Evaluation of suitable reference genes for gene expression studies in the developing mouse cortex using RT-qPCR.

IF 2.4 4区医学 Q3 NEUROSCIENCES

BMC Neuroscience Pub Date : 2025-02-18 DOI:10.1186/s12868-025-00934-y

Ananya Uppalapati, Timothy Wang, Lena H Nguyen

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引用次数: 0

Abstract

Background: Real-time quantitative PCR (RT-qPCR) is a widely used method to investigate gene expression in neuroscience studies. Accurate relative quantification of RT-qPCR requires the selection of reference genes that are stably expressed across the experimental conditions and tissues of interest. While RT-qPCR is often performed to investigate gene expression changes during neurodevelopment, few studies have examined the expression stability of commonly used reference genes in the developing mouse cortex.

Results: Here, we evaluated the stability of five housekeeping genes, Actb, Gapdh, B2m, Rpl13a, and Hprt, in cortical tissue from mice at embryonic day 15 to postnatal day 0 to identify optimal reference genes with stable expression during late corticogenesis. The expression stability was assessed using five computational algorithms: BestKeeper, geNorm, NormFinder, DeltaCt, and RefFinder. Our results showed that B2m, Gapdh, and Hprt, or a combination of B2m/Gapdh and B2m/Hprt, were the most stably expressed genes or gene pairs. In contrast, Actb and Rpl13a were the least stably expressed.

Conclusion: This study identifies B2m, Gapdh, and Hprt as suitable reference genes for relative quantification in RT-qPCR-based cortical development studies spanning the period of embryonic day 15 to postnatal day 0.

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利用RT-qPCR技术评价发育中小鼠皮质基因表达的合适内参基因。

背景：实时定量PCR （RT-qPCR）是神经科学研究中广泛使用的基因表达研究方法。RT-qPCR的准确相对定量需要选择在实验条件和感兴趣的组织中稳定表达的内参基因。虽然RT-qPCR常用于研究神经发育过程中基因表达的变化，但很少有研究检测发育中的小鼠皮层中常用内参基因的表达稳定性。结果：在这里，我们评估了5个管家基因Actb、Gapdh、B2m、Rpl13a和Hprt在胚胎15天至出生后0天小鼠皮质组织中的稳定性，以确定在皮质发生后期稳定表达的最佳内参基因。使用五种计算算法评估表达稳定性：BestKeeper、geNorm、NormFinder、DeltaCt和RefFinder。结果表明，B2m、Gapdh和Hprt或B2m/Gapdh和B2m/Hprt的组合是最稳定表达的基因或基因对。相比之下，Actb和Rpl13a的表达最不稳定。结论：本研究确定B2m、Gapdh和Hprt是基于rt - qpcr的胚胎15天至出生后0天皮质发育研究中相对定量的合适内参基因。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

BMC Neuroscience 医学-神经科学

CiteScore

3.90

自引率

0.00%

发文量

审稿时长

16 months

期刊介绍： BMC Neuroscience is an open access, peer-reviewed journal that considers articles on all aspects of neuroscience, welcoming studies that provide insight into the molecular, cellular, developmental, genetic and genomic, systems, network, cognitive and behavioral aspects of nervous system function in both health and disease. Both experimental and theoretical studies are within scope, as are studies that describe methodological approaches to monitoring or manipulating nervous system function.