Influence of heat stress on reference genes stability in heart and liver of two chickens genotypes

• DOI: 10.1371/journal.pone.0228314
• Publication Date: 2020-02-06T13:28:52Z
• Authors: Juliana Gracielle...; Haniel Cedraz de ...; Daniele Botelho D...; Antônio Amândio P...; Ronaldo Vasconcel...; Caio Fernando Gro...; Teillor Machado S...; Amauri Arias Wenc...
• Keywords: Reference Genes

Introduction Real-time polymerase chain reaction (RT-qPCR) is an important tool for analyzing gene expression. However, before the expression of target genes, it crucial to normalize reference in order find most stable be used as endogenous control. A that remains all samples under different treatments considered a suitable normalizer. In this sense, we aimed identify genes normalization heart and liver tissues from two genetically divergent groups chickens (Cobb 500® commercial line Peloco backyard chickens) comfort acute heat stress environmental conditions. Eight (ACTB, HPRT1, RPL5, EEF1, MRPS27, MRPS30, TFRC LDHA) were analyzed stability. The obtained 24 chickens, 12 Cobb line, exposed conditions (comfort stress). Comfort temperature was 23°C 39.5°C one hour. Subsequently, animals euthanized, tissue fragments collected RNA extraction amplification. To determine stability rate expression, three statistical algorithms applied: BestKeeper, geNorm NormFinder, obtain aggregated list, RankAgregg package R software used. Results using BestKeeper tool, including factors (genetic group condition), LDHA, RPL5 MRPS27 tissue, TFRC, EEF1 tissue. Applying algorithm, best MRPS30 liver. Using NormFinder normalizer LDHA heart, ACTB overall ranking by RankAggreg package, considering algorithms, whereas Conclusion According classification based on (BestKeeper, NormFinder), subjected may vary depending experimental each study, such breeds, stressors, analyzed. Therefore, need perform priori studies assay at outset study emphasized.