MACS: A genetic labeling tool to depict the complete neuroblast lineage of all neurons in individual Drosophila brains

MACS：一种基因标记工具，用于描述单个果蝇大脑中所有神经元的完整神经母细胞谱系

• 批准号: 8831944
• 负责人: Dawen Cai
• 金额: $ 19.43万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-30 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8831944
• 关键词: Address; Adult; Alleles; Animal Behavior; Animal Model; Animals; Arrhythmia; B-Lymphocytes; Behavior; Brain; Cardiac conduction system; Cell Count; Cell Lineage; Cells; Code; Data; Defect; Development; Developmental Process; Disease; Drosophila genus; Embryo; Fishes; Functional disorder; Gene Mutation; Genetic; Genetic Recombination; Goals; Image; In Situ; Individual; Inherited; Label; Lead; Life; Lymphocyte; Maps; Microscopy; Mus; Neural Crest; Neurons; Outcome; Positioning Attribute; Process; Rattus; Research; Resolution; Scheme; Severe Combined Immunodeficiency; Staging; Stem cells; System; Techniques; Technology; Time; TimeLine; Transgenic Animals; Variant; Work; base; brain tissue; cell type; density; design; exhaust; fly; neural circuit; neuroblast; neurogenesis; novel; public health relevance; relating to nervous system; sex; stem; stem cell biology; success; tool; two-photon

 DESCRIPTION: During development, stem/progenitor cells replicate and differentiate into many lineages, which give rise to precise number and subtypes of cells. Defects in lineage development can cause severe developmental diseases. Currently, the state-of-the-art lineage analysis uses mosaic labeling techniques to study one or a few lineages at a time to avoid ambiguity. While the small number of highlighted cells can be investigated extensively, complications in the unlabeled adjacent lineages are hidden from analysis. The ability of unambiguously labeling large number of lineages in situ is highly desired, since it is extremely exhausting, if not impossible to use the available tools to study the precise spatial-temporal relationship of all related lineages in one animal. We propose to develop a two-photon compatible multispectral and subcellular-coding system (MACS), which permits unambiguous labeling of large number of cell lineages in the same animal. We will validate MACS by mapping all of the ~100 neural lineages in single Drosophila central brain and depict the developmental processes of all Drosophila embryo neural lineages precisely in space and time. If success, MACS can be easily adapt to other transgenic animal models, including fish, mouse and rat. MACS will create new opportunities in lineage studies, such as investigating lineage variations among individuals, and between hypomorphic alleles or different sex; as well as cell non-autonomous effects of gene mutations in stem/progenitor cells.

 描述：在发育过程中，干细胞/祖细胞复制和分化为许多谱系，从而产生准确的细胞数量和亚型。谱系发育缺陷会导致严重的发育性疾病。目前，最先进的谱系分析使用镶嵌标记技术一次研究一个或几个谱系，以避免模棱两可。虽然少数突出显示的细胞可以进行广泛的研究，但未标记的相邻谱系中的并发症被隐藏在分析之外。在原位明确地标记大量谱系的能力是非常必要的，因为使用可用的工具来研究一种动物中所有相关谱系的精确时空关系是非常费力的，如果不是不可能的话。我们建议开发一种双光子兼容的多光谱和亚细胞编码系统(MACS)，它允许对同一动物中的大量细胞系进行明确的标记。我们将通过在单个果蝇中央脑中定位所有~100个神经谱系来验证MACs，并在空间和时间上精确地描绘所有果蝇胚胎神经谱系的发育过程。如果成功，Mac可以很容易地适应其他转基因动物模型，包括鱼、小鼠和大鼠。MACs将在谱系研究中创造新的机会，例如调查个体之间的谱系差异，以及亚型等位基因或不同性别之间的谱系差异；以及干细胞/祖细胞基因突变的细胞非自治效应。