Oligodendrocytes - Methods and Protocols
Oligodendrocytes - Methods and Protocols
复制标题
少突胶质细胞 - 方法和方案
DOI：
10.1007/978-1-4939-9072-6_2
复制
发表时间：
2019
期刊：
影响因子：
--
通讯作者：
Davies C
中科院分区：
文献类型：
--
作者：
Davies C
再髓鞘化是神经系统损伤后髓鞘在轴突周围恢复的再生过程，允许恢复电脉冲传导、营养/代谢支持和轴突健康。进行性多发性硬化的髓鞘再生失败被认为是导致轴突丢失的原因，这是临床衰退的相关因素。缺乏批准的MS促再生疗法突出了理解支持成功髓鞘再生的细胞和分子机制的必要性。一种方法是对调节髓鞘再生的细胞类型（如小胶质细胞和单核细胞衍生的巨噬细胞）进行无偏倚的基因表达分析。最近的技术进步解决了小组织样本的RNA测序的挑战，从而允许从离散病变中分离相对少量的细胞用于分析。在这里，我们提出了基于FACS的分离成年小鼠脑的局灶性髓鞘再生病变的细胞和随后的RNA提取测序的方法，以分离小胶质细胞/巨噬细胞为例。
Remyelination is the regenerative process whereby myelin sheaths are restored around axons following nervous system injury, allowing reinstatement of electrical impulse conduction, trophic/metabolic support, and axon health. Failure of remyelination in progressive multiple sclerosis is considered to contribute to axon loss, a correlate of clinical decline. Lack of approved pro-regenerative therapies for MS highlights the need to understand the cellular and molecular mechanisms underpinning successful remyelination. One approach is to conduct nonbiased gene expression analyses of cell types which regulate remyelination, such as microglia and monocyte-derived macrophages. Recent technological advances address the challenges of RNA sequencing of small tissue samples, thus allowing relatively small numbers of cells to be isolated from discrete lesions for analysis. Here, we present methods for FACS-based isolation of cells from focal remyelinating lesions of the adult mouse brain and subsequent RNA extraction for sequencing, using isolation of microglia/macrophages as an example.