Oligodendrocyte Maturation/Myelination in NG2 Null Mice

NG2 无效小鼠的少突胶质细胞成熟/髓鞘形成

• 批准号: 8056780
• 负责人: William B. Stallcup
• 金额: $ 39.84万
• 依托单位: SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-01 至 2012-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8056780
• 关键词: Action Potentials; Axon; Binding; Brain; Budgets; CSPG4 gene; Cell Culture Techniques; Cell Surface Proteins; Cell surface; Cells; Coculture Techniques; Communication; Consultations; Defect; Dimensions; Electrophysiology (science); EphA4 Receptor; Ephrin-A3; Genetic; Goals; Grant; Hand; Heparan Sulfate Proteoglycan; Hippocampus (Brain); Individual; Institutes; Knockout Mice; Knowledge; Left; Letters; Ligands; Membrane; Modeling; Modification; Molecular; Mus; Myelin; Myelin Sheath; Nerve; Neuroglia; Neurons; Oligodendroglia; Postdoctoral Fellow; Preparation; Property; Proteins; Proteoglycan; Ranvier' s Nodes; Research; Research Design; Research Infrastructure; Role; Schwann Cells; Signal Transduction; Signal Transduction Pathway; Slice; Synapses; System; Text; Time; Transgenic Organisms; Work; editorial; in vivo; insight; neuronal excitability; programs; relating to nervous system; response; synaptic function; tissue culture; tool

INTRODUCTION TO REVISED APPLICATION The reviewers of the A1 application considered the Core essential for the successful accomplishment of all the projects in the Program. Discrepancies between the budget and the grant text have been corrected in the current revision. The letter from Ms. Karin Eastham, Vice President and Chief Operating Officer of the Burnham Institute, is enclosed at the end of section II and also in the Appendix. This letter states that the Burnham Institute will provide, as part of its commitment to this Program, two rigs for electrophysiological recording from slices and cell cultures. It is also now clearly stated that both Research Associates, Dr. Hadieh Badie-Mahdavi and Dr. Barbara Fredette, will contribute their expertise to the work in Core A starting in year 1. Their effort has been reduced to 50%, as recommended by the reviewers. This also reflects decreased needs due to elimination of the previous Project 1. Dr. Ranscht's effort, on the other hand, has been increased to 15% due to her ability to dedicate increased effort to the preparation of mixed neuron-glia cultures and also due to the need for her consultation to evaluate the effects of heparan sulfate proteoglycans and NG2 at the nodes of Ranvier, as recommended by the reviewers. The revised portions of the application are indicated with a line on the left margin. The overall editorial modifications introduced to unify the style of the application are not marked. Objectives The goal of this Program is to analyze the molecular signals exchanged between neurons and glia at synapses and in myelinated axons. The program has identified cell surface components implicated in neuron-glia communication and now intends to study the function of these molecules in vivo and in tissue culture models that closely mimic the in vivo interactions. Core A of this Program Project will provide the infrastructure and the expertise to accomplish this goal. The Core will offer support for two aspects of the work, analysis of neuronal function by electrophysiology and modeling neuron-glia interactions in suitable culture systems. The Electrophysiology component adds a new research dimension that will provide the Program with the tools and know-how for functional and activity-dependent studies designed to acquire knowledge of the electrophysiological changes occurring in neurons in response to glial cells or glial-derived ligands. It is now apparent that neuron-glia interactions contribute to the functional properties not only of myelin but also of synapses. Addition of the electrophysiology component to the Program will overcome previous limitations and enable the Program to functionally analyze functional defects resulting from the genetic disruption of neural cell surface proteins and their associated signal transduction pathways. The electrophysiological approach is geared towards the analysis of functional neuron-glia interactions at the level of nerve impulse conduction along axons, as well as synaptic function and plasticity. Electrophysiological recordings will assess the modifications in neuronal membrane properties resulting from changes in the expression or function of glial proteoglycans. The Program will employ transgenic and knockout mice that are already in hand and have a sufficiently long survival time for conducting the electrophysiological analyses. Electrophysiological recordings will be conducted on brain slices, single cells, and isolated nerves. Slice recordings will focus on determining the role of the glial protein ephrin-A3 in synaptic efficacy changes during LTP and LTD. Single-cell recording will assess the contribution of ephrin- A3 to neuronal excitability and plasticity through its neuronal receptor, EphA4. Whole nerve recording will detect conduction velocity changes in the compound action potential caused by malfunction of the myelin sheath. Recordings at these different levels are necessary for gaining insights into the molecular interactions that underlie neuron-glia crosstalk as outlined in the individual projects. The second critical aspect of the proposed work is to probe neuron-glia interactions at the cellular level using material from genetically manipulated mice. This is effectively accomplished using suitable systems of primary neural cultures that mimic specific in vivo interactions. Specifically, Core A will provide the co-cultures of neurons and myelin-forming Schwann cells or oligodendrocytes for studies of myelinogenesis. The use of hippocampal cultures for studies on the influence of glial ephrin-A3 in regulating synaptic function and plasticity will be a continuation of the current Core.

修订申请简介 A1 申请的审核者认为核心对于成功完成所有任务至关重要 该计划中的项目。预算和赠款文本之间的差异已在 当前修订版。副总裁兼首席运营官 Karin Eastham 女士的来信 伯纳姆研究所，附在第二节末尾和附录中。这封信指出 作为该计划承诺的一部分，伯纳姆研究所将提供两台电生理学设备 从切片和细胞培养物中记录。现在还明确指出，两位研究员， Hadieh Badie-Mahdavi 博士和 Barbara Fredette 博士将为核心 A 的工作贡献他们的专业知识 从第一年开始。按照审稿人的建议，他们的工作量已减少到 50%。这也 反映了由于取消之前的项目 1 而导致的需求减少。另一方面，Ranscht 博士的努力 由于她有能力投入更多精力来准备混合材料，因此已增加到 15% 神经元-胶质细胞培养物，也因为需要咨询她以评估硫酸乙酰肝素的效果 根据审稿人的建议，Ranvier 节点处的蛋白聚糖和 NG2。 申请的修订部分在左侧边缘用一条线表示。整体 为统一应用程序风格而引入的编辑修改未标记。 目标 该计划的目标是分析神经元和神经胶质细胞之间交换的分子信号 突触和有髓轴突。该计划已确定了涉及的细胞表面成分 神经元-胶质细胞通讯，现在打算研究这些分子在体内和组织中的功能 密切模仿体内相互作用的培养模型。该计划的核心 A 项目将提供 实现这一目标的基础设施和专业知识。核心将为以下两个方面提供支持 工作，通过电生理学分析神经元功能，并在适当的情况下模拟神经元-胶质细胞相互作用 文化系统。 电生理学部分增加了一个新的研究维度，将为该计划提供 用于功能性和活动依赖性研究的工具和专门知识，旨在获取相关知识 神经元响应神经胶质细胞或神经胶质衍生配体而发生的电生理变化。现在是 显然，神经元-神经胶质细胞的相互作用不仅有助于髓磷脂的功能特性，而且还有助于髓磷脂的功能特性。 突触。在该计划中添加电生理学部分将克服以前的限制 并使该计划能够对因遗传破坏而导致的功能缺陷进行功能分析 神经细胞表面蛋白及其相关信号转导途径。电生理学 该方法旨在分析神经冲动水平上的功能性神经元-胶质细胞相互作用 沿轴突的传导，以及突触功能和可塑性。电生理记录将 评估由表达或变化引起的神经元膜特性的改变 胶质蛋白多糖的功能。该计划将使用已经在 手并有足够长的生存时间来进行电生理分析。 电生理记录将在脑切片、单细胞、 和孤立的神经。切片记录将重点确定神经胶质蛋白 ephrin-A3 在 LTP 和 LTD 期间突触功效发生变化。单细胞记录将评估肝配蛋白的贡献 A3 通过其神经元受体 EphA4 影响神经元的兴奋性和可塑性。整个神经记录将 检测由髓磷脂故障引起的复合动作电位的传导速度变化 鞘。这些不同水平的记录对于深入了解分子是必要的。 正如各个项目中概述的，神经元-神经胶质细胞串扰背后的相互作用。 拟议工作的第二个关键方面是探测细胞中的神经元-胶质细胞相互作用 使用来自基因操纵小鼠的材料达到水平。这是通过使用合适的方法有效地实现的 模仿特定体内相互作用的原代神经培养系统。具体来说，核心A将提供 神经元和髓鞘形成雪旺细胞或少突胶质细胞的共培养物，用于研究 髓磷脂发生。利用海马培养物研究胶质细胞肝配蛋白-A3在调节中的影响 突触功能和可塑性将是当前核心的延续。