Mechanisms regulating Meningeal Development and Function

调节脑膜发育和功能的机制

• 批准号: 10763476
• 负责人: Marina Venero Galanternik
• 金额: $ 24.9万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10763476
• 关键词: Adult; Affect; Age; Aging; Anatomy; Animal Feed; Animals; Bathing; Biochemical; Biological Assay; Blood Vessels; Brain; Brain Injuries; Buffers; Cell Culture Techniques; Cells; Cellular Structures; Central Nervous System; Cerebrospinal Fluid; Circulation; Complex; Connective Tissue; Defect; Development; Developmental Biology; Diet; Disease; Electron Microscopy; Embryo; Endothelium; Environment; Excision; Experimental Models; Fatty acid glycerol esters; Fishes; Functional disorder; Future; Gene Expression; Genes; Genetic; Histology; Homeostasis; Image; Impaired cognition; In Vitro; Infection; Injury; Knowledge; Label; Link; Liquid substance; Lymphatic System; Macrophage; Maintenance; Mechanics; Meningeal; Meninges; Metabolic; Modeling; Molecular; Morphology; Mutagenesis; Nerve Degeneration; Neurocognitive; Neurons; Obesity; Pathogenicity; Pathway interactions; Pericytes; Phase; Play; Population; Positioning Attribute; Process; Property; Recovery; Regulation; Research; Resolution; RiboTag; Role; Shock; Signal Transduction; Site; Structure; System; Tissues; Transgenic Animals; Transgenic Organisms; Transmission Electron Microscopy; Vascular Permeabilities; Vascular System; Vascularization; Zebrafish; age effect; cell type; confocal imaging; design; ependymins; extracellular; grasp; high resolution imaging; in vivo; insight; mutant; novel; pathogen; prevent; programs; research faculty; response; single-cell RNA sequencing; stroke incidence; tenure track; tool; transcriptome sequencing; uptake; wasting

ABSTRACT/PROJECT SUMMARY The meninges, a protective layer ensheathing the Central Nervous System (CNS), is a highly vascularized, complex tissue that serves as the primary site for equilibration of cerebrospinal fluid (CSF). CSF, a transparent, colorless fluid that recirculates throughout the CNS, supports brain buoyancy, prevents vascular and neuronal collapse, and provides buffering against mechanical injury. Given the functional importance of CSF for CNS homeostasis, the clearance of metabolic waste from this fluid compartment is carried out as a nearly constant process. This waste removal process relies heavily on groups of vascular- associated, perivascular cells within the meninges that filter the byproducts from the CSF and transport them into circulation via the lymphatic system for ultimate disposal. Hence, the interaction between meningeal cells and the vascular system is crucial to safeguard brain homeostasis. Some forms of neurodegeneration have been linked with a decline in homeostasis and an increase in metabolic waste accumulation in an age-, diet- or pathogenic infection-dependent manner. Understanding vascular- associated meningeal cells make-up, developmental origin, genetic regulation and function is an important long-term undertaking to fully grasp how neurodegeneration occurs in response to these conditions. The amenability of the zebrafish for live imaging represents a remarkable advantage over other models to study meningeal development in vivo. In Aim 1, I will use newly developed transgenic lines labeling pan- meningeal and meningeal perivascular cell populations in vivo to uncover the developmental origin of these cells using a combination of high resolution confocal imaging for lineage tracing studies, Transmission Electron Microscopy to characterize their cellular structures, and single cell and “RiboTag” RNA- sequencing to identify gene programs regulated in meningeal populations. Aim 2 will uncover the functions that meningeal cells play in supporting CNS homeostasis and maintenance. Analysis will be done utilizing readily available mutants that present defects in meningeal development. In addition, as an unbiased approach, I will utilize a forward genetic mutagenesis screen to uncover genes required for proper meningeal development and function. Lastly, complementary in vivo high resolution confocal imaging and in vitro cell culture assays will be utilized to uncover biochemical changes in meningeal cells resulting from age, diet, and infection in both wild type and mutant zebrafish populations (Aim 3). These aims are designed to expand the current knowledge of meningeal cellular components, genetic signals controlling their development, and a better understanding of their interaction with the vasculature. This proposal offers a foundational niche in the vascular developmental biology field through which I can launch a future tenure- track research faculty position.

摘要/项目摘要 脑膜是包裹中枢神经系统 (CNS) 的保护层，是高度血管化的、 作为脑脊液 (CSF) 平衡主要部位的复杂组织。脑脊液， 透明、无色的液体，在整个中枢神经系统中循环，支持大脑浮力，防止 血管和神经元崩溃，并提供针对机械损伤的缓冲。考虑到函数式 由于脑脊液对于中枢神经系统稳态的重要性，代谢废物从该液体室的清除是 作为一个几乎恒定的过程进行。这种废物清除过程在很大程度上依赖于血管群 脑膜内相关的血管周围细胞，过滤脑脊液和运输中的副产物 它们通过淋巴系统进入循环进行最终处理。因此，之间的相互作用 脑膜细胞和血管系统对于维护大脑稳态至关重要。某些形式的 神经退行性疾病与体内平衡下降和代谢废物增加有关 以年龄、饮食或病原感染依赖性方式积累。了解血管- 相关脑膜细胞的组成、发育起源、遗传调控和功能是重要的 长期致力于充分了解神经退行性变是如何响应这些条件而发生的。这 斑马鱼的实时成像能力比其他研究模型具有显着优势 体内脑膜发育。在目标 1 中，我将使用新开发的转基因品系标记泛- 体内脑膜和脑膜血管周围细胞群，以揭示这些细胞的发育起源 使用高分辨率共聚焦成像组合进行细胞谱系追踪研究、传输 电子显微镜可表征其细胞结构、单细胞和“RiboTag”RNA- 测序以确定脑膜群体中调控的基因程序。目标 2 将揭示功能 脑膜细胞在支持中枢神经系统稳态和维持中发挥作用。分析将利用 容易获得的突变体在脑膜发育中存在缺陷。此外，作为一个公正的 方法，我将利用正向遗传诱变筛选来发现适当的基因所需的基因 脑膜的发育和功能。最后，补充体内高分辨率共焦成像和 体外细胞培养测定将用于揭示脑膜细胞的生化变化 野生型和突变斑马鱼种群的年龄、饮食和感染（目标 3）。这些目标是 旨在扩展脑膜细胞成分、遗传信号控制的当前知识 它们的发育，以及更好地了解它们与脉管系统的相互作用。该提案提供 血管发育生物学领域的一个基础利基市场，通过它我可以启动未来的终身教职- 跟踪研究教员职位。