Department of Biological Chemistry and Department of Neurobiology, David Geffen School of Medicine, University of California, Los Angeles, CA 90095, USA.
Department of Pathology, David Geffen School of Medicine, University of California, Los Angeles, CA 90095, USA.
Neuron. 2017 Oct 11;96(2):313-329.e6. doi: 10.1016/j.neuron.2017.09.026.
Single-cell RNA sequencing offers a promising opportunity for probing cell types mediating specific behavioral functions and the underlying molecular programs. However, this has been hampered by a long-standing issue in transcriptional profiling of dissociated cells, specifically the transcriptional perturbations that are artificially induced during conventional whole-cell dissociation procedures. Here, we develop Act-seq, which minimizes artificially induced transcriptional perturbations and allows for faithful detection of both baseline transcriptional profiles and acute transcriptional changes elicited by behavior/experience-driven activity. Using Act-seq, we provide the first detailed molecular taxonomy of distinct cell types in the amygdala. We further show that Act-seq robustly detects seizure-induced acute gene expression changes in multiple cell types, revealing cell-type-specific activation profiles. Furthermore, we find that acute stress preferentially activates neuronal subpopulations that express the neuropeptide gene Cck. Act-seq opens the way for linking physiological stimuli with acute transcriptional dynamics in specific cell types in diverse complex tissues.
单细胞 RNA 测序为探测介导特定行为功能的细胞类型和潜在的分子程序提供了一个很有前途的机会。然而，这一直受到分离细胞转录谱中长期存在的问题的阻碍，特别是在传统的全细胞分离过程中人为诱导的转录扰动。在这里，我们开发了 Act-seq，它最大限度地减少了人为诱导的转录扰动，并允许忠实检测基线转录谱以及行为/经验驱动活动引起的急性转录变化。使用 Act-seq，我们提供了杏仁核中不同细胞类型的第一个详细的分子分类。我们进一步表明，Act-seq 稳健地检测到多种细胞类型中癫痫发作引起的急性基因表达变化，揭示了细胞类型特异性的激活谱。此外，我们发现急性应激优先激活表达神经肽基因 Cck 的神经元亚群。Act-seq 为将生理刺激与特定细胞类型的急性转录动力学联系起来开辟了道路，这在不同的复杂组织中都适用。
