Selection of Reliable Reference Genes for Real-time qRT-PCR Analysis of Zi Geese (&amp;lt;italic&amp;gt;Anser anser domestica&amp;lt;/italic&amp;gt;) Gene Expression

Ji, Hong; Wang, Jianfa; Liu, Juxiong; Guo, Jiang; Wang, Zhongwei; Zhang, Xu; Guo, Li; Yang, Huanmin

doi:10.5713/ajas.2012.12417

Cited by 10 publications

(9 citation statements)

References 24 publications

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“…Differences in gene ranking were also observed between shell gland and spleen. Similar discrepancies have been observed in our previous study 45 and elsewhere with different species and treatments 48 , 57 , 69 . With different algorithms in different programmes, slight change of gene stability orders can be expected by the analyses using these programmes.…”

Section: Discussionsupporting

confidence: 91%

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Reference gene selection for gene expression study in shell gland and spleen of laying hens challenged with infectious bronchitis virus

Khan

Roberts

2017

Sci Rep

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Ten reference genes were investigated for normalisation of candidate target gene expression data in the shell gland and spleen of laying hens challenged with two strains of infectious bronchitis virus (IBV). Data were analysed with geNorm, NormFinder and BestKeeper, and a comprehensive ranking (geomean) was calculated. In the combined data set of IBV challenged shell gland samples, the comprehensive ranking showed TATA-box binding protein (TBP) and tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein zeta (YWHAZ) as the two most stable, and succinate dehydrogenase complex flavoprotein subunit A (SDHA) and albumin (ALB) as the two least stable reference genes. In the spleen, and in the combined data set of the shell gland and spleen, the two most stable and the two least stable reference genes were TBP and YWHAZ, and ribosomal protein L4 (RPL4) and ALB, respectively. Different ranking has been due to different algorithms. Validation studies showed that the use of the two most stable reference genes produced accurate and more robust gene expression data. The two most and least stable reference genes obtained in the study, were further used for candidate target gene expression data normalisation of the shell gland and spleen under an IBV infection model.

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Section: Discussionsupporting

confidence: 91%

“…In different vertebrate species, different reference genes under different treatments have been validated in the reproductive system. GAPDH and HPRT1 were ranked as the two most stable reference genes in the ovary of geese 57 . In bovines, the two most stable reference genes in the uterus were GAPDH and YWHAZ 58 .…”

Section: Discussionmentioning

confidence: 99%

Reference gene selection for gene expression study in shell gland and spleen of laying hens challenged with infectious bronchitis virus

Khan

Roberts

2017

Sci Rep

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“…g . [ 16 , 69 – 71 ]). Indeed, it has been shown that tissue/cell type can have a larger influence on RG stability than treatment regime [ 72 ].…”

Section: Discussionmentioning

confidence: 99%

With Reference to Reference Genes: A Systematic Review of Endogenous Controls in Gene Expression Studies

2015

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The choice of reference genes that are stably expressed amongst treatment groups is a crucial step in real-time quantitative PCR gene expression studies. Recent guidelines have specified that a minimum of two validated reference genes should be used for normalisation. However, a quantitative review of the literature showed that the average number of reference genes used across all studies was 1.2. Thus, the vast majority of studies continue to use a single gene, with β-actin (ACTB) and/or glyceraldehyde 3-phosphate dehydrogenase (GAPDH) being commonly selected in studies of vertebrate gene expression. Few studies (15%) tested a panel of potential reference genes for stability of expression before using them to normalise data. Amongst studies specifically testing reference gene stability, few found ACTB or GAPDH to be optimal, whereby these genes were significantly less likely to be chosen when larger panels of potential reference genes were screened. Fewer reference genes were tested for stability in non-model organisms, presumably owing to a dearth of available primers in less well characterised species. Furthermore, the experimental conditions under which real-time quantitative PCR analyses were conducted had a large influence on the choice of reference genes, whereby different studies of rat brain tissue showed different reference genes to be the most stable. These results highlight the importance of validating the choice of normalising reference genes before conducting gene expression studies.

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“…Zi goose has high egg production and strong freeze resistance, which is suitable for feeding in North‐east China (Ji et al, ). However, because the goose breeding season is seasonal, their laying period is short (Xu, Wu, & Xu, ).…”

Section: Introductionmentioning

confidence: 99%

Metabonomics analysis of Zi goose follicular granulosa cells using ENO1 gene expression interference

Guo

Niu

et al. 2019

Animal Physiology Nutrition

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The Zi goose is native to North‐east China and is noted for its high egg production. Alpha enolase (ENO1) is a glycolytic enzyme which functions as a plasminogen receptor in follicular granulosa cells (FGCs), with several studies showing that FGCs can support follicular development. By transfecting the ENO1 interfering plasmid (shRNA) into FGCs, ENO1 expression in these cells was downregulated, suggesting the successful knock‐down of ENO1 in these cells. In this knock‐down model, we detected 13 metabolites from FGCs using LC/MS. When compared with the non‐coding shRNA (NC) group, the lower level metabolites were (R)‐(+)‐citronellic acid, altretamine, 3‐hydroxycaproic acid, heptadecanoic acid, cholecalciferol vitamin D3, indole, benzoic acid, capric acid, caffeic acid, azelaic acid, 3,4‐dihydroxyhydrocinnamic acid and cholic acid, while oleic acid was detected at high levels. To further examine the results of metabolomics, six key metabolites were verified by gas chromatography‐mass spectrometry (GC‐MS). We found that vitamin D3, indole, benzoic acid, capric acid and cholic acid were significantly downregulated in the shRNA group, while oleic acid was significantly upregulated. This observation was consistent with the metabolomics data. Through these studies, we found that decreased ENO1 levels altered certain metabolite levels in FGCs.

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Selection of Reliable Reference Genes for Real-time qRT-PCR Analysis of Zi Geese (<italic>Anser anser domestica</italic>) Gene Expression

Cited by 10 publications

References 24 publications

Reference gene selection for gene expression study in shell gland and spleen of laying hens challenged with infectious bronchitis virus

Reference gene selection for gene expression study in shell gland and spleen of laying hens challenged with infectious bronchitis virus

With Reference to Reference Genes: A Systematic Review of Endogenous Controls in Gene Expression Studies

Metabonomics analysis of Zi goose follicular granulosa cells using ENO1 gene expression interference

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