确定可靠的参考基因,用于在老鼠发烧性模型中的基因表达研究
Anna A Kovalenko1, Maria V Zakharova1, Alexander P Schwarz1
1Laboratory of Molecular Mechanisms of Neural Interactions, Sechenov Institute of Evolutionary Physiology and Biochemistry of RAS, 194223 Saint Petersburg, Russia.
International journal of molecular sciences
|October 26, 2024
概括
确定稳定的参考基因对于在发烧研究中准确的基因表达分析至关重要. 在长期新生儿发烧之后,Ppia基因在老鼠大脑区域表达最稳定.
科学领域:
- 神经科学是一个神经科学.
- 分子生物学分子生物学
- 遗传学 遗传学 是一个
背景情况:
- 发烧性发作可能会产生长期的神经后果.
- 基因表达分析,特别是使用量化聚合酶链反应 (RT-qPCR) 的逆转录,对于研究发烧性病原体至关重要.
- 准确的RT-qPCR数据依赖于使用稳定表达的参考基因进行正常化.
研究的目的:
- 在长期新生儿发烧后,在不同大鼠大脑区域中确定最适合用于mRNA表达分析的参考基因.
- 为了评估八个候选家政基因的表达稳定性.
主要方法:
- 候选参考基因表达在经历过长期新生儿发烧的老鼠的背部海马体,腹部海马体,叶皮和中部前额叶皮中进行了分析.
- 用在线工具RefFinder评估基因表达稳定性.
主要成果:
- 普皮亚基因在评估的大脑区域中表达稳定性最高.
- Actb和Ywhaz基因表现出最不稳定的表达,并且不建议在这种发烧性发作模型中进行正常化.
- 对于大多数家政基因,观察到与年龄相关的和特定区域的显著表达波动.
结论:
- 选择适当的参考基因对于在发烧性研究中准确的基因表达概况至关重要.
- 建议Ppia作为稳定的参考基因,用于在长期新生儿发烧之后,在老鼠大脑区域中正常化RT-qPCR数据.
- 由于它们在这个模型中的不稳定性,因此不建议使用Actb和Ywhaz进行正常化,并且考虑年龄和大脑区域是必不可少的.
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