title: Evaluation of reference genes for gene expression studies in human brown adipose tissue.
abstract: Human brown adipose tissue (BAT) has during the last 5 year been subjected to an increasing research interest, due to its putative function as a target for future obesity treatments. The most commonly used method for molecular studies of human BAT is the quantitative polymerase chain reaction (qPCR). This method requires normalization to a reference gene (genes with uniform expression under different experimental conditions, e.g. similar expression levels between human BAT and WAT), but so far no evaluation of reference genes for human BAT has been performed. Two different microarray datasets with samples containing human BAT were used to search for genes with low variability in expression levels. Seven genes (FAM96B, GNB1, GNB2, HUWE1, PSMB2, RING1 and TPT1) identified by microarray analysis, and 8 commonly used reference genes (18S, B2M, GAPDH, LRP10, PPIA, RPLP0, UBC, and YWHAZ) were selected and further analyzed by quantitative PCR in both BAT containing perirenal adipose tissue and subcutaneous adipose tissue. Results were analyzed using 2 different algorithms (Normfinder and geNorm). Most of the commonly used reference genes displayed acceptably low variability (geNorm M-values <0.5) in the samples analyzed, but the novel reference genes identified by microarray displayed an even lower variability (M-values <0.25). Our data suggests that PSMB2, GNB2 and GNB1 are suitable novel reference genes for qPCR analysis of human BAT and we recommend that they are included in future gene expression studies of human BAT.
authors.affiliation: Department of Molecular and Clinical Medicine; Institute of Medicine; The Sahlgrenska Academy; University of Gothenburg ; Gothenburg, Sweden.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Taube
authors.affiliation: Department of Molecular and Clinical Medicine; Institute of Medicine; The Sahlgrenska Academy; University of Gothenburg ; Gothenburg, Sweden.
authors.forename: Johanna C
authors.initials: JC
authors.lastname: Andersson-Assarsson
authors.affiliation: Department of Molecular and Clinical Medicine; Institute of Medicine; The Sahlgrenska Academy; University of Gothenburg ; Gothenburg, Sweden.
authors.forename: Kristin
authors.initials: K
authors.lastname: Lindberg
authors.affiliation: Department of Medical Sciences; Uppsala University ; Uppsala, Sweden.
authors.forename: Maria J
authors.initials: MJ
authors.lastname: Pereira
authors.affiliation: Department of Surgery; Sahlgrenska University Hospital ; Gothenburg, Sweden.
authors.forename: Markus
authors.initials: M
authors.lastname: Gäbel
authors.affiliation: Department of Molecular and Clinical Medicine; Institute of Medicine; The Sahlgrenska Academy; University of Gothenburg ; Gothenburg, Sweden.
authors.forename: Maria K
authors.initials: MK
authors.lastname: Svensson
authors.affiliation: Department of Medical Sciences; Uppsala University ; Uppsala, Sweden.
authors.forename: Jan W
authors.initials: JW
authors.lastname: Eriksson
authors.affiliation: Department of Molecular and Clinical Medicine; Institute of Medicine; The Sahlgrenska Academy; University of Gothenburg ; Gothenburg, Sweden.
authors.forename: Per-Arne
authors.initials: PA
authors.lastname: Svensson
keywords: brown adipose tissue; human; quantitative PCR; reference genes; white adipose tissue