The Experts below are selected from a list of 93 Experts worldwide ranked by ideXlab platform
Junmo Kim - One of the best experts on this subject based on the ideXlab platform.
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effects of a novel p a41p mutation in the swine Myogenic Factor 5 myf5 gene on protein stabilizing muscle fiber characteristics and meat quality
Korean Journal for Food Science of Animal Resources, 2018Co-Authors: Younchul Ryu, Euna Lee, Hanha Chai, Jongeun Park, Junmo KimAbstract:Myogenic Factor 5 (MYF5) plays an important role in regulating skeletal muscle fiber characteristics, consequently affecting meat production and quality. We identified a novel p.A41P mutation in exon1 of the porcine MYF5 gene by direct sequencing. The mutation was predicted to be destabilizing in protein structure based on the resultant amino acid substitution. We estimated the significant substitution effect of p.A41P on the energy stabilization of Myf5 protein structure. Then, we demonstrated that the mutation in Yorkshire population significantly affected muscle fiber type I composition (p<0.05), loin-eye area of lean meat content (p<0.05) and filter-fluid uptake of meat quality (p<0.01). Furthermore, dominant effects significantly influenced total muscle fiber number (p<0.05). This study suggests that the novel p.A41P mutation in porcine MYF5 may be a valuable genetic marker to affect the muscle fiber characteristics and consequently improve meat production quality and quantity.
Jun Mokim - One of the best experts on this subject based on the ideXlab platform.
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effects of a novel p a41p mutation in the swine Myogenic Factor 5 myf5 gene on protein stabilizing muscle fiber characteristics and meat quality
Food Science of Animal Resources, 2018Co-Authors: Youn Chulryu, Eun Alee, Han Hachai, Jong Eunpark, Jun MokimAbstract:Myogenic Factor 5 (MYF5) plays an important role in regulating skeletal muscle fiber characteristics, consequently affecting meat production and...
James A. Thomson - One of the best experts on this subject based on the ideXlab platform.
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Time-dependent Pax3-mediated chromatin remodeling and cooperation with Six4 and Tead2 specify the skeletal Myogenic lineage in developing mesoderm
2019Co-Authors: Alessandro Magli, June Baik, Lauren J. Mills, Il-youp Kwak, Bridget S. Dillon, Ricardo Mondragon Gonzalez, David A. Stafford, Scott A. Swanson, Ron Stewart, James A. ThomsonAbstract:The transcriptional mechanisms driving lineage specification during development are still largely unknown, as the interplay of multiple transcription Factors makes it difficult to dissect these molecular events. Using a cell-based differentiation platform to probe transcription function, we investigated the role of the key paraxial mesoderm and skeletal Myogenic commitment Factors—mesogenin 1 (Msgn1), T-box 6 (Tbx6), forkhead box C1 (Foxc1), paired box 3 (Pax3), Paraxis, mesenchyme homeobox 1 (Meox1), sine oculis-related homeobox 1 (Six1), and Myogenic Factor 5 (Myf5)—in paraxial mesoderm and skeletal myogenesis. From this study, we define a genetic hierarchy, with Pax3 emerging as the gatekeeper between the presomitic mesoderm and the Myogenic lineage. By assaying chromatin accessibility, genomic binding and transcription profiling in mesodermal cells from mouse and human Pax3-induced embryonic stem cells and Pax3-null embryonic day (E)9.5 mouse embryos, we identified conserved Pax3 functions in the activation of the skeletal Myogenic lineage through modulation of Hedgehog, Notch, and bone morphogenetic protein (BMP) signaling pathways. In addition, we demonstrate that Pax3 molecular function involves chromatin remodeling of its bound elements through an increase in chromatin accessibility and cooperation with sine oculis-related homeobox 4 (Six4) and TEA domain family member 2 (Tead2) Factors. To our knowledge, these data provide the first integrated analysis of Pax3 function, demonstrating its ability to remodel chromatin in mesodermal cells from developing embryos and proving a mechanistic footing for the transcriptional hierarchy driving myogenesis.
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Distinct functions of Msgn1, Pax3, and Myf5 during mesoderm specification.
2019Co-Authors: Alessandro Magli, June Baik, Lauren J. Mills, Il-youp Kwak, Bridget S. Dillon, Ricardo Mondragon Gonzalez, David A. Stafford, Scott A. Swanson, Ron Stewart, James A. ThomsonAbstract:(A) Schematic representation of the developmental expression pattern of the TFs analyzed in this study based on published literature [5–15]. (B-C) Graphs show the percentage of PDGFRα+FLK1− cells, measured by FACS, in day 5 EB cultures from A2lox-Pax3, A2lox-FoxC1, A2lox-Meox1, A2lox-Msgn1, A2lox-Myf5, A2lox-Paraxis, and A2lox-Six1 ES cell lines, using both serum (B) and serum-free (C) differentiation protocols. Mean + SD is shown from at least 3 biological replicates. **p < 0.01, ***p < 0.001, ****p < 0.0001. Differences are relative to the Pax3+dox group. (D-E) Immunofluorescence staining for MyoG in serum (D) and serum-free (E) day 10 cultures following 24-hour dox withdrawal. Images are representative of 3 biological replicates. MYOG (red); nuclei (blue). Bar: 100 μm. (F) Quantification of the MYOG+ area of the immunostaining images shown in Fig 1D and 1E. Graph represents mean + SD from at least 3 independent experiments. *p < 0.05, **p < 0.01. (G) Western blot of day 10 cultures (same as panel D-E) from serum- and serum-free differentiation of A2lox-Pax3, A2lox-Myf5, and A2lox-Msgn1 ES cell lines. eMYHC. ACTIN. (H) Live cell imaging of Msgn1-GFP and Pax3-GFP fusion proteins using wide-field microscopy followed by image deconvolution. Images are representative of 3 biological replicates, and the results were similar between serum- and serum-free differentiation. DNA was visualized using Hoechst 33342. Bar: 5 μm. Numerical values are available in S1 Data. dox, doxycycline; EB, embryoid body; eMYHC, embryonic myosin heavy chain; ES, embryonic stem; FACS, fluorescence-activated cell sorting; FoxC1, forkhead box C1; Meox1, mesenchyme homeobox 1; Msgn1, mesogenin 1; Myf5, Myogenic Factor 5; MyoG, myogenin; Pax3, paired box 3; PSM, presomitic mesoderm; Six1, sine oculis-related homeobox 1; Tbx6, T-box 6; TF, transcription Factor.
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Pax3 cooperates with Six4 and Tead2 to activate the Myogenic program.
2019Co-Authors: Alessandro Magli, June Baik, Lauren J. Mills, Il-youp Kwak, Bridget S. Dillon, Ricardo Mondragon Gonzalez, David A. Stafford, Scott A. Swanson, Ron Stewart, James A. ThomsonAbstract:(A) Number of ChIP-seq peaks identified upon PAX3 immunoprecipitation in primary myoblasts, 1-day, and 6-day iPax3-induced cells. Venn diagram indicates the overlap between these 3 datasets. (B) Selected transcription Factor motifs enriched at Pax3-bound loci from 1-day, 6-day, and primary myoblasts. Distribution of the motifs across 500 bp from the peak center is reported below. (C) Six4 and Runx1 bind at a subset of 6-day Pax3 loci. Fewer loci were also bound by Jun, Tead1-4, and Usf1 proteins. k-means clustering was generated using published ChIP-seq data (from Myogenic cell line C2C12). (D) Distribution of ChIP-seq reads for Six4, Jun, Tead1/4, Runx1, and Usf1 1-day and 6-day Pax3 genomic binding data in cluster 1 (from panel C) generated using k-means clustering. Curves show overlapping of Six4 and Runx1 binding at Pax3 peaks (±3 kb from peak center). (E) IGV track displaying genomic occupancy for Pax3, Six4, Runx1, Jun, Tead1/4, and Usf1 at the Cdon locus. Dashed red square indicates Pax3-bound site characterized by Six4, Runx1, Jun, and Tead1/4 occupancy. (F-G) Knockdown of Six4 or Tead2 in iPax3 PDGFRα+FLK1− cells impairs Pax3 transcriptional activity and ultimately Myogenic differentiation. Upon Pax3 induction, day 5 EB cells were analyzed by qRT-PCR with the indicated probes (F) or assayed for terminal differentiation (G). Graph represents mean + SD from at least 3 independent experiments. MYHC (red); nuclei (blue). Bar: 100 μm. *p < 0.05, **p < 0.01, ***p < 0.001. Numerical values are available in S1 Data. AP1, activator protein 1; Cdon, cell adhesion molecule–related/down-regulated by oncogenes; ChIP-seq, chromatin immunoprecipitation sequencing; Ctrl, control; EB, embryoid body; Gapdh, glyceraldehyde 3-phosophate dehydrogenase; IgG, immunoglobulin G; IGV, Integrative Genomics Viewer; iPax3, inducible-Pax3; Lfng, lunatic fringe; Megf10, multiple epidermal growth Factor–like domains protein 10; Myf5, Myogenic Factor 5; MYHC, myosin heavy chain; Pax3, paired box 3; qRT-PCR, quantitative reverse transcription PCR; RFX, regulatory Factor X; Runx1, Runt-related transcription Factor 1; shSCR, scramble control; shSix4, Six4 knockdown; shTead2, Tead2 knockdown; Six4, sine oculis-related homeobox 4; Tead1/2/4, TEA domain family member 1/2/4; Tot, total; Usf1, upstream stimulatory Factor 1.
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, Amanda B Reedmaldonado, Zhihua Lu, Hongxiu Ning, Lia Banie, Tie Zhou, Yajun Ruan, Hsun Shuan Wang, Jun Zhou, 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.
Younchul Ryu - One of the best experts on this subject based on the ideXlab platform.
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effects of a novel p a41p mutation in the swine Myogenic Factor 5 myf5 gene on protein stabilizing muscle fiber characteristics and meat quality
Korean Journal for Food Science of Animal Resources, 2018Co-Authors: Younchul Ryu, Euna Lee, Hanha Chai, Jongeun Park, Junmo KimAbstract:Myogenic Factor 5 (MYF5) plays an important role in regulating skeletal muscle fiber characteristics, consequently affecting meat production and quality. We identified a novel p.A41P mutation in exon1 of the porcine MYF5 gene by direct sequencing. The mutation was predicted to be destabilizing in protein structure based on the resultant amino acid substitution. We estimated the significant substitution effect of p.A41P on the energy stabilization of Myf5 protein structure. Then, we demonstrated that the mutation in Yorkshire population significantly affected muscle fiber type I composition (p<0.05), loin-eye area of lean meat content (p<0.05) and filter-fluid uptake of meat quality (p<0.01). Furthermore, dominant effects significantly influenced total muscle fiber number (p<0.05). This study suggests that the novel p.A41P mutation in porcine MYF5 may be a valuable genetic marker to affect the muscle fiber characteristics and consequently improve meat production quality and quantity.
