Objective: To screen for the most stable reference genes(RGs) in various tissues of rats fed at different dietary concentrations of selenium(Se).
Methods: Twenty-four weaning male SD rats were fed Se deficient diet for 5 weeks, and then randomly divided into 4 groups for<0. 01, 0. 25, 3 and 5 mg Se/kg diet feeding, respectively. After 4 weeks, animals were sacrificed for sample collection of liver, testis, muscle and fat tissue. Twelve candidate RGs of Actb, Atp5f1, B2m, Gapdh, Gusb, Hprt, Pgk1, Ppia, Rplp2, Rps18, Tbp and Ywhaz were tested for their quantitative cycle numbers of mRNA abundances with the quantitative PCR method. The stabilities of the candidate RGs were evaluated by the arithmetic packages of geNorm, NormFinder, BestKeeper, Delta CT and RefFinder.
Results: The top 4 most stable RGs were Ppia >Atp5f1 > Rplp2 >Hprt in liver; Ywhaz > Atp5f1 >Rplp2> Ppia in testis; Tbp > Ppia > B2m > Rps18 in muscle; Hprt>Tbp >Atp5f1>Pgk1 in fat tissue; and Rps18>Hprt> Rplp2>Atp5f1 when all the 4 tissues combined for analysis.
Conclusion: To analyze the expressions of the target genes in rats fed different concentrations of dietary Se, the best RGs should be selected depending on the tissue types.
Keywords: dietary selenium; quantitative PCR; rat; reference gene; various tissues.