Hayley Goullée, Rhonda L Taylor, Alistair R R Forrest, Nigel G Laing, Gianina Ravenscroft, Joshua S Clayton
BACKGROUND: CRISPR/Cas9 is an invaluable tool for studying cell biology and the development of molecular therapies. However, delivery of CRISPR/Cas9 components into some cell types remains a major hurdle. Primary human myoblasts are a valuable cell model for muscle studies, but are notoriously difficult to transfect. There are currently no commercial lipofection protocols tailored for primary myoblasts, and most generic guidelines simply recommend transfecting healthy cells at high confluency...
September 22, 2021: Skeletal Muscle
Robert W Arpke, Ahmed S Shams, Brittany C Collins, Alexie A Larson, Nguyen Lu, Dawn A Lowe, Michael Kyba
BACKGROUND: Although muscle regenerative capacity declines with age, the extent to which this is due to satellite cell-intrinsic changes vs. environmental changes has been controversial. The majority of aging studies have investigated hindlimb locomotory muscles, principally the tibialis anterior, in caged sedentary mice, where those muscles are abnormally under-exercised. METHODS: We analyze satellite cell numbers in 8 muscle groups representing locomotory and non-locomotory muscles in young and 2-year-old mice and perform transplantation assays of low numbers of hind limb satellite cells from young and old mice...
September 4, 2021: Skeletal Muscle
Olga Mucha, Paulina Podkalicka, Katarzyna Kaziród, Emilia Samborowska, Józef Dulak, Agnieszka Łoboda
BACKGROUND: Duchenne muscular dystrophy (DMD) is an incurable disease, caused by the mutations in the DMD gene, encoding dystrophin, an actin-binding cytoskeletal protein. Lack of functional dystrophin results in muscle weakness, degeneration, and as an outcome cardiac and respiratory failure. As there is still no cure for affected individuals, the pharmacological compounds with the potential to treat or at least attenuate the symptoms of the disease are under constant evaluation. The pleiotropic agents, 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors, known as statins, have been suggested to exert beneficial effects in the mouse model of DMD...
September 3, 2021: Skeletal Muscle
Leila Lesanpezeshki, Hiroshi Qadota, Masoud Norouzi Darabad, Karishma Kashyap, Carla M R Lacerda, Nathaniel J Szewczyk, Guy M Benian, Siva A Vanapalli
BACKGROUND: Caenorhabditis elegans has been widely used as a model to study muscle structure and function. Its body wall muscle is functionally and structurally similar to vertebrate skeletal muscle with conserved molecular pathways contributing to sarcomere structure, and muscle function. However, a systematic investigation of the relationship between muscle force and sarcomere organization is lacking. Here, we investigate the contribution of various sarcomere proteins and membrane attachment components to muscle structure and function to introduce C...
August 13, 2021: Skeletal Muscle
Simonetta Andrea Licandro, Luca Crippa, Roberta Pomarico, Raffaella Perego, Gianluca Fossati, Flavio Leoni, Christian Steinkühler
BACKGROUND: In the search of genetic determinants of Duchenne muscular dystrophy (DMD) severity, LTBP4, a member of the latent TGF-β binding protein family, emerged as an important predictor of functional outcome trajectories in mice and humans. Nonsynonymous single-nucleotide polymorphisms in LTBP4 gene associate with prolonged ambulation in DMD patients, whereas an in-frame insertion polymorphism in the mouse LTBP4 locus modulates disease severity in mice by altering proteolytic stability of the Ltbp4 protein and release of transforming growth factor-β (TGF-β)...
July 22, 2021: Skeletal Muscle
Nainita Roy, Swetha Sundar, Malini Pillai, Farah Patell-Socha, Sravya Ganesh, Ajoy Aloysius, Mohammed Rumman, Hardik Gala, Simon M Hughes, Peter S Zammit, Jyotsna Dhawan
BACKGROUND: During skeletal muscle regeneration, satellite stem cells use distinct pathways to repair damaged myofibers or to self-renew by returning to quiescence. Cellular/mitotic quiescence employs mechanisms that promote a poised or primed state, including altered RNA turnover and translational repression. Here, we investigate the role of mRNP granule proteins Fragile X Mental Retardation Protein (Fmrp) and Decapping protein 1a (Dcp1a) in muscle stem cell quiescence and differentiation...
July 8, 2021: Skeletal Muscle
Lin Zhou, Tong Zhang, Wei Shao, Ruohan Lu, Lin Wang, Haisheng Liu, Bin Jiang, Shiqin Li, Huiqin Zhuo, Suheng Wang, Qinxi Li, Caihua Huang, Donghai Lin
BACKGROUND: Cancer cachexia (CAC) reduces patient survival and quality of life. Developments of efficient therapeutic strategies are required for the CAC treatments. This long-term process could be shortened by the drug-repositioning approach which exploits old drugs approved for non-cachexia disease. Amiloride, a diuretic drug, is clinically used for treatments of hypertension and edema due to heart failure. Here, we explored the effects of the amiloride treatment for ameliorating muscle wasting in murine models of cancer cachexia...
July 6, 2021: Skeletal Muscle
Osvaldo Contreras, Fabio M V Rossi, Marine Theret
Striated muscle is a highly plastic and regenerative organ that regulates body movement, temperature, and metabolism-all the functions needed for an individual's health and well-being. The muscle connective tissue's main components are the extracellular matrix and its resident stromal cells, which continuously reshape it in embryonic development, homeostasis, and regeneration. Fibro-adipogenic progenitors are enigmatic and transformative muscle-resident interstitial cells with mesenchymal stem/stromal cell properties...
July 1, 2021: Skeletal Muscle
Yuchun Cai, Hongyue Zhan, Wenci Weng, Yao Wang, Pengxun Han, Xuewen Yu, Mumin Shao, Huili Sun
BACKGROUND: Diabetes-related muscle wasting is one of the devastating complications of diabetes, which is associated with muscle autophagy due to insulin-mediated glucose starvation. However, treatment for diabetes-related muscle wasting is limited. Our previous study already found that niclosamide ethanolamine salt has the therapeutic effects on insulin deficiency of type 1 diabetes mice and muscle wasting induced by doxorubicin. Therefore, we aim to investigate the therapeutic effects of niclosamide ethanolamine salt on diabetes-induced muscle wasting and to explore whether the mechanism is associated with muscle autophagy...
June 9, 2021: Skeletal Muscle
Mitsutoshi Kurosaka, Yuji Ogura, Shuichi Sato, Kazuhisa Kohda, Toshiya Funabashi
BACKGROUND: The 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. METHODS: Myoblasts were isolated from male mice and were differentiated on a culture dish to evaluate the change in STAT6 during myotube formation...
May 29, 2021: Skeletal Muscle
Rohan X Verma, Suraj Kannan, Brian L Lin, Katherine M Fomchenko, Tim O Nieuwenhuis, Arun H Patil, Clarisse Lukban, Xiaoping Yang, Karen Fox-Talbot, Matthew N McCall, Chulan Kwon, David A Kass, Avi Z Rosenberg, Marc K Halushka
BACKGROUND: Skeletal muscle myofibers can be separated into functionally distinct cell types that differ in gene and protein expression. Current single cell expression data is generally based upon single nucleus RNA, rather than whole myofiber material. We examined if a whole-cell flow sorting approach could be applied to perform single cell RNA-seq (scRNA-seq) in a single muscle type. METHODS: We performed deep, whole cell, scRNA-seq on intact and fragmented skeletal myofibers from the mouse fast-twitch flexor digitorum brevis muscle utilizing a flow-gated method of large cell isolation...
May 17, 2021: Skeletal Muscle
Jessica Brunetti, Stéphane Koenig, Arthur Monnier, Maud Frieden
BACKGROUND: In vitro maturation of human primary myoblasts using 2D culture remains a challenging process and leads to immature fibers with poor internal organization and function. This would however represent a valuable system to study muscle physiology or pathophysiology from patient myoblasts, at a single-cell level. METHODS: Human primary myoblasts were cultured on 800-nm wide striated surface between two layers of Matrigel, and in a media supplemented with an inhibitor of TGFβ receptor...
May 5, 2021: Skeletal Muscle
Qiong Lyu, Ya Wen, Xiang Zhang, Alex B Addinsall, Nicola Cacciani, Lars Larsson
BACKGROUND: Old age is associated with a significantly increased mortality in COVID-19 patients exposed to long-term controlled mechanical ventilation (CMV) and suggested to be due to the hyperinflammatory response associated with the viral infection. However, our understanding of age-related differences in the response to CMV in the absence of a viral infection remains insufficient. METHODS: Young (7-8 months) and old (28-32 months) F344 BN hybrid rats were exposed to the ICU condition for 5 days, i...
May 3, 2021: Skeletal Muscle
Badrul Islam, Mohiuddin Ahmed, Zhahirul Islam, S M Begum
BACKGROUND: SARS-CoV2 virus could be potentially myopathic. Serum creatinine phosphokinase (CPK) is frequently found elevated in severe SARS-CoV2 infection, which indicates skeletal muscle damage precipitating limb weakness or even ventilatory failure. CASE PRESENTATION: We addressed such a patient in his forties presented with features of severe SARS-CoV2 pneumonia and high serum CPK. He developed severe sepsis and acute respiratory distress syndrome (ARDS) and received intravenous high dose corticosteroid and tocilizumab to counter SARS-CoV2 associated cytokine surge...
April 21, 2021: Skeletal Muscle
Yahui Peng, Jihong Li, Dixian Luo, Shuai Zhang, Sijia Li, Dayong Wang, Xidi Wang, Zhujun Zhang, Xue Wang, Changhui Sun, Xu Gao, Yang Hui, Rongzhang He
BACKGROUND: ALAS2 (delta-aminolevulinate synthase 2) is one of the two isoenzymes catalyzing the synthesis of delta-aminolevulinic acid (ALA), which is the first precursor of heme synthesis. ALAS2-overexpressing transgenic mice (Tg mice) showed syndrome of porphyria, a series of diseases related to the heme anabolism deficiency. Tg mice showed an obvious decrease in muscle size. Muscle atrophy results from a decrease in protein synthesis and an increase in protein degradation, which ultimately leads to a decrease in myofiber size due to loss of contractile proteins, organelles, nuclei, and cytoplasm...
March 30, 2021: Skeletal Muscle
Adeel Safdar, Konstantin Khrapko, James M Flynn, Ayesha Saleem, Michael De Lisio, Adam P W Johnston, Yevgenya Kratysberg, Imtiaz A Samjoo, Yu Kitaoka, Daniel I Ogborn, Jonathan P Little, Sandeep Raha, Gianni Parise, Mahmood Akhtar, Bart P Hettinga, Glenn C Rowe, Zoltan Arany, Tomas A Prolla, Mark A Tarnopolsky
No abstract text is available yet for this article.
March 30, 2021: Skeletal Muscle
Anna Benedetti, Gianluca Cera, Daniele De Meo, Ciro Villani, Marina Bouche, Biliana Lozanoska-Ochser
Satellite cells (SCs) are muscle stem cells capable of regenerating injured muscle. The study of their functional potential depends on the availability of methods for the isolation and expansion of pure SCs with preserved myogenic properties after serial passages in vitro. Here, we describe the ice-cold treatment (ICT) method, which is a simple, economical, and efficient method for the isolation and in vitro expansion of highly pure mouse and human SCs. It involves a brief (15-30 min) incubation on ice (0 °C) of a dish containing a heterogeneous mix of adherent muscle mononuclear cells, which leads to the detachment of only the SCs, and gives rise to cultures of superior purity compared to other commonly used isolation methods...
March 17, 2021: Skeletal Muscle
Ji Che, Cuidi Xu, Yuanyuan Wu, Peiyu Jia, Qi Han, Yantao Ma, Xiaolei Wang, Yongjun Zheng
BACKGROUND: Sarcopenia is a common skeletal disease related to myogenic disorders and muscle atrophy. Current clinical management has limited effectiveness. We sought to investigate the role of miR-1290 in myoblast differentiation and muscle atrophy. METHODS: By transfecting miR-1290 into C2C12 cells, we investigated whether miR-1290 regulates myogenesis and myotube atrophy via AKT/P70 signaling pathway. MHC staining was performed to assess myoblast differentiation...
March 15, 2021: Skeletal Muscle
Maria Benavente-Diaz, Glenda Comai, Daniela Di Girolamo, Francina Langa, Shahragim Tajbakhsh
BACKGROUND: Myogenin is a transcription factor that is expressed during terminal myoblast differentiation in embryonic development and adult muscle regeneration. Investigation of this cell state transition has been hampered by the lack of a sensitive reporter to dynamically track cells during differentiation. RESULTS: Here, we report a knock-in mouse line expressing the tdTOMATO fluorescent protein from the endogenous Myogenin locus. Expression of tdTOMATO in MyogntdTom mice recapitulated endogenous Myogenin expression during embryonic muscle formation and adult regeneration and enabled the isolation of the MYOGENIN+ cell population...
February 18, 2021: Skeletal Muscle
Jimmy Massenet, Edward Gardner, Bénédicte Chazaud, F Jeffrey Dilworth
In response to muscle injury, muscle stem cells integrate environmental cues in the damaged tissue to mediate regeneration. These environmental cues are tightly regulated to ensure expansion of muscle stem cell population to repair the damaged myofibers while allowing repopulation of the stem cell niche. These changes in muscle stem cell fate result from changes in gene expression that occur in response to cell signaling from the muscle environment.Integration of signals from the muscle environment leads to changes in gene expression through epigenetic mechanisms...
January 11, 2021: Skeletal Muscle
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