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Transcriptional expression study in the central nervous system of rats: what gene should be used as internal control?

机译：大鼠中枢神经系统的转录表达研究：应使用哪种基因作为内部对照？

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Objective A growing number of published articles report the expression of specific genes with different behavior patterns in rats. The levels of messenger ribonucleic acid transcripts are usually analyzed by reverse transcription followed by polymerase chain reaction and quantified after normalization with an internal control or reference gene (housekeeping gene). Nevertheless, housekeeping genes exhibit different expression in the central nervous system, depending on the physiological conditions and the area of the brain to be studied. The choice of a good internal control gene is essential for obtaining reliable results. This study evaluated the expression of three housekeeping genes (beta-actin, cyclophilin A, and ubiquitin C) in different areas of the central nervous system in rats (olfactory bulb, hippocampus, striatum, and prefrontal cortex).Methods Wistar rats (virgin females, n=6) during the diestrum period were used. Total ribonucleic acid was extracted from each region of the brain; the complementary deoxyribonucleic acid was synthesized by reverse transcription and amplified by real-time quantitative polymerase chain reaction using SYBR™ Green and primers specific for each one of the reference genes. The stability of the expression was determined using NormFinder.Results Beta-actin was the most stable gene in the hippocampus and striatum, while cyclophilin A and ubiquitin C showed greater stability in the prefrontal cortex and the olfactory bulb, respectively.Conclusion Based on our study, further studies of gene expression using rats as animal models should take into consideration these results when choosing a reliable internal control gene.
机译：目的越来越多的已发表文章报道了具有不同行为模式的特定基因在大鼠中的表达。信使核糖核酸转录物的水平通常通过反转录，然后进行聚合酶链反应进行分析，并在使用内部对照或参考基因（管家基因）标准化后进行定量。然而，看家基因在中枢神经系统中表现出不同的表达，这取决于生理条件和要研究的大脑区域。选择良好的内部对照基因对于获得可靠的结果至关重要。这项研究评估了三种管家基因（β-肌动蛋白，亲环蛋白A和泛素C）在大鼠中枢神经系统不同区域（嗅球，海马，纹状体和前额叶皮层）的表达。 ，n = 6）在发情期。从大脑的每个区域提取总核糖核酸;互补的脱氧核糖核酸是通过逆转录合成的，并使用SYBR™Green和对每个参考基因特异的引物通过实时定量聚合酶链反应进行扩增。结果表明，β-肌动蛋白是海马和纹状体中最稳定的基因，而亲环蛋白A和泛素C分别在前额叶皮层和嗅球中表现出更大的稳定性。 ，使用大鼠作为动物模型进行基因表达的进一步研究应在选择可靠的内部对照基因时考虑到这些结果。

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