Analysis of Aerobic Respiration in Intact Skeletal Muscle Tissue by Microplate-Based Respirometry

Shintaku, Jonathan; Guttridge, Denis C.

doi:10.1007/978-1-4939-3810-0_23

Cited by 18 publications

(15 citation statements)

References 19 publications

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“…Our previous research using the murine C2C12 myoblast cell line showed that alternative NF-κB signaling is activated during myogenesis to promote OXPHOS in myotubes ( 3 , 48 ). Interestingly, RMS tumors are thought to derive from a skeletal muscle lineage that improperly differentiates and exhibits defects in mitochondrial gene expression, favoring a Warburg effect ( 49 , 50 ).…”

Section: Resultsmentioning

confidence: 99%

Classical NF-κB Metabolically Reprograms Sarcoma Cells Through Regulation of Hexokinase 2

Londhe

Ijiri

et al. 2018

Front. Oncol.

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BackgroundMetabolic reprogramming has emerged as a cancer hallmark, and one of the well-known cancer-associated metabolic alterations is the increase in the rate of glycolysis. Recent reports have shown that both the classical and alternative signaling pathways of nuclear factor κB (NF-κB) play important roles in controlling the metabolic profiles of normal cells and cancer cells. However, how these signaling pathways affect the metabolism of sarcomas, specifically rhabdomyosarcoma (RMS) and osteosarcoma (OS), has not been characterized.MethodsClassical NF-κB activity was inhibited through overexpression of the IκBα super repressor of NF-κB in RMS and OS cells. Global gene expression analysis was performed using Affymetrix GeneChip Human Transcriptome Array 2.0, and data were interpreted using gene set enrichment analysis. Seahorse Bioscience XFe24 was used to analyze oxygen consumption rate as a measure of aerobic respiration.ResultsInhibition of classical NF-κB activity in sarcoma cell lines restored alternative signaling as well as an increased oxidative respiratory metabolic phenotype in vitro. In addition, microarray analysis indicated that inhibition of NF-κB in sarcoma cells reduced glycolysis. We showed that a glycolytic gene, hexokinase (HK) 2, is a direct NF-κB transcriptional target. Knockdown of HK2 shifted the metabolic profile in sarcoma cells away from aerobic glycolysis, and re-expression of HK2 rescued the metabolic shift induced by inhibition of NF-κB activity in OS cells.ConclusionThese findings suggest that classical signaling of NF-κB plays a crucial role in the metabolic profile of pediatric sarcomas potentially through the regulation of HK2.

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Section: Resultsmentioning

confidence: 99%

Classical NF-κB Metabolically Reprograms Sarcoma Cells Through Regulation of Hexokinase 2

Londhe

Ijiri

et al. 2018

Front. Oncol.

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“…For muscle explants, mice were euthanized by CO 2 , and dissected diaphragms were placed in warm (37°C) Seahorse XF medium with 2 mmol/L L-glutamine, 1 mmol/L sodium pyruvate, and 10 mmol/L glucose. Sections (;2 3 2 3 1 mm) were transferred into XF24 islet capture microplates, seeded using islet capture screens, and processed (19).…”

Section: Glucose Metabolismmentioning

confidence: 99%

Regulation of Insulin Receptor Pathway and Glucose Metabolism by CD36 Signaling

et al. 2018

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During reduced energy intake, skeletal muscle maintains homeostasis by rapidly suppressing insulin-stimulated glucose utilization. Loss of this adaptation is observed with deficiency of the fatty acid transporter CD36. A similar loss is also characteristic of the insulin-resistant state where CD36 is dysfunctional. To elucidate what links CD36 to muscle glucose utilization, we examined whether CD36 signaling might influence insulin action. First, we show that CD36 deletion specific to skeletal muscle reduces expression of insulin signaling and glucose metabolism genes. It decreases muscle ceramides but impairs glucose disposal during a meal. Second, depletion of CD36 suppresses insulin signaling in primary-derived human myotubes, and the mechanism is shown to involve functional CD36 interaction with the insulin receptor (IR). CD36 promotes tyrosine phosphorylation of IR by the Fyn kinase and enhances IR recruitment of P85 and downstream signaling. Third, pretreatment for 15 min with saturated fatty acids suppresses CD36-Fyn enhancement of IR phosphorylation, whereas unsaturated fatty acids are neutral or stimulatory. These findings define mechanisms important for muscle glucose metabolism and optimal insulin responsiveness. Potential human relevance is suggested by genome-wide analysis and RNA sequencing data that associate genetically determined low muscle CD36 expression to incidence of type 2 diabetes.

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“…Tissue respirometry. Whole-tissue analysis of basal OCR was conducted adapting reported conditions for intact muscle tissue analysis (54) to the XF96 Extracellular Flux Analyzer platform (Agilent). Immediately after mouse sacrification, target muscle (quadriceps) tissues were quickly collected, rinsed in clean PBS buffer, and dissected into approximately 2 × 2 × 2 mm pieces.…”

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confidence: 99%

Pulsed glucocorticoids enhance dystrophic muscle performance through epigenetic-metabolic reprogramming

Quattrocelli¹,

Zelikovich²,

Jiang³

et al. 2019

JCI Insight

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Analysis of Aerobic Respiration in Intact Skeletal Muscle Tissue by Microplate-Based Respirometry

Cited by 18 publications

References 19 publications

Classical NF-κB Metabolically Reprograms Sarcoma Cells Through Regulation of Hexokinase 2

Classical NF-κB Metabolically Reprograms Sarcoma Cells Through Regulation of Hexokinase 2

Regulation of Insulin Receptor Pathway and Glucose Metabolism by CD36 Signaling

Pulsed glucocorticoids enhance dystrophic muscle performance through epigenetic-metabolic reprogramming

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