The roles of pericyte-derived laminin in neurovascular function and neurodegeneration

项目摘要/摘要 这项应用的长期目标是了解基底膜(BM)、 神经血管单位的非细胞成分，参与阿尔茨海默病的发病机制 该药物可用于治疗阿尔茨海默病和阿尔茨海默病相关痴呆。 这与NIA的使命是一致的。这项建议旨在研究细胞的生物学功能。 周细胞层粘连蛋白：(1)神经血管功能，包括血脑屏障(BBB)的完整性，脑 血流量(CBF)和脑流入/流出功能；(2)神经元存活/功能。在目标1中， 我们将研究周细胞层粘连蛋白在血脑屏障完整性中的作用。第一，损失是否以及损失的程度 将使用不同分子的FITC-葡聚糖来研究周细胞层粘连蛋白对血脑屏障完整性的影响 权重(目标1A)。其次，周细胞层粘连蛋白诱导血脑屏障缺失的分子机制 将研究分解，重点是细胞旁和跨细胞转运的变化。 内皮细胞(目标1B)。此外，介导周细胞层粘连蛋白作用的受体在 将鉴定和检查内皮细胞(目标1C)。在目标2中，周细胞层粘连蛋白的功能 将对CBF进行调查。首先，将使用以下方法研究周细胞层粘连蛋白丢失如何影响脑血流量 定量放射自显影和双光子成像(目标2A)。接下来，降低的CBF是否 将在体外和体内研究周细胞丢失/退化所造成的影响(目标2B)。此外， 将鉴定和检测介导周细胞层粘连蛋白在周细胞层粘连蛋白作用的受体(目标2C)。在……里面 目的研究周细胞层粘连蛋白在脑内流/流出功能中的作用。第一，是否和 周细胞层粘连蛋白的丢失如何影响脑流入/流出功能将通过流入/流出来研究 使用各种荧光标记的大分子进行分析(目标3A)。接下来，无论是弱者 BM损伤及周细胞层粘连蛋白丢失对BM的影响 将探索组成/结构(目标3B)。在目标4中，周细胞层粘连蛋白在神经元中的作用 将对损伤/神经变性进行调查。在这个目标中，周细胞层粘连蛋白的丢失是否会导致 神经元损伤/神经变性将从生化、结构和功能水平进行研究。 此外，还将确定发生神经元损伤/神经变性的年龄和 与发生神经血管功能障碍的情况相比。成功完成这项研究将 阐明周细胞层粘连蛋白在神经血管功能和神经元中的基础作用 生存/功能，并确定阿尔茨海默病治疗潜力的新分子靶点 和阿尔茨海默病相关的痴呆症。此外，这一提议还可能导致 一种用于神经退化的创新小鼠模型，并为新的研究打开了大门。

Project Summary/Abstract The long-term objective of this application is to understand whether the basement membrane (BM), the non-cellular component of the neurovascular unit, is involved in the pathogenesis of Alzheimer’s disease and can be targeted to treat Alzheimer’s disease and Alzheimer’s disease-related dementias. This is consistent with the mission of NIA. This proposal aims to investigate the biological functions of pericytic laminin in: (1) neurovascular function, including blood brain barrier (BBB) integrity, cerebral blood flow (CBF) and brain influx/efflux function, and (2) neuronal survival/function. In Aim 1, the function of pericytic laminin in BBB integrity will be investigated. First, whether and to what extent loss of pericytic laminin affects BBB integrity will be investigated using FITC-Dextrans of various molecular weights (Aim 1A). Next, the molecular mechanism underlying loss of pericytic laminin-induced BBB breakdown will be investigated, with a focus on changes in paracellular and transcellular transport in endothelial cells (Aim 1B). Furthermore, the receptors that mediate pericytic laminin’s effect in endothelial cells will be identified and examined (Aim 1C). In Aim 2, the function of pericytic laminin in CBF will be investigated. First, how loss of pericytic laminin affects CBF will be investigated using quantitative autoradiography and two-photon imaging (Aim 2A). Next, whether the reduced CBF is caused by pericyte loss/degeneration will be investigated in vitro and in vivo (Aim 2B). Furthermore, the receptors that mediate pericytic laminin’s effect in pericytes will be identified and examined (Aim 2C). In Aim 3, the role of pericytic laminin in brain influx/efflux function will be investigated. First, whether and how loss of pericytic laminin affects brain influx/efflux function will be investigated by influx/efflux assays using various fluorescently labeled macromolecules (Aim 3A). Next, whether the impaired influx/efflux function is due to BM damage and how loss of pericytic laminin affects BM composition/structure will be explored (Aim 3B). In Aim 4, the function of pericytic laminin in neuronal injury/neurodegeneration will be investigated. In this aim, whether loss of pericytic laminin leads to neuronal injury/neurodegeneration will be investigated at biochemical, structural, and functional levels. In addition, the age at which neuronal injury/neurodegeneration occurs will be determined and compared to that at which neurovascular dysfunction occurs. Successful completion of this study will elucidate the fundamental roles of pericytic laminin in neurovascular function and neuronal survival/function, and identify novel molecular targets with therapeutic potential in Alzheimer’s disease and Alzheimer’s disease-related dementias. In addition, this proposal may also lead to the generation of an innovative mouse model for neurodegeneration and open doors for new research.