Cellular mechanisms of perivascular fibroblast development and dependence on PDGF signaling

项目总结 血管周围间隙(PVS)是围绕中枢神经系统血管的充满液体的间隙， 被认为在维持大脑健康方面起着重要作用，是各种 血管周围细胞类型。一种特别令人感兴趣的细胞类型是血管周围成纤维细胞(PVF)，这种细胞很差 它们的特性，它们的功能在很大程度上是未知的。已知PVF位于血管的外部 成年小鼠中枢神经系统中中到大直径血管的平滑肌层，并 主要通过胶原-1和血小板衍生生长因子受体(PDGFR)a和 PDGFRb.然而，关于发育起源或细胞和分子方面的知识几乎一无所知。 对PVF发展很重要的机制。初步数据表明，PVF来自于一群细胞 在出生后发育过程中，脑膜和逐渐填充的脑室管瘤，但这还没有 进一步调查。这些知识上的差距是开发新工具来研究这一角色的主要障碍 PVF在大脑动态平衡和疾病中的作用。这项建议的目的是调查发展的 PVF发生的起源、细胞动力学和分子信号机制。在《目标1》中，我将 使用标记PVF的转基因小鼠系的组合、光学组织清除、体外成像和 多色谱系追踪记者，调查发育中的PVF的细胞起源和动力学。在AIM 2、我会用脑池内脑脊液示踪分析来确定时间的关系 PVF开发和PVS功能。最后，在目标3中，我将研究PDGF信号作为一种潜在的机制 控制PVF的发展。总之，这个项目将是第一个全面描述PVF开发方式的项目 是否发生以及是否涉及PDGF信令。

PROJECT SUMMARY Perivascular spaces (PVSs) are fluid-filled spaces surrounding vessels of the central nervous system, are thought to have important roles in maintenance of brain health, and are a unique niche for a variety of perivascular cell types. A cell type of particular interest is perivascular fibroblasts (PVFs) which are poorly characterized, and their functions are largely unknown. PVFs are known to reside on the outside the vascular smooth muscle layer of medium-to large-diameter vessels in the adult mouse central nervous system, and are primarily identified by expression of Collagen-1 and Platelet-derived growth factor receptor (PDGFR)a and PDGFRb. However, nearly nothing is known about the developmental origins, or the cellular and molecular mechanisms important for PVF development. Preliminary data suggests PVFs emerge from a population of cells in the meninges and gradually populate brain PVSs during postnatal development, but this has not yet been investigated further. These gap in knowledge is a major barrier to developing novel tools to investigate the role of PVFs in brain homeostasis and disease. The objective of this proposal is to investigate the developmental origins, cellular dynamics, and molecular signaling mechanisms involved in PVF development. In Aim 1, I will use a combination of transgenic mouse lines that label PVFs, optical tissue clearing, ex vivo imaging, and a multi-color lineage tracing reporter to investigate the cellular origins and dynamics of developing PVFs. In Aim 2, I will use intracisternal cerebrospinal fluid tracer assays to determine the relationship between the timing of PVF development and PVS function. Finally, in Aim 3 I will investigate PDGF signaling as a potential mechanism controlling PVF development. Together, this project will be the first to fully characterize how PVF development occurs and whether or not PDGF signaling is involved.