Adult Myogenesis Research Articles

Lab-grown insect meat – Chemical and biological insights – A comprehensive review

Abstract Lab-grown insect meat is a promising alternative to traditional livestock for sustainable food production. This review paper aims to provide a comprehensive overview of the current state of knowledge regarding lab-grown insect meat, emphasizing key aspects such as life cycle assessment, insect cell culture history, technological advancements, and bio-robotics in insect cell culture. Comparisons and challenges between insect and mammalian/avian cell culture methodologies are presented. The nutritional content of edible insects (proximate, amino acid, mineral, and vitamin content) and the potential health benefits of consuming insect meat are discussed. The paper also explores embryonic and adult myogenesis processes in insect cells, as well as the significance of insect body fat and muscle cells in culture. Applications of insect cell culture in various fields, such as food production and pharmaceutical development, are presented. Moreover, the potential occurrence of mutations in lab-grown insect cells is examined. Lastly, the review addresses the drawbacks and limitations of insect labriculture, discussing factors such as scalability, cost-efficiency, and public acceptance. Overall, this comprehensive review provides essential insights into the chemical and biological aspects of lab-grown insect meat, paving the way for further research and development in this emerging field. This article is the first review article reporting the chemical and biological insights of lab-grown insect meat.

Native collagen VI delays early muscle stem cell differentiation.

Adult muscle stem cells (MuSCs) are critical for muscle homeostasis and regeneration, and their behavior relies on a finely regulated niche made of specific extracellular matrix (ECM) components and soluble factors. Among ECM proteins, collagen VI (Col6) influences the mechanical properties of the niche and, in turn, MuSC self-renewal capabilities. Here, we investigated whether Col6 can exert a direct function as a biochemical signal for regulating the stemness and differentiation of murine MuSCs and myoblasts. Native Col6, but not its pepsin-resistant fragment, counteracts the early differentiation of myogenic cells by reducing the expression of differentiation marker genes and preserving stemness features, with inhibition of the canonical Wnt pathway. Our data indicate that extracellular Col6 acts as a soluble ligand in delaying early myogenic differentiation by regulating intracellular signals involved in adult myogenesis.

Histone methyltransferases and regenerative myogenesis: A focus on SETDB1

Adult skeletal muscle is composed of thousands of fibers (also called myofibers) that contract thus allowing voluntary movements. Following an injury, muscle stem cells, surrounding the myofibers, activate, proliferate, and differentiate to form de novo myofibers. These steps constitute a process called adult (or regenerative) myogenesis. This process is possible thanks to various transcription factors sequentially expressed and regulated by epigenetic factors that modulate the chromatin and therefore lead to the regulation of gene expression. Gene expression changes consequently affect the fate of muscle stem cells. Histone Lysine Methyltransferases methylate some histones involved in the repression of gene expression. We describe here the role of SETDB1 during adult myogenesis, notably its role during muscle stem cell differentiation.

Report and Abstracts of the 19th Meeting of the Interuniversity Institute of Myology: Assisi, October 20-23, 2022

After two years of conferences on a virtual platform due to the COVID-19 pandemic, finally, the 19th annual meeting of the Interuniversity Institute of Myology (IIM) has returned to the heart of central Italy, in Assisi, an important cultural hub, which boasts a wide range of historic buildings and museums. This event brought together scientists from around the world providing a valuable opportunity to discuss scientific issues in the field of myology. Traditionally, the meeting particularly encourages the participation of young trainees, and the panel discussions were moderated by leading international scientists, making this a special event where young researchers had the opportunity to talk to prestigious scientists in a friendly and informal environment. Furthermore, the IIM young researchers’ winners for the best oral and poster presentations, became part of the IIM Young Committee, involved in the scientific organization of sessions and roundtables and for the invitation of a main speaker for the IIM 2023 meeting. The four keynote speakers for the IIM Conference 2022 presented new insights into the role of multinucleation during muscle growth and disease, the long-range distribution of giant mRNAs in skeletal muscle, human skeletal muscle remodelling from type 2 diabetic patients and the genome integrity and cell identity in adult muscle stem cells. The congress hosted young PhD students and trainees and included 6 research sessions, two poster sessions, round tables and socio-cultural events, promoting science outreach and interdisciplinary works that are advancing new directions in the field of myology. All other attendees had the opportunity to showcase their work through poster presentations. The IIM meeting 2022 was also part of an advanced training event, which included dedicated round tables and a training session of Advanced Myology on the morning of 23 October, reserved for students under 35 enrolled in the training school, receiving a certificate of attendance. This course proposed lectures and roundtable discussions coordinated by internationally outstanding speakers on muscle metabolism, pathophysiological regeneration and emerging therapeutic approaches for muscle degenerations. As in past editions, all participants shared their results, opinions, and perspectives in understanding developmental and adult myogenesis with novel insights into muscle biology in pathophysiological conditions. We report here the abstracts of the meeting that describe the basic, translational, and clinical research and certainly contribute to the vast field of myology in an innovative and original way.

Gαi2 regulates the adult myogenesis of masticatory muscle satellite cells

Although similar to trunk and limb skeletal muscles, masticatory muscles are believed as unique in both developmental origins and myogenesis. Gαi2 has been demonstrated to promote muscle hypertrophy and muscle satellite cell differentiation in limb muscles. However, the effect of Gαi2 on masticatory muscles is still unexplored. This study aimed to identify the role of Gαi2 in the proliferation and differentiation of masticatory muscle satellite cells, further exploring the metabolic mechanism of masticatory muscles. The proliferation rate, myotube size, fusion index of masticatory muscle satellite cells and Pax7, Myf5, MyoD, Tcf21 and Musculin expressions were significantly decreased by Gαi2 knockdown, while in cells infected with AdV4-Gαi2, the proliferation rate, myotube size, fusion index and Tbx1 expression were significantly increased. Masticatory muscle satellite cells also displayed phenotype transformation as Gαi2 changed. In addition, Gαi2 altered myosin heavy chain (MyHC) isoforms of myotubes with less MyHC-2A expression in siGαi2 group and more MyHC-slow expression in AdV4-Gαi2 group. In conclusion, Gαi2 could positively affect the adult myogenesis of masticatory muscle satellite cells and maintain the superiority of MyHC-slow. Masticatory muscle satellite cells may have their unique Gαi2-regulated myogenic transcriptional networks, although they may share some common characteristics with trunk and limb muscles.

State of the Art Procedures for the Isolation and Characterization of Mesoangioblasts.

Adult skeletal muscle is a dynamic tissue able to regenerate quite efficiently, thanks to the presence of stem cell machinery. Besides the quiescent satellite cells that are activated upon injury or paracrine factors, other stem cells are described to be directly or indirectly involved in adult myogenesis. Mesoangioblasts (MABs) are vessel-associated stem cells originally isolated from embryonic dorsal aorta and, at later stages, from the adult muscle interstitium expressing pericyte markers. Adult MABs entered clinical trials for the treatment of Duchenne muscular dystrophy and the transcriptome of human fetal MABs has been described. In addition, single cell RNA-seq analyses provide novel information on adult murine MABs and more in general in interstitial muscle stem cells. This chapter provides state-of-the-art techniques to isolate and characterize murine MABs, fetal and adult human MABs.

Carm1 and the Epigenetic Control of Stem Cell Function.

Coactivator-associated arginine methyltransferase 1 (CARM1) is a methyltransferase whose function has been highly studied in the context of nuclear receptor signaling. However, CARM1 is known to epigenetically regulate expression of several myogenic genes involved in differentiation such as Myog and MEF2C. CARM1 also acts to regulate myogenesis through its influence on various cellular processes from embryonic to adult myogenesis. First, CARM1 has a crucial role in establishing polarity-regulated gene expression during an asymmetric satellite cell division by methylating PAX7, leading to the expression of Myf5. Second, satellite cells express the CARM1-FL and CARM1-ΔE15 isoforms. The former has been shown to promote pre-mRNA splicing through its interaction with CA150 and U1C, leading to their methylation and increased activity, while the latter displays a reduction in both metrics, thus, modulating alternative pre-mRNA splice forms in muscle cells. Third, CARM1 is a regulator of autophagy through its positive reinforcement of AMPK activity and gene expression. Autophagy already has known implications in ageing and disease, and CARM1 could follow suite. Thus, CARM1 is a central regulator of several important processes impacting muscle stem cell function and myogenesis.

Transcription factor signal transducer and activator of transcription 6 (STAT6) is an inhibitory factor for adult myogenesis

BackgroundThe signal transducer and activator of transcription 6 (STAT6) transcription factor plays a vitally important role in immune cells, where it is activated mainly by interleukin-4 (IL-4). Because IL-4 is an essential cytokine for myotube formation, STAT6 might also be involved in myogenesis as part of IL-4 signaling. This study was conducted to elucidate the role of STAT6 in adult myogenesis in vitro and in vivo.MethodsMyoblasts were isolated from male mice and were differentiated on a culture dish to evaluate the change in STAT6 during myotube formation. Then, the effects of STAT6 overexpression and inhibition on proliferation, differentiation, and fusion in those cells were studied. Additionally, to elucidate the myogenic role of STAT6 in vivo, muscle regeneration after injury was evaluated in STAT6 knockout mice.ResultsIL-4 can increase STAT6 phosphorylation, but STAT6 phosphorylation decreased during myotube formation in culture. STAT6 overexpression decreased, but STAT6 knockdown increased the differentiation index and the fusion index. Results indicate that STAT6 inhibited myogenin protein expression. Results of in vivo experiments show that STAT6 knockout mice exhibited better regeneration than wild-type mice 5 days after cardiotoxin-induced injury. It is particularly interesting that results obtained using cells from STAT6 knockout mice suggest that this STAT6 inhibitory action for myogenesis was not mediated by IL-4 but might instead be associated with p38 mitogen-activated protein kinase phosphorylation. However, STAT6 was not involved in the proliferation of myogenic cells in vitro and in vivo.ConclusionResults suggest that STAT6 functions as an inhibitor of adult myogenesis. Moreover, results suggest that the IL-4-STAT6 signaling axis is unlikely to be responsible for myotube formation.

Aging of the immune system and impaired muscle regeneration: A failure of immunomodulation of adult myogenesis.

Skeletal muscle regeneration that follows acute injury is strongly influenced by interactions with immune cells that invade and proliferate in the damaged tissue. Discoveries over the past 20years have identified many of the key mechanisms through which myeloid cells, especially macrophages, regulate muscle regeneration. In addition, lymphoid cells that include CD8+ T-cells and regulatory T-cells also significantly affect the course of muscle regeneration. During aging, the regenerative capacity of skeletal muscle declines, which can contribute to progressive loss of muscle mass and function. Those age-related reductions in muscle regeneration are accompanied by systemic, age-related changes in the immune system, that affect many of the myeloid and lymphoid cell populations that can influence muscle regeneration. In this review, we present recent discoveries that indicate that aging of the immune system contributes to the diminished regenerative capacity of aging muscle. Intrinsic, age-related changes in immune cells modify their expression of factors that affect the function of a population of muscle stem cells, called satellite cells, that are necessary for normal muscle regeneration. For example, age-related reductions in the expression of growth differentiation factor-3 (GDF3) or CXCL10 by macrophages negatively affect adult myogenesis, by disrupting regulatory interactions between macrophages and satellite cells. Those changes contribute to a reduction in the numbers and myogenic capacity of satellite cells in old muscle, which reduces their ability to restore damaged muscle. In addition, aging produces changes in the expression of molecules that regulate the inflammatory response to injured muscle, which also contributes to age-related defects in muscle regeneration. For example, age-related increases in the production of osteopontin by macrophages disrupts the normal inflammatory response to muscle injury, resulting in regenerative defects. These nascent findings represent the beginning of a newly-developing field of investigation into mechanisms through which aging of the immune system affects muscle regeneration.

Myogenin is an essential regulator of adult myofibre growth and muscle stem cell homeostasis.

Growth and maintenance of skeletal muscle fibres depend on coordinated activation and return to quiescence of resident muscle stem cells (MuSCs). The transcription factor Myogenin (Myog) regulates myocyte fusion during development, but its role in adult myogenesis remains unclear. In contrast to mice, myog-/-zebrafish are viable, but have hypotrophic muscles. By isolating adult myofibres with associated MuSCs, we found that myog-/- myofibres have severely reduced nuclear number, but increased myonuclear domain size. Expression of fusogenic genes is decreased, Pax7 upregulated, MuSCs are fivefold more numerous and mis-positioned throughout the length of myog-/-myofibres instead of localising at myofibre ends as in wild-type. Loss of Myog dysregulates mTORC1 signalling, resulting in an 'alerted' state of MuSCs, which display precocious activation and faster cell cycle entry ex vivo, concomitant with myod upregulation. Thus, beyond controlling myocyte fusion, Myog influences the MuSC:niche relationship, demonstrating a multi-level contribution to muscle homeostasis throughout life.

Avian Satellite Cell Plasticity.

Simple SummaryAdult muscle regeneration and reconstruction is dependent on a population of adult stem cells, known as satellite cells. These cells were suggested to exhibit a certain degree of plasticity, being able to differentiate into lineages unassociated with muscle cells. In this study, we have used a range of visualization methods, as well as PCR, to identify a population of satellite cells obtained from samples of chicken muscles. Then, the cells, expressing a previously introduced detectable transgene, were introduced into chicken embryos and detected after three and eighteen days of their development. The traces of cell populations derived from the introduced satellite cells were detected in a range of embryonic tissues in both of the studied timeframes. The results of this study give further proof of the plasticity of muscle satellite cells, showing the potential locations of their migration during embryonic development.Adult myogenesis is dependent on a population of precursor cells, located between the sarcolemma and the basal lamina of the muscle fiber. These satellite cells, usually present in a quiescent state, become activated in response to mechanical muscle strain, differentiating and fusing to add new nuclei to enlarging muscles. As their myogenic lineage commitment is induced on demand, muscle satellite cells exhibit a certain amount of plasticity, possibly being able to be directed to differentiate into non-myogenic fates. In this study, myosatellite cells were isolated from chicken muscle samples, characterized in vitro and introduced into developing blastoderms. They were further investigated using fluorescence microscopy, immunohistochemistry and PCR, to determine their location in embryos after three and eighteen days. The results of the in vitro analysis confirmed that the cells obtained from the Pectoralis thoracicus are highly myogenic, based on the expression of Pax7, Myogenin, MyoD, Desmin and the myotube assay. Furthermore, the investigation of satellite cells within the embryo showed their migration to the regions of Pectoralis thoracicus, heart, liver, gizzard, proventriculus, intestine and brain. Overall, the results of the study proved the high myogenicity of chicken Pectoralis thoracicus cell isolates, as well as provided new information about their migration pathways following introduction into the blastocyst. The presence of the introduced LacZ or eGFP transgenes across the embryo, even 20 days after myosatellite cell injection, further supports the notion that satellite cells exhibit significant plasticity, potentially transdifferentiating into non-muscle lineages.

Microfluidic 3D Model of Heterochronic Parabiosis to Study Systemic Regulation of Skeletal Muscle Aging

Muscle satellite cells (MuSCs) play an indispensable role in skeletal muscle regeneration after injury. However, their regenerative capacity declines with aging. Intriguingly, studies using parabiosis, a surgical technique that attaches two animals to share blood circulation, demonstrated that circulating factors from young animals can rejuvenate the regenerative capacity of aged animals. However, the exact identity of these putative youthful factors has yet to be elucidated. Due to the complexity of in vivo parabiosis, reliable identification of systemic rejuvenation factors remains a major hurdle. To overcome this challenge, we employed microengineering technologies and developed a 3D microfluidic circuit with biophysical and biochemical properties similar to in vivo muscle microenvironment. Here, we present a 3D micro‐vascularized muscle‐on‐a‐chip (VMoC) that exhibits key characteristics of the native MuSC niche, where FACS purified quiescent MuSCs are cultured in between myofibers and 3D hydrogel (extracellular matrix mimetic) underneath vascular endothelial cells. With the VMoC, we further integrated it to model parabiosis by co‐culturing cells/sera from young and aged animals. By adding the serum from young, aged, and Sod1−/− (oxidative stress model) mice to VMoCs, we demonstrate distinct myogenic activities of young MuSCs; VMoCs with aged and Sod1−/− sera showed significantly reduced myogenesis as measured by fusion index. Next, we replicated heterochronic parabiosis using our integrated chip‐to‐chip platform to assess the rejuvenating effects of young serum/cells. Interestingly, similar to in vivo parabiosis, when aged VMoCs were continuously exposed to the young systemic environment, the myogenic activity of aged MuSCs were significantly boosted. In addition, genes that regulate different steps of adult myogenesis (Desmin, MyoD, and Myogenin) were up‐regulated in the aged VMoC exposed to the young systemic environment. Furthermore, this rejuvenation effect in myogenesis was correlated with increased VEGF, which has been shown to play a crucial role in MuSC function in parabiosed VMoC. Collectively, our parabiosis‐on‐a‐chip platform will provide a state‐of‐the‐art pre‐clinical testing tool that will facilitate our understanding of the dynamic regulation of circulating humoral factors and potential drug discovery. More importantly, our approach can be translated into human clinical studies by replacing the murine cells with the corresponding human cells, emulating human parabiosis on‐a‐chip that blood transfusion cannot recapitulate.Support or Funding InformationNIH R21AR072287 (YJ), NIH R03AG062976 (YJ), and NIH DP2HL142050 (YK)

Interleukin‐4 administration improves muscle function, adult myogenesis, and lifespan of colon carcinoma‐bearing mice

BackgroundAnorexia, body wasting, inflammation, muscle, and adipose tissue loss are hallmarks of cancer cachexia, a syndrome that affects the majority of cancer patients, impairing their ability to endure chemotherapeutic therapies and reducing their lifespan. In the last 10 years, alterations of protein turnover and impairment of adult myogenesis have been proposed as major contributing factors.MethodsMuscle stem cells, including satellite cells, mesoangioblasts, and fibroadipogenic progenitors, were isolated and characterized from C26 colon carcinoma‐bearing (C26) mice. Circulating levels of interleukin‐4/13 (IL4/IL13) were analysed by ELISA, and the effects of IL4 on muscle mass and function, protein synthesis, muscle regeneration, and myogenic progenitor cell number were analysed at both functional (treadmill and grip test) and molecular levels (qRT–PCR, immunofluorescence analysis, surface sensing of translation, and western blot). The Kaplan–Meier test was used to analyse the survival curve of IL4‐treated and IL4‐untreated C26 mice.ResultsThe administration of IL4 to C26 mice rescued muscle mass by increasing protein synthesis. The IL4 treatment improved performances and prolonged survival of C26 mice. IL4 administration re‐established both number and function of satellite cells and fibroadipogenic progenitors without affecting mesoangioblasts in C26 mice, rescuing myogenesis. Upon IL4 treatment, a high number of cytotoxic lymphocytes and type II macrophages were observed with a subsequent increase in necrotic areas of C26 tumours.ConclusionsThe results here presented shed new light on IL4 signalling during muscle wasting and early stages of muscle regeneration that explain the beneficial effect observed in IL4‐treated C26 mice. These findings might aid to develop therapeutic approaches to improve mobility and quality of life in cachectic patients.

Rbm24 modulates adult skeletal muscle regeneration via regulation of alternative splicing.

Rationale: The adult skeletal muscle can self-repair efficiently following mechanical or pathological damage due to its remarkable regenerative capacity. However, regulatory mechanisms underlying muscle regeneration are complicated and have not been fully elucidated. Alternative splicing (AS) is a major mechanism responsible for post-transcriptional regulation. Many aberrant AS events have been identified in patients with muscular dystrophy which is accompanied by abnormal muscle regeneration. However, little is known about the correlation between AS and muscle regeneration. It has been reported that RNA binding motif protein 24 (Rbm24), a tissue-specific splicing factor, is involved in embryo myogenesis while the role of Rbm24 in adult myogenesis (also called muscle regeneration) is poorly understood.Methods: To investigate the role of Rbm24 in adult skeletal muscle, we generated Rbm24 conditional knockout mice and satellite cell-specific knockout mice. Furthermore, a cardiotoxin (CTX)-induced injury model was utilized to assess the effects of Rbm24 on skeletal muscle regeneration. Genome-wide RNA-Seq was performed to identify the changes in AS following loss of Rbm24.Results: Rbm24 knockout mice displayed abnormal regeneration 4 months after tamoxifen treatment. Using RNA-Seq, we found that Rbm24 regulated a complex network of AS events involved in multiple biological processes, including myogenesis, muscle regeneration and muscle hypertrophy. Moreover, using a CTX-induced injury model, we showed that loss of Rbm24 in skeletal muscle resulted in myogenic fusion and differentiation defects and significantly delayed muscle regeneration. Furthermore, satellite cell-specific Rbm24 knockout mice recapitulated the defects in regeneration seen in the global Rbm24 knockout mice. Importantly, we demonstrated that Rbm24 regulated AS of Mef2d, Naca, Rock2 and Lrrfip1 which are essential for myogenic differentiation and muscle regeneration.Conclusions: The present study demonstrated that Rbm24 regulates dynamic changes in AS and is essential for adult skeletal muscle regeneration.

Heparan Sulfate Mimetics Accelerate Postinjury Skeletal Muscle Regeneration.

Although skeletal muscle is capable of complete recovery after an injury, specific situations require support or acceleration of this process, such as in the elderly and athletes, respectively. Skeletal muscle regeneration is due to muscle stem cells (MuSCs) that undergo adult myogenesis, a process sustained by MuSC environment. Although recognized as important, extracellular matrix (ECM) has been overlooked in this process. Matrix-based therapy aims at improving ECM remodeling to support tissue repair. In this context, we investigated the properties of a single injection of the clinical grade glycosaminoglycan mimetics RGTA® (ReGeneraTing Agents) on skeletal muscle regeneration in a context compatible with a clinical application, that is, 3 days after the injury. Our results show that RGTA-treated muscles showed an increase of the number of myonuclei in regenerating myofibers and an increase of the capillarization of the new myofibers. In vitro experiments showed that RGTA directly acts on MuSCs by stimulating their fusion into myotubes and on endothelial cells by stimulating the formation and maturation of vessels in a 3D culture setup. These results indicate that a single administration of RGTA in regenerating muscle stimulated both myogenesis and angiogenesis, thus accelerating skeletal muscle regeneration. Impact Statement Although highly powerful in normal condition, postinjury skeletal muscle regeneration is less efficient in some situations, such as obese, elderly, or resting people. In other context, such as high-performance sport, skeletal muscle regeneration must be shortened but in a way ensuring a full functional recovery. In this context, our results show that a single injection of the clinical grade glycosaminoglycan mimetics RGTA® (ReGeneraTing Agents), in a context compatible with a clinical application, that is, 3 days after the injury, is beneficial for skeletal muscle regeneration, through the stimulation of both myogenesis and angiogenesis.

MTOR controls embryonic and adult myogenesis via mTORC1.

The formation of multi-nucleated muscle fibers from progenitors requires the fine-tuned and coordinated regulation of proliferation, differentiation and fusion, both during development and after injury in the adult. Although some of the key factors that are involved in the different steps are well known, how intracellular signals are coordinated and integrated is largely unknown. Here, we investigated the role of the cell-growth regulator mTOR by eliminating essential components of the mTOR complexes 1 (mTORC1) and 2 (mTORC2) in mouse muscle progenitors. We show that inactivation of mTORC1, but not mTORC2, in developing muscle causes perinatal death. In the adult, mTORC1 deficiency in muscle stem cells greatly impinges on injury-induced muscle regeneration. These phenotypes are because of defects in the proliferation and fusion capacity of the targeted muscle progenitors. However, mTORC1-deficient muscle progenitors partially retain their myogenic function. Hence, our results show that mTORC1 and not mTORC2 is an important regulator of embryonic and adult myogenesis, and they point to alternative pathways that partially compensate for the loss of mTORC1.This article has an associated 'The people behind the papers' interview.

β-Catenin is essential for differentiation of primary myoblasts via cooperation with MyoD and α-catenin.

Canonical Wnts promote myoblast differentiation; however, the role of β-catenin in adult myogenesis has been contentious, and its mechanism(s) unclear. Using CRISPR-generated β-catenin-null primary adult mouse myoblasts, we found that β-catenin was essential for morphological differentiation and timely deployment of the myogenic gene program. Alignment, elongation and fusion were grossly impaired in null cells, and myogenic gene expression was not coordinated with cytoskeletal and membrane remodeling events. Rescue studies and genome-wide analyses extended previous findings that a β-catenin-TCF/LEF interaction is not required for differentiation, and that β-catenin enhances MyoD binding to myogenic loci. We mapped cellular pathways controlled by β-catenin and defined novel targets in myoblasts, including the fusogenic genes myomaker and myomixer. We also showed that interaction of β-catenin with α-catenin was important for efficient differentiation. Overall the study suggests dual roles for β-catenin: a TCF/LEF-independent nuclear function that coordinates an extensive network of myogenic genes in cooperation with MyoD; and an α-catenin-dependent membrane function that helps control cell-cell interactions. β-Catenin-TCF/LEF complexes may function primarily in feedback regulation to control levels of β-catenin and thus prevent precocious/excessive myoblast fusion.

Sonic hedgehog-regulated myotome formation relies on the myomiR, miR-133

Over the past two decades a great deal of information has been provided concerning myogenesis—the process of skeletal muscle formation. Much is known about the cellular and molecular mechanisms of adult myogenesis—the process through which adult muscle regenerates following acute injury and embryonic myogenesis—the process of skeletal muscle tissue development (1,2); however, the picture of the molecular mechanisms governing myogenesis is far from being complete. A recent paper by Mok et al. (3) adds an important piece of information to the mosaic of molecules/factors driving the ordered sequence of muscle-related events leading to cell fate decisions in developing embryos.

Systematic Screening for Transcriptional Regulators of Adult Myogenesis in Drosophila by RNAi

The muscles of Drosophila show similar structure and pathway of development to the muscles of humans, and allows Drosophila to model human muscle disorders. However, little is known about many of the genes and their potential role in muscle development. Thus, the focus of this project is to efficiently screen for potential transcriptional regulators of muscle development and identify associated disorder phenotypes. Genes were selected based on having a potential role in transcription, which were silenced using RNA interference (RNAi). The 1151‐gal4 driver was utilized to induce RNAi in developing adult muscles. Combined with the driver, the flies harbored enhancers of either Flightin‐LacZ or TpnC41C‐LacZ. The resulting transgenic flies were collected and assayed to measure the effects of this genetic loss. All 101 genes in the latest set have been assayed for a change of enhancer activity of less than 70% or greater than 130%, in Flightin and TpnC41C; the genes selected for immunostaining exhibited semi‐lethality or lethality, and indicated a loss or gain of LacZ. The immunostaining would reveal several factors: muscle morphology, and expansion or loss of enhancer activity associated with specific flight and jump muscle genes. The resulting data was classified into three categories, severe loss, moderate loss, or no change in muscle development. Genes with a severe loss indicates a potential issue with myoblasts fusion, and genes with a moderate loss indicates issues with adult muscle formation and maturation. Therefore, the results demonstrate that the screen is efficient in identifying potential transcriptional regulators and muscle disorder phenotypes.Support or Funding InformationNational Institute of Health ROI GM061738This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Small-RNA sequencing identifies dynamic microRNA deregulation during skeletal muscle lineage progression

Skeletal muscle satellite cells are quiescent adult resident stem cells that activate, proliferate and differentiate to generate myofibres following injury. They harbour a robust proliferation potential and self-renewing capacity enabling lifelong muscle regeneration. Although several classes of microRNAs were shown to regulate adult myogenesis, systematic examination of stage-specific microRNAs during lineage progression from the quiescent state is lacking. Here we provide a genome-wide assessment of the expression of small RNAs during the quiescence/activation transition and differentiation by RNA-sequencing. We show that the majority of small RNAs present in quiescent, activated and differentiated muscle cells belong to the microRNA class. Furthermore, by comparing expression in distinct cell states, we report a massive and dynamic regulation of microRNAs, both in numbers and amplitude, highlighting their pivotal role in regulation of quiescence, activation and differentiation. We also identify a number of microRNAs with reliable and specific expression in quiescence including several maternally-expressed miRNAs generated at the imprinted Dlk1-Dio3 locus. Unexpectedly, the majority of class-switching miRNAs are associated with the quiescence/activation transition suggesting a poised program that is actively repressed. These data constitute a key resource for functional analyses of miRNAs in skeletal myogenesis, and more broadly, in the regulation of stem cell self-renewal and tissue homeostasis.