Search for appropriate reference genes for quantitative reverse transcription PCR studies in somite, prosencephalon and heart of early mouse embryo

Moysés-Oliveira, Mariana; Cabral, Victória; Gigek, Carolina Oliveira; Corrêa, Débora Cabral de Carvalho; Di-Battista, Adriana; Stumpp, Taiza; Melaragno, Maria Isabel

doi:10.1016/j.gene.2019.05.042

Cited by 7 publications

(7 citation statements)

References 28 publications

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“…The previous studies involving reference genes primarily based on myocardial tissues from adult mice or whole hearts [28]. Another limitation of previous studies is that only a limited number of candidate reference genes and a small number of samples were included [20,28,29]. In this study, we investigated the Fig.…”

Section: Discussionmentioning

confidence: 99%

“…The statistical applications for evaluating the expression stability of reference genes were also shown. E = embryonic day; D = postnatal day; M = postnatal month a small number of samples at different developmental stages were included [20]. So far, information about the optimal reference genes sets in murine LV tissues across embryonic and postnatal stages is still inadequate.…”

Section: Introductionmentioning

confidence: 99%

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Identification of reference genes for gene expression studies among different developmental stages of murine hearts

Ren

Sun

et al. 2021

BMC Dev Biol

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Background Real-time quantitative polymerase chain reaction (RT-qPCR) is a widely-used standard assay for assessing gene expression. RT-qPCR data requires reference genes for normalization to make the results comparable. Therefore, the selected reference gene should be highly stable in its expression throughout the experimental datasets. So far, reports about the optimal set of reference genes in murine left ventricle (LV) across embryonic and postnatal stages are few. The objective of our research was to identify the appropriate reference genes in murine LV among different developmental stages. Methods We investigated the gene expression profiles of 21 widely used housekeeping genes in murine LV from 7 different developmental stages (almost throughout the whole period of the mouse lifespan). The stabilities of the potential reference genes were evaluated by five methods: GeNorm, NormFinder, BestKeeper, Delta-Ct and RefFinder. Results We proposed a set of reliable reference genes for normalization of RT-qPCR experimental data in different conditions. Furthermore, our results showed that 6 genes (18S, Hmbs, Ubc, Psmb4, Tfrc and Actb) are not recommended to be used as reference genes in murine LV development studies. The data also suggested that the Rplp0 gene might serve as an optimal reference gene in gene expression analysis. Conclusions Our study investigated the expression stability of the commonly used reference genes in process of LV development and maturation. We proposed a set of optimal reference genes that are suitable for accurate normalization of RT-qPCR data in specific conditions. Our findings may be helpful in future studies for investigating the gene expression patterns and mechanism of mammalian heart development.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Identification of reference genes for gene expression studies among different developmental stages of murine hearts

Ren

Sun

et al. 2021

BMC Dev Biol

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show abstract

“…The number of reference genes studies during heart development and maturation was relatively small. It is the limitations of previous researches that only a limited number of candidate reference genes and a small number of samples with different developmental stages were included [22,27,28]. In this study, we investigated the gene expressions of common 21 candidate housekeeping genes in 7 different developmental stages, almost throughout the whole period of the mouse lifespan.…”

Section: Discussionmentioning

confidence: 99%

“…Previous investigators demonstrated that the expression of some candidate reference genes can signi cantly vary in different condition [22]. For example, gene 18S, Actb and Gapdh held considerably alterations of expression level depending upon developmental stages and experimental conditions [29].…”

Section: Discussionmentioning

confidence: 99%

“…Some housekeeping genes were demonstrated signi cantly altered in heart development or maturation process [21]. A recent study identi ed the appropriate reference genes for normalizing gene expression data of heart with different developmental stages in mouse [22]. So far, there was little optimal set of reference genes had been identi ed in LV tissues across embryonic and postnatal stages in mice.…”

Section: Introductionmentioning

confidence: 99%

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Identification of appropriate reference genes for gene expression studies in mice left ventricles from different developmental stages

Ren

Zhang

et al. 2021

Preprint

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Aims: Real-time quantitative polymerase chain reaction (RT-qPCR) is the standard assay used for revealing the gene expression characteristics. However, the RT-qPCR studies all need reference genes for normalization to make the results comparable, which should hold a high expression stability during all experimental datasets. So far, there was no optimal set of reference genes identified in mice left ventricles (LV) across embryonic and postnatal stages. The objective of our research was to identify the appropriate reference genes in mice LV from different developmental stages.Methods and Results: we investigated the gene expressions of common 21 candidate housekeeping genes in mice LV from 7 different developmental stages, almost throughout the whole period of the mouse lifespan. The expression of some candidate reference genes, such as 18S and Actb, apparently fluctuated. The stability of potential reference genes was evaluated by a number of methods, such as GeNorm, NormFinder, BestKeeper, Delta-Ct and RefFinder method. we identified a set of optimal reference genes that can be reliably used for normalization of RT-qPCR experiments in different developmental stages of mice LV. Our results showed that following genes should not be used as reference genes in mice LV development studies: 18S, Hmbs, Ubc, Psmb4, Tfrc and Actb. And the Rplp0 appeared to represent a good choice. Conclusions: Our study provides the expression stability of the commonly used reference genes in process of LV development and maturation. We also identified a set of optimal reference genes under different conditions. Our findings may be helpful in future studies to investigate the gene expression patterns and mechanism of mammalian heart development.

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Glioma induces atypical depression-like behaviors in mice through the 5-HT and glutamatergic synapse pathways

Li,

Ran,

Jing

et al. 2024

Biochemical and Biophysical Research Communications

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Search for appropriate reference genes for quantitative reverse transcription PCR studies in somite, prosencephalon and heart of early mouse embryo

Cited by 7 publications

References 28 publications

Identification of reference genes for gene expression studies among different developmental stages of murine hearts

Identification of reference genes for gene expression studies among different developmental stages of murine hearts

Identification of appropriate reference genes for gene expression studies in mice left ventricles from different developmental stages

Glioma induces atypical depression-like behaviors in mice through the 5-HT and glutamatergic synapse pathways

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