Identification of reference genes for expression studies in the liver and spleen of laying hens housed in cage and cage-free systems.

María P Herrera-Sánchez, Kelly J Lozano-Villegas, Iang S Rondón-Barragán, Roy Rodríguez-Hernández
Author Information
  1. María P Herrera-Sánchez: Poultry Research Group, Laboratory of Immunology and Molecular Biology, Faculty of Veterinary Medicine and Zootechnics, University of Tolima, Ibagué, Colombia.
  2. Kelly J Lozano-Villegas: Poultry Research Group, Laboratory of Immunology and Molecular Biology, Faculty of Veterinary Medicine and Zootechnics, University of Tolima, Ibagué, Colombia.
  3. Iang S Rondón-Barragán: Poultry Research Group, Laboratory of Immunology and Molecular Biology, Faculty of Veterinary Medicine and Zootechnics, University of Tolima, Ibagué, Colombia.
  4. Roy Rodríguez-Hernández: Poultry Research Group, Laboratory of Immunology and Molecular Biology, Faculty of Veterinary Medicine and Zootechnics, University of Tolima, Ibagué, Colombia.

Abstract

Background: The liver and spleen play a pivotal role in metabolism and immune response. During stress, neuroendocrine response induces changes in gene expression, and its assessment demands the validation of the stability of the reference genes to perform relative gene expression experiments.
Aim: The objective of this study was to determine the expression stability of four reference genes (, and ) in the liver and spleen tissues from laying hens housed in a conventional cage (CC) and cage-free (CF) egg production systems.
Methods: Liver and spleen from Hy-Line Brown hens housed in CC and CF egg production systems were used. mRNA transcript levels were determined by quantitative polymerase chain reaction (qPCR), and the gene expression stability was evaluated using geNorm, BestKeeper, and NormFinder algorithms.
Results: The most stable gene from liver tissue was in CC, CF, and CC-CF groups (overall data). In the spleen, the most stable genes were (CC), (CF), and (CC-CF).
Conclusion: The gene was the most stable gene in the liver, and and genes were stable in spleen tissues that could be used for the normalization in qPCR experiments performed in liver and spleen tissues of laying hens housed CC and CF production systems.

Keywords

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MeSH Term

Animals
Female
Spleen
Chickens
Liver

Word Cloud

Created with Highcharts 10.0.0spleenlivergenegenesexpressionhensCCCFhousedsystemsstablestabilityreferencetissueslayingproductionresponseexperimentscagecage-freeeggLiverusedmRNAqPCRCC-CFBackground:playpivotalrolemetabolismimmunestressneuroendocrineinduceschangesassessmentdemandsvalidationperformrelativeAim:objectivestudydeterminefourconventionalMethods:Hy-LineBrowntranscriptlevelsdeterminedquantitativepolymerasechainreactionevaluatedusinggeNormBestKeeperNormFinderalgorithmsResults:tissuegroupsoveralldataConclusion:normalizationperformedIdentificationstudiesLayingReferenceSpleen

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