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首页> 外文期刊>Acta neurobiologiae experimentalis >Cell?based immunofluorescence assay for screening the neurogenesis potential of new drugs in adult hippocampal neural progenitor cells
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Cell?based immunofluorescence assay for screening the neurogenesis potential of new drugs in adult hippocampal neural progenitor cells

机译：基于细胞的免疫荧光法筛选成年海马神经祖细胞中新药的神经发生潜能

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Preclinical studies have suggested that increased adult neurogenesis in the hippocampus might have potential therapeutic effects for Alzheimer’s disease and depression; therefore, it is a target for the treatment of some brain diseases. In this technical communication,we propose a cell?based fluorescence assay to study the neurogenesis of adult hippocampal progenitor cells that can be used for high?throughput screening of drugs promoting neurogenesis. Three fluorescent dyes (DAPI, Alexa Fluor 488, and Alexa Fluor 594) and a fluorescence spectrophotometry reader were used, which confirmed that the mutual interference of the three fluorescent dyes is very low. We used this cell?based fluorescence assay to evaluate the effects of three neurotrophic factors, ciliary neurotrophic factor (CNTF), insulin?like growth factor 1 (IGF?1), and IGF?2 on the promotion of neurogenesis in adult hippocampal neural progenitor cells. The fluorescence intensity ratio of the neuronal marker, class III β?tubulin, to the housekeeping protein, glyceraldehyde 3?phosphate dehydrogenase, or nuclear staining dye, DAPI, in CNTF?treated cells was significantly higher than in control cells. The ratios in IGF?1? and IGF?2?treated cells were slightly higher under higher cell density conditions. These results are consistent with those in previous reports; therefore, this report proved the efficacy of this method. Taken together, the results showed that this simple, rapid, and economical cell?based immunofluorescence assay could be a powerful tool for the rapid screening of drugs that promote adult neurogenesis.
机译：临床前研究表明，海马中成人神经发生的增加可能对阿尔茨海默氏病和抑郁症具有潜在的治疗作用。因此，它是治疗某些脑部疾病的目标。在这项技术交流中，我们提出了一种基于细胞的荧光测定法，以研究成年海马祖细胞的神经发生，该细胞可用于高通量筛选促进神经发生的药物。使用了三种荧光染料（DAPI，Alexa Fluor 488和Alexa Fluor 594）和荧光分光光度计读取器，证实了这三种荧光染料的相互干扰非常低。我们使用这种基于细胞的荧光测定法来评估三种神经营养因子，睫状神经营养因子（CNTF），胰岛素样生长因子1（IGF？1）和IGF？2对成年海马神经祖细胞促进神经发生的作用。细胞。在CNTF处理过的细胞中，神经元标志物III类β-微管蛋白与管家蛋白，甘油醛3β-磷酸脱氢酶或核染色染料DAPI的荧光强度比显着高于对照细胞。 IGF中的比率为1？在较高的细胞密度条件下，经IGFα2′处理的细胞略高。这些结果与以前的报告一致;因此，本报告证明了该方法的有效性。综上所述，结果表明，这种简单，快速，经济的基于细胞的免疫荧光测定法可能是快速筛选促进成人神经发生的药物的强大工具。

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