Deconstructing the hypothalmic ontogeny and plasticity via clonal analysis

通过克隆分析解构下丘脑个体发育和可塑性

• 批准号: 9126774
• 负责人: Guo-li Ming
• 金额: $ 24.3万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-03-15 至 2018-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9126774
• 关键词: Address; Adult; Animals; Anterior; Behavior; Behavioral; Brain; Brain region; Cell Line; Cell Lineage; Cell Nucleus; Cells; Cerebellum; Cerebral cortex; Chronic; Circadian Rhythms; Complex; Development; Diagnostic; Embryo; Embryonic Development; Endocrine; Environment; Equilibrium; Exposure to; Feeding behaviors; Future; Generations; Genetic; Genetic Models; Goals; Growth; Heterogeneity; Hippocampus (Brain); Homeostasis; Hormonal; Hyperglycemia; Hyperlipidemia; Hypothalamic Diseases; Hypothalamic structure; Individual; Instinct; Intervention; Investigation; Label; Laboratories; Lead; Location; Maintenance; Maps; Modeling; Molecular; Molecular Profiling; Neocortex; Neuraxis; Neuroglia; Neurons; Nuclear; Nuclear Structure; Pattern; Physiological; Physiological Processes; Production; Property; Radial; Research; Resolution; Retina; Siblings; Sorting - Cell Movement; Stem cells; Stereotyping; Stratification; Structure; Structure-Activity Relationship; Techniques; Technology; Telencephalon; Therapeutic; Time; base; cell behavior; diagnostic biomarker; genetic approach; in utero; in vivo; lateral ventricle; learned behavior; migration; mood regulation; nerve stem cell; neurogenesis; newborn neuron; prevent; progenitor; public health relevance; synaptogenesis; targeted treatment; temporal measurement; tool

 DESCRIPTION (provided by applicant): Mammalian brain function critically relies on sophisticated cytoarchitectonic organization during embryonic development. Cell generation, migration, synapse formation and circuit integration often follow highly stereotyped patterns to form complex laminated or nuclear structures in the brain. Recently developed clonal lineage analysis has revealed stem cell behavior giving rise to laminar structures, such as the cerebral cortex, cerebellum and retina, with unprecedented single-cell resolution. However, the formation of nuclear structures by neural stem cells (NSCs) remains unclear and has yet to be systematically investigated. The mammalian hypothalamus is a heterogeneous nuclear structure that is critical for the integration and homeostatic maintenance of endocrine, autonomic and behavioral functions. Reconstructing how hypothalamic neurons are generated from individual NSCs and organized into discrete nuclei during early development is essential to understand the structure-function relationship of different hypothalamic nuclei and the extent to which this can be modulated by environmental conditions. We have developed a genetically-based single-cell lineage tracing-technique that employs MADM (mosaic analysis of double marker) animals to label NSCs in the developing embryo and begin to address these outstanding questions of hypothalamic organization. The goal of the proposed research is to reconstruct and quantify the behavior of individually-labeled NSCs in vivo, decipher the general principles organizing hypothalamic nuclei, decode the ontogeny of individual hypothalamic nuclei and explore the ontogenetic plasticity of nuclear organization in the context of a maternal challenge. The complexity of the anatomical and molecular subdivisions of the hypothalamus, and lack of appropriate genetic tools, has thus far prevented a deep understanding of the organization and ontogeny of this nuclear structure. Successful completion of our study will result in a comprehensive map of single NSCs and their progeny at regional, zonal and nuclear levels, the clonal organization of sibling neurons in a three-dimensional context to determine migratory patterns of newly born neurons, and the capacity of single stem cells to contribute to functionally distinct nuclei. We will also have validated an experimental platform for future mechanistic investigations of hypothalamic dysregulation under pathological conditions, which can lead to targeted diagnostic and therapeutic strategies to preserve critical physiological functions.

 描述（由申请人提供）：哺乳动物脑功能严重依赖于胚胎发育期间复杂的细胞结构组织。细胞生成、迁移、突触形成和电路整合通常遵循高度刻板的模式，在大脑中形成复杂的层压或核结构。最近开发的克隆谱系分析揭示了干细胞行为引起层状结构，如大脑皮层，小脑和视网膜，具有前所未有的单细胞分辨率。然而，神经干细胞（NSCs）的核结构的形成仍然不清楚，尚未进行系统的研究。哺乳动物下丘脑是一种异质核结构，对于内分泌、自主神经和行为功能的整合和稳态维持至关重要。重建下丘脑神经元是如何从单个神经干细胞产生的，并在早期发育过程中组织成离散的核团，对于了解不同下丘脑核团的结构-功能关系以及环境条件对其的调节程度至关重要。 我们已经开发了一种基于遗传学的单细胞谱系追踪技术，该技术采用MADM（双标记镶嵌分析）动物来标记发育中胚胎中的NSC，并开始解决下丘脑组织的这些悬而未决的问题。拟议研究的目标是重建和量化个体标记的神经干细胞在体内的行为，破译组织下丘脑核的一般原则，解码个体下丘脑核的个体发育，并探索在母体挑战的背景下核组织的个体发育可塑性。 下丘脑的解剖学和分子细分的复杂性，以及缺乏适当的遗传工具，迄今为止，阻止了对该核结构的组织和个体发育的深入了解。我们的研究的成功完成将导致在区域，区域和核水平上的单个神经干细胞及其后代的综合地图，在三维背景下的兄弟神经元的克隆组织，以确定新出生的神经元的迁移模式，以及单个干细胞的能力，以促进功能不同的细胞核。我们还将为病理条件下丘脑失调的未来机制研究验证一个实验平台，这可以导致有针对性的诊断和治疗策略，以保护关键的生理功能。