The Experts below are selected from a list of 315 Experts worldwide ranked by ideXlab platform
Steven A Greenberg - One of the best experts on this subject based on the ideXlab platform.
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type 1 interferons inhibit Myotube Formation independently of upregulation of interferon stimulated gene 15
PLOS ONE, 2013Co-Authors: Sara Franzi, Mohammad Salajegheh, Remedios Nazareno, Steven A GreenbergAbstract:INTRODUCTION: Type 1 interferon (IFN)-inducible genes and their inducible products are upregulated in dermatomyositis muscle. Of these, IFN-stimulated gene 15 (ISG15) is one of the most upregulated, suggesting its possible involvement in the pathogenesis of this disease. To test this postulate, we developed a model of type 1 IFN mediated Myotube toxicity and assessed whether or not downregulation of ISG15 expression prevents this toxicity. METHODS: Mouse myoblasts (C2C12 cell line) were cultured in the presence of type 1 or type 2 IFNs and ISG15 expression assessed by microarray analysis. The morphology of newly formed Myotubes was assessed by measuring their length, diameter, and area on micrographs using imaging software. ISG15 expression was silenced through transfection with small interference RNA. RESULTS: Type 1 IFNs, especially IFN-beta, increased ISG15 expression in C2C12 cells and impaired Myotube Formation. Silencing of ISG15 resulted in knockdown of ISG15 protein, but without phenotypic rescue of Myotube Formation. DISCUSSION: IFN-beta affects myoblast differentiation ability and Myotube morphology in vitro.These studies provide evidence that ISG15, which is highly upregulated in dermatomyositis muscle, does not appear to play a key role in IFN-beta-mediated C2C12 myoblast cell fusion.
Miranda D Grounds - One of the best experts on this subject based on the ideXlab platform.
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the absence of myod in regenerating skeletal muscle affects the expression pattern of basement membrane interstitial matrix and integrin molecules that is consistent with delayed Myotube Formation
Acta Histochemica, 2001Co-Authors: Julia Huijbregts, Jason D White, Miranda D GroundsAbstract:Summary MyoD is a member of a skeletal muscle specific family of transcription factors which directs the events of myogenesis during development and regeneration. Muscle cells that lack MyoD show delayed fusion in vivo and in vitro and defects have been observed in vitro in the attachment of MyoD(-/-) myoblasts to complex substrates such as Matrigel. Since interactions with the extracellular matrix (ECM) are important during myoblast fusion (i. e. Myotube Formation), it was hypothesised that expression of ECM molecules or their receptors may be altered in MyoD(-/-) muscle. The production of basement membrane molecules such as collagen type IV and several laminins, the interstitial molecules fibronectin and tenascin-C, and the cell surface molecules integrin α5 and α6 were quantitated in vitro using ELISA on cultured cells from MyoD(-/-) and wild type mice. Differences were observed in the production of fibronectin, tenascin-C, collagen type IV, laminin-1 and integrin α5 between control and MyoD(-/-) Myotubes in vitro . This corresponded with delayed fusion of myoblasts in MyoD(-/-) cultures. On the basis of these findings with respect to matrix expression in vitro , fluorescent immunohistochemistry was carried out on adult whole muscle autografts to examine whether the expression of these molecuies, as well as integrin α7, was altered in the complex in vivo environment. Some minor differences in expression patterns were observed in MyoD(-/-) as compared to normal BALB/c autografts. The overall expression of matrix components was consistent with the delayed onset of Myotube Formation. These resuits suggest that the delay in Myotube Formation in MyoD(-/-) muscle is not a direct result of altered ex-pression of the matrix molecules collagen type IV, laminins, fibronectin, tenascin-C, and integrins α5, α6 or α7.
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Myotube Formation is delayed but not prevented in myod deficient skeletal muscle studies in regenerating whole muscle grafts of adult mice
Journal of Histochemistry and Cytochemistry, 2000Co-Authors: Jason D White, Marilyn Davies, Amelia K Scaffidi, John Mcgeachie, Michael A Rudnicki, Miranda D GroundsAbstract:We compared the time course of myogenic events in vivo in regenerating whole muscle grafts in MyoD(−/−) and control BALB/c adult mice using immunohistochemistry and electron microscopy. Immunohistochemistry with antibodies to desmin and myosin revealed a striking delay by about 3 days in the Formation of Myotubes in MyoD(−/−) autografts compared with BALB/c mice. However, Myotube Formation was not prevented, and autografts in both strains appeared similar by 8 days. Electron microscopy confirmed Myotube Formation in 8- but not 5-day MyoD(−/−) grafts. This pattern was not influenced by cross-transplantation experiments between strains examined at 5 days. Antibodies to proliferating cell nuclear antigen demonstrated an elevated level of replication by MyoD(−/−) myoblasts in autografts, and replication was sustained for about 3 days compared with controls. These data indicate that the delay in the onset of differentiation and hence fusion is related to extended proliferation of the MyoD(−/−) myoblasts. Overa...
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extracellular matrix growth factors genetics their influence on cell proliferation and Myotube Formation in primary cultures of adult mouse skeletal muscle
Experimental Cell Research, 1995Co-Authors: Moira Maley, Marilyn Davies, Miranda D GroundsAbstract:Cell proliferation and Myotube Formation in response to growth factors on various extracellular matrices (ECM) were investigated in primary skeletal muscle cultures from adult SJL/J and BALB/c mice. There was no difference between the rates of proliferation from primary cultures of SJL/J and Balb/c mice measured at 48 h in response to a range of concentrations of PDGF-AA, -AB, -BB, TGFβ1, or LIF (added at 24 h). SJL/J primary cultures were more responsive to bFGF (which was the most potent mitogen) than were BALB/c cultures. Comparison of dose response curves to bFGF and TGFβ1 grown on gelatin or Matrigel showed that the nature of the ECM did not have a significant affect. More Myotubes formed at 4 days in SJL/J than in parallel BALB/c cultures on gelatin or Matrigel (P < 0.05). On gelatin more Myotubes with 4 or more nuclei were formed in cultures from SJL/J than BALB/c muscle (P < 0.05); however, on Matrigel these Myotubes occurred with similar frequency. Myotube Formation examined in BALB/c muscle cultures grown on collagen IV, entactin-free laminin, and fibronectin showed that none of these ECM components alone supported large Myotube Formation (4 or more nuclei) as well as did Matrigel, although fibronectin was as effective as Matrigel with respect to the total number of Myotubes formed. Parallel experiments carried out using the myogenic H-2Kb(27) cell line showed similar effects with the exception of laminin which enhanced large Myotube Formation and desmin expression in the H.2Kb(27) but not in the primary muscle cultures. The greater sensitivity in mitogenic response to bFGF and the more extensive Myotube Formation seen in SJL/J compared with BALB/c cultures in vitro reflects the superior capacity for muscle regeneration of SJL/J mice in vivo.
Tetsuji Naka - One of the best experts on this subject based on the ideXlab platform.
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socs 2 interferes with Myotube Formation and potentiates osteoblast differentiation through upregulation of junb in c2c12 cells
Journal of Cellular Physiology, 2006Co-Authors: Xinshou Ouyang, Minoru Fujimoto, Reiko Nakagawa, Satoshi Serada, Toshio Tanaka, Shintaro Nomura, Ichiro Kawase, Tadamitsu Kishimoto, Tetsuji NakaAbstract:Suppressor of cytokine signaling (SOCS)-2 regulates normal postnatal growth and its deficiency in mice causes gigantism with increased bone length and proportional enlargement in skeletal muscles. Using C2C12 mesenchymal precursor cell line as a model, we investigated a possible role of SOCS-2 in the differentiation process of mesenchymal precursors. Stable transfection of SOCS-2 into C2C12 cells resulted in the acceleration of proliferation and survival, and inhibition of spontaneous Myotube Formation. In addition, SOCS-2 potentiated bone morphogenic protein (BMP)-induced transdifferentiation of C2C12 cells into osteoblast phenotypes. These effects of SOCS-2 on C2C12 cells differed strikingly from that of SOCS-1, another member of SOCS family, and its mechanisms were evaluated. SOCS-2 did not alter BMP-induced phosphorylation and nuclear accumulation of Smad1, nor the expression of inhibitory-Smads mRNA. However, SOCS-2 enhanced BMP-induced transcriptional activation of the Smad-responsive reporter gene, suggesting that the action of SOCS-2 is exerted at the transcriptional level. Interestingly, SOCS-2 overexpression in C2C12 cells increased the endogenous JunB protein, one of the key transcriptional factors in the control of BMP/Smad signaling responsiveness. In addition, the proteasome inhibitor enhanced JunB protein expression in C2C12 cells. Moreover, we found that SOCS-2 reduced JunB ubiquitination in COS-7 cells. Although SOCS-2 is a modulator of growth hormone (GH) signaling, the upregulation of JunB by SOCS-2 did not require GH signaling. Taken together, these results suggest that SOCS-2 positively regulates endogenous JunB protein expression in C2C12 cells through inhibition of JunB destabilization by the ubiquitin–proteasome pathway, and thereby regulates the cell fate of mesenchymal precursors. J. Cell. Physiol. 207: 428–436, 2006. © 2006 Wiley-Liss, Inc.
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SOCS‐2 interferes with Myotube Formation and potentiates osteoblast differentiation through upregulation of JunB in C2C12 cells
Journal of Cellular Physiology, 2006Co-Authors: Xinshou Ouyang, Minoru Fujimoto, Reiko Nakagawa, Satoshi Serada, Toshio Tanaka, Shintaro Nomura, Ichiro Kawase, Tadamitsu Kishimoto, Tetsuji NakaAbstract:Suppressor of cytokine signaling (SOCS)-2 regulates normal postnatal growth and its deficiency in mice causes gigantism with increased bone length and proportional enlargement in skeletal muscles. Using C2C12 mesenchymal precursor cell line as a model, we investigated a possible role of SOCS-2 in the differentiation process of mesenchymal precursors. Stable transfection of SOCS-2 into C2C12 cells resulted in the acceleration of proliferation and survival, and inhibition of spontaneous Myotube Formation. In addition, SOCS-2 potentiated bone morphogenic protein (BMP)-induced transdifferentiation of C2C12 cells into osteoblast phenotypes. These effects of SOCS-2 on C2C12 cells differed strikingly from that of SOCS-1, another member of SOCS family, and its mechanisms were evaluated. SOCS-2 did not alter BMP-induced phosphorylation and nuclear accumulation of Smad1, nor the expression of inhibitory-Smads mRNA. However, SOCS-2 enhanced BMP-induced transcriptional activation of the Smad-responsive reporter gene, suggesting that the action of SOCS-2 is exerted at the transcriptional level. Interestingly, SOCS-2 overexpression in C2C12 cells increased the endogenous JunB protein, one of the key transcriptional factors in the control of BMP/Smad signaling responsiveness. In addition, the proteasome inhibitor enhanced JunB protein expression in C2C12 cells. Moreover, we found that SOCS-2 reduced JunB ubiquitination in COS-7 cells. Although SOCS-2 is a modulator of growth hormone (GH) signaling, the upregulation of JunB by SOCS-2 did not require GH signaling. Taken together, these results suggest that SOCS-2 positively regulates endogenous JunB protein expression in C2C12 cells through inhibition of JunB destabilization by the ubiquitin–proteasome pathway, and thereby regulates the cell fate of mesenchymal precursors. J. Cell. Physiol. 207: 428–436, 2006. © 2006 Wiley-Liss, Inc.
Byung Seok Oh - One of the best experts on this subject based on the ideXlab platform.
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low intensity extracorporeal shock wave therapy promotes myogenesis through perk atf4 pathway
Neurourology and Urodynamics, 2018Co-Authors: Bohan Wang, Jun Zhou, Lia Banie, Amanda B Reedmaldonado, Hongxiu Ning, Zhihua Lu, Yajun Ruan, Tie Zhou, Hsun Shuan Wang, Byung Seok OhAbstract:Aim Stress urinary incontinence (SUI) is a significant health problem for women. Treatments employing muscle derived stem cells (MDSCs) may be a promising approach to this prevalent, bothersome condition, but these treatments are invasive and require collection of cells from one site for injection into another. It is also unknown whether or not these cells establish themselves and function as muscle cells in the target tissues. Alternatively, low-intensity extracorporeal shock wave therapy (Li-ESWT) is non-invasive and has shown positive outcomes in the treatment of multiple musculoskeletal disorders, but the biological effects responsible for clinical success are not yet well understood. The aim of this study is to explore the possibility of employing Li-ESWT for activation of MDSCs in situ and to further elucidate the underlying biological effects and mechanisms of action in urethral muscle. Methods Urethral muscle derived stem cells (uMDSCs) were harvest from Zucker Lean (ZUC-LEAN) (ZUC-Leprfa 186) rats and characterized with flow cytometry. Li-ESWT (0.02 mJ/mm2, 3 Hz, 200 pulses) and GSK2656157, an inhibitor of PERK pathway, were applied to L6 rat myoblast cells. To assess for Myotube Formation, we used immunofluorescence staining and western blot analysis in uMDSCs and L6 cells. Results The results indicate that uMDSCs could form Myotubes. Myotube Formation was significantly increased by the Li-ESWT as was the expression of muscle heavy chain (MHC) and myogenic factor 5 (Myf5) in L6 cells in vitro. Li-ESWT activated protein kinase RNA-like ER kinase (PERK) pathway by increasing the phosphorylation levels of PERK and eukaryotic initiation factor 2a (eIF2α) and by increasing activating transcription factor 4 (ATF4). In addition, GSK2656157, an inhibitor of PERK, effectively inhibited the Myotube Formation in L6 rat myoblast cells. Furthermore, GSK2656157 also attenuated Myotube Formation induced by Li-ESWT. Conclusion In conclusion, this experiment reveals that rat uMDSCs can be isolated successfully and can form Myotubes in vitro. PERK/ATF4 pathway was involved in Myotube Formation, and L6 rat myoblast cells were activated by Li-ESWT to form Myotubes. These findings suggest that PERK/ATF4 pathway is activated by Li-ESWT. This study elucidates one of the biochemical pathways responsible for the clinical improvements seen after Li-ESWT. It is possible that this inFormation will help to establish Li-ESWT as an acceptable treatment modality and may help to further refine the use of Li-ESWT in the clinical practice of medicine.
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Low-intensity extracorporeal shock wave therapy promotes myogenesis through PERK/ATF4 pathway.
Neurourology and Urodynamics, 2017Co-Authors: Bohan Wang, Jun Zhou, Lia Banie, Hongxiu Ning, Zhihua Lu, Yajun Ruan, Tie Zhou, Hsun Shuan Wang, Amanda B. Reed-maldonado, Byung Seok OhAbstract:Aim Stress urinary incontinence (SUI) is a significant health problem for women. Treatments employing muscle derived stem cells (MDSCs) may be a promising approach to this prevalent, bothersome condition, but these treatments are invasive and require collection of cells from one site for injection into another. It is also unknown whether or not these cells establish themselves and function as muscle cells in the target tissues. Alternatively, low-intensity extracorporeal shock wave therapy (Li-ESWT) is non-invasive and has shown positive outcomes in the treatment of multiple musculoskeletal disorders, but the biological effects responsible for clinical success are not yet well understood. The aim of this study is to explore the possibility of employing Li-ESWT for activation of MDSCs in situ and to further elucidate the underlying biological effects and mechanisms of action in urethral muscle. Methods Urethral muscle derived stem cells (uMDSCs) were harvest from Zucker Lean (ZUC-LEAN) (ZUC-Leprfa 186) rats and characterized with flow cytometry. Li-ESWT (0.02 mJ/mm2, 3 Hz, 200 pulses) and GSK2656157, an inhibitor of PERK pathway, were applied to L6 rat myoblast cells. To assess for Myotube Formation, we used immunofluorescence staining and western blot analysis in uMDSCs and L6 cells. Results The results indicate that uMDSCs could form Myotubes. Myotube Formation was significantly increased by the Li-ESWT as was the expression of muscle heavy chain (MHC) and myogenic factor 5 (Myf5) in L6 cells in vitro. Li-ESWT activated protein kinase RNA-like ER kinase (PERK) pathway by increasing the phosphorylation levels of PERK and eukaryotic initiation factor 2a (eIF2α) and by increasing activating transcription factor 4 (ATF4). In addition, GSK2656157, an inhibitor of PERK, effectively inhibited the Myotube Formation in L6 rat myoblast cells. Furthermore, GSK2656157 also attenuated Myotube Formation induced by Li-ESWT. Conclusion In conclusion, this experiment reveals that rat uMDSCs can be isolated successfully and can form Myotubes in vitro. PERK/ATF4 pathway was involved in Myotube Formation, and L6 rat myoblast cells were activated by Li-ESWT to form Myotubes. These findings suggest that PERK/ATF4 pathway is activated by Li-ESWT. This study elucidates one of the biochemical pathways responsible for the clinical improvements seen after Li-ESWT. It is possible that this inFormation will help to establish Li-ESWT as an acceptable treatment modality and may help to further refine the use of Li-ESWT in the clinical practice of medicine.
Leonardo Ricotti - One of the best experts on this subject based on the ideXlab platform.
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proliferation and skeletal Myotube Formation capability of c2c12 and h9c2 cells on isotropic and anisotropic electrospun nanofibrous phb scaffolds
Biomedical Materials, 2012Co-Authors: Leonardo Ricotti, Gianni Ciofani, Alessandro Polini, Giada Graziana Genchi, Donata Iandolo, Helena Vazao, Virgilio Mattoli, Lino Ferreira, Arianna MenciassiAbstract:This study aims at investigating the behavior in terms of the proliferation and skeletal muscle differentiation capability of two myoblastic cell lines, C2C12 and H9c2, on both isotropic and anisotropic electrospun nanofibrous poly(hydroxybutyrate) (PHB) scaffolds, as well as on PHB films and polystyrene controls. After a careful characterization of the matrices in terms of surface morphology, surface roughness and mechanical properties, the proliferation rate and the capability of the two cell lines to form skeletal Myotubes were evaluated. Genetic analyses were also performed in order to assess the differentiation level of the cells on the different substrates. We demonstrated that the aligned nanofibrous mesh decreases the proliferation activity and provides a higher differentiative stimulus. We also clarified how the nanofibrous substrate influences Myotube Formation, and quantified a series of Myotube-related parameters for both C2C12 and H9c2 cells.
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investigation of interactions between poly l lysine coated boron nitride nanotubes and c2c12 cells up take cytocompatibility and differentiation
International Journal of Nanomedicine, 2010Co-Authors: Gianni Ciofani, Stefania Moscato, Delfo Dalessandro, Andrea Pietrabissa, Serena Danti, Dinuccio Dinucci, Federica Chiellini, Claudia Nesti, Leonardo Ricotti, Mario PetriniAbstract:Boron nitride nanotubes (BNNTs) have generated considerable interest within the scientific community by virtue of their unique physical properties, which can be exploited in the biomedical field. In the present in vitro study, we investigated the interactions of poly-l-lysine-coated BNNTs with C2C12 cells, as a model of muscle cells, in terms of cytocompatibility and BNNT internalization. The latter was performed using both confocal and transmission electron microscopy. Finally, we investigated myoblast differentiation in the presence of BNNTs, evaluating the protein synthesis of differentiating cells, Myotube Formation, and expression of some constitutive myoblastic markers, such as MyoD and Cx43, by reverse transcription – polymerase chain reaction and Western blot analysis. We demonstrated that BNNTs are highly internalized by C2C12 cells, with neither adversely affecting C2C12 myoblast viability nor significantly interfering with Myotube Formation.
