Yang Lite, Tochitsky Ivan, Woolf Clifford J, Renthal William
Department of Neurology, Brigham and Women's Hospital and Harvard Medical School, Boston, USA.
F.M. Kirby Neurobiology Center, Boston Children's Hospital, Boston, USA.
Bio Protoc. 2021 Aug 5;11(15):e4102. doi: 10.21769/BioProtoc.4102.
Primary somatosensory neurons, whose cell bodies reside in the dorsal root ganglion (DRG) and trigeminal ganglion, are specialized to transmit sensory information from the periphery to the central nervous system. Our molecular understanding of peripheral sensory neurons has been limited by both their heterogeneity and low abundance compared with non-neuronal cell types in sensory ganglia. We describe a protocol to isolate nuclei from mouse DRGs using iodixanol density gradient centrifugation, which enriches for neuronal nuclei while still sampling non-neuronal cells such as satellite glia and Schwann cells. This protocol is compatible with a range of downstream applications such as single-nucleus transcriptional and epigenomic assays.
初级体感神经元的细胞体位于背根神经节（DRG）和三叉神经节中，专门负责将外周感觉信息传递到中枢神经系统。与感觉神经节中的非神经元细胞类型相比，外周感觉神经元的异质性和低丰度限制了我们对其分子层面的理解。我们描述了一种使用碘克沙醇密度梯度离心法从小鼠背根神经节中分离细胞核的方法，该方法在对卫星神经胶质细胞和施万细胞等非神经元细胞进行采样的同时，能够富集神经元细胞核。该方法与一系列下游应用兼容，如单核转录和表观基因组分析。
