Study of the role of the PGC-1β transcriptional coactivator                 in white adipose tissue and  its implication i...
White AdiposeTissue                             Thiazolidinediones                                                       A...
EXERCISECOLD EXPOSURE                  PGC-1                                                                 mtDNA replic...
Decreased mitochondrial mass and function in diabetic patients is associated with         low PGC-1 expression           ...
Expression of mitochondrial genes and mitochondrial biogenesis is not alteredin WAT of PGC-1-FAT-KO mice                 ...
Rosiglitazone-induced mitochondrial biogenesis and function is independent of                          PGC-1             ...
PGC-1β is highly expressed in mature white adipocytes                   1,4                 PGC-1β     ATGL       Pref-1  ...
Generation of PGC-1β-FAT-KO mice                    1         2         3           4           5                 6 7     ...
Gene expression profiling in WAT of PGC-1β-FAT-KO mice                                      Microarray DNA Wildtype   Knoc...
PGC-1β regulates expression of genes involved in oxidative metabolism in white                           adipose tissue   ...
PGC-1β-FAT-KO mice exhibit reduced levels of TCA cycle and OxPhos proteins          and impaired oxidative capacity in whi...
Rosiglitazone-induced mitochondrial gene expression and function is mediated                  by PGC-1β              4    ...
Lack of PGC-1β in adipocytes does not induce whole body or tissue-specific insulin                         resistance     ...
Conclusions PGC-1β in white adipose tissue specifically regulates the expression of genes  involved in mitochondrial ener...
Thank YouMetabolism and Obesity Laboratory
PGC-1β regulates mitochondrial gene expression in 3T3-L1 adipocytes                                      Gene expression  ...
Estudi de la funció del coactivador transcripcional PGC-1β en el teixit adipós blanc i la seva implicació en el desenvolup...
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Estudi de la funció del coactivador transcripcional PGC-1β en el teixit adipós blanc i la seva implicació en el desenvolupament de resistència a insulina

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Presentació predoctoral de Natàlia Enguix a la 6a Jornada Científica del VHIR.

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Estudi de la funció del coactivador transcripcional PGC-1β en el teixit adipós blanc i la seva implicació en el desenvolupament de resistència a insulina

  1. 1. Study of the role of the PGC-1β transcriptional coactivator in white adipose tissue and its implication in the development of insulin resistance Natàlia Enguix Elena Laboratory of Metabolism and Obesity Unit of Diabetes and Metabolism
  2. 2. White AdiposeTissue Thiazolidinediones Adipogenesis Mitochondrial Biogenesis Oxidative Metabolism Fatty acid uptake Triglycerides accumulation Lipid oxidation Fatty acids Insulin Fatty acids IRS ATP PKC Acyl-CoAs STOP Diacylglycerols Ceramides
  3. 3. EXERCISECOLD EXPOSURE PGC-1 mtDNA replication/transcription PGC-1 NRF1/2 TFAM PGC-1 ERR Oxphos System PGC-1 PPAR Fatty Acid Oxidation Nucleus
  4. 4. Decreased mitochondrial mass and function in diabetic patients is associated with low PGC-1 expression Control Diabetic Skeletal Muscle (Patti et al, PNAS 2003) Control Diabetic PGC-1 mRNA Mitochondrial MassWhite Adipose Tissue(Bogacka et al, Diabetes 2005) No Diabetic Placebo Pioglitazone No Diabetic Placebo Pioglitazone Control Control Diabetic Diabetic
  5. 5. Expression of mitochondrial genes and mitochondrial biogenesis is not alteredin WAT of PGC-1-FAT-KO mice loxP loxP 1 2 3 4 5 6 7 8 9 10 11 12 13 Floxed allele + aP2-Cre 1 2 3 6 7 8 9 10 11 12 13 PGC-1-FAT-KO Gene Expression 1.6 Wildtype mRNA (a.u.) 1.2 PGC-1-FAT-KO 0.8 0.4 ** ** 0 Kidney Liver Testis Spleen Muscle Thymus BAT WAT Gene Expression mtDNA White Adipose Tissue 1,5 1,5 mRNA (a.u.) 1 1,0 a.u. 0,5 0,5 0 0,0 wt ko Pardo et al. PLoS One (2011)
  6. 6. Rosiglitazone-induced mitochondrial biogenesis and function is independent of PGC-1 Wt Cont. Wt Rosig. FAT-KO Cont. FAT-KO Rosig. 3.0 4.5 PGC-1 4.0mRNA expression 2.5 mRNA expression (Relative units) 3.5 (Relative units) 2.0 3.0 2.5 1.5 2.0 1.0 1.5 1.0 0.5 0.5 0 0 Wt FAT-KO COXII COXIV CYCS ATP5b CS ACO2 MCAD 2.5 3.5 Relative mtDNA content mtDNA 3.0 Citrate synthase 2.0 mol/mg.min 2.5 1.5 CS activity 2.0 1.0 1.5 1.0 0.5 0.5 0 0 Cont. Rosig Cont. Rosig Pardo et al. PLoS One (2011)
  7. 7. PGC-1β is highly expressed in mature white adipocytes 1,4 PGC-1β ATGL Pref-1 1 1,2 * ** 0,8(Relative units) 1 (Relative units) 0,8 mRNA 0,6 PGC-1β mRNA 0,6 PPARγ * 0,4 Pref-1 0,4 0,2 0,2 0 0 Pre Conf Post d0 d2 d4 d6 d8 Days during differentation 2,5 Gene Expression 2 (Relative units) 1,5 Wild type + Vehicle mRNA Wild type + Rosiglitazone 1 0,5 0 NRF-2 NRF-1 ERRα ERRγ RIP140 PGC-1β
  8. 8. Generation of PGC-1β-FAT-KO mice 1 2 3 4 5 6 7 8 9 10 11 12 Floxed allele + aP2Cre 1 2 3 6 7 8 9 10 11 12 PGC-1β-FAT-KO PGC-1β Expression 30ºC 1,6 1,4 1,2mRNA (a.u.) 1 0,8 Wildtype 0,6 PGC-1B-FAT-KO 0,4 ** ** ** 0,2 0 BAT iWAT Adipocytes Kidney Lungs Soleus Spleen Testis Thymus Adipose tissues
  9. 9. Gene expression profiling in WAT of PGC-1β-FAT-KO mice Microarray DNA Wildtype Knockout (n=5) (n=5)
  10. 10. PGC-1β regulates expression of genes involved in oxidative metabolism in white adipose tissue Gene Expression 1,4 White Adipose Tissue 1,2 (Relative units) 1 * * * ** * mRNA 0,8 * ** * Wild type 0,6 * PGC-1β-FAT-KO 0,4 0,2 0 NDUFAB1 NDUFB9 SDHB CytC1 CytCs Cox5a Cox5b Cox7a1 ATP5a1 ATP5o CI C II Cyt C C IV CV OXPHOS 1,4 1,2(Relative units) 1 * * mRNA 0,8 * Wild type * ** 0,6 * * * PGC-1β-FAT-KO 0,4 0,2 0 Aco2 Idh3a Idh3b MDH2 LDHB PGK1 Elovl6 Cpt1b Cidea Cideb TCA Cycle Glycolysis Lipid Metabolism / Oxidation
  11. 11. PGC-1β-FAT-KO mice exhibit reduced levels of TCA cycle and OxPhos proteins and impaired oxidative capacity in white adipose tissue White Adipose Tissue Wild type PGC-1β-FAT-KO Citrate Synthase Cytochrome Activity Oxidase ActivityAconitase 0,2 0,03ATPSynβ μmole/mg/min 0,15 μmole/mg/minNDUFB9 * * 0,02COX IV 0,1SDHB 0,01 0,05Cyt c 0 0α-tubulin WT KO WT KO
  12. 12. Rosiglitazone-induced mitochondrial gene expression and function is mediated by PGC-1β 4 Gene Expression White Adipose Tissue 3 Wt + vehiclemRNA (a.u.) KO + vehicle 2 Wt + Rosiglitazone ** * ** * * KO + Rosiglitazone 1 * * ** * 0 CoxIV Cox8b Cox7a1 CytCs ATPSynB Aco2 CS Cpt1b Citrate Synthase Activity mtDNA Content 400 5 nmol/mg·min 300 4 3 a.u. 200 * 2 100 1 0 0 Vehicle Rosiglitazone Vehicle Rosiglitazone
  13. 13. Lack of PGC-1β in adipocytes does not induce whole body or tissue-specific insulin resistance Glucose Tolerance Test 450 Wt + Vehicle KO + Vehicle 350Glucose (mg/dL) Wt + Rosiglitazone KO + Rosiglitazone 250 150 50 0 20 40 60 80 100 120 Time (min) Insulin Tolerance Test 150 Glucose (mg/dL) 100 50 0 0 20 40 60 80 100 120 Time (min)
  14. 14. Conclusions PGC-1β in white adipose tissue specifically regulates the expression of genes involved in mitochondrial energy production. PGC-1β but not PGC-1α mediates the rosiglitazone-induced expression of mitochondrial genes and the oxidative capacity of white adipose tissue. Full mitochondrial oxidative function is not required for the insulin-sensitizing effects of thiazolidinediones.
  15. 15. Thank YouMetabolism and Obesity Laboratory
  16. 16. PGC-1β regulates mitochondrial gene expression in 3T3-L1 adipocytes Gene expression 4 4 PGC-1α PGC-1β 3 3 mRNA (a.u.) mRNA (a.u.) 2 2 1 ** ** 1 ** ** ** ** ** ** 0 0 Oxygen Consumption RatesiControl: + - - - + - - - + - - - + - - - 3T3L1 AdipocytessiPGC-1α: - + - + - + - + - + - + - + - + 7siPGC-1β: - - + + - - + + - - + + - - + + DMSO P=0.16 6 Rosiglitazone P=0.11 DMSO Rosiglitazone DMSO Rosiglitazone 5 6 3 Relative Units CytCs IDH3β 4 mRNA (a.u.) mRNA (a.u.) 4 2 ** ** 3 P=0.09 * ** ** 2 1 ** ** 2 ** ** 1 0 0 0 3 3 + - - - + - - - + - - - + - - - ATPSynβ NDUFB9 siControl: - + - + - + - + - + - + - + - + siPGC-1α: mRNA (a.u.) mRNA (a.u.) 2 siPGC-1β: - - + + - - + + - - + + - - + + 2 ** ** ** Basal CCCP 1 ** ** 1 * ** ** 0 0siControl: + - - - + - - - + - - - + - - -siPGC-1α: - + - + - + - + - + - + - + - +siPGC-1β: - - + + - - + + - - + + - - + + DMSO Rosiglitazone DMSO Rosiglitazone

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