Stem Cell Generation of Transport Epithelia for the Brain's Ventricular System

脑室系统运输上皮的干细胞生成

• 批准号: 9167181
• 负责人: JOANNE C CONOVER
• 金额: $ 23.24万
• 依托单位: UNIVERSITY OF CONNECTICUT STORRS
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-01 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9167181
• 关键词: Address; Affect; Area; Brain; Brain Diseases; Bromodeoxyuridine; Cell Count; Cell Cycle; Cell Lineage; Cell physiology; Cell-Cell Adhesion; Cells; Cerebral Ventricles; Cerebrospinal Fluid; Cerebrospinal fluid shunts procedure; Characteristics; Child; Communicating Hydrocephalus; Complex; Congenital Abnormality; Data; Development; Differentiation and Growth; Disease; Electron Microscopy; Electroporation; Embryonic Development; Ependyma; Ependymal Cell; Epithelium; Fetal Development; Generations; Genetic Models; Gliosis; Goals; Human; Hydrocephalus; Intercellular Fluid; Intercellular Junctions; Intervention; Label; Lesion; Liquid substance; Maps; Measures; Mediating; Modeling; Monitor; Mus; Neuroepithelial; Neuroglia; Neurologic; Neurons; Nutrient; Outcome; Pathology; Pregnancy; Radial; Role; Scanning; Stem cells; Surface; System; Testing; Time; Tissue Model; Tissues; Toxin; Transmission Electron Microscopy; Ventricular; Work; base; brain parenchyma; brain tissue; brain volume; causal model; critical period; design; exhaust; fetal; human tissue; immunocytochemistry; in utero; insight; monolayer; mouse model; neurogenesis; neuropathology; population based; postnatal; postnatal human; pressure; regenerative; relating to nervous system; repaired; research study; response; spatiotemporal; stem; stem cell fate; stem cell niche; stem cell population; tool; ventricular system

The goals of this proposal are to examine how the fetal neuroepithelial stem cells support both neurogenesis and ependymogenesis, and to determine how ventricular enlargement, or hydrocephalus, affects ependymogenesis and associated stem cell functions. Ependymal cells are arranged as a monolayer along the ventricle walls and act as the functional units of transport between the cerebrospinal fluid (CSF) and interstitial fluid (ISF). A healthy ependyma circulates CSF and facilitates the clearance of toxins from the ISF of the brain parenchyma. During mid-gestation, sufficient numbers of ependymal cells are generated from neuroepithelial stem cells to cover the entire ventricle surface, but in hydrocephalus, the periventricular stem cell niche is called upon to generate many more ependymal cells to cover the expanding ventricles. We will test the hypothesis that ventricle expansion exhausts the stem cell pool and results in insufficient coverage of the ventricle surface, periventricular gliosis, and loss of critical transependymal transport/clearance functions. Experiments are designed to document the dynamic supply and demand relationship between stem cells and their progeny ependymal cells, in both human tissue and mouse models of hydrocephalus. In Aim 1, we will create spatiotemporal maps of ependymal cell generation and determine the relationship between the distribution of ependymal cells and stem cell numbers/organization in normal fetal and postnatal human periventricular brain tissue. Similar studies will be performed in mouse, supplemented with time-lapse stem cell lineage tracing using the piggyBac in utero electroporation system, to track stem cell fates and test whether ependymogenesis results in stem cell exit from the cell cycle. The resulting 3D spatiotemporal maps of ependymogenesis will serve as tools for evaluating abnormal ependyma development and stem cell niche changes in hydrocephalus. In Aim 2, we will examine several models of hydrocephalus to determine how the stem cell niche and ependymal cell coverage of the ventricle surface are affected. Cell organization, cell-cell junction complexes, alterations in transependymal flow and clearance mechanisms at the ventricle surface and stem cell dynamics will be elucidated. Data generated will provide insight into the regenerative potential of neuroepithelial stem cells for repair of the ventricle surface and how deficits in stem cell functions may contribute to the multiple neurologic pathologies associated with hydrocephalus.

该提案的目标是检查胎儿神经上皮干细胞如何支持神经发生 和室管膜发生，并确定心室扩大或脑积水如何影响 室管膜发生和相关的干细胞功能。室管膜细胞沿管膜呈单层排列 脑室壁，作为脑脊液 (CSF) 和间质之间运输的功能单位 液体（ISF）。健康的室管膜循环脑脊液并促进大脑脑脊液中毒素的清除 薄壁组织。在妊娠中期，神经上皮细胞产生足够数量的室管膜细胞 干细胞覆盖整个脑室表面，但在脑积水中，脑室周围干细胞生态位是 需要产生更多的室管膜细胞来覆盖扩张的心室。我们将测试 假设心室扩张耗尽干细胞池并导致干细胞覆盖不足 心室表面、脑室周围神经胶质增生和关键的经室管膜转运/清除功能丧失。 实验旨在记录干细胞和干细胞之间的动态供需关系。 在脑积水的人体组织和小鼠模型中，他们的后代室管膜细胞。在目标 1 中，我们将 创建室管膜细胞生成的时空图并确定之间的关系 正常胎儿和出生后人类室管膜细胞和干细胞数量/组织的分布 脑室周围脑组织。类似的研究将在小鼠中进行，并辅以延时茎干 使用piggyBac子宫内电穿孔系统进行细胞谱系追踪，以追踪干细胞命运并进行测试 室管膜发生是否导致干细胞退出细胞周期。由此产生的 3D 时空图 室管膜发生将作为评估异常室管膜发育和干细胞生态位的工具 脑积水的变化。在目标 2 中，我们将检查几种脑积水模型，以确定如何 心室表面的干细胞生态位和室管膜细胞覆盖受到影响。细胞组织、细胞间 连接复合体、心室表面经室管膜血流和清除机制的改变以及 干细胞动力学将得到阐明。生成的数据将提供对再生潜力的深入了解 神经上皮干细胞用于修复心室表面以及干细胞功能缺陷如何可能 导致与脑积水相关的多种神经病理学。