Featured characteristics and pivotal roles of satellite cells in skeletal muscle regeneration Abstract Skeletal muscle, the essential organ for locomotion, as well as energy reservoir and expenditure, has robust regenerative capacity in response to mechanical stress and injury. As muscle-specific stem cells, satellite cells are responsible for providing new myoblasts during the process of muscle growth and regeneration. Self-renewal capacity and the fate of satellite cells are highly regulated and influenced by their surrounding factors, such as extracellular matrix and soluble proteins. The strong myogenic potential of satellite cells makes them a potential resource for stem cell therapy to cure genetic muscle disease and repair injured muscle. Here, we both review key features of satellite cells during skeletal muscle development and regeneration and summarize recent outcomes of satellite cell transplantation studies.

Effect of PGC1-beta ablation on myonuclear organisation Abstract Skeletal muscle fibres are large, elongated multinucleated cells. Each nucleus within a myofibre is responsible for generating gene products for a finite volume of cytoplasm—the myonuclear domain (MND). Variation in MND sizes during atrophy, hypertrophy and disease states, are common. The factors that contribute to definitive MND sizes are not yet fully understood. Previous work has shown that peroxisome proliferator-activated receptor gamma coactivator 1α (PGC1-α) modulates MND volume, presumably to support increased biogenesis of mitochondria. The transcriptional co-regulator peroxisome proliferator-activated receptor gamma coactivator 1β (PGC1-β) is a homologue of PGC1-α with overlapping functions. To investigate the role of this protein in MND size regulation, we studied a mouse skeletal muscle specific knockout (cKO). Myofibres were isolated from the fast twitch extensor digitorum longus (EDL) muscle, membrane-permeabilised and analysed in 3 dimensions using confocal microscopy. PGC1-β ablation resulted in no significant difference in MND size between cKO and wild type (WT) mice, however, subtle differences in nuclear morphology were observed. To determine whether these nuclear shape changes were associated with alterations in global transcriptional activity, acetyl histone H3 immunostaining was carried out. We found there was no significant difference in nuclear fluorescence intensity between the two genotypes. Overall, the results suggest that PGC-1α and PGC-1β play different roles in regulating nuclear organisation in skeletal muscle; however, further work is required to pinpoint their exact functions.

Effects of adrenaline on contractility and endurance of isolated mammalian soleus with different calcium concentrations Abstract The β-adrenergic receptor stimulation improves endurance in fast twitch muscles and these effects are sensitive to extracellular Ca2+ influx. Present study is aimed to determine the effects of adrenaline, with different concentrations of extracellular Ca2+ \(\left( {{\text{Ca}}_{\text{ECF}}^{ 2+ } } \right)\) , on the contractility and endurance of slow twitch muscles during high frequency stimulations (HFS). Isolated soleus of rabbit was electrically stimulated (strength; 50 Hz, duration; 0.5 ms) in the presence (Test) of adrenaline (1 × 10−7 mM) or without adrenaline (CTL). Fatigue was induced with HFS (80 Hz) for the duration of 20 s. Contractions were recorded through isometric transducer connected with Powerlab. Kreb’s buffer was used with three compositions: standard with 2.5 mM Ca2+ (Ca-S), Ca2+ free buffer (Ca-F) and buffer with raised Ca2+ i.e., 10 mM (Ca-R). Muscles endurance was assessed by measuring the decline in tetanic tension in the terms of percentage (%Pmax) and rate of decline in tetanic tension (dP/dt). During 20 s, %Pmax showed reduction of only 10% in Ca-S. This decline was enhanced in Ca-F (50%) and reduced in Ca-R (6%). Effect of adrenaline was observed only in Ca-F where %Pmax was about 20% greater in Test than CTL. These effects were not observed in both Ca-S and Ca-R during 20 s. However, when duration of stimulation was increased to 120 or 150 s in Ca-S and Ca-R respectively, decline in %Pmax was less in Test as compared to CTL. Thus, \({\text{Ca}}_{\text{ECF}}^{ 2+ }\) plays protective role against fatigue during continuous HFS in slow twitch muscles. In addition, adrenaline improves the muscles endurance during fatiguing contraction but these effects are not mediated through \({\text{Ca}}_{\text{ECF}}^{ 2+ }\) influx.

Skeletal muscle cell transplantation: models and methods Abstract Xenografts of skeletal muscle are used to study muscle repair and regeneration, mechanisms of muscular dystrophies, and potential cell therapies for musculoskeletal disorders. Typically, xenografting involves using an immunodeficient host that is pre-injured to create a niche for human cell engraftment. Cell type and method of delivery to muscle depend on the specific application, but can include myoblasts, satellite cells, induced pluripotent stem cells, mesangioblasts, immortalized muscle precursor cells, and other multipotent cell lines delivered locally or systemically. Some studies follow cell engraftment with interventions to enhance cell proliferation, migration, and differentiation into mature muscle fibers. Recently, several advances in xenografting human-derived muscle cells have been applied to study and treat Duchenne muscular dystrophy and Facioscapulohumeral muscular dystrophy. Here, we review the vast array of techniques available to aid researchers in designing future experiments aimed at creating robust muscle xenografts in rodent hosts.

Biotoxins in muscle regeneration research Abstract Skeletal muscles are characterized by their unique regenerative capacity following injury due to the presence of muscle precursor cells, satellite cells. This characteristic allows researchers to study muscle regeneration using experimental injury models. These injury models should be stable and reproducible. Variety of injury models have been used, among which the intramuscular injection of myotoxic biotoxins is considered the most common and widespread method in muscle regeneration research. By using isolated biotoxins, researchers could induce acute muscle damage and regeneration in a controlled and reproducible manner. Therefore, it is considered an easy method for inducing muscle injury in order to understand the different mechanisms involved in muscle injuries and tissue response following injury. However, different toxins and venoms have different compositions and subsequently the possible effects of these toxins on skeletal muscle vary according to their composition. Moreover, regeneration of injured muscle by venoms and toxins varies according to the target of toxin or venom. Therefore, it is essential for researcher to be aware of the mechanism and possible target of toxin-induced injury. The current paper provides an overview of the biotoxins used in skeletal muscle research.

Components of activation heat in skeletal muscle Abstract Activation heat (qA) production by muscle is the thermal accompaniment of the release of Ca2+ from the sarcoplasmic reticulum (SR) into the cytoplasm, its interactions with regulatory proteins and other cytoplasmic Ca2+ buffers and its return to the SR. The contribution of different Ca2+-related reactions to qA is difficult to determine empirically and therefore, for this study, a mathematical model was developed to describe Ca2+ movements and accompanying thermal changes in muscle fibres in response to stimulation. The major sources of heat within a few milliseconds of the initiation of Ca2+ release are Ca2+ binding to Tn and Pv. Ca2+ binding to ATP produces a relatively small amount of heat. Ca2+ dissociation from ATP and Tn, with heat absorption, are of similar time course to the decline of force. In muscle lacking Pv (e.g. mouse soleus), Ca2+ is then rapidly pumped into the SR. In muscles with Pv, Ca2+ that dissociates from Tn and ATP binds to Pv and then dissociates slowly (over 10 s of seconds) and is then pumped into the SR; the net effect of these two processes is heat absorption. It is proposed that this underlies Hill’s “negative delayed heat”. After all the Ca2+ is returned to the SR, qA is proportional to the amount of Ca2+ released into the cytoplasm. In muscles with Pv this is 20–60 s after Ca2+ release; in muscles without Pv, all Ca2+ is returned to the SR soon after the end of force relaxation.

ADF/cofilin regulation from a structural viewpoint Abstract ADF/cofilins disassemble the actin filament and contribute to a wide range of actin dynamics. These proteins are regulated by several factors, including phosphorylation, pH and mechanical forces. Although the cofilin-decorated actin filament structure was published recently, the mechanisms of these regulating factors remain unclear. Models that describe regulation based on the latest structural data and research will be discussed. Aspects about the interaction between actin and cofilin that require further investigation are also discussed.

Cardiomyocyte nuclearity and ploidy: when is double trouble? Abstract Considerable effort has gone into investigating mechanisms that underlie the developmental transition in which mammalian cardiomyocytes (CMs) switch from being able to proliferate during development, to essentially having lost that ability at maturity. This problem is interesting not only for scientific curiosity, but also for its clinical relevance because controlling the ability of mature CMs to replicate would provide a much-needed approach for restoring cardiac function in damaged hearts. In this review, we focus on the propensity of mature mammalian CMs to be multinucleated and polyploid, and the extent to which this may be necessary for normal physiology yet possibly disadvantageous in some circumstances. In this context, we explore whether the concept of the myonuclear domain (MND) in multinucleated skeletal muscle fibers might apply to cardiomyocytes, and whether cardio-MND size might be related to the transition of CMs to become multinuclear. Nuclei in CMs are almost certainly integrators of not only biochemical, but also—because of their central location within the myofibrils—mechanical information, and this multimodal, integrative function in adult CMs—involving molecules that have been extensively studied along with newly identified possibilities—could influence both gene expression as well as replication of the genome and the nuclei themselves.

Mitochondria and autophagy in adult stem cells: proliferate or differentiate Abstract Adult stem cells are undifferentiated cells that are found in many different tissues after development. They are responsible for regenerating and repairing tissues after injury, as well as replacing cells when needed. Adult stem cells maintain a delicate balance between self-renewal to prevent depletion of the stem cell pool and differentiation to continually replenish downstream lineages. The important role of mitochondria in generating energy, calcium storage and regulating cell death is well established. However, new research has linked mitochondria to stem cell maintenance and fate. In addition, efficient mitochondrial quality control is critical for stem cell homeostasis to ensure their long-term survival in tissues. In this review, we discuss the latest evidence linking mitochondrial function, remodeling and turnover via autophagy to regulation of adult stem cell self-renewal and differentiation.

Quantitative high-precision imaging of myosin-dependent filamentous actin dynamics Abstract Over recent decades, considerable effort has been made to understand how mechanical stress applied to the actin network alters actin assembly and disassembly dynamics. However, there are conflicting reports concerning the issue both in vitro and in cells. In this review, we discuss concerns regarding previous quantitative live-cell experiments that have attempted to evaluate myosin regulation of filamentous actin (F-actin) turnover. In particular, we highlight an error-generating mechanism in quantitative live-cell imaging, namely convection-induced misdistribution of actin-binding probes. Direct observation of actin turnover at the single-molecule level using our improved electroporation-based Single-Molecule Speckle (eSiMS) microscopy technique overcomes these concerns. We introduce our recent single-molecule analysis that unambiguously demonstrates myosin-dependent regulation of F-actin stability in live cells. We also discuss the possible application of eSiMS microscopy in the analysis of actin remodeling in striated muscle cells.