title: Selection and evaluation of reference genes for analysis of mouse(Mus musculus) sex-dimorphic brain development.
abstract: The development of the brain is sex-dimorphic, and as a result so are many neurological disorders. One approach for studying sex-dimorphic brain development is to measure gene expression in biological samples using RT-qPCR. However, the accuracy and consistency of this technique relies on the reference gene(s) selected. We analyzed the expression of ten reference genes in male and female samples over three stages of brain development, using popular algorithms NormFinder, GeNorm and Bestkeeper. The top ranked reference genes at each time point were further used to quantify gene expression of three sex-dimorphic genes (Wnt10b,XistandCYP7B1). When comparing gene expression between the sexes expression at specific time points the best reference gene combinations are:Sdha/Pgk1at E11.5,RpL38/SdhaE12.5, andActb/RpL37at E15.5. When studying expression across time, the ideal reference gene(s) differs with sex. For XY samples a combination ofActb/Sdha. In contrast, when studying gene expression across developmental stage with XX samples,Sdha/Gapdhwere the top reference genes. Our results identify the best combination of two reference genes when studying male and female brain development, and emphasize the importance of selecting the correct reference genes for comparisons between developmental stages.
authors.affiliation: Department of Anatomy, University of Otago , Dunedin , New Zealand.
authors.forename: Tanya T
authors.initials: TT
authors.lastname: Cheung
authors.affiliation: Department of Anatomy, University of Otago , Dunedin , New Zealand.
authors.forename: Mitchell K
authors.initials: MK
authors.lastname: Weston
authors.affiliation: Department of Anatomy, University of Otago , Dunedin , New Zealand.
authors.forename: Megan J
authors.initials: MJ
authors.lastname: Wilson
keywords: Brain; Development; Mice; RT-qpcr; Reference gene; Sex-dimorphic