title: Adipocyte nuclei captured from VAT and SAT.
abstract: BACKGROUND
  Obesity-related comorbidities are thought to result from the reprogramming of the epigenome in numerous tissues and cell types, and in particular, mature adipocytes within visceral and subcutaneous adipose tissue, VAT and SAT. The cell-type specific chromatin remodeling of mature adipocytes within VAT and SAT is poorly understood, in part, because of the difficulties of isolating and manipulating large fragile mature adipocyte cells from adipose tissues.
  
  
  METHODS
  We constructed MA-INTACT (Mature Adipocyte-Isolation of Nuclei TAgged in specific Cell Types) mice using the adiponectin (ADIPOQ) promoter (ADNp) to tag the surface of mature adipocyte nuclei with a reporter protein. The SUN1mRFP1Flag reporter is comprised of a fragment of the nuclear transmembrane protein SUN1, the fluorescent protein mRFP1, and three copies of the Flag epitope tag.
  
  
  RESULTS
  Mature adipocyte nuclei were rapidly and efficiently immuno-captured from VAT and SAT (MVA and MSA nuclei, respectively), of MA-INTACT mice. MVA and MSA nuclei contained 1,000 to 10,000-fold higher levels of adipocyte-specific transcripts, ADIPOQ, PPARg2, EDNRB, and LEP, relative to uncaptured nuclei, while the latter expressed higher levels of leukocyte and endothelial cell markers IKZF1, RETN, SERPINF1, SERPINE1, ILF3, and TNFA. MVA and MSA nuclei differentially expressed several factors linked to adipogenesis or obesity-related health risks including CEBPA, KLF2, RETN, SERPINE1, and TNFA. The various nuclear populations dramatically differentially expressed transcripts encoding chromatin remodeler proteins regulating DNA cytosine methylation and hydroxymethylation (TETs, DNMTs, TDG, GADD45s) and nucleosomal histone modification (ARID1A, KAT2B, KDM4A, PRMT1, PRMT5, PAXIP1). Remarkably, MSA and MVA nuclei expressed 200 to 1000-fold higher levels of thermogenic marker transcripts PRDM16 and UCP1.
  
  
  CONCLUSIONS
  The MA-INTACT mouse enables a simple way to perform cell-type specific analysis of highly purified mature adipocyte nuclei from VAT and SAT and increases the statistical significance of data collected on adipocytes. Isolated VAT and SAT adipocyte nuclei expressed distinct patterns of transcripts encoding chromatin remodeling factors and proteins relevant to diabetes, cardiovascular disease, and thermogenesis. The MA-INTACT mouse is an useful model to test the impact of caloric intake, dietary nutrients, exercise, and pharmaceuticals on the epigenome-induced health risks of obesity.
authors.affiliation: Department of Genetics, University of Georgia, Athens, GA USA.
authors.forename: Suresh
authors.initials: S
authors.lastname: Ambati
authors.affiliation: Department of Genetics, University of Georgia, Athens, GA USA.
authors.forename: Ping
authors.initials: P
authors.lastname: Yu
authors.affiliation: Department of Genetics, University of Georgia, Athens, GA USA.
authors.forename: Elizabeth C
authors.initials: EC
authors.lastname: McKinney
authors.affiliation: Department of Genetics, University of Georgia, Athens, GA USA.
authors.forename: Muthugapatti K
authors.initials: MK
authors.lastname: Kandasamy
authors.affiliation: Department of Foods and Nutrition, University of Georgia, Athens, GA USA ; Department of Animal and Dairy Science, University of Georgia, Athens, GA USA.
authors.forename: Diane
authors.initials: D
authors.lastname: Hartzell
authors.affiliation: Department of Foods and Nutrition, University of Georgia, Athens, GA USA ; Department of Animal and Dairy Science, University of Georgia, Athens, GA USA.
authors.forename: Clifton A
authors.initials: CA
authors.lastname: Baile
authors.affiliation: Department of Genetics, University of Georgia, Athens, GA USA.
authors.forename: Richard B
authors.initials: RB
authors.lastname: Meagher
keywords: Cell-type specific; Epigenetics; INTACT transgenic mouse; Visceral and subcutaneous adipose tissue