44 results on '"Skeletal muscle cell proliferation"'
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2. A MIOSTATINA NA COORDENAÇÃO DA PROLIFERAÇÃO E DIFERENCIAÇÃO DE CÉLULAS MUSCULARES ADIPOSAS E ESQUELÉTICAS E NO EQUILÍBRIO DO METABOLISMO ENERGÉTICO COM BASE NA TECNOLOGIA DE CHIPS GENÉTICOS
- Author
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Qijun Ren and Rihua Cong
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Peroxisome proliferator-activated receptor gamma ,Adipose tissue ,Physical Therapy, Sports Therapy and Rehabilitation ,Myostatin ,030204 cardiovascular system & hematology ,Miostatina ,Myoblasts ,03 medical and health sciences ,0302 clinical medicine ,Mioblastos ,RNA Antissenso ,medicine ,Myocyte ,RNA, Antisense ,Orthopedics and Sports Medicine ,RNA Antisentido ,Gene ,Metabolismo ,biology ,Skeletal muscle ,030229 sport sciences ,Antisense RNA ,Cell biology ,Metabolism ,medicine.anatomical_structure ,Skeletal muscle cell proliferation ,Sports medicine ,biology.protein ,RC1200-1245 ,Miooblastos - Abstract
Myoblasts fuse into multinucleated muscle fibers to form and promote the growth of skeletal muscle. In order to analyze the role of myostatin (MSTN) in body fat, skeletal muscle cell proliferation and differentiation and energy metabolism, this study will use the antisense RNA technology of gene chip technology to study it. The results showed that the MSTN gene regulated the growth and proliferation of myoblasts and affected the development of skeletal muscle by affecting the expression of Cdc42, bnip2, p38 and other genes; knockout or overexpression of the MSTN gene would lead to a trend of fat-related genes from fat synthesis to fat decomposition; after the MSTN gene was knocked down, the expression levels of cpti-b, PPARG and other genes in the cells were corresponding after MSTN overexpression, the relative expression of the PPARG gene decreased. It is suggested that the knockout or overexpression of MSTN may affect lipid accumulation, and cpti-b and PPARG may directly regulate lipid level. It is hoped that this experiment can provide a reference for the study of MSTN effect on fat deposition. RESUMO Os mioblastos se fundem eM fibras musculares multinucleadas para formar e promover o crescimento do músculo esquelético. A fim de analisar o papel da miostatina (MSTN) na gordura corporal, proliferação de células musculares esqueléticas e diferenciação e metabolismo energético, este estudo utilizará a tecnologia anti-RNA de chips genéticos para estudá-la. Os resultados mostraram que o gene MSTN regulava o crescimento e a proliferação de mioblastos e afetava o desenvolvimento do músculo esquelético, afetando a expressão de Cdc42, bnip2, p38 e outros genes; a eliminação ou sobrexpressão do gene MSTN conduziria a uma tendência de os genes adiposos sintetizarem a gordura até sua decomposição; após a eliminação do gene MSTN, os níveis de expressão de cpti-b, PPARG e outros genes nas células mostraram-se correspondentes após a sobrexpressão do gene MSTN, e a expressão relativa do gene PPARG diminuiu. Sugere-se que a eliminação ou sobrexpressão da MSTN possa afetar a acumulação de lipídeos, e o cpti-b e o PPARG podem regular diretamente o nível lipídico. Espera-se que esta experiência possa fornecer uma referência para o estudo do efeito da MSTN sobre a deposição de gordura. RESUMEN Los mioblastos se funden en fibras musculares multinucleadas para formar y promover el crecimiento del músculo esquelético. A fin de analizar el papel de la miostatina (MSTN) en la grasa corporal, proliferación de células musculares esqueléticas y diferenciación y metabolismo energético, este estudio utilizará la tecnología anti-RNA de chips genéticos para estudiarla. Los resultados mostraron que el gen MSTN regulaba el crecimiento y la proliferación de mioblastos y afectaba el desarrollo del músculo esquelético, afectando la expresión de Cdc42, bnip2, p38 y otros genes; la eliminación o sobreexpresión del gen MSTN conduciría a una tendencia de que los genes adiposos sinteticen la grasa hasta su descomposición; después de la eliminación del gen MSTN, los niveles de expresión de cpti-b, PPARG y otros genes en las células se mostraron correspondientes después de la sobreexpresión del gen MSTN, y la expresión relativa del gen PPARG disminuyó. Se sugiere que la eliminación o sobreexpresión de la MSTN pueda afectar la acumulación de lipídos, y el cpti-b y el PPARG pueden regular directamente el nivel lipídico. Se espera que esta experiencia pueda proveer una referencia para el estudio del efecto de la MSTN sobre el depósito de grasa.
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- 2021
3. Beef extract supplementation promotes myoblast proliferation and myotube growth in C2C12 cells
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Yusuke Sato, Yoshihide Ikeuchi, Mitsuhiro Furuse, Jun ichiro Wakamatsu, Wataru Mizunoya, Yuka Sonoda, Shoko Sawano, Ryuichi Tatsumi, Shozo Tomonaga, and Kento Baba
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0301 basic medicine ,Myoblast proliferation ,Muscle Fibers, Skeletal ,Medicine (miscellaneous) ,030209 endocrinology & metabolism ,Muscle hypertrophy ,Myoblasts ,Andrology ,Mice ,03 medical and health sciences ,0302 clinical medicine ,medicine ,Animals ,Myocyte ,Muscle, Skeletal ,Protein kinase B ,PI3K/AKT/mTOR pathway ,Cell Proliferation ,030109 nutrition & dietetics ,Nutrition and Dietetics ,Chemistry ,food and beverages ,Skeletal muscle ,Cell Differentiation ,Rats ,medicine.anatomical_structure ,Skeletal muscle cell proliferation ,Dietary Supplements ,Cattle ,C2C12 - Abstract
We previously determined that the intake of beef extract for 4 weeks increases skeletal muscle mass in rats. Thus, this study aimed to clarify whether beef extract has a hypertrophic effect on muscle cells and to determine the signaling pathway underlying beef extract-induced myotube hypertrophy. We assessed the effects of beef extract supplement on mouse C2C12 skeletal muscle cell proliferation and differentiation and myotube growth. In addition, the phosphorylation of Akt, ERK1/2, and mTOR following beef extract supplementation was examined by western blotting. Furthermore, the bioactive constituents of beef extract were examined using amino acid analysis and dialysis. In the proliferative stage, beef extract significantly increased myoblast proliferation. In the differentiation stage, beef extract supplementation did not promote myoblast differentiation. In mature myotubes, beef extract supplementation increased myotube diameter and promoted protein synthesis. Although Akt and ERK1/2 levels were not affected, beef extract supplementation increased mTOR phosphorylation, which indicated that the mTOR pathway mediates beef extract-induced myotube hypertrophy. The hypertrophic activity was observed in fractions of > 7000 Da. Beef extract promoted C2C12 myoblast proliferation and C2C12 myotube hypertrophy. Myotube hypertrophy was potentially induced by mTOR activation and active components in beef extract were estimated to be > 7000 Da.
- Published
- 2020
4. Designed Functional Dispersion for Insulin Protection from Pepsin Degradation and Skeletal Muscle Cell Proliferation: In Silico and In Vitro Study
- Author
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Veera C. S. R. Chittepu, Poonam Kalhotra, Tzayhri Gallardo-Velázquez, Raúl René Robles-de la Torre, and Guillermo Osorio-Revilla
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PEGylated SWCNTs ,functional dispersion ,pharmaceutical nanotechnology ,skeletal muscle cell proliferation ,insulin therapy ,diabetes ,glucose metabolism ,Chemistry ,QD1-999 - Abstract
Functionalized single-walled carbon nanotubes with polyethylene glycol (PEGylated SWCNTs) are a promising nanomaterial that recently has emerged as the most attractive “cargo” to deliver chemicals, peptides, DNA and RNAs into cells. Insulin therapy is a recommended therapy to treat diabetes mellitus despite its side effects. Recently, functional dispersion made up of bioactive peptides, bioactive compounds and functionalized carbon nanomaterials such as PEGylated SWCNTs have proved to possess promising applications in nanomedicine. In the present study, molecular modeling simulations are utilized to assist in designing insulin hormone-PEGylated SWCNT composites, also called functional dispersion; to achieve this experimentally, an ultrasonication tool was utilized. Enzymatic degradation assay revealed that the designed functional dispersion protects about 70% of free insulin from pepsin. In addition, sulforhodamine B (SRB) assay, the quantification of insulin and glucose levels in differentiated skeletal muscle cell supernatants, reveals that functional dispersion regulates glucose and insulin levels to promote skeletal muscle cell proliferation. These findings offer new perspectives for designed functional dispersion, as potential pharmaceutical preparations to improve insulin therapy and promote skeletal muscle cell health.
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- 2018
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5. The Functional Roles of RNAs Cargoes Released by Neutrophil-Derived Exosomes in Dermatomyositis
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Liya Li, Xiaoxia Zuo, Di Liu, Hui Luo, and Honglin Zhu
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MAPK/ERK pathway ,AMPK ,Pharmacology ,dermatomyositis ,PI3K-Akt ,RM1-950 ,Biology ,MAPK ,Microvesicles ,Cell biology ,lncRNA ,Downregulation and upregulation ,Skeletal muscle cell proliferation ,microRNA ,Pharmacology (medical) ,Therapeutics. Pharmacology ,FoxO ,KEGG ,Gene ,neutrophil-derived exosome ,Original Research ,miRNA - Abstract
Dermatomyositis (DM) is an idiopathic inflammatory myopathy characterized by cutaneous manifestations. We first identified the profiles of noncoding RNAs (lncRNAs and miRNAs) in peripheral neutrophil exosomes (EXOs) of DM patients and explored their potential functional roles. Bioinformatics analyses were performed with R packages. Real-time quantitative PCR was used to validate the altered RNAs in DM neutrophil EXO-stimulated human dermal microvascular endothelial cells (HDMECs) and human skeletal muscle myoblasts (HSkMCs). In DM neutrophil EXOs, 124 upregulated lncRNAs (with 1,392 target genes), 255 downregulated lncRNAs (with 1867 target genes), 17 upregulated miRNAs (with 2,908 target genes), and 15 downregulated miRNAs (with 2,176 target genes) were identified. GO analysis showed that the differentially expressed (DE) lncRNAs and DE miRNAs participated in interleukin-6 and interferon-beta production, skeletal muscle cell proliferation and development, and endothelial cell development and differentiation. KEGG analysis suggested that DE lncRNAs and DE miRNAs were enriched in the PI3K–Akt, MAPK, AMPK and FoxO signalling pathways. Many novel and valuable DE lncRNAs and DE miRNAs interacted and cotargeted in the PI3K–Akt, MAPK, AMPK and FoxO signalling pathways. Our study suggests that neutrophil EXOs participate in DM pathogenesis through lncRNAs and miRNAs in the PI3K–Akt, MAPK, AMPK and FoxO signalling pathways.
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- 2021
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6. Genetic effects of the EIF5A2 gene on chicken growth and skeletal muscle development
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Guihuan Li, Zhijun Wang, Hongjia Ouyang, Xiaolan Chen, Qinghua Nie, and Biao Chen
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0301 basic medicine ,Hypusine ,Myoblast proliferation ,General Veterinary ,Myogenesis ,0402 animal and dairy science ,Skeletal muscle ,04 agricultural and veterinary sciences ,Biology ,040201 dairy & animal science ,Cell biology ,03 medical and health sciences ,chemistry.chemical_compound ,030104 developmental biology ,medicine.anatomical_structure ,chemistry ,Skeletal muscle cell proliferation ,medicine ,Myocyte ,Polyamine homeostasis ,Animal Science and Zoology ,Myogenin - Abstract
Eukaryotic translation initiation factor 5A2 (EIF5A2) is an essential protein closely related to cellular polyamine homeostasis. It is the only cellular protein that contains the unusual amino acid hypusine [Nɛ-(4-amino-2-hydroxybutyl) lysine] which is necessary for the biochemical activity of EIF5A2 and cellular proliferation. In this study, we aim to characterize chicken EIF5A2 and explore its potential effects on skeletal muscle cell proliferation and differentiation. Here, we found that chicken EIF5A2 has four variant transcripts. The single nucleotide polymorphisms (SNPs) in the 5′-flanking region of EIF5A2 were significantly associated with the chicken growth and fatness traits. Expression pattern of EIF5A2 in skeletal muscle revealed that it was highly expressed at an early embryonic age and then decreased sharply after d 15 of the embryonic age. The luciferase reporter assays confirmed that EIF5A2 was directly targeted by miRNA-223. EIF5A2 was negatively regulated by miR-223 in myoblast. Both 5-Ethynyl-2′-deoxyuridine staining assays and flow cytometry analyses of the cell cycle showed that down-regulation of EIF5A2 could markedly inhibit myoblast proliferation. We found that down-regulation of EIF5A2 could promote myoblast differentiation through promoting the expression of Myogenin (MYOG). MYOG is a muscle-specific transcription factor involved in myogenesis. Altogether, the results indicated that EIF5A2 is an important candidate gene in chicken growth, and down-regulation of EIF5A2 could suppress myoblast proliferation and promote myoblast differentiation by targeting miR-223.
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- 2019
7. Expression and localization of heat-shock proteins during skeletal muscle cell proliferation and differentiation and the impact of heat stress
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Victoria L Chhen, James G. Ryall, Savant S Thakur, Janine L. James, Gordon S. Lynch, Nicola J Cranna, and Kristy Swiderski
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0301 basic medicine ,Cellular differentiation ,Muscle Fibers, Skeletal ,Muscle Development ,Biochemistry ,Myoblasts ,03 medical and health sciences ,0302 clinical medicine ,Heat shock protein ,Animals ,Cells, Cultured ,Heat-Shock Proteins ,Cell Proliferation ,Original Paper ,Myogenesis ,Chemistry ,Cell Differentiation ,Cell Biology ,Cell biology ,030104 developmental biology ,Proteostasis ,Cytoplasm ,Skeletal muscle cell proliferation ,030220 oncology & carcinogenesis ,HSP60 ,C2C12 ,Heat-Shock Response - Abstract
Skeletal myogenesis is a coordinated sequence of events associated with dramatic changes in cell morphology, motility, and metabolism, which causes cellular stress and alters proteostasis. Chaperones, such as heat-shock proteins (HSPs), play important roles in limiting cellular stresses and maintaining proteostasis, but whether HSPs are specifically involved in myogenesis is not well understood. Here, we characterized gene and protein expression and subcellular localization of various HSPs in proliferating C2C12 myoblasts and differentiating myotubes under control conditions and in response to heat stress. Hsp25, Hsp40, and Hsp60 protein expression declined by 48, 35, and 83%, respectively, during differentiation. In contrast, Hsp70 protein levels doubled during early differentiation. Hsp25 was predominantly localized to the cytoplasm of myoblasts and myotubes but formed distinct aggregates in perinuclear spaces of myoblasts after heat-shock. Hsp40 was distributed diffusely throughout the cytoplasm and nucleus and, after heat-shock, translocated to the nucleus of myoblasts but formed aggregates in myotubes. Hsp60 localized to the perinuclear space in myoblasts but was distributed more diffusely across the cytoplasm in myotubes. Hsp70 was expressed diffusely throughout the cytoplasm and nucleus and translocated to the nucleus after heat-shock in myoblasts, but not in myotubes. Hsp90 was expressed diffusely across the cytoplasm in both myoblasts and myotubes under control conditions and did not change in response to heat-shock. These findings reveal distinct and different roles for HSPs in the regulation of myogenic cell proliferation and differentiation. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s12192-019-01001-2) contains supplementary material, which is available to authorized users.
- Published
- 2019
8. Diphlorethohydroxycarmalol (DPHC) Isolated from the Brown Alga Ishige okamurae Acts on Inflammatory Myopathy as an Inhibitory Agent of TNF-α
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Kil-Nam Kim, Jun-Geon Je, Hyun-Soo Kim, Ginnae Ahn, You-Jin Jeon, and Seo-Young Kim
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inflammatory myopathy ,Anti-Inflammatory Agents ,Down-Regulation ,Muscle Proteins ,Pharmaceutical Science ,Inflammation ,Pharmacology ,Phaeophyta ,pro-inflammatory cytokines ,01 natural sciences ,Article ,Proinflammatory cytokine ,Myoblasts ,Mice ,03 medical and health sciences ,Muscular Diseases ,Drug Discovery ,medicine ,Animals ,Myocyte ,RNA, Messenger ,Myopathy ,lcsh:QH301-705.5 ,Pharmacology, Toxicology and Pharmaceutics (miscellaneous) ,030304 developmental biology ,0303 health sciences ,phlorotannin ,Tumor Necrosis Factor-alpha ,010405 organic chemistry ,Chemistry ,Myogenesis ,Skeletal muscle ,Muscle atrophy ,0104 chemical sciences ,RAW 264.7 Cells ,medicine.anatomical_structure ,lcsh:Biology (General) ,Gene Expression Regulation ,Skeletal muscle cell proliferation ,inflammation ,marine algae ,Cytokines ,medicine.symptom ,Heterocyclic Compounds, 3-Ring ,myopathy - Abstract
Inflammation affects various organs of the human body, including skeletal muscle. Phlorotannins are natural biologically active substances found in marine brown algae and exhibit anti-inflammatory activities. In this study, we focused on the effects of phlorotannins on anti-inflammatory activity and skeletal muscle cell proliferation activity to identify the protective effects on the inflammatory myopathy. First, the five species of marine brown algal extracts dramatically inhibited nitric oxide (NO) production in lipopolysaccharide (LPS)-induced RAW 264.7 cells without toxicity at all the concentrations tested. Moreover, the extracts collected from Ishige okamurae (I. okamurae) significantly increased cell proliferation of C2C12 myoblasts compared to the non-treated cells with non-toxicity. In addition, as a result of finding a potential tumor necrosis factor (TNF)-&alpha, inhibitor that regulates the signaling pathway of muscle degradation in I. okamurae-derived natural bioactive compounds, Diphlorethohydroxycarmalol (DPHC) is favorably docked to the TNF-&alpha, with the lowest binding energy and docking interaction energy value. Moreover, DPHC down-regulated the mRNA expression level of pro-inflammatory cytokines and suppressed the muscle RING-finger protein (MuRF)-1 and Muscle Atrophy F-box (MAFbx)/Atrgoin-1, which are the key protein muscle atrophy via nuclear factor-&kappa, B (NF-&kappa, B), and mitogen-activated protein kinase (MAPKs) signaling pathways in TNF-&alpha, stimulated C2C12 myotubes. Therefore, it is expected that DPHC isolated from IO would be developed as a TNF-&alpha, inhibitor against inflammatory myopathy.
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- 2020
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9. Microcarriers for Upscaling Cultured Meat Production
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Vincent Bodiou, Panagiota Moutsatsou, and Mark J. Post
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0301 basic medicine ,Computer science ,Endocrinology, Diabetes and Metabolism ,SKELETAL-MUSCLE CELLS ,030209 endocrinology & metabolism ,lcsh:TX341-641 ,Review ,03 medical and health sciences ,Global population ,Cultured meat ,0302 clinical medicine ,cultivated meat ,MAMMALIAN-CELLS ,EXTRACELLULAR-MATRIX ,Production (economics) ,bovine myoblasts ,Bioprocess ,LARGE-SCALE PRODUCTION ,Nutrition ,satellite cells ,030109 nutrition & dietetics ,Nutrition and Dietetics ,CURRENT TECHNOLOGIES ,SHEAR-STRESS ,bioprocessing ,Final product ,Microcarrier ,HYDROXYBUTYL CHITOSAN ,ENDOTHELIAL-CELLS ,Skeletal muscle cell proliferation ,clean meat ,microbeads ,Biochemical engineering ,Sustainable production ,EMBRYONIC STEM-CELLS ,lcsh:Nutrition. Foods and food supply ,cell expansion ,SATELLITE CELL ,Food Science - Abstract
Due to the considerable environmental impact and the controversial animal welfare associated with industrial meat production, combined with the ever-increasing global population and demand for meat products, sustainable production alternatives are indispensable. In 2013, the world's first laboratory grown hamburger made from cultured muscle cells was developed. However, coming at a price of $300.000, and being produced manually, substantial effort is still required to reach sustainable large-scale production. One of the main challenges is scalability. Microcarriers (MCs), offering a large surface/volume ratio, are the most promising candidates for upscaling muscle cell culture. However, although many MCs have been developed for cell lines and stem cells typically used in the medical field, none have been specifically developed for muscle stem cells and meat production. This paper aims to discuss the MCs' design criteria for skeletal muscle cell proliferation and subsequently for meat production based on three scenarios: (1) MCs are serving only as a temporary substrate for cell attachment and proliferation and therefore they need to be separated from the cells at some stage of the bioprocess, (2) MCs serve as a temporary substrate for cell proliferation but are degraded or dissolved during the bioprocess, and (3) MCs are embedded in the final product and therefore need to be edible. The particularities of each of these three bioprocesses will be discussed from the perspective of MCs as well as the feasibility of a one-step bioprocess. Each scenario presents advantages and drawbacks, which are discussed in detail, nevertheless the third scenario appears to be the most promising one for a production process. Indeed, using an edible material can limit or completely eliminate dissociation/degradation/separation steps and even promote organoleptic qualities when embedded in the final product. Edible microcarriers could also be used as a temporary substrate similarly to scenarios 1 and 2, which would limit the risk of non-edible residues.
- Published
- 2020
10. Effects of MicroRNA-27a on Myogenin Expression and Akt/FoxO1 Signal Pathway during Porcine Myoblast Differentiation
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Junqiu Luo, Jun He, Bing Yu, Shurun Zhang, Hong Chen, Daiwen Chen, Ping Zheng, Zhiqing Huang, Jie Yu, Yuheng Luo, and Xiaoling Chen
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Male ,0301 basic medicine ,Swine ,Bioengineering ,FOXO1 ,Biology ,Myoblasts ,03 medical and health sciences ,0302 clinical medicine ,microRNA ,Transcriptional regulation ,Animals ,Myocyte ,Transcription factor ,Protein kinase B ,Myogenin ,Forkhead Box Protein O1 ,Gene Expression Regulation, Developmental ,Cell Differentiation ,musculoskeletal system ,Cell biology ,MicroRNAs ,030104 developmental biology ,Skeletal muscle cell proliferation ,030220 oncology & carcinogenesis ,Animal Science and Zoology ,Proto-Oncogene Proteins c-akt ,tissues ,Biotechnology - Abstract
Skeletal myoblast differentiation is controlled by a multitude of transcription factors and signal pathways. Myogenin is a critical transcriptional regulator in the initiation and maintenance of myoblast differentiation. The Akt/FoxO1 signal pathway plays an important role in myoblast differentiation. MicroRNAs are a kind of small noncoding RNAs that have been regarded as important regulators in skeletal muscle cell proliferation and differentiation. The objective of this study was to investigate the effects of microRNA-27a (miR-27a) on myogenin expression and Akt/FoxO1 signal pathway during porcine myoblast differentiation. Here, we found that the expression of miR-27a was gradually diminished at the early differentiation stage and then rebounded. Overexpression of miR-27a suppressed the mRNA and protein expression levels of myogenin during porcine myoblast differentiation, whereas inhibition of miR-27a promoted the mRNA and protein expression levels of myogenin. In addition, overexpression of miR-27a decreased the level of P-Akt/Akt and increased the protein level of FoxO1; however, inhibition of miR-27a increased the level of P-Akt/Akt and decreased the protein level of FoxO1. The present study demonstrated that miR-27a could inhibit myogenin expression and Akt/FoxO1 signal pathway during porcine myoblast differentiation.
- Published
- 2017
11. Histone methyltransferase Setd2 is critical for the proliferation and differentiation of myoblasts
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Xuejun Jiang, Yuan Dai, Ye Tao, Ding-Sheng Jiang, Xiaojing Yue, Tingli Yang, Jiang Chang, Xi Lin, Xiaoyan Li, Xin Yi, and Kelsey C. Andrade
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Cyclin-Dependent Kinase Inhibitor p21 ,0301 basic medicine ,Myoblast proliferation ,Muscle Fibers, Skeletal ,Biology ,Article ,Cell Line ,Histones ,Myoblasts ,Mice ,03 medical and health sciences ,Cyclins ,Transcriptional regulation ,Animals ,Cyclin D1 ,Gene Silencing ,Phosphorylation ,Molecular Biology ,Myogenin ,Cell Proliferation ,Gene Editing ,Base Sequence ,Myosin Heavy Chains ,Myogenesis ,Cyclin-Dependent Kinase 4 ,Cell Differentiation ,Cell Cycle Checkpoints ,Cyclin-Dependent Kinase 6 ,Histone-Lysine N-Methyltransferase ,Cell Biology ,Molecular biology ,Chromatin ,030104 developmental biology ,Cyclin E2 ,Skeletal muscle cell proliferation ,Histone methyltransferase ,CRISPR-Cas Systems ,C2C12 - Abstract
Skeletal muscle cell proliferation and differentiation are tightly regulated. Epigenetic regulation is a major component of the regulatory mechanism governing these processes. Histone modification is part of the epigenetic code used for transcriptional regulation of chromatin through the establishment of an active or repressive state for genes involved in myogenesis in a temporal manner. Here, we uncovered the function of SET domain containing 2 (Setd2), an essential histone 3 lysine 36 trimethyltransferase, in regulating the proliferation and differentiation of myoblasts. Setd2 was silenced in the skeletal muscle myoblast cell line, C2C12, using the CRISPR/CAS9 system. The mutant cells exhibited defect in myotube formation. The myotube formation marker, myosin heavy chain (MHC), was downregulated earlier in Setd2 silenced cells compared to wild-type myoblasts during differentiation. The deficiency in Setd2 also resulted in repression of Myogenin (MyoG) expression, a key myogenic regulator during differentiation. In addition to the myoblast differentiation defect, decreased proliferation rate with significantly reduced levels of histone 3 phosphorylation, indicative of cell proliferation defect, were observed in the Setd2 silenced cells; suggesting an impaired proliferation phenotype. Furthermore, compromised G1/S- and G2/M-phase transition and decreased expression levels of major regulators of cell cycle G1/S checkpoints, cyclin D1, CDK4, CDK6, and cyclin E2 were detected in Setd2 silenced cells. Consistent with the cell cycle arrested phenotype, cyclin-dependent kinase inhibitor p21 was upregulated in Setd2 silenced cells. Together, this study demonstrates an essential role of Setd2 in myoblast proliferation and differentiation, and uncovers Setd2-mediated molecular mechanism through regulating MyoG and p21.
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- 2017
12. Separation and purification of the bovine milk fat globule membrane protein and its effect on improvement of C2C12mouse skeletal muscle cell proliferation
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Shaobo Zhou, He Li, Weili Xu, and Ying Ma
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0301 basic medicine ,Gel electrophoresis ,medicine.diagnostic_test ,Cell growth ,Chemistry ,General Chemistry ,Molecular biology ,Catalysis ,Flow cytometry ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,Biochemistry ,Skeletal muscle cell proliferation ,030220 oncology & carcinogenesis ,Materials Chemistry ,Fluorescence microscope ,medicine ,MTT assay ,Protein kinase B ,PI3K/AKT/mTOR pathway - Abstract
A novel method to improve the proliferation activity of C2C12 cells by the bovine milk fat globule membrane (MFGM) protein was established in this study. The MFGM protein was extracted and isolated into 4 fractions using an electric cream separator, and purified by a cellulose DEAE-52 column. Fraction 2 accounted for 57.8% of the total MFGM protein, and was used in the following study. The MTT assay showed that it induced cell proliferation activity, increased the cell survival rate and the cell number using flow cytometry and fluorescence microscopy analysis. There were only subtle changes in the morphology as observed using confocal scanning laser microscopy, but the number of mitochondria was significantly increased as observed using transmission electron microscopy analysis. Furthermore, the mRNA expression of MyoD, cyclin D1, p70S6K and mTOR was up-regulated as determined utilizing the quantitative real-time PCR assay, and the activation of Akt and mTOR phosphorylation was up regulated as determined using the Western blot assay. The main protein in fraction 2, assayed by 1-D gel electrophoresis and MALDI TOF-TOF, was identified as milk fat globule-EGF factor 8, the content was 65.6% of the total protein in fraction 2. The results elucidate a new molecular mechanism of the MFGM protein fraction 2: the activation of the Akt signal pathway in promoting cell proliferation.
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- 2017
13. Phytochemicals in Chinese Chive (Allium tuberosum) Induce the Skeletal Muscle Cell Proliferation via PI3K/Akt/mTOR and Smad Pathways in C2C12 Cells
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Mira Oh, SeonJu Park, Seung Hyun Kim, Kil-Nam Kim, and Seo-Young Kim
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Flavonoid ,SMAD ,Catalysis ,lcsh:Chemistry ,Inorganic Chemistry ,Allium tuberosum ,skeletal muscle differentiation ,medicine ,Physical and Theoretical Chemistry ,Chinese chive ,lcsh:QH301-705.5 ,Molecular Biology ,Protein kinase B ,Spectroscopy ,PI3K/AKT/mTOR pathway ,chemistry.chemical_classification ,Chemistry ,Cell growth ,Organic Chemistry ,food and beverages ,Skeletal muscle ,General Medicine ,Computer Science Applications ,cell proliferation ,medicine.anatomical_structure ,lcsh:Biology (General) ,lcsh:QD1-999 ,Biochemistry ,Skeletal muscle cell proliferation ,flavonoid glycoside ,C2C12 - Abstract
Chinese chive (Allium tuberosum) is a medicinal food that is cultivated and consumed mainly in Asian countries. Its various phytochemicals and physiological effects have been reported, but only a few phytochemicals are available for skeletal muscle cell proliferation. Herein, we isolated a new compound, kaempferol-3-O-(6″-feruloyl)-sophoroside (1), along with one known flavonoid glycoside (2) and six amino acid (3–8) compounds from the water-soluble fraction of the shoot of the Chinese chive. The isolated compounds were identified using extensive spectroscopic methods, including 1D and 2D NMR, and evaluated for their proliferation activity on skeletal muscle cells. Among the tested compounds, newly isolated flavonoid (1) and 5-aminouridine (7) up-regulated PI3K/Akt/mTOR pathways, which implies a positive effect on skeletal muscle growth and differentiation. In particular, compound 1 down-regulated the Smad pathways, which are negative regulators of skeletal muscle growth. Collectively, we suggest that major constituents of Chinese chive, flavonoids and amino acids, might be used in dietary supplements that aid skeletal muscle growth.
- Published
- 2021
14. Phytochemicals in Garlic Extract Inhibit Therapeutic Enzyme DPP-4 and Induce Skeletal Muscle Cell Proliferation: A Possible Mechanism of Action to Benefit the Treatment of Diabetes Mellitus
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Tzayhri Gallardo-Velázquez, Veera C S R Chittepu, Poonam Kalhotra, and Guillermo Osorio-Revilla
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endocrine system diseases ,serine protease inhibition ,030309 nutrition & dietetics ,Dipeptidyl Peptidase 4 ,Phytochemicals ,Cell Culture Techniques ,lcsh:QR1-502 ,Pharmacology ,Biochemistry ,Article ,Antioxidants ,lcsh:Microbiology ,03 medical and health sciences ,dipeptidyl peptidase-4 inhibition ,Diabetes mellitus ,medicine ,Animals ,Hypoglycemic Agents ,Vildagliptin ,Garlic ,Muscle, Skeletal ,Mexico ,Molecular Biology ,Dipeptidyl peptidase-4 ,Cell Proliferation ,030304 developmental biology ,Dipeptidyl-Peptidase IV Inhibitors ,garlic extract ,0303 health sciences ,Plant Extracts ,Chemistry ,ultrasonic-assisted extraction ,Ascorbic acid ,medicine.disease ,Rats ,Molecular Docking Simulation ,Diabetes Mellitus, Type 2 ,Mechanism of action ,Skeletal muscle cell proliferation ,Sitagliptin ,diabetes mellitus ,medicine.symptom ,Alogliptin ,medicine.drug - Abstract
Diabetes mellitus is a severe health problem in Mexico, and its prevalence is increasing exponentially every year. Recently, DPP-4 (dipeptidyl peptidase-4) inhibitors have become attractive oral anti-hyperglycemic agents to reduce the pathology of diabetes. Gliptin&rsquo, s family, such as sitagliptin, vildagliptin, and alogliptin, are in clinical use to treat diabetes mellitus but possess side effects. Therefore, there is a specific need to look for new therapeutic scaffolds (biomolecules). Garlic bulb is widely used as a traditional remedy for the treatment of diabetes. The garlic extracts are scientifically proven to control glucose levels in patients with diabetes, despite the unknown mechanism of action. The aim of the study is to investigate the antidiabetic effects of ultrasonication assisted garlic bulb extract. To achieve this, in-vitro assays such as DPP-4 inhibitory and antioxidant activities were investigated. Further, functional group analysis using FTIR and identification of phytochemicals using mass spectrometry analysis was performed. The results showed that 70.9 µ, g/mL of garlic bulb extract inhibited 50% DPP-4 activity. On top of that, the garlic extract exhibited a 20% scavenging activity, equivalent to 10 µ, g/mL of ascorbic acid. Molecular docking simulations on identified phytochemicals using mass spectrometry revealed their potential binding at the DPP-4 druggable region, and therefore the possible DPP-4 inhibition mechanism. These results suggest that prepared garlic extract contains phytochemicals that inhibit DPP-4 and have antioxidant activity. Also, the prepared extract induces skeletal muscle cell proliferation that demonstrates the antidiabetic effect and its possible mechanism of action.
- Published
- 2020
15. Designed Functional Dispersion for Insulin Protection from Pepsin Degradation and Skeletal Muscle Cell Proliferation: In Silico and In Vitro Study
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Raúl René Robles de la Torre, Veera C S R Chittepu, Tzayhri Gallardo-Velázquez, Poonam Kalhotra, and Guillermo Osorio-Revilla
- Subjects
0301 basic medicine ,pharmaceutical nanotechnology ,General Chemical Engineering ,Sonication ,medicine.medical_treatment ,In silico ,glucose metabolism ,Sulforhodamine B ,PEGylated SWCNTs ,functional dispersion ,02 engineering and technology ,Polyethylene glycol ,Carbohydrate metabolism ,Article ,lcsh:Chemistry ,03 medical and health sciences ,chemistry.chemical_compound ,medicine ,General Materials Science ,skeletal muscle cell proliferation ,diabetes ,Insulin ,021001 nanoscience & nanotechnology ,030104 developmental biology ,lcsh:QD1-999 ,Biochemistry ,chemistry ,Skeletal muscle cell proliferation ,insulin therapy ,Nanomedicine ,0210 nano-technology - Abstract
Functionalized single-walled carbon nanotubes with polyethylene glycol (PEGylated SWCNTs) are a promising nanomaterial that recently has emerged as the most attractive &ldquo, cargo&rdquo, to deliver chemicals, peptides, DNA and RNAs into cells. Insulin therapy is a recommended therapy to treat diabetes mellitus despite its side effects. Recently, functional dispersion made up of bioactive peptides, bioactive compounds and functionalized carbon nanomaterials such as PEGylated SWCNTs have proved to possess promising applications in nanomedicine. In the present study, molecular modeling simulations are utilized to assist in designing insulin hormone-PEGylated SWCNT composites, also called functional dispersion, to achieve this experimentally, an ultrasonication tool was utilized. Enzymatic degradation assay revealed that the designed functional dispersion protects about 70% of free insulin from pepsin. In addition, sulforhodamine B (SRB) assay, the quantification of insulin and glucose levels in differentiated skeletal muscle cell supernatants, reveals that functional dispersion regulates glucose and insulin levels to promote skeletal muscle cell proliferation. These findings offer new perspectives for designed functional dispersion, as potential pharmaceutical preparations to improve insulin therapy and promote skeletal muscle cell health.
- Published
- 2018
16. Fibroblast growth factor 23 does not directly influence skeletal muscle cell proliferation and differentiation or ex vivo muscle contractility
- Author
-
Kun Wang, Michael J. Wacker, Keith G. Avin, Sarah L. Dallas, Chad D. Touchberry, Sharon M. Moe, Neal X. Chen, Julian Vallejo, and Marco Brotto
- Subjects
0301 basic medicine ,Fibroblast growth factor 23 ,medicine.medical_specialty ,Physiology ,Endocrinology, Diabetes and Metabolism ,Muscle Fibers, Skeletal ,Gene Expression ,urologic and male genital diseases ,Muscle Development ,Cell Line ,Contractility ,Myoblasts ,03 medical and health sciences ,Mice ,Physiology (medical) ,Internal medicine ,medicine ,Animals ,Muscle, Skeletal ,Cell Proliferation ,Chronic Kidney Disease-Mineral and Bone Disorder ,business.industry ,Myogenesis ,Skeletal muscle ,Deoxyguanosine ,Cell Differentiation ,medicine.disease ,Myocardial Contraction ,Rats ,Fibroblast Growth Factors ,Hypophosphatemic Rickets ,stomatognathic diseases ,Fibroblast Growth Factor-23 ,Oxidative Stress ,030104 developmental biology ,Endocrinology ,medicine.anatomical_structure ,Muscle Fibers, Slow-Twitch ,Skeletal muscle cell proliferation ,8-Hydroxy-2'-Deoxyguanosine ,Muscle Fibers, Fast-Twitch ,Calcium ,business ,Ex vivo ,Kidney disease ,Muscle Contraction ,Research Article - Abstract
Skeletal muscle dysfunction accompanies the clinical disorders of chronic kidney disease (CKD) and hereditary hypophosphatemic rickets. In both disorders, fibroblast growth factor 23 (FGF23), a bone-derived hormone regulating phosphate and vitamin D metabolism, becomes chronically elevated. FGF23 has been shown to play a direct role in cardiac muscle dysfunction; however, it is unknown whether FGF23 signaling can also directly induce skeletal muscle dysfunction. We found expression of potential FGF23 receptors ( Fgfr1–4) and α-Klotho in muscles of two animal models (CD-1 and Cy/+ rat, a naturally occurring rat model of chronic kidney disease-mineral bone disorder) as well as C2C12 myoblasts and myotubes. C2C12 proliferation, myogenic gene expression, oxidative stress marker 8-OHdG, intracellular Ca2+ ([Ca2+]i), and ex vivo contractility of extensor digitorum longus (EDL) or soleus muscles were assessed after treatment with various amounts of FGF23. FGF23 (2–100 ng/ml) did not alter C2C12 proliferation, expression of myogenic genes, or oxidative stress after 24- to 72-h treatment. Acute or prolonged FGF23 treatment up to 6 days did not alter C2C12 [Ca2+]i handling, nor did acute treatment with FGF23 (9–100 ng/ml) affect EDL and soleus muscle contractility. In conclusion, although skeletal muscles express the receptors involved in FGF23-mediated signaling, in vitro FGF23 treatments failed to directly alter skeletal muscle development or function under the conditions tested. We hypothesize that other endogenous substances may be required to act in concert with FGF23 or apart from FGF23 to promote muscle dysfunction in hereditary hypophosphatemic rickets and CKD.
- Published
- 2018
17. miR-34b Modulates Skeletal Muscle Cell Proliferation and Differentiation
- Author
-
Lan Luo, Zhixiong Tang, Huiling Qiu, Deming Gou, Kang Kang, Nian Liu, and Jiasheng Zhong
- Subjects
0301 basic medicine ,Proteomics ,Myoblast proliferation ,Cell type ,Muscle Development ,Biochemistry ,Cell Line ,Myoblasts ,03 medical and health sciences ,Mice ,Myocyte ,Animals ,Humans ,Muscle, Skeletal ,Molecular Biology ,3' Untranslated Regions ,Cell Proliferation ,Chemistry ,Myogenesis ,Muscle cell proliferation ,Adenosylhomocysteinase ,RNA-Binding Proteins ,Cell Differentiation ,Cell Biology ,Phosphoproteins ,Cell biology ,MicroRNAs ,030104 developmental biology ,14-3-3 Proteins ,Gene Expression Regulation ,Skeletal muscle cell proliferation ,Models, Animal ,C2C12 ,Nucleolin - Abstract
Myogenesis involves myoblast proliferation and differentiation to myocytes, followed by fusion and hypertrophy to form myotubes during muscle development. Increasing evidence showed that microRNAs (miRNAs) play important roles in the regulation of myogenesis. We have previously revealed that miR-34b is steadily increased during this process. This miRNA regulates differentiation in various cell types, though its function in myogenesis remains to be elucidated. In this study, we show that miR-34b represses muscle cell proliferation and promotes myotube formation. Our quantitative iTRAQ-based proteomic analysis reveals 97 proteins are regulated by miR-34b in mouse myoblast C2C12. We identified that miR-34b targets 14-3-3 protein gamma, adenosylhomocysteinase and nucleolin by binding to their 3'UTR. Further analysis of these proteins expression patterns show that nucleolin is a cognate target of miR-34b during myogenic differentiation. Here, we proved that a moderate reduction of nucleolin in cells enhanced the myotube formation. However, nucleolin is required for myogenesis, as cells with low levels of nucleolin reduced cell proliferation rate and are unable to differentiate. Our data demonstrated that nucleolin regulates myogenesis in a protein-abundance-dependent manner. J. Cell. Biochem. 118: 4285-4295, 2017. © 2017 Wiley Periodicals, Inc.
- Published
- 2017
18. Effects of vitamin D on primary human skeletal muscle cell proliferation, differentiation, protein synthesis and bioenergetics
- Author
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Vinicius Fernandes Cruzat, Philip Newsholme, Rodrigo Carlessi, and Karina Romeu Montenegro
- Subjects
0301 basic medicine ,Myoblast proliferation ,Calcitriol ,Myoblasts, Skeletal ,Endocrinology, Diabetes and Metabolism ,Muscle Fibers, Skeletal ,Clinical Biochemistry ,Muscle Proteins ,Biochemistry ,Muscle hypertrophy ,03 medical and health sciences ,Oxygen Consumption ,0302 clinical medicine ,Endocrinology ,medicine ,Humans ,Insulin ,Myocyte ,Vitamin D ,Molecular Biology ,Cells, Cultured ,Cell Proliferation ,Myogenesis ,Chemistry ,TOR Serine-Threonine Kinases ,Skeletal muscle ,Cell Differentiation ,Vitamins ,Cell Biology ,Mitochondria ,Cell biology ,Glucose ,030104 developmental biology ,medicine.anatomical_structure ,Skeletal muscle cell proliferation ,030220 oncology & carcinogenesis ,Myogenic regulatory factors ,Molecular Medicine ,Energy Metabolism ,medicine.drug - Abstract
The active form of Vitamin D (1,25(OH)2D), has been suggested to have a regulatory role in skeletal muscle function and metabolism, however, the effects and mechanisms of vitamin D (VitD) action in this tissue remain to be fully established. In this study, we have used primary human skeletal muscle myoblast (HSMM) cells that display typical characteristics of human skeletal muscle function and protein levels, to investigate the effects of the active form of VitD on proliferation, differentiation, protein synthesis and bioenergetics. Myoblast cells were treated with 100 nM of VitD for 24 h, 48 h, 72 h and five days (cells were differentiated into myotubes) and then analyses were performed. We report that VitD inhibits myoblast proliferation and enhances differentiation by altering the expression of myogenic regulatory factors. In addition, we found that protein synthesis signaling improved in myotubes after VitD treatment in the presence of insulin. We also report an increase in oxygen consumption rate after 24 h of treatment in myoblasts and after 5 days of treatment in myotubes after VitD exposure. VitD significantly impacted HSMM myogenesis, as well as protein synthesis in the presence of insulin.
- Published
- 2019
19. Mast cells can regulate skeletal muscle cell proliferation by multiple mechanisms
- Author
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Patrice Bouchard, Marie-Pier Roussel, Claude H. Côté, and Élise Duchesne
- Subjects
medicine.medical_specialty ,Myoblast proliferation ,Physiology ,Cell growth ,Muscle cell proliferation ,Regeneration (biology) ,Skeletal muscle ,Biology ,MyoD ,Cell biology ,Cellular and Molecular Neuroscience ,Endocrinology ,medicine.anatomical_structure ,Skeletal muscle cell proliferation ,Physiology (medical) ,Internal medicine ,medicine ,Neurology (clinical) ,Myogenin - Abstract
Introduction: Mast cells (MCs) can stimulate cell proliferation, but their specific contribution to skeletal muscle regeneration is not well defined. Methods: L6 myoblast proliferation was assessed in coculture with MCs or when grown with MC-conditioned media. To address the in vivo implication of MCs in regeneration, rats were treated with cromolyn, and myoblast proliferation, immune cell accumulation, and myogenic factors were assessed in bupivacaine-injured muscles. Results: In vitro, both procedures increased the L6 cell proliferation rate, and this was tryptase-dependent. In vivo, MC stabilization increased myoblast proliferation and accumulation of macrophages CD68 and CD163 after injury. This correlated with a sequential increase in MyoD and myogenin protein level expression. Conclusions: MCs can directly stimulate muscle cell proliferation via tryptase. MCs can influence myoblast proliferation in vivo, but this effect seems to be predominantly related to their modulation of macrophage recruitment. The MC is a potential actor in the early stages of muscle healing. Muscle Nerve 48: 403–414, 2013
- Published
- 2013
20. Overexpression of Striated Muscle Activator of Rho Signaling (STARS) Increases C2C12 Skeletal Muscle Cell Differentiation
- Author
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Paula A Della Gatta, Séverine Lamon, Bilal Ahmad Mir, Greg M. Kowalski, Aaron P. Russell, Malcom J McConville, Marita A. Wallace, and Joachim Kloehn
- Subjects
0301 basic medicine ,Physiology ,proliferation ,striated activator of Rho signalling ,Biology ,Bioinformatics ,lcsh:Physiology ,03 medical and health sciences ,Skeletal muscle cell differentiation ,Physiology (medical) ,medicine ,Myocyte ,skeletal muscle ,Original Research ,lcsh:QP1-981 ,Cardiac muscle ,Skeletal muscle ,differentiation ,Cell biology ,Myotube differentiation ,030104 developmental biology ,medicine.anatomical_structure ,Skeletal muscle cell proliferation ,regeneration ,MYF5 ,myogenesis ,ITGA7 - Abstract
Background: Skeletal muscle growth and regeneration depend on the activation of satellite cells, which leads to myocyte proliferation, differentiation and fusion with existing muscle fibres. Skeletal muscle cell proliferation and differentiation are tightly coordinated by a continuum of molecular signalling pathways. The striated muscle activator of Rho signalling (STARS) is an actin binding protein that regulates the transcription of genes involved in muscle cell growth, structure and function via the stimulation of actin polymerization and activation of serum-response factor (SRF) signalling. While STARS mediates cell proliferation in smooth and cardiac muscle models; however, whether STARS overexpression enhances cell proliferation and differentiation has not been investigated in skeletal muscle cells. Results: We demonstrate for the first time that STARS overexpression enhances differentiation but not proliferation in C2C12 mouse skeletal muscle cells. Increased differentiation was associated with an increase in the gene levels of the myogenic differentiation markers Ckm, Ckmt2 and Myh4, the differentiation factor Igf2 and the myogenic regulatory factors (MRFs) Myf5 and Myf6. Exposing C2C12 cells to CCG-1423, a pharmacological inhibitor of SRF preventing the nuclear translocation of its co-factor MRTF-A, had no effect on myotube differentiation rate, suggesting that STARS regulates differentiation via a MRTF-A independent mechanism. Conclusion: These findings position STARS as an important regulator of skeletal muscle growth and regeneration.
- Published
- 2016
- Full Text
- View/download PDF
21. Regulation of Skeletal Muscle Myoblast Differentiation and Proliferation by Pannexins
- Author
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Kyle N. Cowan and Stéphanie Langlois
- Subjects
0301 basic medicine ,Myoblast proliferation ,Myogenesis ,Skeletal muscle ,Body movement ,Pannexin ,Biology ,Cell biology ,03 medical and health sciences ,030104 developmental biology ,Skeletal muscle cell differentiation ,medicine.anatomical_structure ,Skeletal muscle cell proliferation ,medicine ,Myocyte - Abstract
Pannexins are newly discovered channels that are now recognized as mediators of adenosine triphosphate release from several cell types allowing communication with the extracellular environment. Pannexins have been associated with various physiological and pathological processes including apoptosis, inflammation, and cancer. However, it is only recently that our work has unveiled a role for Pannexin 1 and Pannexin 3 as novel regulators of skeletal muscle myoblast proliferation and differentiation. Myoblast differentiation is an ordered multistep process that includes withdrawal from the cell cycle and the expression of key myogenic factors leading to myoblast differentiation and fusion into multinucleated myotubes. Eventually, myotubes will give rise to the diverse muscle fiber types that build the complex skeletal muscle architecture essential for body movement, postural behavior, and breathing. Skeletal muscle cell proliferation and differentiation are crucial processes required for proper skeletal muscle development during embryogenesis, as well as for the postnatal skeletal muscle regeneration that is necessary for muscle repair after injury or exercise. However, defects in skeletal muscle cell differentiation and/or deregulation of cell proliferation are involved in various skeletal muscle pathologies. In this review, we will discuss the expression of pannexins and their post-translational modifications in skeletal muscle, their known functions in various steps of myogenesis, including myoblast proliferation and differentiation, as well as their possible roles in skeletal muscle development, regeneration, and diseases such as Duchenne muscular dystrophy.
- Published
- 2016
22. Skeletal muscle cell proliferation and differentiation on polypyrrole substrates doped with extracellular matrix components
- Author
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Amy Gelmi, Magdalena Kita, Graeme M. Clark, Michael J. Higgins, Kerry J. Gilmore, Yao Han, Simon E. Moulton, Robert M. I. Kapsa, and Gordon G. Wallace
- Subjects
Male ,Materials science ,Polymers ,Surface Properties ,Cellular differentiation ,Muscle Fibers, Skeletal ,Biophysics ,Mice, Transgenic ,Bioengineering ,Biomaterials ,Extracellular matrix ,Mice ,Coated Materials, Biocompatible ,Tissue engineering ,Materials Testing ,Cell Adhesion ,medicine ,Animals ,Myocyte ,Pyrroles ,Cell adhesion ,Cells, Cultured ,Cell Proliferation ,Tissue Engineering ,Cell growth ,Stem Cells ,Skeletal muscle ,Cell Differentiation ,Extracellular Matrix ,Cell biology ,Mice, Inbred C57BL ,medicine.anatomical_structure ,Mechanics of Materials ,Skeletal muscle cell proliferation ,Ceramics and Composites ,Biomedical engineering - Abstract
Conducting polymers have been developed as substrates for in vitro studies with a range of cell types including electrically-excitable cells such as nerve and smooth muscle. The goal of this study was to optimise and characterise a range of polypyrrole materials to act as substrates for electrical stimulation of differentiating skeletal myoblasts. Although all of the polymer materials provided suitable substrates for myoblast adhesion and proliferation, significant differences became apparent under the low-serum conditions used for differentiation of primary myoblasts. The significance of the work lies in the design and control of polymer materials to facilitate different stages of skeletal muscle cell proliferation and/or differentiation, opening up opportunities for engineering of this tissue. This paper therefore constitutes not just a biocompatibility assessment but a comprehensive study of how synthesis conditions affect the final outcome in terms of cell response.
- Published
- 2009
23. Skeletal muscle Kv7 (KCNQ) channels in myoblast differentiation and proliferation
- Author
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Meritxell Roura-Ferrer, Laura Solé, Ramón Martínez-Mármol, Antonio Felipe, and Núria Villalonga
- Subjects
medicine.medical_specialty ,Myoblasts, Skeletal ,Biophysics ,Biology ,Muscle Development ,Biochemistry ,Cell Line ,Internal medicine ,medicine ,Animals ,Myocyte ,Molecular Biology ,Cell Proliferation ,Membrane potential ,KCNQ Potassium Channels ,Myogenesis ,Cell growth ,Cell Cycle ,Skeletal muscle ,Cell Differentiation ,Cell Biology ,Cell cycle ,Potassium channel ,Rats ,Cell biology ,Endocrinology ,medicine.anatomical_structure ,Skeletal muscle cell proliferation ,KCNQ1 Potassium Channel - Abstract
Voltage-dependent K(+) channels (Kv) are involved in myocyte proliferation and differentiation by triggering changes in membrane potential and regulating cell volume. Since Kv7 channels may participate in these events, the purpose of this study was to investigate whether skeletal muscle Kv7.1 and Kv7.5 were involved during proliferation and myogenesis. Here we report that, while myotube formation did not regulate Kv7 channels, Kv7.5 was up-regulated during cell cycle progression. Although, Kv7.1 mRNA also increased during the G(1)-phase, pharmacological evidence mainly involves Kv7.5 in myoblast growth. Our results indicate that the cell cycle-dependent expression of Kv7.5 is involved in skeletal muscle cell proliferation.
- Published
- 2008
24. Cell cycle-dependent expression of Kv1.5 is involved in myoblast proliferation
- Author
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Carmen Valenzuela, Miren David, Antonio Felipe, Meritxell Roura-Ferrer, Núria Villalonga, Concepció Soler, and Ramón Martínez-Mármol
- Subjects
Cyclin-Dependent Kinase Inhibitor p21 ,Myoblast proliferation ,Cell cycle checkpoint ,Myoblasts, Skeletal ,Proliferation ,Cyclin A ,Cell ,Gene Expression ,Skeletal muscle ,Cell cycle ,Cell Line ,Potassium channels ,Myoblasts ,Kv1.5 Potassium Channel ,medicine ,Animals ,Myocyte ,Molecular Biology ,Cell Proliferation ,Kv1.3 Potassium Channel ,biology ,Cell growth ,Cell Biology ,Rats ,Cell biology ,medicine.anatomical_structure ,Skeletal muscle cell proliferation ,biology.protein ,Cyclin-Dependent Kinase Inhibitor p27 - Abstract
Voltage-dependent K+ channels (Kv) are involved in the proliferation of many types of cells, but the mechanisms by which their activity is related to cell growth remain unclear. Kv antagonists inhibit the proliferation of mammalian cells, which is of physiological relevance in skeletal muscle. Although myofibres are terminally differentiated, some resident myoblasts may re-enter the cell cycle and proliferate. Here we report that the expression of Kv1.5 is cell-cycle dependent during myoblast proliferation. In addition to Kv1.5 other Kv, such as Kv1.3, are also up-regulated. However, pharmacological evidence mainly implicates Kv1.5 in myoblast growth. Thus, the presence of S0100176, a Kv antagonist, but not margatoxin and dendrotoxin, led to cell cycle arrest during the G1-phase. The use of selective cell cycle blockers showed that Kv1.5 was transiently accumulated during the early G1-phase. Furthermore, while myoblasts treated with S0100176 expressed low levels of cyclin A and D1, the expression of p21cip-1 and p27kip1, two cyclin-dependent kinase inhibitors, increased. Our results indicate that the cell cycle-dependent expression of Kv1.5 is involved in skeletal muscle cell proliferation. © 2008 Elsevier B.V. All rights reserved., This work was supported by the Ministerio de Educación y Ciencia (MEC), Spain (BFI2002-00764 and BFU2005-00695 to AF, SAF2007-65868 to CV, and BFU2006-06076 to CS), Generalitat de Catalunya 2005SGR00308 to AF and FIS D06/0014/0006 to CV. NV and RMM hold fellowships from the MEC and MRF from the Generalitat de Catalunya.
- Published
- 2008
25. Effect of Syndecan-1, Syndecan-4, and Glypican-1 on Turkey Muscle Satellite Cell Proliferation, Differentiation, and Responsiveness to Fibroblast Growth Factor 2
- Author
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Douglas C. McFarland, C. S. Coy, and Sandra G. Velleman
- Subjects
Male ,Turkeys ,medicine.medical_specialty ,Time Factors ,animal structures ,Satellite Cells, Skeletal Muscle ,Biology ,Transfection ,Fibroblast growth factor ,Syndecan 1 ,Mice ,Glypicans ,Internal medicine ,medicine ,Animals ,Receptor ,Cells, Cultured ,Glypican-1 ,Cell Proliferation ,Dose-Response Relationship, Drug ,Cell growth ,Muscle cell proliferation ,Cell Differentiation ,General Medicine ,Cell biology ,carbohydrates (lipids) ,Endocrinology ,Gene Expression Regulation ,Skeletal muscle cell proliferation ,embryonic structures ,Fibroblast Growth Factor 2 ,Syndecan-4 ,Animal Science and Zoology ,Syndecan-1 - Abstract
The membrane-associated heparan sulfate proteoglycan families consisting of the syndecans and glypicans are low-affinity receptors for fibroblast growth factor 2 (FGF2). Fibroblast growth factor 2 is a potent stimulator of skeletal muscle cell proliferation and a strong inhibitor of differentiation. Because syndecan-1, syndecan-4, and glypican-1 potentially play unique, but pivotal, roles in muscle cell proliferation and differentiation, these proteoglycans were examined for their effect on muscle cell proliferation and differentiation and FGF2 responsiveness. In the present study, turkey Randombred Control 2 line myogenic satellite cells were transfected with expression vector constructions of syndecan-1, syndecan-4, or glypican-1 to assay their role during muscle development and the effect on FGF2 responsiveness. During proliferation, only syndecan-1 increased proliferation. Both syndecan-4 and glypican-1 decreased proliferation at 72 h but generally did not affect the proliferation process. There was no interaction between the transfected gene and cell proliferation response to FGF2. Glypican-1 increased differentiation early in the process (24 h), and at later times differentiation was decreased by glypican-1. Both syndecan-1 and syndecan-4 overexpression decreased differentiation. During differentiation, except for glypican-1 at 48 h of differentiation, there was no interaction between gene treatment and FGF2 responsiveness. This result indicates that FGF2 responsiveness was not affected by the overexpression of syndecan-1, syndecan-4, and glypican-1 during differentiation. These data demonstrate that syndecan-1, syndecan-4, or glypican-1 differentially affect the processes of turkey muscle cell proliferation and differentiation, and can regulate these developmental stages in an FGF2-independent manner.
- Published
- 2007
26. Differentiation of bovine satellite cell-derived myoblasts under different culture conditions
- Author
-
Steffen Maak, Katja Will, Lisa Schering, Elke Albrecht, and Claudia Kalbe
- Subjects
Satellite Cells, Skeletal Muscle ,Cellular differentiation ,Cell Culture Techniques ,Skeletal muscle ,Cell Differentiation ,Cell Biology ,General Medicine ,Biology ,Cell biology ,Extracellular matrix ,Myoblasts ,Skeletal muscle cell differentiation ,medicine.anatomical_structure ,Biochemistry ,Skeletal muscle cell proliferation ,Cell culture ,medicine ,Myocyte ,Animals ,Cattle ,Muscle, Skeletal ,Fetal bovine serum ,Developmental Biology - Abstract
The aim of this study was to develop adequate in vitro conditions for the differentiation of bovine skeletal muscle cells. Therefore, satellite cells isolated from the left foreleg of a Holstein-Friesian fetus at 4.5 mo of gestation were seeded on 24-well plates coated with extracellular matrix gel. Cells were cultured for 5 d in growth medium containing 10% fetal bovine serum. After reaching confluence, several differentiation media were tested for inducing myotube formation. The highest fusion rate of approximately 30% was achieved with a serum-free medium containing 1 μM dexamethasone, 1 μg/ml linoleic acid, and 0.1 μM insulin after a differentiation phase of 72 h. Two different culture conditions (serum-free and serum-containing) appropriate for bovine skeletal muscle cell differentiation are described in detail which allow the investigation of bovine skeletal muscle cell proliferation and differentiation in general as well as in response to bioactive compounds.
- Published
- 2015
27. Designed Functional Dispersion for Insulin Protection from Pepsin Degradation and Skeletal Muscle Cell Proliferation: In Silico and In Vitro Study.
- Author
-
Chittepu, Veera C. S. R., Kalhotra, Poonam, Gallardo-Velázquez, Tzayhri, Robles-de la Torre, Raúl René, and Osorio-Revilla, Guillermo
- Subjects
- *
PEPSIN , *CELL proliferation , *INSULIN - Abstract
Functionalized single-walled carbon nanotubes with polyethylene glycol (PEGylated SWCNTs) are a promising nanomaterial that recently has emerged as the most attractive "cargo" to deliver chemicals, peptides, DNA and RNAs into cells. Insulin therapy is a recommended therapy to treat diabetes mellitus despite its side effects. Recently, functional dispersion made up of bioactive peptides, bioactive compounds and functionalized carbon nanomaterials such as PEGylated SWCNTs have proved to possess promising applications in nanomedicine. In the present study, molecular modeling simulations are utilized to assist in designing insulin hormone-PEGylated SWCNT composites, also called functional dispersion; to achieve this experimentally, an ultrasonication tool was utilized. Enzymatic degradation assay revealed that the designed functional dispersion protects about 70% of free insulin from pepsin. In addition, sulforhodamine B (SRB) assay, the quantification of insulin and glucose levels in differentiated skeletal muscle cell supernatants, reveals that functional dispersion regulates glucose and insulin levels to promote skeletal muscle cell proliferation. These findings offer new perspectives for designed functional dispersion, as potential pharmaceutical preparations to improve insulin therapy and promote skeletal muscle cell health. [ABSTRACT FROM AUTHOR]
- Published
- 2018
- Full Text
- View/download PDF
28. Dexamethasone enhances insulin-like growth factor-I effects on skeletal muscle cell proliferation. Role of specific intracellular signaling pathways
- Author
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R J Smith and Francesco Giorgino
- Subjects
medicine.medical_specialty ,Insulin Receptor Substrate Proteins ,medicine.medical_treatment ,Gene Expression ,Phosphatidylinositol 3-Kinases ,Biology ,Dexamethasone ,Cell Line ,Receptor, IGF Type 1 ,Insulin-like growth factor ,chemistry.chemical_compound ,Insulin-Like Growth Factor II ,Internal medicine ,medicine ,Animals ,Phosphatidylinositol ,Insulin-Like Growth Factor I ,Phosphorylation ,Muscle, Skeletal ,Phosphotyrosine ,Autocrine signalling ,Receptor Protein-Tyrosine Kinases ,Drug Synergism ,Tyrosine phosphorylation ,DNA ,General Medicine ,Blotting, Northern ,Phosphoproteins ,Rats ,Phosphotransferases (Alcohol Group Acceptor) ,Endocrinology ,chemistry ,Skeletal muscle cell proliferation ,Tyrosine ,Proto-Oncogene Proteins c-fos ,Cell Division ,Research Article ,Signal Transduction - Abstract
IGF-I stimulation of cell proliferation and c-Fos expression in skeletal muscle cells is markedly enhanced by dexamethasone. The effect of dexamethasone is not mediated by changes in IGF-binding proteins, as evidenced by similar effects of dexamethasone on the actions of insulin, PDGF-BB, and the IGF-I analogue long R3IGF-I. Dexamethasone also does not alter autocrine IGF-II secretion by muscle cells. To investigate the mechanism of the augmentation of IGF-I action, the effects of dexamethasone on intracellular IGF-I signaling pathways were determined. In dexamethasone-treated cells, the levels of IGF-I receptor tyrosine phosphorylation and receptor-associated phosphatidylinositol 3-kinase activity were increased. Dexamethasone-treated cells also showed increased and prolonged tyrosine phosphorylation of the Shc proteins. In contrast, dexamethasone decreased both tyrosine phosphorylation and expression of insulin receptor substrate 1 (IRS-1) and IRS-1-associated phosphatidylinositol 3-kinase activity. Thus, distinct signaling pathways activated by the IGF-I receptor in skeletal muscle cells are differentially regulated by dexamethasone. Potentiation of IGF-I action correlates with increased IGF-I receptor-associated phosphatidylinositol 3-kinase activity and tyrosine phosphorylation of Shc, but appears to be independent of activation of the IRS-1/phosphatidylinositol 3-kinase signaling pathway.
- Published
- 1995
29. The role of insulin-like-growth factor binding protein 2 (IGFBP2) and phosphatase and tensin homologue (PTEN) in the regulation of myoblast differentiation and hypertrophy
- Author
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Mark P. Lewis, Nasser Al-Shanti, Claire E. Stewart, Adam P. Sharples, and David C. Hughes
- Subjects
medicine.medical_specialty ,MAP Kinase Signaling System ,Endocrinology, Diabetes and Metabolism ,Myoblasts, Skeletal ,Blotting, Western ,Real-Time Polymerase Chain Reaction ,Immunoenzyme Techniques ,Mice ,Endocrinology ,Internal medicine ,medicine ,Tensin ,PTEN ,Myocyte ,Animals ,RNA, Messenger ,Phosphorylation ,RNA, Small Interfering ,Protein kinase B ,Creatine Kinase ,Cells, Cultured ,biology ,Reverse Transcriptase Polymerase Chain Reaction ,PTEN Phosphohydrolase ,Skeletal muscle ,Cell Differentiation ,Hypertrophy ,musculoskeletal system ,Flow Cytometry ,Cell biology ,Insulin-Like Growth Factor Binding Protein 2 ,medicine.anatomical_structure ,Phenotype ,Skeletal muscle cell proliferation ,Mitogen-activated protein kinase ,biology.protein ,C2C12 - Abstract
The complex actions of the insulin-like-growth factor binding proteins (IGFBPs) in skeletal muscle are becoming apparent, with IGFBP2 being implicated in skeletal muscle cell proliferation and differentiation (Ernst et al., 1992; Sharples et al., 2010). Furthermore, PTEN signalling has been linked to IGFBP2 action in other cell types by co-ordinating downstream Akt signalling, a known modulator of myoblast differentiation. The present study therefore aimed to determine the interaction between IGFBP2 and PTEN on myoblast differentiation. It has previously been established that C2C12 cells have high IGFBP2 gene expression upon transfer to low serum media, and that expression reduces rapidly as cells differentiate over 72 h [1]. Wishing to establish a potential role for IGFBP2 in this model, a neutralising IGFBP2 antibody was administered to C2C12 myoblasts upon initiation of differentiation. Myoblasts subsequently displayed reduced morphological differentiation (myotube number), biochemical differentiation (creatine kinase) and myotube hypertrophy (myotube area) with an early reduction in Akt phosphorylation. Knock-down of phosphatase and tensin homologue (PTEN) using siRNA in the absence of the neutralising antibody did not improve differentiation or hypertrophy vs. control conditions, however, in the presence of the neutralising IGFBP2 antibody, differentiation was restored and importantly hypertrophy exceeded that of control levels. Overall, these data suggest that; 1) reduced early availability of IGFBP2 can inhibit myoblast differentiation at later time points, 2) knock-down of PTEN levels can restore myoblast differentiation in the presence of neutralising IGFBP2 antibody, and 3) PTEN inhibition acts as a potent inducer of myotube hypertrophy when the availability of IGFBP2 is reduced in C2C12 myoblasts.
- Published
- 2012
30. Different effects of omega-3 fatty acids on the cell cycle in C2C12 myoblast proliferation
- Author
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Yunsheng Zhang, Jie Zhao, Yu Zheng, Ning Li, Xiaoxiang Hu, Tianyu Lu, Qiuyan Li, Ran Zhang, Yunqian Peng, and Fei Chang
- Subjects
MAPK/ERK pathway ,medicine.medical_specialty ,Myoblast proliferation ,Linolenic acid ,Cell Survival ,MAP Kinase Signaling System ,Clinical Biochemistry ,Gene Expression ,Cell Cycle Proteins ,Biology ,Cell Line ,Myoblasts ,Mice ,Internal medicine ,Cyclin E ,Fatty Acids, Omega-3 ,medicine ,Animals ,Cyclin D1 ,Phosphorylation ,Extracellular Signal-Regulated MAP Kinases ,Molecular Biology ,Cell Shape ,Cell Proliferation ,chemistry.chemical_classification ,Cell growth ,Cyclin-Dependent Kinase 2 ,food and beverages ,Cell Biology ,General Medicine ,Cell cycle ,G1 Phase Cell Cycle Checkpoints ,Endocrinology ,chemistry ,Biochemistry ,Skeletal muscle cell proliferation ,lipids (amino acids, peptides, and proteins) ,C2C12 ,Protein Processing, Post-Translational ,Polyunsaturated fatty acid - Abstract
Polyunsaturated fatty acids (PUFAs) are important molecules for human health. We investigated the effects of three major omega-3 PUFAs on C2C12 myoblast proliferation. Both docosahexaenoic (DHA) and eicosapentaenoic (EPA) acids decreased cell growth, whereas linolenic (ALA) acid did not, compared with the control. Cell cycle analysis showed that G(1) phase duration was increased markedly and S-phase duration was decreased by DHA and EPA. In contrast, there was no change in the G(1) or S-phase duration when the cells were treated with linolenic acid. To determine how DHA and EPA affected the cell cycle, cyclins and MAPK proteins were investigated. Western blotting and real-time quantitative PCR showed that DHA and EPA decreased cyclin E and CDK2 levels at both the protein and mRNA level. Also, MAPK phosphorylation levels were decreased by treatment with DHA and EPA. Our results indicated that different kinds of n-3 PUFA differentially affected myoblast cell proliferation. DHA and EPA decreased skeletal muscle cell proliferation through a mechanism involving MAPK-ERK.
- Published
- 2011
31. The effect of fibroblast growth factor 2 on the in vitro expression of syndecan-4 and glypican-1 in turkey satellite cells
- Author
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Sandra G. Velleman, Douglas C. McFarland, C. S. Coy, and X. Li
- Subjects
Turkeys ,animal structures ,Time Factors ,Satellite Cells, Skeletal Muscle ,Cellular differentiation ,medicine.medical_treatment ,Biology ,Fibroblast growth factor ,Polymerase Chain Reaction ,Cell Line ,Glypicans ,medicine ,Animals ,Cell Proliferation ,integumentary system ,Dose-Response Relationship, Drug ,Fibroblast growth factor receptor 2 ,Growth factor ,Cell Differentiation ,General Medicine ,Fibroblast growth factor receptor 4 ,Fibroblast growth factor receptor 3 ,Molecular biology ,Cell biology ,carbohydrates (lipids) ,Gene Expression Regulation ,Skeletal muscle cell proliferation ,Fibroblast growth factor receptor ,embryonic structures ,RNA ,Animal Science and Zoology ,Fibroblast Growth Factor 2 ,Syndecan-4 - Abstract
The membrane-associated heparan sulfate proteoglycan families, consisting of the syndecans and glypicans, are low-affinity receptors for fibroblast growth factor 2 (FGF2) that are essential in regulating the cellular response to FGF2. Fibroblast growth factor 2 is a potent stimulator of skeletal muscle cell proliferation and a strong inhibitor of differentiation. The regulation of the expression of the syndecans and glypicans will likely play a role in modulating the effects of FGF2 on cellular growth properties. In the present study, the effect of FGF2 on the expression of syndecan-4 and glypican-1 was measured by real-time PCR during turkey myogenic satellite cell proliferation and differentiation in vitro. Both syndecan-4 and glypican-1 transcription were influenced by the addition of exogenous FGF2. Syndecan-4 mRNA expression was reduced only during proliferation, whereas glypican-1 expression was reduced during both proliferation and differentiation. These results suggest that FGF2 growth factor signaling is, in part, regulated by an autoregulatory loop involving FGF2 regulation of syndecan-4 and glypican-1 expression and will affect the growth of skeletal muscle by modulating the proliferation and differentiation of satellite cells.
- Published
- 2008
32. Effects of dietary isoflavones on proliferation and DNA integrity of myoblasts derived from newborn piglets
- Author
-
Charlotte Rehfeldt, Torsten Viergutz, Marcus Mau, Claudia Kalbe, and Gerd Nürnberg
- Subjects
DNA Replication ,medicine.medical_specialty ,Time Factors ,Diet therapy ,Estrone ,Swine ,Myoblasts, Skeletal ,Genistein ,Apoptosis ,Phytoestrogens ,Biology ,chemistry.chemical_compound ,Internal medicine ,medicine ,Animals ,Cells, Cultured ,Cell Proliferation ,DNA synthesis ,Dose-Response Relationship, Drug ,Estradiol ,Cell growth ,Daidzein ,Cell Cycle ,food and beverages ,Isoflavones ,Endocrinology ,chemistry ,Animals, Newborn ,Skeletal muscle cell proliferation ,Protein Biosynthesis ,Pediatrics, Perinatology and Child Health ,DNA Damage - Abstract
Soy-based formulas are consumed by growing numbers of infants and used as regular food supplements in livestock production. Moreover, constituent dietary phytoestrogens may compete with endogenous estrogens and affect individual growth. This study aimed to investigate the in vitro effects of isoflavones in comparison with estrogens on the proliferation of porcine satellite cells derived from neonatal muscle. After 7 h of exposure in serum-free medium, 17beta-estradiol (1 nM, 1 microM), estrone (1 microM), and daidzein (1, 100 microM) slightly decreased whereas 100 microM genistein substantially lowered DNA synthesis. Declines in DNA amount were observed with genistein (1, 100 microM) and daidzein (100 microM). After 26 h of exposure, 100 microM genistein reduced DNA synthesis, whereas it was increased by 10 microM genistein and 10 and 100 microM daidzein. In the case of 10 microM genistein and 100 microM daidzein, these increases apparently resulted from the repair of damaged DNA. Genistein and daidzein (100 microM) reduced protein synthesis, caused a G2/M phase block, and decreased DNA amount in association with higher rates of cell death partially resulting from apoptosis. Conclusively, isoflavones at concentrations of greater than 1 muM act as inhibitors of porcine skeletal muscle cell proliferation.
- Published
- 2007
33. Myostatin induces cyclin D1 degradation to cause cell cycle arrest through a phosphatidylinositol 3-kinase/AKT/GSK-3 beta pathway and is antagonized by insulin-like growth factor 1
- Author
-
Zhenguo Wu, Wei Yang, Yong Zhang, Yanfeng Li, and Dahai Zhu
- Subjects
MAP Kinase Signaling System ,Cyclin D ,Cyclin A ,Cyclin B ,Apoptosis ,Myostatin ,Biochemistry ,Cell Line ,Glycogen Synthase Kinase 3 ,Mice ,Phosphatidylinositol 3-Kinases ,Cyclin D1 ,Transforming Growth Factor beta ,Animals ,Insulin-Like Growth Factor I ,Molecular Biology ,Protein kinase B ,PI3K/AKT/mTOR pathway ,Cells, Cultured ,Phosphoinositide-3 Kinase Inhibitors ,Mice, Knockout ,Glycogen Synthase Kinase 3 beta ,biology ,Chemistry ,Cell Cycle ,Cell Biology ,musculoskeletal system ,Skeletal muscle cell proliferation ,biology.protein ,Cancer research ,Proto-Oncogene Proteins c-akt ,Signal Transduction - Abstract
Myostatin is a transforming growth factor beta superfamily member and is known as an inhibitor of skeletal muscle cell proliferation and differentiation. Exposure to myostatin induces G1 phase cell cycle arrest. In this study, we demonstrated that myostatin down-regulates Cdk4 activity via promotion of cyclin D1 degradation. Overexpression of cyclin D1 significantly blocked myostatin-induced proliferation inhibition. We further showed that phosphorylation at threonine 286 by GSK-3beta was required for myostatin-stimulated cyclin D1 nuclear export and degradation. This process is dependent upon the activin receptor IIB and the phosphatidylinositol 3-kinase/Akt pathway but not Smad3. Insulin-like growth factor 1 (IGF-1) treatment or Akt activation attenuated the myostatin-stimulated cyclin D1 degradation as well as the associated cell proliferation repression. In contrast, attenuation of IGF-1 signaling caused C2C12 cells to undergo apoptosis in response to myostatin treatment. The observation that IGF-1 treatment increases myostatin expression through a phosphatidylinositol 3-kinase pathway suggests a possible feedback regulation between IGF-1 and myostatin. These findings uncover a novel role for myostatin in the regulation of cell growth and cell death in concert with IGF-1.
- Published
- 2006
34. Effects of glypican-1 on turkey skeletal muscle cell proliferation, differentiation and fibroblast growth factor 2 responsiveness
- Author
-
Caini Liu, Douglas C. McFarland, Sandra G. Velleman, and C. S. Coy
- Subjects
Male ,medicine.medical_specialty ,Turkeys ,Glypican ,Satellite Cells, Skeletal Muscle ,Cellular differentiation ,Biology ,Fibroblast growth factor ,Muscle Development ,Transfection ,Internal medicine ,medicine ,Animals ,Cell Proliferation ,integumentary system ,Fibroblast growth factor receptor 2 ,Cell Differentiation ,Cell Biology ,Fibroblast growth factor receptor 4 ,Fibroblast growth factor receptor 3 ,Cell biology ,Endocrinology ,Skeletal muscle cell proliferation ,Fibroblast growth factor receptor ,embryonic structures ,Fibroblast Growth Factor 2 ,Heparan Sulfate Proteoglycans ,Developmental Biology - Abstract
The heparan sulfate proteoglycan, glypican-1, is a low affinity receptor for fibroblast growth factor 2 (FGF2). Fibroblast growth factor 2 is a potent stimulator of skeletal muscle cell proliferation and an inhibitor of differentiation. Heparan sulfate proteoglycans like glypican-1 are required for FGF2 to transduce an intracellular signal. Understanding the role of glypican-1 in the regulation of FGF2-mediated signaling is important in furthering the understanding of the biological processes involved in muscle development and growth. In the current study, a turkey glypican-1 expression vector construct was transfected into turkey myogenic satellite cells resulting in the overexpression of glypican-1. The proliferation, differentiation, and responsiveness to FGF2 were measured in control and transfected cell cultures. The overexpression of glypican-1 in turkey myogenic satellite cells increased both satellite cell proliferation and FGF2 responsiveness, but decreased the rate of differentiation. The current data support glypican-1 modulation of both proliferation and differentiation through an FGF2-mediated pathway.
- Published
- 2006
35. In ovo exposure to monochromatic green light promotes skeletal muscle cell proliferation and affects myofiber growth in posthatch chicks
- Author
-
Y. Piestun, Orna Halevy, Zipora Yablonka-Reuveni, and Israel Rozenboim
- Subjects
medicine.medical_specialty ,animal structures ,Light ,Physiology ,Muscle Fibers, Skeletal ,Chick Embryo ,Biology ,Myosins ,In ovo ,Muscle Development ,Myoblasts ,Physiology (medical) ,Internal medicine ,medicine ,Myocyte ,Animals ,Muscle, Skeletal ,Myogenin ,Cell Proliferation ,Muscle Cells ,Antibodies, Monoclonal ,Gene Expression Regulation, Developmental ,PAX7 Transcription Factor ,Embryo ,Cell biology ,Endocrinology ,Skeletal muscle cell proliferation ,embryonic structures ,Muscle weight ,Monochromatic color ,PAX7 ,Chickens ,Biomarkers ,Photic Stimulation - Abstract
Our previous studies demonstrated that illumination of chicken embryos with monochromatic green light results in enhanced body and muscle weight at later posthatch stages. In the present study, we investigated the cellular and molecular basis of this phenomenon. First, we showed that on day 6 posthatch, myofibers were more uniform in the in ovo illuminated group than in the control group incubated in the dark, with respect to the number of myofibers displaying diameter values within the range of the mean value. Second, we tested the hypothesis that in ovo illumination causes an increase in the number of myoblasts; this in turn can promote posthatch muscle growth. Indeed, a significant increase in the number of skeletal muscle cells isolated from pectoralis muscle was observed in the in ovo illuminated group on days 1 and 3 posthatch relative to the control group. This increased cell number was accompanied by higher expression levels of Pax7 and myogenin proteins on posthatch days 1 and 3, respectively. A parallel analysis of proliferating cells in the intact muscle further demonstrated a significant increase in the number of cells positive for proliferating cell nuclear antigen in muscle from the in ovo illuminated group. Third, we demonstrated that the transition from fetal- to adult-type myoblasts, normally occurring in late stages of chicken embryogenesis, is initiated earlier in embryos subjected to in ovo green-light illumination. We suggest that the stimulatory effect of in ovo illumination on posthatch muscle growth is the result of enhanced proliferation and differentiation of adult myoblasts and myofiber synchronization.
- Published
- 2005
36. Effects of posthatch feed deprivation on heparan sulfate proteoglycan, syndecan-1, and glypican expression: implications for muscle growth potential in chickens
- Author
-
Sandra G. Velleman and Paul Mozdziak
- Subjects
Muscle tissue ,Male ,medicine.medical_specialty ,animal structures ,Glypican ,Syndecans ,Gene Expression ,Biology ,Fibroblast growth factor ,Syndecan 1 ,Muscle hypertrophy ,Internal medicine ,medicine ,Animals ,Receptor ,Muscle, Skeletal ,Messenger RNA ,Membrane Glycoproteins ,General Medicine ,carbohydrates (lipids) ,Endocrinology ,medicine.anatomical_structure ,Biochemistry ,Skeletal muscle cell proliferation ,embryonic structures ,Animal Science and Zoology ,Proteoglycans ,Food Deprivation ,Chickens ,Heparan Sulfate Proteoglycans - Abstract
The heparan sulfate proteoglycans, syndecan-1 and glypican-1 (glypican), are low affinity receptors for fibroblast growth factor 2 (FGF2). Because FGF2 stimulates skeletal muscle cell proliferation but inhibits differentiation, changes in FGF2 signaling due to early posthatch feed deprivation may play a significant role in modulating muscle growth. To study the effect of early posthatch feed deprivation in chickens on heparan sulfate proteoglycan relative protein concentration, syndecan-1 expression, and glypican mRNA expression, pectoralis major muscle tissue was isolated from pretreatment d 0 chicks and chicks fed or feed deprived for 3 d, and after d 3 feeding was resumed in the feed-deprived birds until d 7. Heparan sulfate proteoglycan protein concentration was measured by ELISA analysis and was significantly decreased in the feed-deprived birds beginning at d 2 (P0.05). The expression of syndecan-1 and glypican was measured by semi-quantitative reverse transcription PCR. Syndecan-1 expression was unaffected by feed withdrawal and refeeding (P0.05). Glypican mRNA expression was decreased in the muscle tissue from feed-deprived birds at d 3 (P0.05), but by d 7, after initiating feeding on d 4, it was significantly elevated compared with in muscle tissue from chicks maintained on feed (P0.05). The results from the present study demonstrate that the heparan sulfate proteoglycan protein concentration and syndecan-1 and glypican mRNA expressions are differentially affected by early posthatch feed deprivation, which may alter signaling events associated with muscle growth.
- Published
- 2005
37. Effects of syndecan-1 and glypican on muscle cell proliferation and differentiation: implications for possible functions during myogenesis
- Author
-
Xiaosong Liu, Sandra G. Velleman, C. S. Coy, and Douglas C. McFarland
- Subjects
Turkeys ,animal structures ,Glypican ,Syndecans ,Biology ,Fibroblast growth factor ,Transfection ,Syndecan 1 ,Mice ,Animals ,Muscle, Skeletal ,Cells, Cultured ,Membrane Glycoproteins ,Myogenesis ,Reverse Transcriptase Polymerase Chain Reaction ,Muscle cell proliferation ,Cell Differentiation ,General Medicine ,Molecular biology ,carbohydrates (lipids) ,Skeletal muscle cell proliferation ,Cell culture ,embryonic structures ,Animal Science and Zoology ,Fibroblast Growth Factor 2 ,Proteoglycans ,Syndecan-1 ,Chickens ,Cell Division ,Heparan Sulfate Proteoglycans ,Signal Transduction - Abstract
The heparan sulfate proteoglycans, syndecan-1 and glypican, are low-affinity receptors for fibroblast growth factor 2 (FGF2). Because FGF2 is a potent stimulator of skeletal muscle cell proliferation and a strong inhibitor of differentiation, it is likely that changes in syndecan-1 and glypican expression will affect myogenesis as both, in part, regulate FGF-dependent signaling. In the current study, expression vector constructs containing either syndecan-1 or glypican were transfected into turkey myogenic satellite cells resulting in the overexpression of these genes. The amount of expression of each of these genes was measured by semiquantitative reverse transcriptase polymerase chain reaction. The satellite cell cultures overexpressing syndecan-1 were unable to fuse to form multinucleated myotubes after differentiation was induced. The syndecan-1-transfected cells maintained a rounded morphology typical of cells during proliferation. In contrast, the satellite cells transfected with glypican formed larger myotubes. These results suggest that both syndecan-1 and glypican play pivotal, but different, roles in both muscle cell proliferation and differentiation.
- Published
- 2004
38. Developmental regulated expression of syndecan-1 and glypican in pectoralis major muscle in turkeys with different growth rates
- Author
-
Xiaosong Liu, Sandra G. Velleman, Douglas C. McFarland, and Karl E. Nestor
- Subjects
Male ,medicine.medical_specialty ,Turkeys ,animal structures ,Glypican ,Syndecans ,Molecular Sequence Data ,In situ hybridization ,Biology ,Fibroblast growth factor ,Syndecan 1 ,Sex Factors ,Internal medicine ,medicine ,Animals ,Receptor ,Muscle, Skeletal ,Cells, Cultured ,In Situ Hybridization ,DNA Primers ,Membrane Glycoproteins ,Base Sequence ,Reverse Transcriptase Polymerase Chain Reaction ,Skeletal muscle ,Gene Expression Regulation, Developmental ,Cell Biology ,Cell biology ,carbohydrates (lipids) ,Reverse transcription polymerase chain reaction ,Endocrinology ,medicine.anatomical_structure ,Skeletal muscle cell proliferation ,embryonic structures ,Female ,Fibroblast Growth Factor 2 ,Proteoglycans ,Sequence Alignment ,Heparan Sulfate Proteoglycans ,Developmental Biology - Abstract
Heparan sulfate proteoglycans, syndecan-1 and glypican, are low-affinity receptors for fibroblast growth factor 2 (FGF2). Since FGF2 stimulates skeletal muscle cell proliferation but inhibits differentiation, differences in syndecan-1 and glypican expression might affect muscle development and growth by changing the intensity of FGF2 signaling. In the present study, the pectoralis major muscle from 14 to 24-day-old-embryos, and from 1 to 16-week-old birds from a turkey line (F) selected for increased 16-week bodyweight and its genetic control line (RBC2), were used to address how syndecan-1 and glypican are expressed during skeletal muscle formation. The expression of syndecan-1 and glypican was measured by semiquantitative reverse transcription polymerase chain reaction. For males, the F-line embryos expressed more syndecan-1 (days 14 and 16) and glypican (days 14 and 18) than the RBC2 line. Similar line differences for males were observed during posthatch development. The male embryos from both lines expressed more syndecan-1 at days 18 through 22 and more glypican at days 20 and 22 than the corresponding females. The temporal and spatial distribution of syndecan-1 and glypican was detected by in situ hybridization. Syndecan-1 was identified in all muscle fibers at all embryonic stages studied, whereas the glypican was detected from embryonic day 18. The data from the current study provided new information about the expression of syndecan-1 and glypican as it relates to skeletal muscle growth properties.
- Published
- 2004
39. The transient expression of miR-203 and its inhibiting effects on skeletal muscle cell proliferation and differentiation
- Author
-
L Kong, X Zheng, Hao Wu, Xiquan Zhang, Qinghua Nie, S Lin, Wen Luo, Zhenhui Li, Y Ye, and S Hao
- Subjects
Cancer Research ,Myoblast proliferation ,Proto-Oncogene Proteins c-jun ,Cellular differentiation ,Immunology ,Down-Regulation ,Chick Embryo ,Biology ,Myoblasts ,Cellular and Molecular Neuroscience ,medicine ,Animals ,Myocyte ,Muscle, Skeletal ,Cells, Cultured ,Cell Proliferation ,Gene knockdown ,MEF2 Transcription Factors ,Cell growth ,Myogenesis ,Skeletal muscle ,Cell Differentiation ,Cell Biology ,Cell biology ,MicroRNAs ,medicine.anatomical_structure ,Skeletal muscle cell proliferation ,Original Article ,Chickens - Abstract
Previous studies have shown that miR-203 is a skin-specific microRNA (miRNA) with a profound role in skin cell differentiation. However, emerging microarray and deep sequencing data revealed that miR-203 is also expressed in embryonic skeletal muscle and myoblasts. In this study, we found that miR-203 was transiently upregulated in chicken embryos on days 10 to 16 (E10–E16) and was sharply downregulated and even not expressed after E16 in chicken embryonic skeletal muscle. Histological profiles and weight variations of embryo skeletal muscle revealed that miR-203 expression is correlated with muscle development. In vitro experiments showed that miR-203 exhibited downregulated expression during myoblast differentiation into myotubes. miR-203 overexpression inhibited myoblast proliferation and differentiation, whereas its loss-of-function increased myoblast proliferation and differentiation. During myogenesis, miR-203 can target and inhibit the expression of c-JUN and MEF2C, which were important for cell proliferation and muscle development, respectively. The overexpression of c-JUN significantly promoted myoblast proliferation. Conversely, knockdown of c-JUN by siRNA suppressed myoblast proliferation. In addition, the knockdown of MEF2C by siRNA significantly inhibited myoblast differentiation. Altogether, these data not only suggested that the expression of miR-203 is transitory during chicken skeletal muscle development but also showed a novel role of miR-203 in inhibiting skeletal muscle cell proliferation and differentiation by repressing c-JUN and MEF2C, respectively.
- Published
- 2014
40. EPO-receptor is present in mouse C2C12 and human primary skeletal muscle cells but EPO does not influence myogenesis
- Author
-
Andrew N. Stephens, Evelyn Zacharewicz, Séverine Lamon, and Aaron P. Russell
- Subjects
medicine.medical_specialty ,Physiology ,Myogenesis ,Skeletal muscle ,erythropoietin‐receptor ,Biology ,Cell biology ,Erythropoietin receptor ,medicine.anatomical_structure ,Endocrinology ,Skeletal muscle cell proliferation ,Erythropoietin ,hemic and lymphatic diseases ,Physiology (medical) ,Internal medicine ,medicine ,Myocyte ,erythropoietin ,myogenesis ,skeletal muscle ,Cytokine ,Protein kinase B ,C2C12 ,Original Research ,medicine.drug - Abstract
The role and regulation of the pleiotropic cytokine erythropoietin (EPO) in skeletal muscle are controversial. EPO exerts its effects by binding its specific receptor (EPO‐R), which activates intracellular signaling and gene transcription in response to internal and external stress signals. EPO is suggested to play a direct role in myogenesis via the EPO‐R, but several studies have questioned the effect of EPO treatment in muscle in vitro and in vivo. The lack of certainty surrounding the use of nonspecific EPO‐R antibodies contributes to the ambiguity of the field. Our study demonstrates that the EPO‐R gene and protein are expressed at each stage of mouse C2C12 and human skeletal muscle cell proliferation and differentiation and validates a specific antibody for the detection of the EPO‐R protein. However, in our experimental conditions, EPO treatment had no effect on mouse C2C12 and human muscle cell proliferation, differentiation, protein synthesis or EPO‐R expression. While an increase in Akt and MAPK phosphorylation was observed, we demonstrate that this effect resulted from the stress caused by changing medium and not from EPO treatment. We therefore suggest that skeletal muscle EPO‐R might be present in a nonfunctional form, or too lowly expressed to play a role in muscle cell function., The EPO‐R is expressed at the gene and protein level in mouse and human myoblasts and myotubes. However, EPO treatment does not seem to activate the EPO‐R and its downstream signaling pathways in skeletal muscle cells, questioning its functionality.
- Published
- 2014
41. Ethanol inhibits skeletal muscle cell proliferation and delays its differentiation in cell culture
- Author
-
Judit Garriga, Alvaro Urbano-Márquez, J Fernández-Solà, Roser Cussó, and E. Adanero
- Subjects
Nervous system ,endocrine system ,medicine.medical_specialty ,Muscle Proteins ,Biology ,Isozyme ,Rats, Sprague-Dawley ,Internal medicine ,mental disorders ,Gene expression ,medicine ,Animals ,Muscle, Skeletal ,Creatine Kinase ,reproductive and urinary physiology ,Cells, Cultured ,Ethanol ,Skeletal muscle ,Central Nervous System Depressants ,Cell Differentiation ,General Medicine ,DNA ,In vitro ,Rats ,Isoenzymes ,Endocrinology ,medicine.anatomical_structure ,Skeletal muscle cell proliferation ,Toxicity ,biology.protein ,Creatine kinase ,Cell Division - Abstract
Chronic ingestion of ethanol (EtOH) produces physiological and morphological alterations in skeletal muscle. The effects of EtOH on skeletal muscle have been studied in experimental animals or on biopsies from alcoholic patients. However, alterations in skeletal muscle from alcoholic patients could be secondary to the effects of EtOH on the nervous system. In this study, by assaying the action of EtOH on primary skeletal muscle cell cultures, we provide evidence of its direct effect on skeletal muscle proliferation and differentiation. The results indicate that EtOH: (1) significantly inhibits skeletal muscle cell proliferation at the beginning of the proliferation phase; (2) delays skeletal muscle differentiation, shown by the significant changes in the evolution of the percentage of the creatine kinase isozymes; (3) has no significant effect on skeletal muscle DNA or protein content during the proliferation phase.
- Published
- 2000
42. The effects of bFGF, IGF-I, and TGF-beta on RMo skeletal muscle cell proliferation and differentiation
- Author
-
Ronald E. Allen and Sally E. Johnson
- Subjects
DNA Replication ,medicine.medical_specialty ,medicine.medical_treatment ,Cellular differentiation ,Basic fibroblast growth factor ,Biology ,Fibroblast growth factor ,Tritium ,Cell Line ,Immunoenzyme Techniques ,chemistry.chemical_compound ,Somatomedins ,Internal medicine ,medicine ,Myocyte ,Animals ,Insulin-Like Growth Factor I ,Growth factor ,Muscles ,Skeletal muscle ,Cell Differentiation ,Cell Biology ,Rats ,Fibroblast Growth Factors ,Kinetics ,Endocrinology ,medicine.anatomical_structure ,chemistry ,Skeletal muscle cell proliferation ,Transforming Growth Factors ,Cell Division ,Transforming growth factor ,Thymidine - Abstract
A new skeletal muscle cell line, rat myoblast omega or RMo, has been characterized with regard to the effects of three growth factors: basic fibroblast growth factor (bFGF), insulin-like growth factor I (IGF-I), and transforming growth factor beta (TGF-beta). Results indicate a differential response of these factors on both cell proliferation and differentiation. Exposure to bFGF and IGF-I stimulate proliferation, while TGF-beta has no effect on cell number. RMo cell differentiation, as indicated by skeletal myosin synthesis, is enhanced by IGF-I, whereas both bFGF and TGF-beta suppress differentiation. These responses are in agreement with the effects of bFGF, IGF-I, and TGF-beta on myogenic cells cultured from fetal and postnatal muscle, thereby suggesting that RMo cells can serve as a model system for the study of growth factor effects on skeletal muscle cells.
- Published
- 1990
43. 004.Control of skeletal muscle cell proliferation and differentiation
- Author
-
M. Grounds
- Subjects
Cell growth ,Myogenesis ,Skeletal muscle ,NFAT ,Biology ,musculoskeletal system ,MyoD ,Cell biology ,Endocrinology ,medicine.anatomical_structure ,Reproductive Medicine ,Skeletal muscle cell proliferation ,Immunology ,Genetics ,medicine ,Myocyte ,Animal Science and Zoology ,Molecular Biology ,C2C12 ,Developmental Biology ,Biotechnology - Abstract
Skeletal muscle is formed by mononucleated precursor cells (myoblasts) that cease cell proliferation to start differentiation; this results in fusion between the myoblasts to form multinucleated cells (myotubes) that continue to differentiate (and fuse with more muscle cells) and mature into myofibres. Myogenesis has been widely used as a model to study in vitro factors controlling cell proliferation and differentiation. Condition in vitro may not reflect what happens in the more complex in vivo environment. Some of the key issues are what activates quiescent myoblasts in mature skeletal muscle in vivo, and what controls the switch between proliferation and differentiation? The role of the matrix, and molecules such as MyoD, p53, NFAT and IGF-1 will be considered.
- Published
- 2005
44. EFFECTS OF CHRONIC EXPOSURE TO SIMULATED MICROGRAVITY ON SKELETAL MUSCLE CELL PROLIFERATION AND DIFFERENTIATION
- Author
-
William E. Kraus, George A. Truskey, and Dorothy H. Slentz
- Subjects
Cellular differentiation ,Skeletal muscle ,Cell Biology ,General Medicine ,Cell cycle ,Biology ,Cell biology ,medicine.anatomical_structure ,Cell culture ,Skeletal muscle cell proliferation ,Myosin ,medicine ,Myocyte ,Stem cell ,Developmental Biology - Abstract
Cell culture models that mimic long-term exposure to microgravity provide important insights into the cellular biological adaptations of human skeletal muscle to long-term residence in space. We developed insert scaffolding for the NASA-designed rotating cell culture system (RCCS) in order to study the effects of time-averaged microgravity on the proliferation and differentiation of anchorage-dependent skeletal muscle myocytes. We hypothesized that prolonged microgravity exposure would result in the retardation of myocyte differentiation. Microgravity exposure in the RCCS resulted in increased cellular proliferation. Despite shifting to media conditions promoting cellular differentiation, 5 d later, there was an increase in cell number of approximately 62%, increases in total cellular protein (52%), and cellular proliferating cell nuclear antigen (PCNA) content (2.7 times control), and only a modest (insignificant) decrease (10%) in sarcomeric myosin protein expression. We grew cells in an inverted orientation on membrane inserts. Changes in cell number and PCNA content were the converse to those observed for cells in the RCCS. We also grew cells on inserts at unit gravity with constant mixing. Mixing accounted for part, but not all, of the effects of microgravity exposure on skeletal muscle cell cultures (53% of the RCCS effect on PCNA at 4–6 d). In summary, the mechanical effects of simulated microgravity exposure in the RCCS resulted in the maintenance of cellular proliferation, manifested as increases in cell number and expression of PCNA relative to control conditions, with only a modest reciprocal inhibition of cellular differentiation. Therefore, this model provides conditions wherein cellular differentiation and proliferation appear to be uncoupled.
- Published
- 2001
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