电压-Seq:全光学后突触连接体引导的单细胞转录组学
Veronika Csillag1, Marianne Hiriart Bizzozzero1, J C Noble2
1Department of Neuroscience, Karolinska Institute, Stockholm, Sweden.
Nature methods
|July 20, 2023
概括
电压Seq通过分类 postsynaptic 响应类型 (PRTs) 来实现单细胞RNA 测序的目标神经元选择. 这种方法将神经元连接与基因表达联系起来,推进了连接组研究.
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
- 系统神经科学 系统神经科学
背景情况:
- 描述神经元连接对于理解信息流动至关重要.
- 后突触反应类型 (PRT) 多样化,但由于低通量电生理学和缺乏scRNA-seq选择标准,很难与基因表达联系起来.
研究的目的:
- 基于特定的PRT开发一种针对神经元的方法,用于随后的单细胞RNA测序 (scRNA-seq).
- 克服传统电生理学的局限性,并将功能反应与遗传特征进行整合.
主要方法:
- 开发了Voltage-Seq,一种全光电压成像技术.
- 记录了 8,347 个神经元的 PRT,这些神经元在小鼠的周周导管灰色 (PAG) 中通过光遗传学激活 ventromedial hypothalamic (VMH) 终端而引起.
- 创建了VoltView,一个现场分析工具与分类器,用于使用VMH-PAG连接组数据库向目标PRT导航索马收获.
主要成果:
- 在VMH-PAG连接组中分类和空间解析的PRT.
- 通过在PAG中识别VMH驱动的γ-aminobutyric acid-ergic神经元,成功地证明了Voltage-Seq,仅基于现场VoltView分类.
- 建立了功能神经元活动 (PRTs) 和遗传身份 (scRNA-seq) 之间的联系.
结论:
- 电压-Seq提供了一个高通量方法,以功能性地描述神经元群体.
- 这种技术可以为scRNA-seq进行向细胞采集,弥合神经元连接,功能和基因表达之间的差距.
- 电压-Seq显著推进神经电路和连接学研究.
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