Stability of reference genes for real-time PCR analyses in channel catfish (Ictalurus punctatus) tissues under varying physiological conditions

Small, Brian C.; Murdock, Christopher A.; Bilodeau‐Bourgeois, A. Lelania; Peterson, Brian; Waldbieser, Geoffrey C.

doi:10.1016/j.cbpb.2008.07.010

Cited by 120 publications

(70 citation statements)

References 43 publications

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“…In this study, ranking stabilities of eight candidate genes exhibited various degrees of expression among ten tissues during juvenile and adult stages no matter which methods were used for evaluation. Similar observations were found in other fish species, such as channel catfish (Small et al, 2008), Atlantic halibut (Fernandes et al, 2008) and fathead minnow (Filby & Tyler, 2007).…”

Section: Discussionsupporting

confidence: 85%

“…However, ACTB is not suitable for using as reference gene due to high variation in different tissues at adult fish. Other studies involve in zebrafish and channel catfish have a similar result to rejected ACTB as a reference gene, especially in adult fish (McCurley & Callard, 2008;Small et al, 2008). Although many studies used G6PD as one of candidate genes in the process of selecting reliable reference genes as we do, few results recommended it because of its high variation (Hibbeler et al, 2008;Aursnes et al, 2011).…”

Section: Discussionmentioning

confidence: 80%

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Selection of Reliable Reference Genes for Quantitative Real-Time PCR in Golden-Line Barbell (Sinocyclocheilus grahami) During Juvenile and Adult Stages

yuanwei¹,

Pan²,

Wang³

et al. 2017

FisheriesSciences.com

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Abstract:In order to obtain reliable results of gene expression, appropriate internal control gene(s) were required for normalization before real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). Here, we selected eight candidate housekeeping genes as the potential reference genes in normalizing qRT-PCR data in ten tissues of the juvenile and adult stages of a naturally tetraploid cyprinid fish Sinocyclocheilus grahami. Before qRT-PCR, appropriate primers were designed and compared for distinguishing different copies in some duplicate genes. Candidate reference genes were evaluated for their stability using online software RefFinder, which is integrated by four normal software of the comparative delta-C T , BestKeeper, NormFinder and GeNorm. According to the rankings produced by these analyses, Eef2, ACTB and G6PD were the most stable reference genes as internal controls for qRT-PCR in juvenile, while B2M, Eef2 and RPL17 were shown to be most appropriate in adult fish. All the analyses show that GAPDH was the least stable expression gene which would be unsuitable as reference gene at both stages. This is the first report about reference gene selection in Sinocyclocheilus and we think that is beneficial for the future gene expression studies in golden-line barbel.

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Section: Discussionsupporting

confidence: 85%

Section: Discussionmentioning

confidence: 80%

Selection of Reliable Reference Genes for Quantitative Real-Time PCR in Golden-Line Barbell (Sinocyclocheilus grahami) During Juvenile and Adult Stages

yuanwei¹,

Pan²,

Wang³

et al. 2017

FisheriesSciences.com

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“…GAPDH was shown to be a stable gene under heat shock in Ditylum brightwellii (Guo et al, 2012), but was described as the least stable gene in salmons infected by virus (Jorgensen et al, 2006). The different performance of GAPDH might be due to its multiple functions in glycolysis cycle, DNA replication and repair, translational regulation and phosphotransferase activity (Bustin, 2002), which might make it sensitive to many perturbations in cellular homeostasis (Small et al, 2008).…”

Section: Discussionmentioning

confidence: 99%

Reference gene selection for quantitative gene expression studies during biological invasions: A test on multiple genes and tissues in a model ascidian Ciona savignyi

Huang

Gao

Jiang

et al. 2016

Gene

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“…ACTB, GAPDH and 18S). In catfish, Ictalurus punctatus, 18S gene was found to be the best for normalization under different physiological conditions (LHRH injection, fasting, and low water stress response) (Small et al 2008). However, there were concerns for the use of 18S gene as a reference, as its much greater level of expression relative to target mRNA (Tang et al 2007).…”

Section: Discussionmentioning

confidence: 99%

Identification of candidate reference genes for quantitative expression analysis by real-time PCR for hypoxic stress in Indian catfish, Clarias batrachus (Linnaeus, 1758)

et al. 2014

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The selection of most appropriate reference gene(s) is a crucial step in studies quantifying gene expression by quantitative real-time PCR (qPCR). As a need reference gene(s) should be unaffected at transcription level by experimental conditions and/or tissue types. In this study, 11 candidate reference genes were tested in Clarias batrachus for their expression stability using the geNorm, NormFinder and BestKeeper statistical algorithms, and were compared in different tissues (brain, heart, liver, muscle, spleen and head kidney) and treatments (before and after hypoxia exposure). The results indicated that in brain, heart, liver, muscle, spleen and head kidney, respectively, 28S rRNA/TUB, 28S rRNA/TUB, RPL30/ 28S rRNA, RPL30/TUB, ELF-1A/28S rRNA and ELF-1A/TUB gene pairs were highly stable and were suitable as reference genes to study oxidative stress, while ACTB and B2M were the least stable genes in examined tissues under normoxic and/or hypoxic conditions. The observations suggested the consideration of tissue types and use of at least two reference genes, instead of one in accurate normalization of qPCR data.

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Stability of reference genes for real-time PCR analyses in channel catfish (Ictalurus punctatus) tissues under varying physiological conditions

Cited by 120 publications

References 43 publications

Selection of Reliable Reference Genes for Quantitative Real-Time PCR in Golden-Line Barbell (Sinocyclocheilus grahami) During Juvenile and Adult Stages

Selection of Reliable Reference Genes for Quantitative Real-Time PCR in Golden-Line Barbell (Sinocyclocheilus grahami) During Juvenile and Adult Stages

Reference gene selection for quantitative gene expression studies during biological invasions: A test on multiple genes and tissues in a model ascidian Ciona savignyi

Identification of candidate reference genes for quantitative expression analysis by real-time PCR for hypoxic stress in Indian catfish, Clarias batrachus (Linnaeus, 1758)

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