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）：我们建议研究羊水（AF）和新生脑脊液（CSF）中的信号如何在神经管闭合期间的早期大脑发育阶段指导神经干细胞行为（E8.5-E10） .5).在这些早期阶段，神经干数量的快速变化和增长将产生成人大脑中的所有神经元和神经胶质细胞，但关于驱动这些早期阶段的内在基因程序或基于外在流体的信号的研究相对较少。发展，很大程度上是由于技术限制。我们和其他人已经证明了脑脊液和血管液体生态位在指导大脑发育后期的广泛影响。然而，在脉络丛和脉管系统尚未形成的早期大脑发育过程中，对于 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 和新生脑脊液中的分泌信号对于指导和同步沐浴在这些液体中的胚胎 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 AF、早期 CSF 和缓冲培养基对 E8.5 和 E10.5 龄干细胞外植体的影响来检验这一假设。我们预测，与后期发育一样，早期外植体只有在沐浴在适合年龄的液体中时才能正常发育，这表明 AF/CSF 蛋白质组的变化反映在神经干细胞顶端表面相关受体表达的变化。因此，我们预计正常干细胞功能应因神经干细胞顶端表面纤毛和膜的遗传扰动而受损，该顶端表面与 AF/CSF 直接接触。然后，将使用定量质谱和 RNAseq 技术以及免疫染色来推断出近乎完整的 CSF 蛋白及其顶膜上相关受体的列表。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的贡献，以及这些 CSF 信号的胚胎来源。这一提议具有重要的临床意义：我们目前对早期发育障碍（神经管错误闭合、脑积水和致畸剂浸润导致的缺陷）的起源了解相对较少，早期诊断或干预的能力也不足。该提案应该为询问这些早期扰动如何最终破坏正常大脑发育的部分或全部方面提供基础，并为开发用于疾病诊断和神经干细胞重新编程的微创 AF/CSF 采样和替代策略提供基础。大脑发育回到正轨。