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利用双标记马赛克分析进行神经元和神经胶质细胞的形态分析

Morphological Analysis of Neurons and Glia Using Mosaic Analysis with Double Markers.

机构信息

Institute of Science and Technology Austria (ISTA), Klosterneuburg, Austria.

出版信息

Methods Mol Biol. 2024;2831:283-299. doi: 10.1007/978-1-0716-3969-6_19.

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

马赛克分析双标记(MADM)是一种强大的遗传方法,通常用于谱系追踪和单细胞分辨率解析候选基因的细胞自主和组织范围作用。鉴于相对稀疏的标记,根据选择的 19 个 MADM 染色体之一,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)中形态异常和表型的报告。对于任何候选基因的 MADM 都可以轻松地将形态分析纳入实验工作流程中。在这里,我们描述了我们在最近的各种 MADM 研究中开发的形态细胞分析方法。本章将特别侧重于定量 CNS 中神经元和星形胶质细胞形态的方法,但这些方法可以广泛应用于整个生物体中任何 MADM 标记的细胞。我们将涵盖两种分析 - 体感皮层中锥体神经元的体体积和树突特征化,以及两种分析 - 体积和 Sholl 分析 - 星形胶质细胞形态。

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