Analysis Tools
adipose tissue
| N |
• chow-fed young adult mice are grossly normal and show no differences in brown adipose tissue (BAT) weight, epididymal and inguinal white adipose tissue (WAT) weight, or gross appearance of inguinal WAT (iWAT) and visceral adipose tissue (VAT) relative to controls
|
|
• quantitative proteomics analysis of ex vivo iWAT adipocytes indicates upregulation of genes involved in adipogenesis
|
|
• ex vivo iWAT and VAT adipocytes isolated from HFD-fed, but not from chow-fed, mice show a 56%-59% decrease in cytoplasmic mitochondrial DNA (mtDNA) despite a 9%-17% increase in total cellular mtDNA
|
|
• ex vivo iWAT and VAT adipocytes isolated from HFD-fed, but not from chow-fed, mice show a 9%-17% increase in total cellular mtDNA
|
|
• ex vivo iWAT adipocytes show upregulation of genes involved in oxidative phosphorylation, adipogenesis, and fatty acid metabolism, with strong induction of the Ppargc1a activity signature, including genes involved in oxidative phosphorylation and adipose thermogenesis
• some genes involved in thermogenesis and oxidative metabolism are also upregulated in ex vivo VAT adipocytes but to a lesser extent than in iWAT adipocytes
• ex vivo VAT adipocytes isolated from HFD-fed mice show an increase in the Ppargc1a activity signature along with increased oxidative phosphorylation complex subunit expression, with similar trends seen in iWAT adipocytes from HFD-fed mice
• strikingly, ex vivo VAT adipocytes from HFD-fed mice show downregulation of obesity-related pathways (e.g. NF-kappaB- and IFN-stimulated inflammatory signaling and epithelial-mesenchymal transition (EMT)), indicating impaired pro-inflammatory adipocyte remodeling
• ex vivo both iWAT and VAT adipocytes from HFD-fed, but not from chow-fed, mice contain significantly lower levels of the cGAS product 23-cGAMP, express lower levels of cGAS and STING proteins, and show decreased phosphorylation of downstream signaling components TBK1 and IRF3, indicating protection against cGAS-STING activation in the context of obesity
• however, BAT shows no changes in mRNAs involved in thermogenesis and oxidative metabolism at either 22 degrees or 30 degrees Celsius
|
homeostasis/metabolism
|
• inguinal WAT (iWAT) shows a significant increase in the protein level of PPARGC1 [PPARG coactivator 1 alpha (peroxisome proliferative activated receptor, gamma, coactivator 1 alpha)] with no significant change in Ppargc1a mRNA level, suggesting altered post-transcriptional regulation
• polysome profiles of ex vivo iWAT adipocytes, in which sucrose density gradients resolve untranslated mRNAs (low fractions) from highly translated mRNAs (heavy fractions), show a shift in the distribution of Ppargc1a mRNA towards higher-density fractions, indicating an increase in its translation
|
|
• after administration of beta3-adrenergic receptor agonist CL-316,243, mice show significantly greater energy expenditure in metabolic cages than similarly treated controls
|
|
• in an intraperitoneal glucose tolerance test, both male and female mice fed a high-fat diet (HFD) show markedly improved glucose tolerance relative to sex- and diet-matched control mice
• however, chow-fed mice exhibit largely unchanged glucose tolerance
|
|
• inguinal WAT (iWAT) shows a significant increase in PPARGC1 protein levels but no change in Ppargc1a mRNA levels
• ex vivo VAT adipocytes isolated from HFD-fed mice show increased PPARGC1 protein levels without a change in Ppargc1a mRNA
|
|
• quantitative proteomics analysis of ex vivo iWAT adipocytes indicates upregulation of genes involved in fatty acid metabolism
|
cellular
|
• ex vivo iWAT and VAT adipocytes isolated from HFD-fed, but not from chow-fed, mice show a 56%-59% decrease in cytoplasmic mitochondrial DNA (mtDNA) despite a 9%-17% increase in total cellular mtDNA
|
|
• ex vivo iWAT and VAT adipocytes isolated from HFD-fed, but not from chow-fed, mice show a 9%-17% increase in total cellular mtDNA
|
|
• ex vivo minced iWAT tissue, but not perigonadal VAT, shows a significantly higher oxygen consumption (respiration per unit mass), as measured by a Seahorse bioanalyzer
• ex vivo VAT adipocytes from HFD-fed mice show a significant increase in oxygen consumption, with a similar trend seen in iWAT adipocytes from HFD-fed mice
|
|
• quantitative proteomics analysis of ex vivo iWAT adipocytes indicates upregulation of genes involved in oxidative phosphorylation with a >10-fold increase in UCP1 (uncoupling protein 1) and a1.2- to 2-fold increase of individual oxidative phosphorylation complex components
• ex vivo VAT adipocytes isolated from HFD-fed mice show increased oxidative phosphorylation complex subunit expression, with similar trends seen in iWAT adipocytes from HFD-fed mice
|
|
• inguinal WAT (iWAT) shows a significant increase in the protein level of PPARGC1 [PPARG coactivator 1 alpha (peroxisome proliferative activated receptor, gamma, coactivator 1 alpha)] with no significant change in Ppargc1a mRNA level, suggesting altered post-transcriptional regulation
• polysome profiles of ex vivo iWAT adipocytes, in which sucrose density gradients resolve untranslated mRNAs (low fractions) from highly translated mRNAs (heavy fractions), show a shift in the distribution of Ppargc1a mRNA towards higher-density fractions, indicating an increase in its translation
|
immune system
|
• immunohistochemical staining for macrophage marker F4/80 shows a significant reduction in the number of crown-like structures per mm2 of visceral adipose tissue (VAT) in HFD-fed mice
|
|
• ex vivo iWAT and VAT adipocytes isolated from HFD-fed, but not from chow-fed, mice show suppressed activation of the cGAS-STING innate immune pathway
|
growth/size/body
| N |
• surprisingly, mice fed either a chow diet or a high-fat diet (HFD) show no significant changes in body weight or body (fat and lean mass) composition relative to diet-matched controls
|
