title: Identification of Reference Genes for Quantitative Real Time PCR Assays in Aortic Tissue of Syrian Hamsters with Bicuspid Aortic Valve.
abstract: Bicuspid aortic valve (BAV) is the most frequent congenital cardiac malformation in humans, and appears frequently associated with dilatation of the ascending aorta. This association is likely the result of a common aetiology. Currently, a Syrian hamster strain with a relatively high (∼40%) incidence of BAV constitutes the only spontaneous animal model of BAV disease. The characterization of molecular alterations in the aorta of hamsters with BAV may serve to identify pathophysiological mechanisms and molecular markers of disease in humans. In this report, we evaluate the expression of ten candidate reference genes in aortic tissue of hamsters in order to identify housekeeping genes for normalization using quantitative real time PCR (RT-qPCR) assays. A total of 51 adult (180-240 days old) and 56 old (300-440 days old) animals were used. They belonged to a control strain of hamsters with normal, tricuspid aortic valve (TAV; n = 30), or to the affected strain of hamsters with TAV (n = 45) or BAV (n = 32). The expression stability of the candidate reference genes was determined by RT-qPCR using three statistical algorithms, GeNorm, NormFinder and Bestkeeper. The expression analyses showed that the most stable reference genes for the three algorithms employed were Cdkn1β, G3pdh and Polr2a. We propose the use of Cdkn1β, or both Cdkn1β and G3pdh as reference genes for mRNA expression analyses in Syrian hamster aorta.
authors.affiliation: UGC del Corazón, Hospital Universitario Virgen de la Victoria, Universidad de Málaga, Red de Investigación Cardiovascular (RIC), Málaga, Spain.
authors.forename: Carmen
authors.initials: C
authors.lastname: Rueda-Martínez
authors.affiliation: Departamento de Biología Animal, Facultad de Ciencias, Universidad de Málaga, Málaga, Spain.
authors.forename: M Carmen
authors.initials: MC
authors.lastname: Fernández
authors.affiliation: Departamento de Biología Animal, Facultad de Ciencias, Universidad de Málaga, Málaga, Spain.
authors.forename: María Teresa
authors.initials: MT
authors.lastname: Soto-Navarrete
authors.affiliation: UGC del Corazón, Hospital Universitario Virgen de la Victoria, Universidad de Málaga, Red de Investigación Cardiovascular (RIC), Málaga, Spain.|Departamento de Medicina, Facultad de Medicina, Universidad de Málaga, Málaga, Spain.|Instituto de Investigación Biomédica de Málaga (IBIMA), Málaga, Spain.
authors.forename: Manuel
authors.initials: M
authors.lastname: Jiménez-Navarro
authors.affiliation: Departamento de Biología Animal, Facultad de Ciencias, Universidad de Málaga, Málaga, Spain.|Instituto de Investigación Biomédica de Málaga (IBIMA), Málaga, Spain.
authors.forename: Ana Carmen
authors.initials: AC
authors.lastname: Durán
authors.affiliation: Departamento de Biología Animal, Facultad de Ciencias, Universidad de Málaga, Málaga, Spain.|Instituto de Investigación Biomédica de Málaga (IBIMA), Málaga, Spain.
authors.forename: Borja
authors.initials: B
authors.lastname: Fernández
mesh_terms: D000818:Animals; D001011:Aorta / Q000378:metabolism* / Q000473:pathology; D001021:Aortic Valve / Q000002:abnormalities*; D000082882:Bicuspid Aortic Valve Disease; D006224:Cricetinae; D005786:Gene Expression Regulation; D006349:Heart Valve Diseases / Q000235:genetics*; D008647:Mesocricetus; D060888:Real-Time Polymerase Chain Reaction / Q000592:standards*; D012015:Reference Standards