Brain-Derived Neurotrophic Factor (BDNF)

Brain-Derived Neurotrophic Factor (BDNF)
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脑源性神经营养因子 (BDNF)

DOI:
10.1007/7657_2017_4
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发表时间:
2019
期刊:
--
影响因子:
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通讯作者:
Nath A
Nath A
中科院分区:
--
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作者:
Nath A

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高通量下一代测序技术是深入了解大群细胞转录状态的宝贵工具。这些数据可以帮助阐明正常和病理条件下组织和通路的组织。在我们的案例中,我们正在使用上述技术来破译纹状体的脑啡能介质棘神经元(msn)如何在存在或不存在BDNF/TrkB信号的情况下适应衰老。然而,必须对测序数据进行验证,理想情况下,使用另一种方法来询问细胞的转录状态,并且能够在罕见的单细胞或大细胞中以高灵敏度检测基因表达。因此,我们优化了一种高通量微流控分析方案[Fluidigm动态阵列集成微流控电路(IFC)],以验证来自有限数量的成年和老年分类神经元的RNA测序数据。以下是该协议的详细说明。
High-throughput next generation sequencing technologies are an invaluable tool to gain insight into the transcriptional states of large cohorts of cells. Such data can help to shed light on the organization of tissues and pathways under normal and pathological conditions. In our case, we are using the above technology to decipher how the enkephalinergic medium spiny neurons (MSNs) of the striatum adapt to aging in the presence or absence of BDNF/TrkB signaling. However, sequencing data must be validated, ideally with an alternative method that interrogates the transcriptional state of cells, and is able to detect gene expression in rare single cells or bulk cells with high sensitivity. Thus, we have optimized a protocol for high-throughput microfluidic analysis [Fluidigm Dynamic Array Integrated Microfluidic Circuit (IFC)] to validate RNA sequencing data from a limited number of adult and aged sorted neurons. Here is a detailed description of this protocol.
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