Control of neural stem cells by the nascent cerebrospinal fluid

新生脑脊液对神经干细胞的控制

• 批准号: 8765073
• 负责人: MARIA LEHTINEN
• 金额: $ 38.41万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-05-15 至 2019-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8765073
• 关键词: Adult; Age; Amniotic Fluid; Apical; Automobile Driving; Back; Bathing; Biochemical Genetics; Blood Vessels; Brain; Buffers; Cell Maintenance; Cell physiology; Cells; Cerebrospinal Fluid; Cerebrospinal Fluid Proteins; Choroid; Cilia; Clinical; Complex; Congenital Hydrocephalus; Data; Defect; Development; Diagnosis; Disease; ES Cell Line; Early Diagnosis; Early Intervention; Embryo; Embryonic Development; Foundations; Future; Genetic; Genetic Programming; Health; Hydrocephalus; In Vitro; Individual; Infiltration; Intraventricular; Liquid substance; Maintenance; Mediating; Membrane; Methods; Mus; Nervous system structure; Neural Tube Closure; Neuroglia; Neurons; Pathway interactions; Population; Prosencephalon; Proteins; Proteome; Proteomics; Receptor Signaling; Regulation; Research; Role; Sampling; Signal Pathway; Signal Transduction; Source; Staging; Stem cells; Structure of choroid plexus; Surface; Technology; Teratogens; Testing; Time; Work; age related; apical membrane; base; cell behavior; clinically significant; cytokine; developmental disease; disease diagnosis; genetic manipulation; immunocytochemistry; in vivo; leukemia inhibitory factor; leukemia inhibitory factor receptor; minimally invasive; nerve stem cell; programs; protein expression; public health relevance; receptor; relating to nervous system; repaired; self-renewal; stem; stem cell fate

DESCRIPTION (provided by applicant): We propose to investigate how signals in the amniotic fluid (AF) and nascent cerebrospinal fluid (CSF) instruct neural stem cell behavior during early brain developmental stages surrounding the time of neural tube closure (E8.5-E10.5). The rapidly changing and growing population of neural stems at these early stages will give rise to all neurons and glia in the adult brain, yet comparatively few studies exist on the intrinsic genetc programs or the extrinsic fluid-based signals involved in driving these early stages of development, largely due to technical limitations. We and others have demonstrated broad influences of CSF and vascular fluid niches in instructing later stages of brain development. Yet virtually nothing is known about the protein composition of AF and early CSF during early brain development, at a time when the choroid plexus and vasculature have yet to form. Our overarching hypothesis is that secreted signals in the AF and nascent CSF are critical in instructing and synchronizing the proliferation and fate of embryonic E8.5-E10.5 neural stem cells bathed in these fluids. We will first test this hypothesis by comparing the effects of AF, early CSF and buffered media on stem cell explants at ages E8.5 and E10.5. We predict that, as in later development, early explants only develop normally when bathed in the age-appropriate fluid, suggesting that changes in the AF/CSF proteome are mirrored by changes in expression of associated receptors at the apical surface of neural stem cells. As such, we expect that normal stem cell function should be impaired by genetic perturbations to the cilia and membrane of neural stems cells at their apical surface, which is in direct contact with the AF/CSF. A near- complete list of CSF proteins and their associated receptors on apical membranes will then be deduced using quantitative mass-spec and RNAseq technologies, together with immunostaining. We will then determine the contributions of CSF-LIF and other top candidate proteins in instructing specific aspects of E10.5 stem cell behavior, and the embryonic sources of these CSF signals. This proposal has important clinical significance: We currently have relatively little understanding of the origins of early developmental disorders (defects due to errant neural tube closure, hydrocephalus, and infiltration by teratogens), and scant capacities for early diagnosis or intervention. This proposal should provide a foundation for asking how these early perturbations ultimately derail some or all aspects of normal brain development, and for developing minimally invasive AF/CSF sampling and replacement strategies for disease diagnosis and for reprogramming of neural stem cells in order to bring the development brain back on track.

描述（由申请人提供）：我们建议调查羊水（AF）和新生脑脊液（CSF）中的信号如何指示神经管闭合时围绕脑部发育早期发育阶段的神经干细胞行为（E8.5-E10.5）。在这些早期阶段，神经茎的快速变化和增长将引起成人大脑中的所有神经元和神经胶质，但在很大程度上涉及推动这些早期发展阶段的基于内在流体的信号上，对成人大脑的所有神经元和神经胶质的存在相对较少，这很大程度上是由于技术限制。我们和其他人在指导大脑发育的后期阶段表现出了CSF和血管液生态位的广泛影响。然而，在早期大脑发育过程中，AF和早期CSF的蛋白质组成几乎一无所知。我们的总体假设是，AF和新生CSF中的分泌信号对于指导和同步胚胎中的胚胎E8.5-E10.5神经干细胞的增殖和命运至关重要。我们将首先通过比较E8.5和E10.5的AF，早期CSF和缓冲培养基对干细胞外植体的影响来检验这一假设。我们预测，与以后的发育一样，早期外植体仅在沐浴在适合年龄的液体中时正常发展，这表明AF/CSF蛋白质组的变化是通过神经干细胞顶端表面表达的表达的变化反映的。因此，我们期望正常的干细胞功能受到对纤毛和神经干膜的遗传扰动在其顶部表面的纤毛和膜，该纤毛和膜与AF/CSF直接接触。然后，将使用定量质谱和RNASEQ技术来推导CSF蛋白及其相关受体的接近完整列表，并进行免疫染色。然后，我们将确定CSF-LIF和其他顶级候选蛋白在指导E10.5干细胞行为的特定方面以及这些CSF信号的胚胎来源方面的贡献。该提议具有重要的临床意义：目前，我们对早期发育障碍的起源的了解相对较少（由于神经管闭合，脑积水和Teratogens的渗透导致的缺陷），以及早期诊断或干预的能力很少。该提案应为询问这些早期扰动最终如何最终使正常脑发育的某些或所有方面脱轨，并为疾病诊断和重新编程神经干细胞的最小侵入性AF/CSF抽样和替代策略而列出一些或所有方面，以使发育脑的发育脑恢复正轨。