Shen Hung-Chang, Hsu Tsai-Chi, Chung Pei-Chi, Yu Hung-Hsiang
Institute of Cellular and Organismic Biology, Academia Sinica; Graduate Institute of Life Sciences, National Defense Medical Center.
Institute of Cellular and Organismic Biology, Academia Sinica.
J Vis Exp. 2017 Mar 2(121):55278. doi: 10.3791/55278.
Mosaic analysis with a repressible cell marker (MARCM) is a positive mosaic labeling system that has been widely applied in Drosophila neurobiological studies to depict intricate morphologies and to manipulate the function of genes in subsets of neurons within otherwise unmarked and unperturbed organisms. Genetic mosaics generated in the MARCM system are mediated through site-specific recombination between homologous chromosomes within dividing precursor cells to produce both marked (MARCM clones) and unmarked daughter cells during mitosis. An extension of the MARCM method, called twin-spot MARCM (tsMARCM), labels both of the twin cells derived from a common progenitor with two distinct colors. This technique was developed to enable the retrieval of useful information from both hemi-lineages. By comprehensively analyzing different pairs of tsMARCM clones, the tsMARCM system permits high-resolution neural lineage mapping to reveal the exact birth-order of the labeled neurons produced from common progenitor cells. Furthermore, the tsMARCM system also extends gene function studies by permitting the phenotypic analysis of identical neurons of different animals. Here, we describe how to apply the tsMARCM system to facilitate studies of neural development in Drosophila.
利用可抑制细胞标记的镶嵌分析（MARCM）是一种正向镶嵌标记系统，已广泛应用于果蝇神经生物学研究中，用于描绘复杂的形态，并在其他未标记和未受干扰的生物体中操纵神经元亚群中基因的功能。MARCM系统中产生的遗传镶嵌体是通过分裂前体细胞内同源染色体之间的位点特异性重组介导的，从而在有丝分裂过程中产生标记的（MARCM克隆）和未标记的子细胞。MARCM方法的一个扩展，称为双斑MARCM（tsMARCM），用两种不同颜色标记来自共同祖细胞的两个孪生细胞。开发这项技术是为了能够从两个半谱系中获取有用信息。通过全面分析不同的tsMARCM克隆对，tsMARCM系统允许进行高分辨率的神经谱系图谱绘制，以揭示由共同祖细胞产生的标记神经元的确切出生顺序。此外，tsMARCM系统还通过允许对不同动物的相同神经元进行表型分析来扩展基因功能研究。在这里，我们描述了如何应用tsMARCM系统来促进果蝇神经发育的研究。
