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 这些脑膜是一种揭示中枢神经系统（CNS）的保护层，是高度血管化的， 复杂的组织是脑脊液（CSF）平衡的主要部位。 CSF，a 透明，无色的流体在整个中枢神经系统中循环，支持脑浮力，可防止 血管和神经元塌陷，并提供防止机械损伤的缓冲。给定功能 CSF对于中枢神经系统稳态的重要性，该液体室的代谢废物清除率是 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取决于血管 相关的脑膜周细胞中的脑血管周细胞过滤了CSF的副产品并运输 他们通过淋巴系统进入循环以进行最终处置。因此， 脑膜细胞和血管系统对于保护大脑稳态至关重要。某些形式 神经变性与稳态下降和代谢废物的增加有关 以年龄，饮食或致病感染依赖性方式积累。了解血管 - 相关的脑膜细胞构成，发育起源，遗传调节和功能是重要的 长期的承诺完全抓住神经变性是如何响应这些条件的。 斑马鱼对实时成像的适用性代表了与其他模型的显着优势 体内脑膜发展。在AIM 1中，我将使用新开发的转基因线标记板 - 体内脑膜和脑膜周围细胞群体揭示了这些的发育起源 使用高分辨率共聚焦成像进行谱系跟踪研究的组合，传播 电子显微镜以表征其细胞结构，以及单细胞和“ Ribotag” RNA- 测序以识别在脑膜种群中调节的基因程序。 AIM 2将揭示功能 该脑膜细胞在支持CNS稳态和维护方面发挥作用。分析将使用 易于使用的突变体，这些突变体存在脑膜发育中的缺陷。另外，作为公正 方法，我将利用正向遗传诱变筛选来发现适当的基因 脑膜开发和功能。最后，体内高分辨率共焦成像和 体外细胞培养分析将用于发现由 野生型和突变斑马鱼种群中的年龄，饮食和感染（AIM 3）。这些目标是 旨在扩大当前脑膜细胞成分的知识，控制的遗传信号 他们的发展，以及对他们与脉管系统的互动的更好理解。该建议提供 血管发育生物学领域的基本利基市场，我可以通过该领域发起未来的任期 - 跟踪研究教师职位。