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[PMID]:29278025
[Au] Autor:Kataoka H; Nakano J; Kondo Y; Honda Y; Sakamoto J; Origuchi T; Okita M
[Ad] Endereço:1 Department of Locomotive Rehabilitation Science, Unit of Rehabilitation Sciences, Nagasaki University Graduate School of Biomedical Sciences , Nagasaki, Japan.
[Ti] Título:The influence of aging on the effectiveness of heat stress in preventing disuse muscle atrophy.
[So] Source:Physiol Int;104(4):316-328, 2017 Dec 01.
[Is] ISSN:2498-602X
[Cp] País de publicação:Hungary
[La] Idioma:eng
[Ab] Resumo:This study examined the aging effect on disuse muscle atrophy prevention using heat stress. Wistar rats aged 7 and 60 weeks were divided into three groups as follows: control, immobilized (Im), and immobilized and heat stressed (ImH). Heat stress was given by immersing the hindlimbs in hot water (42 °C) for 60 min, once in every 3 days and the gastrocnemius (GAS) and soleus (SOL) muscles were extracted after 14 days. Muscle-fiber types were classified using ATPase staining. Heat shock protein 70 (HSP70) was assessed through Western blotting. In GAS muscle of both groups and SOL muscle of 7-week-old rats, the fiber diameter of each muscle type in the ImH group significantly increased compared with that in the Im group. However, this could not be observed in the SOL muscle of the 60-week-old rats. The increased percentage of type-I fibers and variability of types I and II muscle-fiber diameter were evident in the SOL muscle of the 60-week rats. HSP70 was significantly elevated in the ImH group compared with in the Im group in both muscle types of both age groups. Thus, effectiveness of heat stress in the prevention of disuse muscle atrophy appears unsatisfactory in aging muscle fibers.
[Mh] Termos MeSH primário: Envelhecimento
Proteínas de Choque Térmico HSP70/metabolismo
Hipertermia Induzida/métodos
Músculo Esquelético/fisiopatologia
Transtornos Musculares Atróficos/prevenção & controle
Transtornos Musculares Atróficos/fisiopatologia
[Mh] Termos MeSH secundário: Animais
Resposta ao Choque Térmico
Masculino
Fibras Musculares Esqueléticas/patologia
Músculo Esquelético/patologia
Transtornos Musculares Atróficos/diagnóstico
Ratos
Ratos Wistar
Resultado do Tratamento
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (HSP70 Heat-Shock Proteins)
[Em] Mês de entrada:1803
[Cu] Atualização por classe:180309
[Lr] Data última revisão:
180309
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171227
[St] Status:MEDLINE
[do] DOI:10.1556/2060.104.2017.4.1


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[PMID]:28376674
[Au] Autor:Rubio-Arias JÁ; Ramos-Campo DJ; Esteban P; Martínez F; Jiménez JF
[Ad] Endereço:a Department of Physical Activity and Sports Sciences, Faculty of Sports, UCAM , Catholic University San Antonio , Murcia , Spain.
[Ti] Título:Effect of 6-weeks WBVT on the behaviour of the lower limb muscle fibres during vertical jumping.
[So] Source:J Sports Sci;36(4):398-406, 2018 Feb.
[Is] ISSN:1466-447X
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:The purpose of the current study was to examine the effect of 6 weeks of whole body vibration training (WBVT) on body composition, muscle activity of the gastrocnemius and vastus lateralis, gastrocnemius muscle architecture (static and dynamic) and ground reaction forces (performance jump) during the take-off phase of a countermovement jump in young healthy adult males. A total of 33 men (23.31 ± 5.62 years) were randomly assigned to a whole body vibration group (experimental group, EG : n = 17; 22.11 ± 4.97 years) or a control group (CG: n = 16; 24.5 ± 6.27 years). The total duration of the intervention phase (WBVT) was 6 weeks with a frequency of 3 sessions per week. Statistically significant differences were observed (P ≤ 0.05) between pre- and post-test in the power peak (Δ 1.91 W · kg ; P = 0.001), take-off velocity (0.1 cm · s ; P = 0.002) and jump height (Δ 0.4 cm; P = 0.002) for EG . There were no statistically significant differences in any of the body composition and muscle architecture variables. Moreover, no significant differences were found between EG and CG nor changes in muscle activity during take-off phase of the gastrocnemius and vastus lateralis pre- versus post-training. This study suggests that a 6-week WBVT programme with increasing intensity improves jump performance but does not alter muscle activity nor muscle architecture in healthy young men.
[Mh] Termos MeSH primário: Composição Corporal
Extremidade Inferior/fisiologia
Fibras Musculares Esqueléticas/fisiologia
Músculo Esquelético/fisiologia
Condicionamento Físico Humano/métodos
Exercício Pliométrico
Músculo Quadríceps/fisiologia
Vibração
[Mh] Termos MeSH secundário: Adulto
Eletromiografia
Seres Humanos
Masculino
Músculo Esquelético/anatomia & histologia
Músculo Esquelético/diagnóstico por imagem
Músculo Quadríceps/anatomia & histologia
Músculo Quadríceps/diagnóstico por imagem
Ultrassonografia
Adulto Jovem
[Pt] Tipo de publicação:JOURNAL ARTICLE; RANDOMIZED CONTROLLED TRIAL
[Em] Mês de entrada:1803
[Cu] Atualização por classe:180302
[Lr] Data última revisão:
180302
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170406
[St] Status:MEDLINE
[do] DOI:10.1080/02640414.2017.1309059


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[PMID]:29459024
[Au] Autor:Fang P; Yu M; Min W; Wan D; Han S; Shan Y; Wang R; Shi M; Zhang Z; Bo P
[Ad] Endereço:Department of Physiology, Nanjing University of Chinese Medicine Hanlin College, Taizhou, Jiangsu 225300, China; Department of Endocrinology, Clinical Medical College, Yangzhou University, Yangzhou, Jiangsu 225001, China.
[Ti] Título:Effect of baicalin on GLUT4 expression and glucose uptake in myotubes of rats.
[So] Source:Life Sci;196:156-161, 2018 Mar 01.
[Is] ISSN:1879-0631
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:AIMS: Although baicalin could attenuate obesity-induced insulin resistance, the detailed mechanism of baicalin on glucose uptake has not been sufficiently explored as yet. The aim of this study was to survey if baicalin might facilitate glucose uptake and to explore its signal mechanisms in L6 myotubes. MATERIALS AND METHODS: L6 myotubes were treated with 100, 200, 400 µM baicalin for 6 h, 12 h and 24 h in this study. Then 2-NBDG and insulin signal protein levels in myotubes of L6 cells were examined. KEY FINDINGS: We discovered that administration of baicalin enhanced GLUT4, PGC-1α, pP38MAPK, pAKT and pAS160 contents, as well as GLUT4 mRNA and PGC-1α mRNA levels in L6 myotubes. The beneficial metabolic changes elicited by baicalin were abrogated in myotubes of L6 by P38MAPK or AKT inhibitors. SIGNIFICANCE: These results suggest that baicalin promoted glucose uptake in myotubes by differential regulation on P38MAPK and AKT activity. In conclusion, these data provide insight that baicalin is a powerful and promising agent for the treament of hyperglycemia via AKT/AS160/GLUT4 and P38MAPK/PGC1α/GLUT4 pathway.
[Mh] Termos MeSH primário: Flavonoides/farmacologia
Transportador de Glucose Tipo 4/biossíntese
Glucose/metabolismo
Hipoglicemiantes/farmacologia
Fibras Musculares Esqueléticas/metabolismo
[Mh] Termos MeSH secundário: 4-Cloro-7-nitrobenzofurazano/análogos & derivados
4-Cloro-7-nitrobenzofurazano/metabolismo
Animais
Células Cultivadas
Desoxiglucose/análogos & derivados
Desoxiglucose/metabolismo
Insulina/metabolismo
Fibras Musculares Esqueléticas/efeitos dos fármacos
Músculo Esquelético/efeitos dos fármacos
Músculo Esquelético/metabolismo
Proteína Oncogênica v-akt/efeitos dos fármacos
Proteína Oncogênica v-akt/metabolismo
Coativador 1-alfa do Receptor gama Ativado por Proliferador de Peroxissomo/efeitos dos fármacos
Coativador 1-alfa do Receptor gama Ativado por Proliferador de Peroxissomo/metabolismo
Ratos
Transdução de Sinais/efeitos dos fármacos
Frações Subcelulares/efeitos dos fármacos
Frações Subcelulares/metabolismo
Proteínas Quinases p38 Ativadas por Mitógeno/efeitos dos fármacos
Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Flavonoids); 0 (Glucose Transporter Type 4); 0 (Hypoglycemic Agents); 0 (Insulin); 0 (Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha); 0 (Ppargc1a protein, rat); 0 (Slc2a4 protein, rat); 347Q89U4M5 (baicalin); 9G2MP84A8W (Deoxyglucose); EC 2.7.11.1 (Oncogene Protein v-akt); EC 2.7.11.24 (p38 Mitogen-Activated Protein Kinases); EQF2794IRE (4-Chloro-7-nitrobenzofurazan); IY9XDZ35W2 (Glucose); JE4F4P486R (2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxyglucose)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180226
[Lr] Data última revisão:
180226
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:180221
[St] Status:MEDLINE


  4 / 13517 MEDLINE  
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[PMID]:29329354
[Au] Autor:Perry BD; Rahnert JA; Xie Y; Zheng B; Woodworth-Hobbs ME; Price SR
[Ad] Endereço:Department of Medicine, Renal Division, Emory University, Atlanta, GA, United States of America.
[Ti] Título:Palmitate-induced ER stress and inhibition of protein synthesis in cultured myotubes does not require Toll-like receptor 4.
[So] Source:PLoS One;13(1):e0191313, 2018.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Saturated fatty acids, such as palmitate, are elevated in metabolically dysfunctional conditions like type 2 diabetes mellitus. Palmitate has been shown to impair insulin sensitivity and suppress protein synthesis while upregulating proteolytic systems in skeletal muscle. Increased sarco/endoplasmic reticulum (ER) stress and subsequent activation of the unfolded protein response may contribute to the palmitate-induced impairment of muscle protein synthesis. In some cell types, ER stress occurs through activation of the Toll-like receptor 4 (TLR4). Given the link between ER stress and suppression of protein synthesis, we investigated whether palmitate induces markers of ER stress and protein synthesis by activating TLR4 in cultured mouse C2C12 myotubes. Myotubes were treated with vehicle, a TLR4-specific ligand (lipopolysaccharides), palmitate, or a combination of palmitate plus a TLR4-specific inhibitor (TAK-242). Inflammatory indicators of TLR4 activation (IL-6 and TNFα) and markers of ER stress were measured, and protein synthesis was assessed using puromycin incorporation. Palmitate substantially increased the levels of IL-6, TNF-α, CHOP, XBP1s, and ATF 4 mRNAs and augmented the levels of CHOP, XBP1s, phospho-PERK and phospho-eIF2α proteins. The TLR4 antagonist attenuated both acute palmitate and LPS-induced increases in IL-6 and TNFα, but did not reduce ER stress signaling with either 6 h or 24 h palmitate treatment. Similarly, treating myotubes with palmitate for 6 h caused a 43% decline in protein synthesis consistent with an increase in phospho-eIF2α, and the TLR4 antagonist did not alter these responses. These results suggest that palmitate does not induce ER stress through TLR4 in muscle, and that palmitate impairs protein synthesis in skeletal muscle in part by induction of ER stress.
[Mh] Termos MeSH primário: Estresse do Retículo Endoplasmático/efeitos dos fármacos
Fibras Musculares Esqueléticas/citologia
Fibras Musculares Esqueléticas/metabolismo
Palmitatos/farmacologia
Biossíntese de Proteínas/efeitos dos fármacos
[Mh] Termos MeSH secundário: Animais
Células Cultivadas
Camundongos
Fibras Musculares Esqueléticas/efeitos dos fármacos
Fosforilação/efeitos dos fármacos
Proteínas Serina-Treonina Quinases/metabolismo
RNA Mensageiro/genética
RNA Mensageiro/metabolismo
Transdução de Sinais/efeitos dos fármacos
Receptor 4 Toll-Like/antagonistas & inibidores
Receptor 4 Toll-Like/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, N.I.H., EXTRAMURAL; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Palmitates); 0 (RNA, Messenger); 0 (Toll-Like Receptor 4); EC 2.7.11.1 (Protein-Serine-Threonine Kinases); EC 2.7.11.1 (eIF2alpha kinase, mouse)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180226
[Lr] Data última revisão:
180226
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:180113
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0191313


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[PMID]:29317646
[Au] Autor:Rao L; Qian Y; Khodabukus A; Ribar T; Bursac N
[Ad] Endereço:Department of Biomedical Engineering, Duke University, Durham, NC, 27708, USA.
[Ti] Título:Engineering human pluripotent stem cells into a functional skeletal muscle tissue.
[So] Source:Nat Commun;9(1):126, 2018 01 09.
[Is] ISSN:2041-1723
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:The generation of functional skeletal muscle tissues from human pluripotent stem cells (hPSCs) has not been reported. Here, we derive induced myogenic progenitor cells (iMPCs) via transient overexpression of Pax7 in paraxial mesoderm cells differentiated from hPSCs. In 2D culture, iMPCs readily differentiate into spontaneously contracting multinucleated myotubes and a pool of satellite-like cells endogenously expressing Pax7. Under optimized 3D culture conditions, iMPCs derived from multiple hPSC lines reproducibly form functional skeletal muscle tissues (iSKM bundles) containing aligned multi-nucleated myotubes that exhibit positive force-frequency relationship and robust calcium transients in response to electrical or acetylcholine stimulation. During 1-month culture, the iSKM bundles undergo increased structural and molecular maturation, hypertrophy, and force generation. When implanted into dorsal window chamber or hindlimb muscle in immunocompromised mice, the iSKM bundles survive, progressively vascularize, and maintain functionality. iSKM bundles hold promise as a microphysiological platform for human muscle disease modeling and drug development.
[Mh] Termos MeSH primário: Músculo Esquelético/citologia
Mioblastos/citologia
Células-Tronco Pluripotentes/citologia
Engenharia Tecidual/métodos
[Mh] Termos MeSH secundário: Animais
Diferenciação Celular
Células Cultivadas
Células HEK293
Seres Humanos
Células-Tronco Pluripotentes Induzidas/citologia
Células-Tronco Pluripotentes Induzidas/metabolismo
Camundongos Endogâmicos NOD
Camundongos Knockout
Camundongos Nus
Camundongos SCID
Fibras Musculares Esqueléticas/citologia
Fibras Musculares Esqueléticas/metabolismo
Músculo Esquelético/metabolismo
Mioblastos/metabolismo
Fator de Transcrição PAX7/metabolismo
Células-Tronco Pluripotentes/metabolismo
Transplante de Células-Tronco/métodos
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, N.I.H., EXTRAMURAL
[Nm] Nome de substância:
0 (PAX7 Transcription Factor)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180226
[Lr] Data última revisão:
180226
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:180111
[St] Status:MEDLINE
[do] DOI:10.1038/s41467-017-02636-4


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Registro de Ensaios Clínicos
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[PMID]:28743757
[Au] Autor:Nielsen J; Christensen AE; Nellemann B; Christensen B
[Ad] Endereço:Department of Sports Science and Clinical Biomechanics, SDU Muscle Research Cluster (SMRC), University of Southern Denmark, Odense M, Denmark; jnielsen@health.sdu.dk.
[Ti] Título:Lipid droplet size and location in human skeletal muscle fibers are associated with insulin sensitivity.
[So] Source:Am J Physiol Endocrinol Metab;313(6):E721-E730, 2017 12 01.
[Is] ISSN:1522-1555
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:In skeletal muscle, an accumulation of lipid droplets (LDs) in the subsarcolemmal space is associated with insulin resistance, but the underlying mechanism is not clear. We aimed to investigate how the size, number, and location of LDs are associated with insulin sensitivity and muscle fiber types and are regulated by aerobic training and treatment with an erythropoiesis-stimulating agent (ESA) in healthy young untrained men. LD analyses were performed by quantitative transmission electron microscopy, and insulin sensitivity was assessed by a hyperinsulinemic-euglycemic clamp. At baseline, we found that only the diameter (and not the number) of individual subsarcolemmal LDs was negatively associated with insulin sensitivity ( = 0.20, = 0.03, = 29). Despite 34% ( = 0.004) fewer LDs, the diameter of individual subsarcolemmal LDs was 20% ( = 0.0004) larger in type 2 fibers than in type 1 fibers. Furthermore, aerobic training decreased the size of subsarcolemmal LDs in the type 2 fibers, and ESA treatment lowered the number of both intermyofibrillar and subsarcolemmal LDs in the type 1 fibers. In conclusion, the size of individual subsarcolemmal LDs may be involved in the mechanism by which LDs are associated with insulin resistance in skeletal muscle.
[Mh] Termos MeSH primário: Resistência à Insulina
Gotículas Lipídicas/metabolismo
Metabolismo dos Lipídeos/fisiologia
Fibras Musculares Esqueléticas/metabolismo
Músculo Esquelético/metabolismo
[Mh] Termos MeSH secundário: Adolescente
Adulto
Exercício/fisiologia
Técnica Clamp de Glucose
Seres Humanos
Gotículas Lipídicas/patologia
Masculino
Tamanho da Partícula
Resistência Física
Adulto Jovem
[Pt] Tipo de publicação:JOURNAL ARTICLE; RANDOMIZED CONTROLLED TRIAL; RESEARCH SUPPORT, NON-U.S. GOV'T
[Em] Mês de entrada:1712
[Cu] Atualização por classe:180223
[Lr] Data última revisão:
180223
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170727
[Cl] Clinical Trial:ClinicalTrial
[St] Status:MEDLINE
[do] DOI:10.1152/ajpendo.00062.2017


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[PMID]:28465283
[Au] Autor:Pinto SK; Lamon S; Stephenson EJ; Kalanon M; Mikovic J; Koch LG; Britton SL; Hawley JA; Camera DM
[Ad] Endereço:Centre for Exercise and Nutrition, Mary MacKillop Institute for Health Research, Australian Catholic University, Melbourne, Victoria, Australia.
[Ti] Título:Expression of microRNAs and target proteins in skeletal muscle of rats selectively bred for high and low running capacity.
[So] Source:Am J Physiol Endocrinol Metab;313(3):E335-E343, 2017 09 01.
[Is] ISSN:1522-1555
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Impairments in mitochondrial function and substrate metabolism are implicated in the etiology of obesity and Type 2 diabetes. MicroRNAs (miRNAs) can degrade mRNA or repress protein translation and have been implicated in the development of such disorders. We used a contrasting rat model system of selectively bred high- (HCR) or low- (LCR) intrinsic running capacity with established differences in metabolic health to investigate the molecular mechanisms through which miRNAs regulate target proteins mediating mitochondrial function and substrate oxidation processes. Quantification of select miRNAs using the rat miFinder miRNA PCR array revealed differential expression of 15 skeletal muscles (musculus tibialis anterior) miRNAs between HCR and LCR rats (14 with higher expression in LCR; < 0.05). Ingenuity Pathway Analysis predicted these altered miRNAs to collectively target multiple proteins implicated in mitochondrial dysfunction and energy substrate metabolism. Total protein abundance of citrate synthase (CS; miR-19 target) and voltage-dependent anion channel 1 (miR-7a target) were higher in HCR compared with LCR cohorts (~57 and ~26%, respectively; < 0.05). A negative correlation was observed for miR-19a-3p and CS ( = 0.32, = 0.015) protein expression. To determine whether miR-19a-3p can regulate CS in vitro, we performed luciferase reporter and transfection assays in C2C12 myotubes. MiR-19a-3p binding to the CS untranslated region did not change luciferase reporter activity; however, miR-19a-3p transfection decreased CS protein expression (∼70%; < 0.05). The differential miRNA expression targeting proteins implicated in mitochondrial dysfunction and energy substrate metabolism may contribute to the molecular basis, mediating the divergent metabolic health profiles of LCR and HCR rats.
[Mh] Termos MeSH primário: Tolerância ao Exercício/genética
MicroRNAs/metabolismo
Mitocôndrias Musculares/metabolismo
Músculo Esquelético/metabolismo
Corrida
[Mh] Termos MeSH secundário: Animais
Western Blotting
Linhagem Celular
Citrato (si)-Sintase/metabolismo
Metabolismo Energético/genética
Técnicas In Vitro
Camundongos
Fibras Musculares Esqueléticas/metabolismo
RNA Mensageiro/metabolismo
Ratos
Ratos Endogâmicos
Reação em Cadeia da Polimerase em Tempo Real
Reação em Cadeia da Polimerase Via Transcriptase Reversa
Canal de Ânion 1 Dependente de Voltagem/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, N.I.H., EXTRAMURAL; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (MIRN103 microRNA, rat); 0 (MIRN181 microRNA, rat); 0 (MIRN19 microRNA, rat); 0 (MIRN194 microRNA, rat); 0 (MIRN223 microRNA, rat); 0 (MIRN24 microRNA, rat); 0 (MIRN26 microRNA, rat); 0 (MIRN30 microRNA, rat); 0 (MicroRNAs); 0 (RNA, Messenger); EC 1.6.- (Voltage-Dependent Anion Channel 1); EC 2.3.3.1 (Citrate (si)-Synthase)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:180222
[Lr] Data última revisão:
180222
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170504
[St] Status:MEDLINE
[do] DOI:10.1152/ajpendo.00043.2017


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[PMID]:28470530
[Au] Autor:Buttgereit A
[Ad] Endereço:Institute of Medical Biotechnology, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany. Andreas.Buttgereit@t-online.de.
[Ti] Título:Second Harmonic Generation Microscopy of Muscle Cell Morphology and Dynamics.
[So] Source:Methods Mol Biol;1601:229-241, 2017.
[Is] ISSN:1940-6029
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Microscopy in combination with contrast-increasing dyes allows the visualization and analysis of organs, tissues, and various cells. Because of their better resolution, the development of confocal and laser microscopes enables the investigations of cell components, which are labeled with fluorescent dyes. The imaging of living cells on subcellular level (also in vivo) needs a labeling by gene transfection of GFP or similar labeled proteins. We present a method for visualization of cell structure in skeletal and heart muscle by label-free Second Harmonic Generation (SHG) microscopy and describe analytic methods for quantitative measurements of morphology and dynamics in skeletal muscle fibers.
[Mh] Termos MeSH primário: Fibras Musculares Esqueléticas/ultraestrutura
Miócitos Cardíacos/ultraestrutura
Microscopia de Geração do Segundo Harmônico/métodos
[Mh] Termos MeSH secundário: Animais
Corantes Fluorescentes/química
Proteínas de Fluorescência Verde/química
Imagem Tridimensional
Microscopia Confocal
Fibras Musculares Esqueléticas/química
Miócitos Cardíacos/química
Miosinas/química
Fótons
Sarcômeros/química
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Fluorescent Dyes); 147336-22-9 (Green Fluorescent Proteins); EC 3.6.4.1 (Myosins)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180219
[Lr] Data última revisão:
180219
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170505
[St] Status:MEDLINE
[do] DOI:10.1007/978-1-4939-6960-9_18


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[PMID]:28470526
[Au] Autor:Friedrich O; Head SI
[Ad] Endereço:Friedrich-Alexander University (FAU) Erlangen-Nürnberg, Institute of Medical Biotechnology, Paul-Gordan-Street 3, Erlangen, 91052, Germany. oliver.friedrich@fau.de.
[Ti] Título:Quantitative Ratiometric Ca Imaging to Assess Cell Viability.
[So] Source:Methods Mol Biol;1601:171-193, 2017.
[Is] ISSN:1940-6029
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Viability of cells is strongly related to their Ca homeostasis. Ca signal fluctuations can be on a slow time scale, e.g., in non-excitable cells, but also in the range of tens of milliseconds for excitable cells, such as nerve and muscle. Muscle fibers respond to electrical stimulation with Ca transients that exceed their resting basal level about 100 times. Fluorescent Ca dyes have become an indispensable means to monitor Ca fluctuations in living cells online. Fluorescence intensity of such "environmental dyes" relies on a buffer-ligand interaction which is not only governed by laws of mass action but also by binding and unbinding kinetics that have to be considered for proper Ca kinetics and amplitude validation. The concept of Ca dyes including the different approaches using ratiometric and non-ratiometric dyes, the way to correctly choose dyes according to their low-/high-affinity properties and kinetics as well as staining techniques, and in situ calibration are reviewed and explained. We provide detailed protocols to apply ratiometric Fura-2 imaging of resting Ca and Ca fluctuations during field-stimulation in single isolated skeletal muscle cells and how to translate fluorescence intensities into absolute Ca concentrations using appropriate calibration techniques.
[Mh] Termos MeSH primário: Sinalização do Cálcio
Cálcio/metabolismo
Sobrevivência Celular
Imagem Molecular/métodos
[Mh] Termos MeSH secundário: Animais
Cálcio/análise
Quelantes de Cálcio/química
Calibragem
Estimulação Elétrica
Corantes Fluorescentes/química
Fura-2/química
Cinética
Microscopia de Fluorescência
Fibras Musculares Esqueléticas/citologia
Imagem Óptica/métodos
Análise de Célula Única
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Calcium Chelating Agents); 0 (Fluorescent Dyes); SY7Q814VUP (Calcium); TSN3DL106G (Fura-2)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180219
[Lr] Data última revisão:
180219
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170505
[St] Status:MEDLINE
[do] DOI:10.1007/978-1-4939-6960-9_14


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[PMID]:28470519
[Au] Autor:Smolina N; Bruton J; Kostareva A; Sejersen T
[Ad] Endereço:Karolinska Institutet, Stockholm, Sweden. natalia.smolina@ki.se.
[Ti] Título:Assaying Mitochondrial Respiration as an Indicator of Cellular Metabolism and Fitness.
[So] Source:Methods Mol Biol;1601:79-87, 2017.
[Is] ISSN:1940-6029
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Mitochondrial respiration is the most important generator of cellular energy under most circumstances. It is a process of energy conversion of substrates into ATP. The Seahorse equipment allows measuring oxygen consumption rate (OCR) in living cells and estimates key parameters of mitochondrial respiration in real-time mode. Through use of mitochondrial inhibitors, four key mitochondrial respiration parameters can be measured: basal, ATP production-linked, maximal, and proton leak-linked OCR. This approach requires application of mitochondrial inhibitors-oligomycin to block ATP synthase, FCCP-to make the inner mitochondrial membrane permeable for protons and allow maximum electron flux through the electron transport chain, and rotenone and antimycin A-to inhibit complexes I and III, respectively. This chapter describes the protocol of OCR assessment in the culture of primary myotubes obtained upon satellite cell fusion.
[Mh] Termos MeSH primário: Trifosfato de Adenosina/metabolismo
Bioensaio/instrumentação
Mitocôndrias/metabolismo
Fosforilação Oxidativa
Consumo de Oxigênio
[Mh] Termos MeSH secundário: Animais
Antimicina A/farmacologia
Carbonil Cianeto p-Trifluormetoxifenil Hidrazona/farmacologia
Respiração Celular
Sobrevivência Celular
Complexo I de Transporte de Elétrons/antagonistas & inibidores
Complexo III da Cadeia de Transporte de Elétrons/antagonistas & inibidores
Camundongos
Mitocôndrias/efeitos dos fármacos
Fibras Musculares Esqueléticas/efeitos dos fármacos
Fibras Musculares Esqueléticas/metabolismo
Oligomicinas/farmacologia
Cultura Primária de Células
Ionóforos de Próton/farmacologia
Rotenona/farmacologia
Células Satélites de Músculo Esquelético/efeitos dos fármacos
Células Satélites de Músculo Esquelético/metabolismo
Desacopladores/farmacologia
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Oligomycins); 0 (Proton Ionophores); 0 (Uncoupling Agents); 03L9OT429T (Rotenone); 370-86-5 (Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone); 642-15-9 (Antimycin A); 8L70Q75FXE (Adenosine Triphosphate); EC 1.10.2.2 (Electron Transport Complex III); EC 1.6.5.3 (Electron Transport Complex I)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180219
[Lr] Data última revisão:
180219
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170505
[St] Status:MEDLINE
[do] DOI:10.1007/978-1-4939-6960-9_7



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