title: Selection of appropriate reference genes for RT-qPCR analysis in a streptozotocin-induced Alzheimer's disease model of cynomolgus monkeys (Macaca fascicularis).
abstract: Reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) has been widely used to quantify relative gene expression because of the specificity, sensitivity, and accuracy of this technique. In order to obtain reliable gene expression data from RT-qPCR experiments, it is important to utilize optimal reference genes for the normalization of target gene expression under varied experimental conditions. Previously, we developed and validated a novel icv-STZ cynomolgus monkey model for Alzheimer's disease (AD) research. However, in order to enhance the reliability of this disease model, appropriate reference genes must be selected to allow meaningful analysis of the gene expression levels in the icv-STZ cynomolgus monkey brain. In this study, we assessed the expression stability of 9 candidate reference genes in 2 matched-pair brain samples (5 regions) of control cynomolgus monkeys and those who had received intracerebroventricular injection of streptozotocin (icv-STZ). Three well-known analytical programs geNorm, NormFinder, and BestKeeper were used to choose the suitable reference genes from the total sample group, control group, and icv-STZ group. Combination analysis of the 3 different programs clearly indicated that the ideal reference genes are RPS19 and YWHAZ in the total sample group, GAPDH and RPS19 in the control group, and ACTB and GAPDH in the icv-STZ group. Additionally, we validated the normalization accuracy of the most appropriate reference genes (RPS19 and YWHAZ) by comparison with the least stable gene (TBP) using quantification of the APP and MAPT genes in the total sample group. To the best of our knowledge, this research is the first study to identify and validate the appropriate reference genes in cynomolgus monkey brains. These findings provide useful information for future studies involving the expression of target genes in the cynomolgus monkey.
authors: National Primate Research Center, Korea Research Institute of Bioscience and Biotechnology, Ochang, Chungbuk, Republic of Korea.
authors: Sang-Je
authors: SJ
authors: Park
authors: Young-Hyun
authors: YH
authors: Kim
authors: Youngjeon
authors: Y
authors: Lee
authors: Kyoung-Min
authors: KM
authors: Kim
authors: Heui-Soo
authors: HS
authors: Kim
authors: Sang-Rae
authors: SR
authors: Lee
authors: Sun-Uk
authors: SU
authors: Kim
authors: Sang-Hyun
authors: SH
authors: Kim
authors: Ji-Su
authors: JS
authors: Kim
authors: Kang-Jin
authors: KJ
authors: Jeong
authors: Kyoung-Min
authors: KM
authors: Lee
authors: Jae-Won
authors: JW
authors: Huh
authors: Kyu-Tae
authors: KT
authors: Chang
mesh_terms: D000544:Alzheimer Disease / Q000139:chemically induced / Q000235:genetics*; D000818:Animals; D001921:Brain / Q000378:metabolism*; D020869:Gene Expression Profiling / Q000592:standards*; D005796:Genes; D008252:Macaca fascicularis / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000592:standards*; D012015:Reference Standards; D013311:Streptozocin
chemical_list: D013311:Streptozocin