Disentangling glial diversity in peripheral nerves at single-nuclei resolution [bulk and RiboTag RNA-seq]

The peripheral nerve contains diverse cell types that support its proper function and maintenance. We analyzed multiple peripheral nerves using single-nuclei RNA-sequencing (snRNA-seq), which allowed us to circumvent difficulties encountered in analyzing cells with complex morphologies via conventional single cell methods. The resultant mouse peripheral nerve cell atlas highlights a diversity of cell types, including multiple subtypes of SCs, immune cells, and stromal cells. We identified a distinct myelinating-SC subtype that expresses Cldn14, Adamtsl1, and Pmp2 and preferentially ensheathes motor axons. The number of these motor-associated, Pmp2+ SCs is reduced in both ALS SOD1G93A mouse model and human ALS nerve samples. Our findings reveal the diversity of SCs and other cell types in peripheral nerve and serve as a reference for future studies of nerve biology and disease. Comparison between Mpz-RiboTag enriched RNA-seq and bulk RNA-seq data for both sciatic and vagus nerves外周神经拥有多种细胞类型，这些细胞对于维持其正常功能与稳态不可或缺。我们采用单细胞核RNA测序（single-nuclei RNA-sequencing, snRNA-seq）技术对多种外周神经进行分析，该方法可规避传统单细胞技术在解析具有复杂形态细胞时遇到的难题。最终构建的小鼠外周神经细胞图谱展现了丰富的细胞类型多样性，涵盖施万细胞（Schwann cells, SCs）的多个亚型、免疫细胞与基质细胞。我们鉴定出一种独特的有髓施万细胞亚型，该亚型表达Cldn14、Adamtsl1与Pmp2基因，并优先包裹运动轴突。在肌萎缩侧索硬化（Amyotrophic Lateral Sclerosis, ALS）SOD1G93A小鼠模型与人类ALS神经样本中，这类与运动神经元相关的Pmp2阳性施万细胞的数量均出现减少。本研究揭示了外周神经中施万细胞及其他细胞类型的多样性，可为未来神经生物学与神经疾病相关研究提供重要参考依据。同时还开展了针对坐骨神经与迷走神经的Mpz-RiboTag富集RNA测序数据与批量RNA测序数据的对比分析。