akirin loss of function mutants exhibit defects in muscle development in the embryo, as well as defects in movement and maintenance of muscle integrity in the C. elegans adult. We also have determined that Akirin acts downstream of the TGF-β Sma/Mab signaling pathway in controlling body size. Moreover, we found that the loss of Akirin resulted in an increase in autophagy markers, similar to mutants in the TGF-β Sma/Mab signaling pathway. In contrast to what is known in rodent and fly models, C. elegans Akirin does not act with the SWI/SNF chromatin-remodeling complex, and is instead involved with the NuRD chromatin remodeling complex in both movement and regulation of body size. Our studies define a novel developmental role (body size) and a new pathway (TGF-β Sma/Mab) for Akirin function, and confirmed its evolutionarily conserved function in muscle development in a new organism.

A minimal constraint device for imaging nuclei in live Drosophila contractile larval muscles reveals novel nuclear mechanical dynamics.

Muscle contractions produce reiterated cytoplasmic mechanical variations, which potentially influence nuclear mechanotransduction, however information regarding the dynamics of muscle nuclei (myonuclei) in the course of muscle contraction is still missing. Towards that end, a minimal constraint device was designed in which intact live Drosophila larva is imaged, while its muscles still contract. The device is placed under spinning disc confocal microscope enabling imaging of fluorescently labeled sarcomeres and nuclei during muscle contraction, without any external stimulation. As a proof of principle we studied myonuclei dynamics in wild-type, as well as in Nesprin/klar mutant larvae lacking proper nuclear-cytoskeletal connections. Myonuclei in control larvae exhibited comparable dynamics in the course of multiple contractile events, independent of their position along the muscle fiber. In contrast, myonuclei of mutant larvae displayed differential dynamics at distinct positions along individual myofibers. Moreover, we identified a linear link between myonuclear volume and its acceleration values during muscle contraction which, in Nesprin/klar mutants exhibited an opposite tendency relative to control. Estimation of the drag force applied on individual myonuclei revealed that force fluctuations in time, but not the average force, differed significantly between control and Nesprin/klar mutant, and were considerably higher in the mutant myonuclei. Taken together these results imply significant alterations in the mechanical dynamics of individual myonuclei in the Nesprin/klar myonuclei relative to control. Such differences provide novel mechanical insight into Nesprin function in contractile muscles, and might reveal the mechanical basis underlying Nesprin-related human diseases.

Novel “omega muscle units” in superficial body-wall myotomes during metamorphosis in the northern brook lamprey (Ichthyomyzonfossor)

Lampreys transform from sedentary filter feeders to more mobile adults through a dramatic metamorphosis that includes remodeling of head muscle and skeletal systems. Metamorphic modifications of body-wall myotomes that could support changes in swimming behavior from larvae to adults have not been previously reported. Thus, transverse sections of northern brook lamprey (Ichthyomyzon fossor Reighard and Cummins, 1916) in larval (n = 4), metamorphosing (n = 3), and adult (n = 2) stages were used to investigate the architecture of body-wall muscle and to detect whether Pax7 and MyoD, proteins important in myogenesis, were co-localized in any muscle nuclei. In addition to myotomal complexity of muscle units composed of parietal and central fibers, there was a novel pattern of omega-shaped muscle units with curves of muscle fibers in the superficial mid-body myotome in metamorphosing lamprey. Small satellite-like cells were identified on central fibers in metamorphosing and adult lamprey muscle using routine histology and immunolocalization of Pax7 and MyoD with antibodies that specifically detect mammalian and teleost proteins. Transient “omega muscle units” may be a marker for impending myotomal growth and increasing swimming efficiency during maturation, possibly restricted to metamorphosis. Finding satellite-like cells suggests that Pax7 and MyoD may have distinctive roles in lamprey myogenesis.

Intermittent application of external positive pressure helps to preserve organ viability during ex vivo perfusion and culture

The perfusion of medium through blood vessels allows the preservation of donor organs and culture of bioengineered organs. However, tissue damage due to inadequate perfusion remains a problem. We evaluated whether intermittent external pressurization would improve the perfusion and viability of organs in culture. A bioreactor system was used to perfuse and culture rat small intestine and femoral muscle preparations. Intermittent positive external pressure (10 mmHg) was applied for 20 s at intervals of 20 s. Intermittent pressurization resulted in uniform perfusion of small intestine preparations and minimal tissue damage after 20 h of perfusion, whereas non-pressurized (control) preparations exhibited significantly worse perfusion of the upper surface than the lower surface and histologic evidence of tissue damage. Longer term studies were undertaken in luciferase-expressing rat femoral muscle preparations. Compared with non-pressurized controls, intermittent pressurization led to better perfusion throughout the 14-day experimental period, improved organ viability as indicated by a higher bioluminescence intensity after perfusion with luciferin, and reduced levels of tissue necrosis with better preservation of vascular structures and skeletal muscle nuclei (histologic analyses). Therefore, intermittent application of external positive pressure improved the perfusion of small intestine and skeletal muscle preparations and enhanced tissue viability when compared with controls. We anticipate that this innovative perfusion technique could be used to improve the preservation of donor organs and culture of bioengineered organs.

Nesprin-1-alpha2 associates with kinesin at myotube outer nuclear membranes, but is restricted to neuromuscular junction nuclei in adult muscle

Nesprins, nuclear envelope spectrin-repeat proteins encoded by the SYNE1 and SYNE2 genes, are involved in localization of nuclei. The short isoform, nesprin-1-alpha2, is required for relocation of the microtubule organizer function from centromeres to the nuclear rim during myogenesis. Using specific antibodies, we now show that both nesprin-1-alpha2 and nesprin-1-giant co-localize with kinesin at the junctions of concatenated nuclei and at the outer poles of nuclear chains in human skeletal myotubes. In adult muscle, nesprin-1-alpha2 was found, together with kinesin, only on nuclei associated with neuromuscular junctions, whereas all adult cardiomyocyte nuclei expressed nesprin-1-alpha2. In a proteomics study, kinesin heavy and light chains were the only significant proteins in myotube extracts pulled down by nesprin-1-alpha2, but not by a mutant lacking the highly-conserved STAR domain (18 amino-acids, including the LEWD motif). The results support a function for nesprin-1-alpha2 in the specific localization of skeletal muscle nuclei mediated by kinesins and suggest that its primary role is at the outer nuclear membrane.

P6503Impact of ascending aortic disease categories on the histopathological degenerative changes in relation to aorta dimensions

Abstract Background Recently a consensus statement on surgical pathology of the aorta was published to improve pathological assessment of non-inflammatory aortic diseases. Purpose We used data of the ascending aorta surgical specimens for aneurysm or dissection to better understand the relationship between the histological medial degenerative changes (MDC) and aortic size assessed by computed tomography. Methods In this retrospective single center study we retrieved 719 ascending aorta surgical specimens from January 2010 until June 2018 and analyzed them according to the consensus statement and scored MDC [elastic fiber fragmentation and/or loss (EFFL), smooth muscle nuclei loss (SMNL), mucoid extracellular matrix accumulation (MEMA); intralamellar (I) or translamellar (T)] and measured medial wall thickness on correlation with imaging data and the status [thoracic aorta aneurysm (TAA), dissection (TAD), bicuspid aortic valve (BAV) or non-BAV]. Results We analyzed 517 patients with degenerative aortic diseases (mean age, 61 years) whose imaging data were obtained, with BAV in 203 (TAD 4%, TAA 96%) and with non-BAV in 314 (TAD 44%, TAA 56%). In TAA subset, scores of EFFL, SMNL and MEMA-T were lower in BAV than in non-BAV (p<0.01). Maximum aortic diameters averaged 50 mm in TAD and 53 mm in TAA. In relation to the aortic diameter, the scores of EFFL, SMNL and MEMA-T were more important in non-BAV subset than in BAV, and in TAD subset than in TAA particularly at the small aortic diameters (<50mm) (Figure). Independent predictors of aortic dissection included thicker medial wall (odds ratio [OR], 6.5; 95% confidence interval [CI], 2.6 to 17.6; p<0.0001) and greater SMNL (OR, 1.2; 95% CI, 1.1 to 1.3; p=0.003). Conclusions Non-BAV aortas were associated with higher scores of MDC than BAV aortas. Advanced MDC was correlated with increased aortic diameter in the ascending aortic diseases. However, in even smaller aortic diameters, MDC in patients with TAD was important.

Aortic Oxidative Stress, Inflammation and DNA Damage Following Pulmonary Exposure to Cerium Oxide Nanoparticles in a Rat Model of Vascular Injury.

Pulmonary exposure to cerium oxide nanoparticles (CeO2 NPs) can occur either at the workplace, or due to their release in the environment. Inhaled CeO2 NPs are known to cross the alveolar–capillary barrier and reach various parts of the body, including the vasculature. The anticancer drug cisplatin (CP) causes vascular damage. However, the effects CeO2 NPs on vascular homeostasis in a rat model of CP-induced vascular injury remain unclear. Here, we assessed the impact and underlying mechanism of pulmonary exposure to CeO2 NPs on aorta in rats given a single intraperitoneal injection of cisplatin (CP, 6 mg/kg) to induce vascular damage. Six days later, the rats were intratracheally instilled with either CeO2 NPs (1 mg/kg) or saline (control), and various variables were studied 24 h thereafter in the aortic tissue. The concentration of reduced glutathione and the activity of catalase were significantly increased in the CP + CeO2 NPs group compared with both the CP + saline and the CeO2 NPs groups. The activity of superoxide dismutase was significantly decreased in the CP + CeO2 NPs group compared with both the CP + saline and CeO2 NPs groups. The expression of nuclear factor erythroid-derived 2-like 2 (Nrf2) by the nuclei of smooth muscles and endocardial cells assessed by immunohistochemistry was significantly augmented in CeO2 NPs versus saline, in CP + saline versus saline, and in CP + CeO2 NPs versus CeO2 NPs. Moreover, the concentrations of total nitric oxide, lipid peroxidation and 8-hydroxy-2-deoxyguanosine were significantly elevated in the CP + CeO2 NPs group compared with both the CP + saline and the CeO2 NPs groups. Similarly, compared with both the CP + saline and CeO2 NPs groups, the combination of CP and CeO2 NPs significantly elevated the concentrations of interleukin-6 and tumour necrosis factor-α. Additionally, aortic DNA damage assessed by Comet assay was significantly increased in CeO2 NPs compared with saline, and in CP + saline versus saline, and all these effects were significantly aggravated by the combination of CP and CeO2 NPs. We conclude that pulmonary exposure to CeO2 NPs aggravates vascular toxicity in animal model of vascular injury through mechanisms involving oxidative stress, Nrf2 expression, inflammation and DNA damage.

A NOVEL ROLE FOR POSTSYNAPTIC γ-SECRETASE IN PROMOTING WNT-MEDIATED SYNAPTIC MATURATION

The normal developmental role of γ-secretase in neurons and at synapses is incompletely understood despite considerable study. We sought to determine if there was a role for γ-secretase components Presenilin and Nicastrin in early synapse development. Using Drosophila null mutants for psn and nct, we examined synaptic structure and organization at the larval neuromuscular junction (NMJ) using immunocytochemistry, confocal microscopy, genetics, tissue-specific rescue experiments, and protein biochemistry. We find that synaptic structure is largely normal in both psn and nct mutants, but there is a severe defect in synaptic maturation. The incidence of pre- to postsynaptic apposition failures is increased 4- to 7-fold and markers associated with a mature synapse are decreased by 85-93%. This is due to disruption in postsynaptic, not presynaptic, γ-secretase. Genetic and biochemical analyses suggest that these defects are due to faulty Wnt signaling at the NMJ – normally, the Wnt receptor Fz2 is cleaved and the C-terminus is imported into muscle nuclei to ensure maturation. This cleavage is absent in psn and nct mutants, suggesting a novel mechanism for γ-secretase by which it cleaves Fz2 to promote synaptic maturation and early development. γ-secretase has a novel role in synaptic maturation, via developmental cleavage of a postsynaptic Wnt receptor. This raises the possibility that early events in synaptic maturation can be linked to later-onset neurodegenerative diseases.

Lactate Stimulates a Potential for Hypertrophy and Regeneration of Mouse Skeletal Muscle

The effects of lactate on muscle mass and regeneration were investigated using mouse skeletal muscle tissue and cultured C2C12 cells. Male C57BL/6J mice were randomly divided into (1) control, (2) lactate (1 mol/L in distilled water, 8.9 mL/g body weight)-administered, (3) cardio toxin (CTX)-injected (CX), and (4) lactate-administered after CTX-injection (LX) groups. CTX was injected into right tibialis anterior (TA) muscle before the oral administration of sodium lactate (five days/week for two weeks) to the mice. Oral lactate administration increased the muscle weight and fiber cross-sectional area, and the population of Pax7-positive nuclei in mouse TA skeletal muscle. Oral administration of lactate also facilitated the recovery process of CTX-associated injured mouse TA muscle mass accompanied with a transient increase in the population of Pax7-positive nuclei. Mouse myoblast-derived C2C12 cells were differentiated for five days to form myotubes with or without lactate administration. C2C12 myotube formation with an increase in protein content, fiber diameter, length, and myo-nuclei was stimulated by lactate. These observations suggest that lactate may be a potential molecule to stimulate muscle hypertrophy and regeneration of mouse skeletal muscle via the activation of muscle satellite cells.

Salmeterol with Liver Depot Gene Therapy Enhances the Skeletal Muscle Response in Murine Pompe Disease.

Gene therapy for Pompe disease with adeno-associated virus (AAV) vectors has advanced into early phase clinical trials; however, the paucity of cation-independent mannose-6-phosphate receptor (CI-MPR) in skeletal muscle, where it is needed to take up acid α-glucosidase (GAA), has impeded the efficacy of Pompe disease gene therapy. Long-acting selective β2 receptor agonists previously enhanced the CI-MPR expression in muscle. In this study we have evaluated the selective β2 agonist salmeterol in GAA knockout mice in combination with an AAV vector expressing human GAA specifically in the liver. Quadriceps glycogen content was significantly decreased by administration of the AAV vector with salmeterol, in comparison with the AAV vector alone (p < 0.01). Importantly, glycogen content of the quadriceps was reduced to its lowest level by the combination of AAV vector and salmeterol administration. Rotarod testing revealed significant improvement following treatment, in comparison with untreated mice, and salmeterol improved wirehang performance. Salmeterol treatment decreased abnormalities of autophagy in the quadriceps, as shown be lower LC3 and p62. Vector administration reduced the abnormal vacuolization and accumulation of nuclei in skeletal muscle. Thus, salmeterol could be further developed as adjunctive therapy to improve the efficacy of liver depot gene therapy for Pompe disease.

Cul4 ubiquitin ligase cofactor DCAF12 promotes neurotransmitter release and homeostatic plasticity.

We genetically characterized the synaptic role of the Drosophila homologue of human DCAF12, a putative cofactor of Cullin4 (Cul4) ubiquitin ligase complexes. Deletion of Drosophila DCAF12 impairs larval locomotion and arrests development. At larval neuromuscular junctions (NMJs), DCAF12 is expressed presynaptically in synaptic boutons, axons, and nuclei of motor neurons. Postsynaptically, DCAF12 is expressed in muscle nuclei and facilitates Cul4-dependent ubiquitination. Genetic experiments identified several mechanistically independent functions of DCAF12 at larval NMJs. First, presynaptic DCAF12 promotes evoked neurotransmitter release. Second, postsynaptic DCAF12 negatively controls the synaptic levels of the glutamate receptor subunits GluRIIA, GluRIIC, and GluRIID. The down-regulation of synaptic GluRIIA subunits by nuclear DCAF12 requires Cul4. Third, presynaptic DCAF12 is required for the expression of synaptic homeostatic potentiation. We suggest that DCAF12 and Cul4 are critical for normal synaptic function and plasticity at larval NMJs.

Silencing of ADAM33 restrains proliferation and induces apoptosis of airway smooth muscle cells in ovalbumin-induced asthma model.

A defibrinogen and metalloproteinase 33 (ADAM33) was reported to play an important role in asthma. Furthermore, ADAM33 may play a possible role in airway remodeling due to its high expression in myo-/fibroblasts, epithelium, as well as the airway smooth muscle cells (ASMCs). Thus, the study is supposed to investigate the effect of the downregulation of ADAM33 on the proliferation and apoptosis of ASMCs in allergic asthma. An ovalbumin-induced asthma model in rats was established for investigating the function of the silencing of ADAM33. ASMCs were cultured and divided into four groups after transfection. The messenger RNA and protein expressions of ADAM33 were measured by reverse transcription quantitative polymerase chain reaction and Western blot analysis. Cell proliferation was tested by cell counting kit-8 and cell apoptosis by TdT-mediated dUTP nick-end labeling. The allergic asthma rats showed a large number of inflammatory cell infiltration, airway smooth muscle hypertrophy and hyperplasia, and increased WA t , WA m , and numbers of bronchial smooth muscle nucleus. Additionally, increased numbers of eosinophils and neutrophils, expressions of immunoglobulin E and interleukin-4, content of airway air pressure, and NO, although decreased in expression of interferon-γ, were exhibited in rats with allergic asthma. In our study, upregulated ADAM33 was found, and after the silencing of ADAM33, decreased proliferation and increased apoptosis of ASMCs were observed. The study evidences that silencing of ADAM33 can decrease the proliferation and increase the apoptosis of ASMCs in a rat model of allergic asthma, suggesting ADAM33 represents a potential investigative focus target aiding allergic asthma.

Mathematical Modeling of Nuclear Trafficking of FOXO Transcription Factors.

Nuclear cytoplasmic flux of Foxo transcription factors is paramount in cellular gene regulation. For example, excluding Foxo from skeletal muscle nuclei is necessary to avoid muscle wasting through elevated protein breakdown. Constructing a mathematical model of the signaling process leading to alteration of Foxo nuclear cytoplasm ratio is useful in predicting and interpreting such ratio changes. In this chapter we derive a general mathematical model for nuclear cytoplasmic flux. We apply this model to Foxo flux and take advantage of rapid phosphorylation approximation and conservation conditions to reduce the Foxo flux model. We constrain our model with data from mouse skeletal muscle with applied IGF. This procedure provides an example of what might be called the central approach of mathematical modeling: The cycling of a biological question through mathematical formulation and back to biological interpretation.

Specialized Positioning of Myonuclei Near Cell-Cell Junctions.

Skeletal muscles are large cells with multiple nuclei that are precisely positioned. The importance of the correct nuclear position is highlighted by the correlation between mispositioned nuclei and muscle disease (Spiro et al., 1966; Gueneau et al., 2009). Myonuclei are generally considered to be equivalent and therefore how far nuclei are from their nearest neighbor is the primary measurement of nuclear positioning. However, skeletal muscles have two specialized cell-cell contacts, the neuromuscular (NMJ) and the myotendinous junction (MTJ). Using these cell-cell contacts as reference points, we have determined that there are at least two distinct populations of myonuclei whose position is uniquely regulated. The post-synaptic myonuclei (PSMs) near the NMJ, and the myonuclei near the myotendinous junction myonuclei (MJMs) have different spacing requirements compared to other myonuclei. The correct positioning of pairs of PSMs depends on the specific action of dynein and kinesin. Positions of the PSMs and MJMs relative to the junctions that define them depend on the KASH-domain protein, Klar. We also found that MJMs are positioned close to the MTJ as a consequence of muscle stretching. Our study defines for the first time that nuclei in skeletal muscles are not all equally positioned, and that subsets of distinct myonuclei have specialized rules that dictate their spacing.

PL - 032 Physiological triggers involved in reduced slow myosin expression in disused postural muscle

PL - 032 Physiological triggers involved in reduced slow myosin expression in disused postural muscle

Rapid tissue histology using multichannel confocal fluorescence microscopy with focus tracking.

Simplified hematoxylin and eosin (H&E) staining followed by cryo-sectioning enables rapid identification of cancerous tissue within the procedure room during Mohs micrographic surgery. Yet, a faster evaluation method is desirable as the staining protocol requires physically sectioning of the tissue after freezing, which leads to prolonged sectioning time along with the frozen artifacts that may occur in frozen sectioning. We present a multichannel confocal microscopy system to rapidly evaluate cancerous tissue. Using the optical sectioning capability of the confocal microscope, optically sectioned images of the freshly excised mouse tissue were acquired and converted into images resembling H&E histology. To show details of the nuclei and structure of the tissue, we applied a newly developed rapid tissue staining method using Hoechst 33342 and Eosin-Y. Line scanning and stitching was performed to overcome the limited field of view of the confocal microscope. Unlike previous confocal systems requiring an additional mechanical device to tilt the sample and match the focus of the objective lens, we developed a focus tracking method to rapidly scan large sample area. The focus tracking provides an effective means of keeping the image of the thick tissue in focus without additional devices. We then evaluated the performance of the confocal microscope to obtain optically sectioned images in thick tissue by comparing fluorescence stained slide images. We also obtained the corresponding H&E histology image to assess the potential of the system as a diagnostic tool. We successfully imaged freshly excised mouse organs including stomach, tumor, and heart within a few minutes using the developed multichannel confocal microscopy and the tissue staining method. Using the pseudocolor method, colors of the acquired confocal grayscale images are converted to furthermore resemble Hematoxylin and Eosin histology. Due to the focus tracking and the line scanning, optically sectioned images were obtained over the large field of view. Comparisons with H&E histology have shown that the confocal images can acquire large details such as the ventricle as well as small details such as muscle fibers and nuclei. This study confirms the use of confocal fluorescence microscopy technique to acquire rapid pathology results using optical sectioning, line scanning and focus tracking. We anticipate that the presented method will enable intraoperative histology and significantly reduce stress on patients undergoing surgery requiring repeated histology examinations.

Angiotensin‐converting enzyme 2 deficiency accelerates and angiotensin 1‐7 restores age‐related muscle weakness in mice

BackgroundA pharmacologic strategy for age‐related muscle weakness is desired to improve mortality and disability in the elderly. Angiotensin‐converting enzyme 2 (ACE2) cleaves angiotensin II into angiotensin 1‐7, a peptide known to protect against acute and chronic skeletal muscle injury in rodents. Since physiological aging induces muscle weakness via mechanisms distinct from other muscle disorders, the role of ACE2‐angiotensin 1‐7 in age‐related muscle weakness remains undetermined. Here, we investigated whether deletion of ACE2 alters the development of muscle weakness by aging and whether angiotensin 1‐7 reverses muscle weakness in older mice.MethodsAfter periodic measurement of grip strength and running distance in male ACE2KO and wild‐type mice until 24 months of age, we infused angiotensin 1‐7 or vehicle for 4 weeks, and measured grip strength, and excised tissues. Tissues were also excised from younger (3‐month‐old) and middle‐aged (15‐month‐old) mice. Microarray analysis of RNA was performed using tibialis anterior (TA) muscles from middle‐aged mice, and some genes were further tested using RT‐PCR.ResultsGrip strength of ACE2KO mice was reduced at 6 months and was persistently lower than that of wild‐type mice (p < 0.01 at 6, 12, 18, and 24‐month‐old). Running distance of ACE2KO mice was shorter than that of wild‐type mice only at 24 months of age [371 ± 26 vs. 479 ± 24 (m), p < 0.01]. Angiotensin 1‐7 improved grip strength in both types of older mice, with larger effects observed in ACE2KO mice (% increase, 3.8 ± 1.5 and 13.3 ± 3.1 in wild type and ACE2KO mice, respectively). Older, but not middle‐aged ACE2KO mice had higher oxygen consumption assessed by a metabolic cage than age‐matched wild‐type mice. Angiotensin 1‐7 infusion modestly increased oxygen consumption in older mice. There was no difference in a wheel‐running activity or glucose tolerance between ACE2KO and wild‐type mice and between mice with vehicle and angiotensin 1‐7 infusion. Analysis of TA muscles revealed that p16INK4a, a senescence‐associated gene, and central nuclei of myofibers increased in middle‐aged, but not younger ACE2KO mice. p16INK4a and central nuclei increased in TA muscles of older wild‐type mice, but the differences between ACE2KO and wild‐type mice remained significant (p < 0.01). Angiotensin 1‐7 did not alter the expression of p16INK4a or central nuclei in TA muscles of both types of mice. Muscle ACE2 expression of wild‐type mice was the lowest at middle age (2.6 times lower than younger age, p < 0.05).ConclusionsDeletion of ACE2 induced the early manifestation of muscle weakness with signatures of muscle senescence. Angiotensin 1‐7 improved muscle function in older mice, supporting future application of the peptide or its analogues in the treatment of muscle weakness in the elderly population.

Mechanical positioning of multiple nuclei in muscle cells.

Many types of large cells have multiple nuclei. In skeletal muscle fibers, the nuclei are distributed along the cell to maximize their internuclear distances. This myonuclear positioning is crucial for cell function. Although microtubules, microtubule associated proteins, and motors have been implicated, mechanisms responsible for myonuclear positioning remain unclear. We used a combination of rough interacting particle and detailed agent-based modeling to examine computationally the hypothesis that a force balance generated by microtubules positions the muscle nuclei. Rather than assuming the nature and identity of the forces, we simulated various types of forces between the pairs of nuclei and between the nuclei and cell boundary to position the myonuclei according to the laws of mechanics. We started with a large number of potential interacting particle models and computationally screened these models for their ability to fit biological data on nuclear positions in hundreds of Drosophila larval muscle cells. This reverse engineering approach resulted in a small number of feasible models, the one with the best fit suggests that the nuclei repel each other and the cell boundary with forces that decrease with distance. The model makes nontrivial predictions about the increased nuclear density near the cell poles, the zigzag patterns of the nuclear positions in wider cells, and about correlations between the cell width and elongated nuclear shapes, all of which we confirm by image analysis of the biological data. We support the predictions of the interacting particle model with simulations of an agent-based mechanical model. Taken together, our data suggest that microtubules growing from nuclear envelopes push on the neighboring nuclei and the cell boundaries, which is sufficient to establish the nearly-uniform nuclear spreading observed in muscle fibers.

The UNC-83/UNC-84 LINC members are required for body wall muscle nuclei positioning in &lt;em&gt;C. elegans&lt;/em&gt;

The UNC-83/UNC-84 LINC members are required for body wall muscle nuclei positioning in &lt;em&gt;C. elegans&lt;/em&gt;

The Protective Effect of Melatonin Against Age-Associated, Sarcopenia-Dependent Tubular Aggregate Formation, Lactate Depletion, and Mitochondrial Changes.

To gain insight into the mechanism of sarcopenia and the protective effect of melatonin, the gastrocnemius muscles of young (3-4 months), early-aged (12 months), and old-aged (24 months) wild-type C57BL/6J female mice were examined by magnetic resonance and microscopy. Locomotor activity, lactate production, and nuclear apoptosis were also assessed. The results support the early onset of sarcopenia at 12 months of age, with reduction of muscle fiber number, muscle weight/body weight ratio, lactate, and locomotor activity. Lipid droplet infiltration and autophagosomes were also detected. These changes driven little effects on the early-aged muscle, but they got worse in old-aged animals by the progressive damage of the muscle. Old-aged muscle showed a reduction of the mitochondrial number, a destruction of the mitochondrial cristae, and swelling. Tubular aggregates and nucleic acid fragmentation were the most striking findings in old-aged muscle, reflecting a broad damage with loss of autophagy efficacy. Oral melatonin administration conserved the normal muscular architecture, weight, muscle fiber number, and activity in the old age; it stimulated lactate production, prevented mitochondrial damage and tubular aggregates, and reduced the percentage of apoptotic nuclei in aged muscles. Altogether, gastrocnemius muscle showed age-mediated signs of sarcopenia that were reduced by melatonin treatment.