Project description:To understand differences in microRNA (miRNA) signatures between two different diets with and without EPA in brown, subcutaneous, and viscerl tissue from C57BL/6 mice to understand mechanistic insight regarding their contribution to metabolic disorders in obesity. We performed small RNA-sequencing of brown, subcutaneous adipose from high fat diet (45% kcal from fat) and high fat diet supplemented with EPA (45% Kcal from fat, 6.75% EPA). Using the Gunaratne Next Generation pipeline (published in Creighton et al. 2009) miRNA expression profiles were identified. Counts of each unique read were normalized to total usable reads, and had 40 counts added. We mapped about 13.8 million sequence reads per sample to the Mus musculus genome (build mm 10). AS a total 1251 miRNAs were identified in three adipose tissue and out of which in bown adipose tissue 15 showd differential expression between BF-HF and BF-EPA .IN subcutaneous adipose tissue 3 miRNAs showed differntial expression between SUB-HF and SUB-EPA. EPA differentially regulate specific miRNAs expression in brown, subcutaneous, and visceral adipose tissue.
Project description:The goal of this subproject is to identify microRNAs (miRNAs) expressed in visceral (VIS) adipose tissue (gonadal fat from male C57BL/6 mice) and regulated by eicosapentaenoic acid (EPA). This will provide insight into microRNAs regulated by EPA and their potential role in obesity-associated inflammation. We performed small RNA-sequencing of white (VIS) adipose tissue from high-fat diet (45% kcal from fat) supplemented with EPA (45% Kcal from fat, 6.75% EPA). Using the Gunaratne Next-Generation pipeline (published in Creighton et al. 2009), miRNA expression profiles were identified. Counts of each unique read were normalized to total usable reads, and had 40 counts added. We mapped about 13.8 million sequence reads per sample to the Mus musculus genome (build mm 10). We are specifically interested in those miRNAs expressed in VIS fat from EPA-fed mice compared to HF-fed mice.
Project description:Assessment of secreted proteins from EDL and soleus muscle, as well as visceral and subcutaneous adipose tissue of sedentary and exercise-trained obese male mice.
Project description:Diabetes and obesity are widespread diseases with signifciant socioeconomic implications. We used three different types of human adipose tissue (epigastric, visceral, and subcutaneous) in order to determine differences in global gene expression between these adipose depots in severely obese patients. In this dataset, we include the expression data obtained from three types of adipose tissue; epigastric, subcutaneous, and visceral all obtained through open gastric bypass surgery. 18 total samples were analyzed. Tissues were paired together to run on one genechip, with three pairs of epigastric, three pairs of subcutaneous, and three pairs of visceral were ran on nine genechips. Comparisons of gene expression in the form of fold changes between pairs of adipose types (i.e., subcutaneous/epigastric, visceral/epigastric, and subcutaneous/visceral) were completed by Spotfire Analysis.
Project description:Three different progenitor cell subsets in subcutaneous and visceral adipose tissues derived from 5 obese patients were subjected to AmpliSeq transcriptome profiling. Transcriptomic profiles were analyzed to compare progenitor cell subsets and the impact of subcutaneous and visceral adipose tissue location.
Project description:While dysregulation of adipocyte endocrine function plays a central role in obesity and its complications, the vast majority of adipokines remain uncharacterized. We employed bio-orthogonal non-canonical amino acid tagging (BONCAT) and mass spectrometry to comprehensively characterize the secretome of murine visceral and subcutaneous white and interscapular brown adipocytes. Over 600 proteins were identified, the majority of which showed cell type-specific enrichment. We here describe a metabolic role for leucine-rich α-2 glycoprotein 1 (LRG1) as an obesity-regulated adipokine secreted by mature adipocytes. LRG1 overexpression significantly improved glucose homeostasis in diet-induced and genetically obese mice. This was associated with markedly reduced white adipose tissue macrophage accumulation and systemic inflammation. Mechanistically, we found LRG1 binds cytochrome c in circulation to dampen its pro-inflammatory effect. These data support a new role for LRG1 as an insulin sensitizer with therapeutic potential given its immunomodulatory function at the nexus of obesity, inflammation, and associated pathology.
Project description:To investigate the proteomic profiles of paired subcutaneous adipose tissue (SAT) and visceral adipose tissue (VAT) samples, as well as their correlations with clinical traits in severely obese patients, and to identify potential serum protein markers associated with tissue expression or metabolic states.
Project description:To investigate the proteomic profiles of paired subcutaneous adipose tissue (SAT) and visceral adipose tissue (VAT) samples, as well as their correlations with clinical traits in severely obese patients, and to identify potential serum protein markers associated with tissue expression or metabolic states.
Project description:To further analyze the effect of aging and caloric restriction in the microRNA expression, we have employed microarray expression profiling as a discovery platform to identify differentially expressed microRNAs in middle-aged animals and the impact of caloric restriction in the microRNA expression profile. Subcutaneous and visceral adipose tissue were extracted from 3 groups of mice: 3 month-old, 12 month-old fed ad libitum and 12 month-old fed with a caloric restricted diet. Comparisons between young and middle-aged animals in subcutaneous and visceral adipose tissue, and between the 12 month old ad libitum and 12 month old caloric restricted diet in both adipose depots were made.
Project description:94 human adipocyte samples isolated from whole adipose tissues using collagenase digestion of tissue and flotation of lipid-laden adipocytes, followed by RNA isolation and RNA sequencing (SMARTer Stranded Total RNA-Seq library preparation, HiSeq 4000 100-bp paired-end reads). Adipocyte samples comprise subcutaneous and visceral adipocytes isolated from obese and lean people (N=24 obese-subcutaneous, N=24 obese-visceral, N=22 control-subcutaneous, N=24 control-visceral). Human adipocyte RNA sequencing data are provided as BAM files.