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: 9592762
• 负责人: Guo-li Ming
• 金额: $ 20.13万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-12-12 至 2019-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9592762
• 关键词: Deconstructing; hypothalmic; ontogeny; plasticity; via

 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（应用程序提供）：哺乳动物的大脑功能严重依赖于胚胎发育过程中复杂的细胞结构组织。细胞的产生，迁移，突触形成和电路整合通常遵循高度定型的模式，以形成大脑中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产生了层状结构，例如大脑皮层，小脑和视网膜，并具有前所未有的单细胞分辨率。但是，神经元干细胞（NSC）核结构的形成尚不清楚，尚待系统地研究。哺乳动物下丘脑是一种异质性核结构，对于内分泌，自主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NSC产生的下丘脑神经元，并在早期发育过程中组织成离散的核，对于了解不同下丘脑核的结构 - 功能关系以及可以通过环境条件来调节的结构 - 功能连接的程度。我们已经开发了一种基于遗传的单细胞谱系追踪技术，该谱系技术采用MADM（对双标记）动物的动物标记NSC在发育中的胚胎中标记，并开始解决下丘脑组织的这些杰出问题。拟议的研究的目的是重建和量化体内标记的NSC的行为，解释组织下丘脑核的一般原则，解释了个体下丘脑核的个体，并探索在母体挑战的背景下，探索了核组织的个体遗传学可变性。到目前为止，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的复杂性，无法深入了解这种核结构的组织和个体发育。成功完成我们的研究将导致单个NSC的全面图及其在区域，区域和核水平上的进展，在三维环境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，有助于有效地有效。我们还将验证一个实验平台，以在病理状况下对下丘脑失调的未来机械研究，这可能导致有针对性的诊断和治疗策略来保留关键的生理功能。