Abstract:
Mosaic Analysis with Double Markers (MADM) is a powerful genetic method typically used for lineage tracing and to disentangle cell autonomous and tissue-wide roles of candidate genes with single cell resolution. Given the relatively sparse labeling, depending on which of the 19 MADM chromosomes one chooses, the MADM approach represents the perfect opportunity for cell morphology analysis. Various MADM studies include reports of morphological anomalies and phenotypes in the central nervous system (CNS). MADM for any candidate gene can easily incorporate morphological analysis within the experimental workflow. Here, we describe the methods of morphological cell analysis which we developed in the course of diverse recent MADM studies. This chapter will specifically focus on methods to quantify aspects of the morphology of neurons and astrocytes within the CNS, but these methods can broadly be applied to any MADM-labeled cells throughout the entire organism. We will cover two analyses-soma volume and dendrite characterization-of physical characteristics of pyramidal neurons in the somatosensory cortex, and two analyses-volume and Sholl analysis-of astrocyte morphology.
摘要:
具有双标记的马赛克分析(MADM)是一种强大的遗传方法,通常用于谱系追踪和解开具有单细胞分辨率的候选基因的细胞自主和组织范围的作用。鉴于标签相对稀疏,根据一个人选择的19条MADM染色体中的哪一条,MADM方法代表了细胞形态分析的绝佳机会。各种MADM研究包括中枢神经系统(CNS)形态异常和表型的报告。任何候选基因的MADM都可以轻松地将形态学分析纳入实验工作流程。这里,我们描述了我们在最近的各种MADM研究过程中开发的形态细胞分析方法。本章将特别关注量化中枢神经系统内神经元和星形胶质细胞形态方面的方法,但是这些方法可以广泛应用于整个生物体中的任何MADM标记的细胞。我们将涵盖两个分析-体细胞体积和树突表征-体感皮层中锥体神经元的物理特征,和两个分析-体积和Sholl分析-星形胶质细胞形态。