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1.
bioRxiv ; 2023 Aug 07.
Artigo em Inglês | MEDLINE | ID: mdl-37609130

RESUMO

A key challenge of the modern genomics era is developing data-driven representations of gene function. Here, we present the first unbiased morphology-based genome-wide perturbation atlas in human cells, containing three genome-scale genotype-phenotype maps comprising >20,000 single-gene CRISPR-Cas9-based knockout experiments in >30 million cells. Our optical pooled cell profiling approach (PERISCOPE) combines a de-stainable high-dimensional phenotyping panel (based on Cell Painting1,2) with optical sequencing of molecular barcodes and a scalable open-source analysis pipeline to facilitate massively parallel screening of pooled perturbation libraries. This approach provides high-dimensional phenotypic profiles of individual cells, while simultaneously enabling interrogation of subcellular processes. Our atlas reconstructs known pathways and protein-protein interaction networks, identifies culture media-specific responses to gene knockout, and clusters thousands of human genes by phenotypic similarity. Using this atlas, we identify the poorly-characterized disease-associated transmembrane protein TMEM251/LYSET as a Golgi-resident protein essential for mannose-6-phosphate-dependent trafficking of lysosomal enzymes, showing the power of these representations. In sum, our atlas and screening technology represent a rich and accessible resource for connecting genes to cellular functions at scale.

2.
Mol Metab ; 66: 101643, 2022 12.
Artigo em Inglês | MEDLINE | ID: mdl-36400401

RESUMO

OBJECTIVE: Skeletal muscle is a pivotal organ for the coordination of systemic metabolism, constituting one of the largest storage site for glucose, lipids and amino acids. Tight temporal orchestration of protein breakdown in times of fasting has to be balanced with preservation of muscle mass and function. However, the molecular mechanisms that control the fasting response in muscle are poorly understood. METHODS: We now have identified a role for the peroxisome proliferator-activated receptor γ coactivator 1ß (PGC-1ß) in the regulation of catabolic pathways in this context in muscle-specific loss-of-function mouse models. RESULTS: Muscle-specific knockouts for PGC-1ß experience mitigated muscle atrophy in fasting, linked to reduced expression of myostatin, atrogenes, activation of AMP-dependent protein kinase (AMPK) and other energy deprivation signaling pathways. At least in part, the muscle fasting response is modulated by a negative effect of PGC-1ß on the nuclear factor of activated T-cells 1 (NFATC1). CONCLUSIONS: Collectively, these data highlight the complex regulation of muscle metabolism and reveal a new role for muscle PGC-1ß in the control of proteostasis in fasting.


Assuntos
Músculo Esquelético , Fatores de Transcrição , Animais , Camundongos , Jejum/fisiologia , Músculo Esquelético/metabolismo , Músculo Esquelético/patologia , Atrofia Muscular/metabolismo , Miofibrilas/patologia , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
3.
Sci Rep ; 12(1): 11477, 2022 07 07.
Artigo em Inglês | MEDLINE | ID: mdl-35798791

RESUMO

Non-alcoholic fatty liver disease is a continuum of disorders among which non-alcoholic steatohepatitis (NASH) is particularly associated with a negative prognosis. Hepatocyte lipotoxicity is one of the main pathogenic factors of liver fibrosis and NASH. However, the molecular mechanisms regulating this process are poorly understood. The main aim of this study was to dissect transcriptional mechanisms regulated by lipotoxicity in hepatocytes. We achieved this aim by combining transcriptomic, proteomic and chromatin accessibility analyses from human liver and mouse hepatocytes. This integrative approach revealed several transcription factor networks deregulated by NASH and lipotoxicity. To validate these predictions, genetic deletion of the transcription factors MAFK and TCF4 was performed, resulting in hepatocytes that were better protected against saturated fatty acid oversupply. MAFK- and TCF4-regulated gene expression profiles suggest a mitigating effect against cell stress, while promoting cell survival and growth. Moreover, in the context of lipotoxicity, some MAFK and TCF4 target genes were to the corresponding differentially regulated transcripts in human liver fibrosis. Collectively, our findings comprehensively profile the transcriptional response to lipotoxicity in hepatocytes, revealing new molecular insights and providing a valuable resource for future endeavours to tackle the molecular mechanisms of NASH.


Assuntos
Hepatopatia Gordurosa não Alcoólica , Animais , Hepatócitos/patologia , Fígado/patologia , Cirrose Hepática/patologia , Camundongos , Camundongos Endogâmicos C57BL , Hepatopatia Gordurosa não Alcoólica/patologia , Proteômica
4.
Proc Natl Acad Sci U S A ; 118(36)2021 09 07.
Artigo em Inglês | MEDLINE | ID: mdl-34465622

RESUMO

Plasticity of cells, tissues, and organs is controlled by the coordinated transcription of biological programs. However, the mechanisms orchestrating such context-specific transcriptional networks mediated by the dynamic interplay of transcription factors and coregulators are poorly understood. The peroxisome proliferator-activated receptor γ coactivator 1α (PGC-1α) is a prototypical master regulator of adaptive transcription in various cell types. We now uncovered a central function of the C-terminal domain of PGC-1α to bind RNAs and assemble multiprotein complexes including proteins that control gene transcription and RNA processing. These interactions are important for PGC-1α recruitment to chromatin in transcriptionally active liquid-like nuclear condensates. Notably, such a compartmentalization of active transcription mediated by liquid-liquid phase separation was observed in mouse and human skeletal muscle, revealing a mechanism by which PGC-1α regulates complex transcriptional networks. These findings provide a broad conceptual framework for context-dependent transcriptional control of phenotypic adaptations in metabolically active tissues.


Assuntos
Núcleo Celular/metabolismo , Regulação da Expressão Gênica/fisiologia , Coativador 1-alfa do Receptor gama Ativado por Proliferador de Peroxissomo/fisiologia , RNA/metabolismo , Animais , Linhagem Celular , Cromatina/metabolismo , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Coativador 1-alfa do Receptor gama Ativado por Proliferador de Peroxissomo/metabolismo , Domínios Proteicos , Domínios e Motivos de Interação entre Proteínas
5.
Sci Rep ; 8(1): 636, 2018 01 12.
Artigo em Inglês | MEDLINE | ID: mdl-29330505

RESUMO

SRP-35 is a short-chain dehydrogenase/reductase belonging to the DHRS7C dehydrogenase/ reductase family 7. Here we show that its over-expression in mouse skeletal muscles induces enhanced muscle performance in vivo, which is not related to alterations in excitation-contraction coupling but rather linked to enhanced glucose metabolism. Over-expression of SRP-35 causes increased phosphorylation of AktS473, triggering plasmalemmal targeting of GLUT4 and higher glucose uptake into muscles. SRP-35 signaling involves RARα and RARγ (non-genomic effect), PI3K and mTORC2. We also demonstrate that all-trans retinoic acid, a downstream product of the enzymatic activity of SRP-35, mimics the effect of SRP-35 in skeletal muscle, inducing a synergistic effect with insulin on AKTS473 phosphorylation. These results indicate that SRP-35 affects skeletal muscle metabolism and may represent an important target for the treatment of metabolic diseases.


Assuntos
Glucose/metabolismo , Alvo Mecanístico do Complexo 2 de Rapamicina/metabolismo , Músculo Esquelético/fisiologia , Oxirredutases/genética , Oxirredutases/metabolismo , Animais , Expressão Gênica , Transportador de Glucose Tipo 4/metabolismo , Sistema de Sinalização das MAP Quinases , Masculino , Camundongos , Camundongos Transgênicos , Fosforilação , Receptores do Ácido Retinoico , Receptor alfa de Ácido Retinoico/metabolismo
6.
Sci Rep ; 7(1): 13485, 2017 10 18.
Artigo em Inglês | MEDLINE | ID: mdl-29044196

RESUMO

Enhanced coverage and sensitivity of next-generation 'omic' platforms has allowed the characterization of gene, metabolite and protein responses in highly metabolic tissues, such as, skeletal muscle. A limitation, however, is the capability to determine interaction between dynamic biological networks. To address this limitation, we applied Weighted Analyte Correlation Network Analysis (WACNA) to RNA-seq and metabolomic datasets to identify correlated subnetworks of transcripts and metabolites in response to a high-fat diet (HFD)-induced obesity and/or exercise. HFD altered skeletal muscle lipid profiles and up-regulated genes involved in lipid catabolism, while decreasing 241 exercise-responsive genes related to skeletal muscle plasticity. WACNA identified the interplay between transcript and metabolite subnetworks linked to lipid metabolism, inflammation and glycerophospholipid metabolism that were associated with IL6, AMPK and PPAR signal pathways. Collectively, this novel experimental approach provides an integrative resource to study transcriptional and metabolic networks in skeletal muscle in the context of health and disease.


Assuntos
Dieta Hiperlipídica , Metaboloma , Músculo Esquelético/metabolismo , Esforço Físico , Transcriptoma , Quinases Proteína-Quinases Ativadas por AMP , Animais , Glicerofosfolipídeos/metabolismo , Interleucina-6/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Músculo Esquelético/fisiologia , Receptores Ativados por Proliferador de Peroxissomo/metabolismo , Proteínas Quinases/metabolismo , Transdução de Sinais
7.
Am J Physiol Cell Physiol ; 313(3): C257-C261, 2017 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-28659288

RESUMO

Although the signal pathways mediating muscle protein synthesis and degradation are well characterized, the transcriptional processes modulating skeletal muscle mass and adaptive growth are poorly understood. Recently, studies in mouse models of muscle wasting or acutely exercised human muscle have suggested a potential role for the transcription factor signal transducer and activator of transcription 3 (STAT3), in adaptive growth. Hence, in the present study we sought to define the contribution of STAT3 to skeletal muscle adaptive growth. In contrast to previous work, two different resistance exercise protocols did not change STAT3 phosphorylation in human skeletal muscle. To directly address the role of STAT3 in load-induced (i.e., adaptive) growth, we studied the anabolic effects of 14 days of synergist ablation (SA) in skeletal muscle-specific STAT3 knockout (mKO) mice and their floxed, wild-type (WT) littermates. Plantaris muscle weight and fiber area in the nonoperated leg (control; CON) was comparable between genotypes. As expected, SA significantly increased plantaris weight, muscle fiber cross-sectional area, and anabolic signaling in WT mice, although interestingly, this induction was not impaired in STAT3 mKO mice. Collectively, these data demonstrate that STAT3 is not required for overload-mediated hypertrophy in mouse skeletal muscle.


Assuntos
Músculo Esquelético/fisiopatologia , Miofibrilas/metabolismo , Miofibrilas/patologia , Treinamento Resistido/efeitos adversos , Fator de Transcrição STAT3/genética , Fator de Transcrição STAT3/metabolismo , Animais , Técnicas de Inativação de Genes , Hipertrofia/etiologia , Hipertrofia/genética , Hipertrofia/patologia , Hipertrofia/fisiopatologia , Masculino , Camundongos , Camundongos Knockout , Músculo Esquelético/patologia , Tamanho do Órgão
8.
Eur J Pharmacol ; 765: 429-36, 2015 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-26364538

RESUMO

Cyclic nucleotide phosphodiesterase (PDE)3 and PDE4 provide the major PDE activity in cardiac myocytes and shape ß1-adrenoceptor-dependent cardiac cAMP signaling but their role in regulating ß2-adrenoceptor-mediated responses is less well known. We investigated potential differences in PDE3 and PDE4 activities between right (RV) and left (LV) ventricular myocardium, and their role in regulating ß2-adrenoceptor effects. PDE3 activity in the microsomal fraction was lower in RV than in LV but was the same in the cytosolic fraction. However, no significant difference between RV and LV was found when the PDE4 activity was studied. ß2-adrenoceptor activation increased inotropism and lusitropism in LV when measured in the presence of either the PDE3 inhibitor cilostamide, the PDE4 inhibitor rolipram or a non-selective PDE inhibitor IBMX. However, the joint inhibition of both PDE3 and PDE4 was necessary in RV to uncover ß2-adrenoceptor-induced inotropic and lusitropic effects. Our results indicate different regulation of ß2-adrenoceptor-mediated contractility by PDE3 and PDE4 in RV and LV of the rat heart. In the case of PDE3 due to a different contribution of the enzyme in the microsomal fraction whereas in the case of PDE4 it can be attributed to differences in the intracellular distribution and coupling to ß2-adrenoceptors.


Assuntos
Contração Miocárdica/fisiologia , Inibidores da Fosfodiesterase 3/farmacologia , Inibidores da Fosfodiesterase 4/farmacologia , Receptores Adrenérgicos beta 2/fisiologia , Função Ventricular Esquerda/fisiologia , Função Ventricular Direita/fisiologia , 3',5'-AMP Cíclico Fosfodiesterases/fisiologia , Animais , Nucleotídeo Cíclico Fosfodiesterase do Tipo 3/fisiologia , Relação Dose-Resposta a Droga , Masculino , Contração Miocárdica/efeitos dos fármacos , Técnicas de Cultura de Órgãos , Ratos , Ratos Sprague-Dawley , Função Ventricular Esquerda/efeitos dos fármacos , Função Ventricular Direita/efeitos dos fármacos
9.
J Physiol ; 593(18): 4275-84, 2015 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-26227152

RESUMO

The present study aimed to investigate the role of the mechanistic target of rapamycin complex 1 (mTORC1) in the regulation of myofibrillar (MyoPS) and mitochondrial (MitoPS) protein synthesis following endurance exercise. Forty-two female C57BL/6 mice performed 1 h of treadmill running (18 m min(-1) ; 5° grade), 1 h after i.p. administration of rapamycin (1.5 mg · kg(-1) ) or vehicle. To quantify skeletal muscle protein fractional synthesis rates, a flooding dose (50 mg · kg(-1) ) of l-[ring-(13) C6 ]phenylalanine was administered via i.p. injection. Blood and gastrocnemius muscle were collected in non-exercised control mice, as well as at 0.5, 3 and 6 h after completing exercise (n = 4 per time point). Skeletal muscle MyoPS and MitoPS were determined by measuring isotope incorporation in their respective protein pools. Activation of the mTORC1-signalling cascade was measured via direct kinase activity assay and immunoblotting, whereas genes related to mitochondrial biogenesis were measured via a quantitative RT-PCR. MyoPS increased rapidly in the vehicle group post-exercise and remained elevated for 6 h, whereas this response was transiently blunted (30 min post-exercise) by rapamycin. By contrast, MitoPS was unaffected by rapamycin, and was increased over the entire post-exercise recovery period in both groups (P < 0.05). Despite rapid increases in both MyoPS and MitoPS, mTORC1 activation was suppressed in both groups post-exercise for the entire 6 h recovery period. Peroxisome proliferator activated receptor-γ coactivator-1α, pyruvate dehydrogenase kinase 4 and mitochondrial transcription factor A mRNA increased post-exercise (P < 0.05) and this response was augmented by rapamycin (P < 0.05). Collectively, these data suggest that endurance exercise stimulates MyoPS and MitoPS in skeletal muscle independently of mTORC1 activation.


Assuntos
Mitocôndrias/fisiologia , Miofibrilas/fisiologia , Condicionamento Físico Animal/fisiologia , Biossíntese de Proteínas/efeitos dos fármacos , Biossíntese de Proteínas/fisiologia , Sirolimo/farmacologia , Animais , Terapia por Exercício/métodos , Feminino , Camundongos , Camundongos Endogâmicos C57BL , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Músculo Esquelético/efeitos dos fármacos , Músculo Esquelético/metabolismo , Miofibrilas/efeitos dos fármacos , Miofibrilas/metabolismo , Biogênese de Organelas , Proteínas Quinases/metabolismo , RNA Mensageiro/metabolismo , Transdução de Sinais/efeitos dos fármacos , Serina-Treonina Quinases TOR/metabolismo
10.
Clin Sci (Lond) ; 129(7): 589-99, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26186742

RESUMO

Skeletal muscle metabolism is highly dependent on mitochondrial function, with impaired mitochondrial biogenesis associated with the development of metabolic diseases such as insulin resistance and type 2 diabetes. Mitochondria display substantial plasticity in skeletal muscle, and are highly sensitive to levels of physical activity. It is thought that physical activity promotes mitochondrial biogenesis in skeletal muscle through increased expression of genes encoded in both the nuclear and the mitochondrial genome; however, how this process is co-ordinated at the cellular level is poorly understood. Nuclear receptors (NRs) are key signalling proteins capable of integrating environmental factors and mitochondrial function, thereby providing a potential link between exercise and mitochondrial biogenesis. The aim of this review is to highlight the function of NRs in skeletal muscle mitochondrial biogenesis and discuss the therapeutic potential of NRs for the management and treatment of chronic metabolic disease.


Assuntos
Regulação da Expressão Gênica , Mitocôndrias/fisiologia , Músculo Esquelético/fisiologia , Receptores Citoplasmáticos e Nucleares/fisiologia , Animais , Galinhas , Diabetes Mellitus Tipo 2/metabolismo , Exercício Físico , Genoma Mitocondrial , Humanos , Camundongos , Mitocôndrias Musculares/fisiologia , Doenças Musculares/metabolismo , Ovalbumina/metabolismo , Receptores Ativados por Proliferador de Peroxissomo/fisiologia , Condicionamento Físico Animal , Ratos , Receptores dos Hormônios Tireóideos/fisiologia , Fatores de Transcrição/fisiologia
11.
Exp Biol Med (Maywood) ; 240(9): 1205-13, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-25432985

RESUMO

The important regulator of cardiac function, cAMP, is hydrolyzed by different cyclic nucleotide phosphodiesterases (PDEs), whose expression and activity are not uniform throughout the heart. Of these enzymes, PDE2 shapes ß1 adrenoceptor-dependent cardiac cAMP signaling, both in the right and left ventricular myocardium, but its role in regulating ß2 adrenoceptor-mediated responses is less well known. Our aim was to investigate possible differences in PDE2 transcription and activity between right (RV) and left (LV) rat ventricular myocardium, as well as its role in regulating ß2 adrenoceptor effects. The free walls of the RV and the LV were obtained from Sprague-Dawley rat hearts. Relative mRNA for PDE2 (quantified by qPCR) and PDE2 activity (evaluated by a colorimetric procedure and using the PDE2 inhibitor EHNA) were determined in RV and LV. Also, ß2 adrenoceptor-mediated effects (ß2-adrenoceptor agonist salbutamol + ß1 adrenoceptor antagonist CGP-20712A) on contractility and cAMP concentrations, in the absence or presence of EHNA, were studied in the RV and LV. PDE2 transcript levels were less abundant in RV than in LV and the contribution of PDE2 to the total PDE activity was around 25% lower in the microsomal fraction of the RV compared with the LV. ß2 adrenoceptor activation increased inotropy and cAMP levels in the LV when measured in the presence of EHNA, but no such effects were observed in the RV, either in the presence or absence of EHNA. These results indicate interventricular differences in PDE2 transcript and activity levels, which may distinctly regulate ß2 adrenoceptor-mediated contractility and cAMP concentrations in the RV and in the LV of the rat heart.


Assuntos
Nucleotídeo Cíclico Fosfodiesterase do Tipo 2/metabolismo , Miocárdio/metabolismo , Receptores Adrenérgicos beta 2/metabolismo , Adenina/análogos & derivados , Adenina/farmacologia , Agonistas de Receptores Adrenérgicos beta 2/farmacologia , Antagonistas Adrenérgicos beta/farmacologia , Albuterol/farmacologia , Animais , AMP Cíclico/metabolismo , Nucleotídeo Cíclico Fosfodiesterase do Tipo 2/antagonistas & inibidores , Nucleotídeo Cíclico Fosfodiesterase do Tipo 2/genética , Ventrículos do Coração/efeitos dos fármacos , Ventrículos do Coração/metabolismo , Imidazóis/farmacologia , Técnicas In Vitro , Contração Miocárdica/efeitos dos fármacos , Inibidores de Fosfodiesterase/farmacologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley
12.
Eur J Sport Sci ; 15(1): 41-52, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25159707

RESUMO

Concurrent training (the combination of endurance exercise to resistance training) is a common practice for athletes looking to maximise strength and endurance. Over 20 years ago, it was first observed that performing endurance exercise after resistance exercise could have detrimental effects on strength gains. At the cellular level, specific protein candidates have been suggested to mediate this training interference; however, at present, the physiological reason(s) behind the concurrent training effect remain largely unknown. Even less is known regarding the optimal nutritional strategies to support concurrent training and whether unique nutritional approaches are needed to support endurance and resistance exercise during concurrent training approaches. In this review, we will discuss the importance of protein supplementation for both endurance and resistance training adaptation and highlight additional nutritional strategies that may support concurrent training. Finally, we will attempt to synergise current understanding of the interaction between physiological responses and nutritional approaches into practical recommendations for concurrent training.


Assuntos
Condicionamento Físico Humano/métodos , Condicionamento Físico Humano/fisiologia , Resistência Física/fisiologia , Treinamento Resistido , Fenômenos Fisiológicos da Nutrição Esportiva/fisiologia , Animais , Ciclismo , Humanos , Masculino , Ratos , Ratos Wistar , Transdução de Sinais , Levantamento de Peso
13.
Am J Physiol Cell Physiol ; 307(9): C763-73, 2014 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-25186010

RESUMO

It is well established that exercise elicits a finely tuned adaptive response in skeletal muscle, with contraction frequency, duration, and recovery shaping skeletal muscle plasticity. Given the power of physical activity to regulate metabolic health, numerous research groups have focused on the molecular mechanisms that sense, interpret, and translate this contractile signal into postexercise adaptation. While our current understanding is that contraction-sensitive allosteric factors (e.g., Ca(2+), AMP, NAD(+), and acetyl-CoA) initiate signaling changes, how the muscle translates changes in these factors into the appropriate adaptive response remains poorly understood. During the past decade, systems biology approaches, utilizing "omics" screening techniques, have allowed researchers to define global processes of regulation with incredible sensitivity and specificity. As a result, physiologists are now able to study substrate flux with stable isotope tracers in combination with metabolomic approaches and to coordinate these functional changes with proteomic and transcriptomic analysis. In this review, we highlight lysine acetylation as an important posttranslational modification in skeletal muscle. We discuss the evolution of acetylation research and detail how large proteomic screens in diverse metabolic systems have led to the current hypothesis that acetylation may be a fundamental mechanism to fine-tune metabolic adaptation in skeletal muscle.


Assuntos
Acetiltransferases/metabolismo , Processamento de Proteína Pós-Traducional , Acetilação , Histona Acetiltransferases/metabolismo , Humanos , Lisina/metabolismo , Proteínas Mitocondriais/metabolismo , Músculo Esquelético/enzimologia , Proteômica
14.
Diabetologia ; 57(11): 2405-12, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-25116175

RESUMO

AIMS/HYPOTHESIS: Physical activity improves oxidative capacity and exerts therapeutic beneficial effects, particularly in the context of metabolic diseases. The peroxisome proliferator-activated receptor (PPAR) γ coactivator-1α (PGC-1α) and the nuclear receptor PPARß/δ have both been independently discovered to play a pivotal role in the regulation of oxidative metabolism in skeletal muscle, though their interdependence remains unclear. Hence, our aim was to determine the functional interaction between these two factors in mouse skeletal muscle in vivo. METHODS: Adult male control mice, PGC-1α muscle-specific transgenic (mTg) mice, PPARß/δ muscle-specific knockout (mKO) mice and the combination PPARß/δ mKO + PGC-1α mTg mice were studied under basal conditions and following PPARß/δ agonist administration and acute exercise. Whole-body metabolism was assessed by indirect calorimetry and blood analysis, while magnetic resonance was used to measure body composition. Quantitative PCR and western blot were used to determine gene expression and intracellular signalling. The proportion of oxidative muscle fibre was determined by NADH staining. RESULTS: Agonist-induced PPARß/δ activation was only disrupted by PPARß/δ knockout. We also found that the disruption of the PGC-1α-PPARß/δ axis did not affect whole-body metabolism under basal conditions. As expected, PGC-1α mTg mice exhibited higher exercise performance, peak oxygen consumption and lower blood lactate levels following exercise, though PPARß/δ mKO + PGC-1α mTg mice showed a similar phenotype. Similarly, we found that PPARß/δ was dispensable for PGC-1α-mediated enhancement of an oxidative phenotype in skeletal muscle. CONCLUSIONS/INTERPRETATION: Collectively, these results indicate that PPARß/δ is not an essential partner of PGC-1α in the control of skeletal muscle energy metabolism.


Assuntos
Músculo Esquelético/metabolismo , PPAR delta/metabolismo , PPAR beta/metabolismo , Fatores de Transcrição/metabolismo , Animais , Western Blotting , Calorimetria Indireta , Masculino , Camundongos , PPAR delta/genética , PPAR beta/genética , Coativador 1-alfa do Receptor gama Ativado por Proliferador de Peroxissomo , Condicionamento Físico Animal/fisiologia , Comportamento Sedentário , Fatores de Transcrição/genética
15.
Proc Natl Acad Sci U S A ; 110(50): 20314-9, 2013 Dec 10.
Artigo em Inglês | MEDLINE | ID: mdl-24277823

RESUMO

Skeletal muscle mass loss and dysfunction have been linked to many diseases. Conversely, resistance exercise, mainly by activating mammalian target of rapamycin complex 1 (mTORC1), promotes skeletal muscle hypertrophy and exerts several therapeutic effects. Moreover, mTORC1, along with peroxisome proliferator-activated receptor γ coactivator 1α (PGC-1α), regulates skeletal muscle metabolism. However, it is unclear whether PGC-1α is required for skeletal muscle adaptations after overload. Here we show that although chronic overload of skeletal muscle via synergist ablation (SA) strongly induces hypertrophy and a switch toward a slow-contractile phenotype, these effects were independent of PGC-1α. In fact, SA down-regulated PGC-1α expression and led to a repression of energy metabolism. Interestingly, however, PGC-1α deletion preserved peak force after SA. Taken together, our data suggest that PGC-1α is not involved in skeletal muscle remodeling induced by SA.


Assuntos
Metabolismo Energético/fisiologia , Complexos Multiproteicos/metabolismo , Músculo Esquelético/metabolismo , Serina-Treonina Quinases TOR/metabolismo , Fatores de Transcrição/metabolismo , Animais , Western Blotting , Primers do DNA/genética , Hipertrofia/metabolismo , Imuno-Histoquímica , Masculino , Alvo Mecanístico do Complexo 1 de Rapamicina , Camundongos , Camundongos Transgênicos , Coativador 1-alfa do Receptor gama Ativado por Proliferador de Peroxissomo , Fosfofrutoquinases/metabolismo , Reação em Cadeia da Polimerase , Succinato Desidrogenase/metabolismo
16.
Proc Natl Acad Sci U S A ; 110(21): 8738-43, 2013 May 21.
Artigo em Inglês | MEDLINE | ID: mdl-23650363

RESUMO

The peroxisome proliferator-activated receptor-γ coactivator 1α (PGC-1α) controls metabolic adaptations. We now show that PGC-1α in skeletal muscle drives the expression of lactate dehydrogenase (LDH) B in an estrogen-related receptor-α-dependent manner. Concomitantly, PGC-1α reduces the expression of LDH A and one of its regulators, the transcription factor myelocytomatosis oncogene. PGC-1α thereby coordinately alters the composition of the LDH complex and prevents the increase in blood lactate during exercise. Our results show how PGC-1α actively coordinates lactate homeostasis and provide a unique molecular explanation for PGC-1α-mediated muscle adaptations to training that ultimately enhance exercise performance and improve metabolic health.


Assuntos
Receptor alfa de Estrogênio/metabolismo , Regulação Enzimológica da Expressão Gênica/fisiologia , Homeostase/fisiologia , L-Lactato Desidrogenase/biossíntese , Ácido Láctico/sangue , Proteínas Musculares/metabolismo , Músculo Esquelético/metabolismo , Transativadores/metabolismo , Animais , Linhagem Celular , Receptor alfa de Estrogênio/genética , Isoenzimas/biossíntese , Isoenzimas/genética , L-Lactato Desidrogenase/genética , Lactato Desidrogenase 5 , Camundongos , Camundongos Knockout , Proteínas Musculares/genética , Músculo Esquelético/citologia , Coativador 1-alfa do Receptor gama Ativado por Proliferador de Peroxissomo , Condicionamento Físico Animal , Transativadores/genética , Fatores de Transcrição
17.
Eur J Pharmacol ; 698(1-3): 39-47, 2013 Jan 05.
Artigo em Inglês | MEDLINE | ID: mdl-23183106

RESUMO

ß-adrenoceptors are members of the G protein-coupled receptor superfamily which play a key role in the regulation of myocardial function. Their activation increases cardiac performance but can also induce deleterious effects such as cardiac arrhythmias or myocardial apoptosis. In fact, inhibition of ß-adrenoceptors exerts a protective effect in patients with sympathetic over-stimulation during heart failure. Although ß(2)-adrenoceptor is not the predominant subtype in the heart, it seems to importantly contribute to the cardiac effects of adrenergic stimulation; however, the mechanism by which this occurs is not fully understood. This review summarizes the current knowledge on the role of ß(2)-adrenoceptors in the regulation of cardiac contractility, metabolism, cardiomyocyte survival and cardiac arrhythmias. In addition, therapeutic considerations relating to stimulation of the ß(2)-adrenoceptor such as an increase in cardiac contractility with low arrythmogenic effect, protection of the myocardium again apoptosis or positive regulation of heart metabolism are discussed.


Assuntos
Coração/efeitos dos fármacos , Coração/fisiopatologia , Terapia de Alvo Molecular/métodos , Miocárdio/metabolismo , Receptores Adrenérgicos beta 2/metabolismo , Animais , Arritmias Cardíacas/tratamento farmacológico , Arritmias Cardíacas/metabolismo , Arritmias Cardíacas/patologia , Arritmias Cardíacas/fisiopatologia , Humanos , Miocárdio/patologia , Miócitos Cardíacos/efeitos dos fármacos , Miócitos Cardíacos/metabolismo , Miócitos Cardíacos/patologia
18.
Mol Cell Biol ; 32(24): 4913-24, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-23028049

RESUMO

Skeletal muscle exhibits a high plasticity and accordingly can quickly adapt to different physiological and pathological stimuli by changing its phenotype largely through diverse epigenetic mechanisms. The nuclear receptor corepressor 1 (NCoR1) has the ability to mediate gene repression; however, its role in regulating biological programs in skeletal muscle is still poorly understood. We therefore studied the mechanistic and functional aspects of NCoR1 function in this tissue. NCoR1 muscle-specific knockout mice exhibited a 7.2% higher peak oxygen consumption (VO(2peak)), a 11% reduction in maximal isometric force, and increased ex vivo fatigue resistance during maximal stimulation. Interestingly, global gene expression analysis revealed a high overlap between the effects of NCoR1 deletion and peroxisome proliferator-activated receptor gamma (PPARγ) coactivator 1α (PGC-1α) overexpression on oxidative metabolism in muscle. Importantly, PPARß/δ and estrogen-related receptor α (ERRα) were identified as common targets of NCoR1 and PGC-1α with opposing effects on the transcriptional activity of these nuclear receptors. In fact, the repressive effect of NCoR1 on oxidative phosphorylation gene expression specifically antagonizes PGC-1α-mediated coactivation of ERRα. We therefore delineated the molecular mechanism by which a transcriptional network controlled by corepressor and coactivator proteins determines the metabolic properties of skeletal muscle, thus representing a potential therapeutic target for metabolic diseases.


Assuntos
Músculo Esquelético/fisiologia , Correpressor 1 de Receptor Nuclear/metabolismo , Receptores de Estrogênio/metabolismo , Transativadores/metabolismo , Animais , Masculino , Camundongos , Camundongos Knockout , Camundongos Transgênicos , Modelos Biológicos , Contração Muscular/genética , Contração Muscular/fisiologia , Músculo Esquelético/metabolismo , Correpressor 1 de Receptor Nuclear/deficiência , Correpressor 1 de Receptor Nuclear/genética , Fosforilação Oxidativa , Consumo de Oxigênio , PPAR delta/metabolismo , PPAR beta/metabolismo , Coativador 1-alfa do Receptor gama Ativado por Proliferador de Peroxissomo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Receptores de Estrogênio/antagonistas & inibidores , Receptores de Estrogênio/genética , Transativadores/antagonistas & inibidores , Transativadores/genética , Fatores de Transcrição , Receptor ERRalfa Relacionado ao Estrogênio
19.
Life Sci ; 88(19-20): 892-7, 2011 May 09.
Artigo em Inglês | MEDLINE | ID: mdl-21466811

RESUMO

AIMS: Cardiac function is modulated by the sympathetic nervous system through ß-adrenergic receptor (ß-AR) activity and this represents the main regulatory mechanism for cardiac performance. To date, however, the metabolic and molecular responses to ß(2)-agonists are not well characterized. Therefore, we studied the inotropic effect and signaling response to selective ß(2)-AR activation by tulobuterol. MAIN METHODS: Strips of rat right ventricle were electrically stimulated (1Hz) in standard Tyrode solution (95% O(2), 5% CO(2)) in the presence of the ß(1)-antagonist CGP-20712A (1µM). A cumulative dose-response curve for tulobuterol (0.1-10µM), in the presence or absence of the phosphodiesterase (PDE) inhibitor IBMX (30µM), or 10min incubation (1µM) with the ß(2)-agonist tulobuterol was performed. KEY FINDINGS: ß(2)-AR stimulation induced a positive inotropic effect (maximal effect=33±3.3%) and a decrease in the time required for half relaxation (from 45±0.6 to 31±1.8ms, -30%, p<0.001) after the inhibition of PDEs. After 10min of ß(2)-AR stimulation, p-AMPKα(T172) (54%), p-PKB(T308) (38%), p-AS160(T642) (46%) and p-CREB(S133) (63%) increased, without any change in p-PKA(T197). SIGNIFICANCE: These results suggest that the regulation of ventricular contractility is not the primary function of the ß(2)-AR. Rather, ß(2)-AR could function to activate PKB and AMPK signaling, thereby modulating muscle mass and energetic metabolism of rat ventricular muscle.


Assuntos
Ventrículos do Coração/metabolismo , Contração Miocárdica/fisiologia , Receptores Adrenérgicos beta 2/fisiologia , Transdução de Sinais/fisiologia , Proteínas Quinases Ativadas por AMP/fisiologia , Antagonistas de Receptores Adrenérgicos beta 2/farmacologia , Animais , Ventrículos do Coração/efeitos dos fármacos , Masculino , Contração Miocárdica/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacos
20.
Am J Physiol Cell Physiol ; 299(2): C240-50, 2010 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-20410436

RESUMO

Sodium pyruvate can increase mitochondrial biogenesis in C2C12 myoblasts in a peroxisome proliferator-activated receptor-gamma coactivator-1alpha (PGC1alpha)-independent manner. The present study examined the effect of 72-h treatment with sodium pyruvate (5-50 mM) or sodium chloride (50 mM) as an osmotic control on the regulation of mitochondrial substrate metabolism and biogenesis in C2C12 myotubes. Pyruvate (50 mM) increased the levels of fatty acid oxidation enzymes (CD36, 61%, and beta-oxidative enzyme 3-hydroxyacyl-CoA dehydrogenase, 54%) and the expression of cytochrome-c oxidase subunit I (220%) and cytochrome c (228%), consistent with its previous described role as a promoter of mitochondrial biogenesis. However, in contrast, pyruvate treatment reduced glucose transporter 4 (42%), phosphofructokinase (57%), and PGC1alpha (72%) protein content as well as PGC1alpha (48%) and PGC1beta (122%) mRNA. The decrease in PGC1alpha was compensated for by an increase in the PGC1alpha-related coactivator (PRC; 187%). Pyruvate treatment reduced basal and insulin-stimulated glucose uptake (41% and 31%, respectively) and palmitate uptake and oxidation (24% and 31%, respectively). The addition of the pyruvate dehydrogenase activator dichloroacetate (DCA) and the TCA precursor glutamine increased PGC1alpha expression (368%) and returned PRC expression to basal. Glucose uptake increased by 4.2-fold with DCA and glutamine and palmitate uptake increased by 18%. Coupled to this adaptation was an 80% increase in oxygen consumption. The data suggest that supraphysiological doses of pyruvate decrease mitochondrial function despite limited biogenesis and that anaplerotic agents can reverse this effect.


Assuntos
Regulação para Baixo/fisiologia , Transporte de Elétrons/fisiologia , Fibras Musculares Esqueléticas/metabolismo , Ácido Pirúvico/farmacologia , Transativadores/antagonistas & inibidores , Regulação para Cima/fisiologia , Animais , Linhagem Celular , Células Cultivadas , Ciclo do Ácido Cítrico/efeitos dos fármacos , Ciclo do Ácido Cítrico/fisiologia , Regulação para Baixo/efeitos dos fármacos , Transporte de Elétrons/efeitos dos fármacos , Camundongos , Fibras Musculares Esqueléticas/efeitos dos fármacos , Coativador 1-alfa do Receptor gama Ativado por Proliferador de Peroxissomo , Especificidade por Substrato/efeitos dos fármacos , Especificidade por Substrato/fisiologia , Transativadores/biossíntese , Fatores de Transcrição , Regulação para Cima/efeitos dos fármacos
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