Evaluation of potential reference genes in real-time RT-PCR studies of Atlantic salmon

Olsvik, Pål A; Lie, Kai K; Jordal, Ann-Elise O; Nilsen, Tom O; Hordvik, Ivar

Transcription levels of the examined genes and the coefficient of variance (CV) in different tissues varied considerably. mRNA levels in tissues are regulated by numerous endogenous and exogenous stimuli [16] . Transcription rates in metabolic active tissues might be up-regulated compared to those of less active tissues, whereas inter-tissue variation in degradation rates of mRNAs, for example, might affect mRNA stability [25] . The results revealed that muscle had the lowest CVs of the studied genes, compared to higher CVs in more active tissues like thymus, head kidney and spleen. In thymus, intestine, head kidney, gills, brain, liver and spleen, the 18S and S20 genes had the lowest CVs, based on raw Ct values. In all tissues, except intestine, the GAPDH gene had the highest CV. Except for thymus, the two elongation factor genes had relatively similar expression in all eight examined tissues. Their expression are most likely co-expressed in the examined tissues, and therefore favored in geNorm calculations [22] . The results also demonstrated that assays optimized for one tissue of an organism do not necessarily work equally well in other tissues. Of the tissues studied in this work, intestine, head kidney and spleen were the most troublesome.