title: Selection of internal reference genes for normalization of quantitative reverse transcription polymerase chain reaction (qRT-PCR) analysis in the canine brain and other organs.
abstract: Quantitative reverse transcription polymerase chain reaction (qRT-PCR) is a specific and sensitive technique for quantifying gene expression. To analyze qRT-PCR data accurately, suitable reference genes that show consistent expression patterns across different tissues and experimental conditions should be selected. The objective of this study was to obtain the most stable reference genes in dogs, using samples from 13 different brain tissues and 10 other organs. 16 well-known candidate reference genes were analyzed by the geNorm, NormFinder, and BestKeeper programs. Brain tissues were derived from several different anatomical regions, including the forebrain, cerebrum, diencephalon, hindbrain, and metencephalon, and grouped accordingly. Combination of the three different analyses clearly indicated that the ideal reference genes are ribosomal protien S5 (RPS5) in whole brain, RPL8 and RPS5 in whole body tissues, RPS5 and RPS19 in the forebrain and cerebrum, RPL32 and RPS19 in the diencephalon, GAPDH and RPS19 in the hindbrain, and MRPS7 and RPL13A in the metencephalon. These genes were identified as ideal for the normalization of qRT-PCR results in the respective tissues. These findings indicate more suitable and stable reference genes for future studies of canine gene expression.
authors.affiliation: National Primate Research Center, Korea Research Institute of Bioscience and Biotechnology, Ochang, Chungbuk 363-883, Republic of Korea.
authors.forename: Sang-Je
authors.initials: SJ
authors.lastname: Park
authors.forename: Jae-Won
authors.initials: JW
authors.lastname: Huh
authors.forename: Young-Hyun
authors.initials: YH
authors.lastname: Kim
authors.forename: Sang-Rae
authors.initials: SR
authors.lastname: Lee
authors.forename: Sang-Hyun
authors.initials: SH
authors.lastname: Kim
authors.forename: Sun-Uk
authors.initials: SU
authors.lastname: Kim
authors.forename: Heui-Soo
authors.initials: HS
authors.lastname: Kim
authors.forename: Min Kyu
authors.initials: MK
authors.lastname: Kim
authors.forename: Kyu-Tae
authors.initials: KT
authors.lastname: Chang
mesh_terms: D000818:Animals; D001921:Brain / Q000378:metabolism*; D004285:Dogs; D005069:Evaluation Studies as Topic; D015870:Gene Expression; D012015:Reference Standards; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012269:Ribosomal Proteins / Q000378:metabolism*; D014018:Tissue Distribution
chemical_list: D012269:Ribosomal Proteins