Name: imaging analysis of neuron to glia interaction in microfluidic culture platform (mcp)-based neuronal axon and glia co-culture system
Uploaded: 2012-10-14T14:00:00
Description: Watch this Scientific Journal Video about Imaging Analysis of Neuron to Glia Interaction in Microfluidic Culture Platform MCP-based Neuronal Axon and Glia Co-culture System at JoVE.com

我们想感谢Jeffrey罗斯坦提供BAC GLT1的绿色荧光蛋白的小鼠和GLT1抗体; Tufts中心神经科学研究所（NIH P30 NS047243 PI，罗布·杰克逊）提供有价值的核心设备，招聘新教师津贴（NIH P30 5P30NS069254-02 PI，菲尔·海登）在塔夫茨大学神经科学系。

1。大会的微流控文化厅（MCP） <ol><li> MCP（ 图1）是开放的腔室隔间培养不同类型的单元4设计。它典型地具有被连接的两个隔室，通过中央通道（直径为3微米）。大会MCP玻璃底的菜肴是必须的为编制文化和后续的图像分析。 </li><li>首先，外套的无菌玻璃有底菜肴用多聚鸟氨酸（Sigma-Aldrich公司，1毫克/毫升）溶解在四硼酸钠（Sigma-Aldrich公司，10mM的pH为8.5）中，并在37℃下孵?...

     

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MCP基于神经元和星形胶质细胞共培养体系可以详细的神经元，星形胶质细胞信号转导通路的解剖允许唯一的轴突通过中央通道，并与星形胶质细胞。此共培养体系可以方便地设置与传统的神经元，星形胶质细胞的培养程序。我们也采用的绿色荧光蛋白记者展示轴突取决于GLT1的启动子活性在星形胶质细胞共培养系统的实际应用。 这种MCP基于神经元和星形胶质细胞共培养体系提...

<table border="1"><tbody><tr><td>胎牛血清</td><td> Hyclone公司</td><td> SH30070.03 </td><td>电镀神经元神经元cutlure培养基</td></tr><tr><td>胎牛血清</td><td> Sigma-Aldrich公司</td><td> F4135 </td><td>星形胶质细胞培养基</td></tr><tr><td>胶质细胞源性神经因子</td><td> R＆D系统</td><td> 212-GD </td><td> 10-20毫微克/毫升侧腔的神经元</td></tr><tr><td> Dulbecco改良Eagle培养基高糖</td><td> Sigma-Aldrich公司</td><td> 11995 </td><td></td></tr><tr><td> 70毫米电池过滤器， </td><td> BD猎鹰</td><td> 352350 </td><td></td></tr><tr><td>无菌玻璃底菜</td><td> MatTek公司</td><td></td><td></td></tr><tr><td>微流控文化平台</td><td> XONA微流体有限责任公司</td><td> SND150 </td><td></td></tr><tr><td> 6个孔的培养板</TD&gt; <td>蜂星</td><td> 657 160 </td><td></td></tr><tr><td></td><td></td><td></td><td> 神经元培养基 <ul><li> Neurobasal培养基</li><li> 2％B27的Neurobasal补充</li><li>通过添加1％的100倍GlutaMAX 2mM的谷氨酸</li><li> 1％青霉素 - 链霉素</li></ul></td></tr><tr><td></td><td></td><td></td><td> 的镀敷细胞的神经元的培养基 <ul><li> Neurobasal培养基</li><li> 2％B27的Neurobasal补充</li><li>通过添加1％的100倍GlutaMAX 2mM的谷氨酸</li><li> 1％青霉素 - 链霉素</li><li> 5％胎牛血清SH30070.03 </li></ul></td></tr><tr><td></td><td></td><td></td><td> 星形胶质细胞培养基 <ul><li> Dulbecco改良EAGLE中高血糖</li><li> 10％胎牛血清F4135 </li><li> 1％青霉素 - 链霉素</li></ul></td></tr><tr><td></td><td></td><td></td><td> 表1中。材料中使用的微流体培养基于平台的神经元轴突和神经胶质细胞的共培养系统。 </td></tr></tbody></table>

imaging analysis of neuron to glia interaction in microfluidic culture platform (mcp)-based neuronal axon and glia co-culture system

适当的神经元，神经胶质细胞相互作用的中枢神经系统（CNS）的生理功能是至关重要的。这种双向通信老练介导的神经元和神经胶质细胞1,2之间通过特定的信号转导通路。这些信号通路的识别和表征如何神经元的神经胶质细胞相互作用的形状中枢神经系统生理学的理解是必不可少的。在此之前，神经元和神经胶质细胞混合培养已被广泛用于测试和表征神经元和神经胶质细胞之间的信号传导通路。我们已经从这些制剂及其他体内的工具，但是，已经表明，神经元和神经胶质细胞相互之间的信号往往发生在特定的车厢内神经元（ 即轴突，树突或胞体）3。这使得重要的是要开发一种新的文化系统，让神经车厢分离，专门研究神经胶质细胞和神经之间的相互作用NAL轴突/树突。此外，以往的混合培养系统是不能够分化的可溶性因子和神经元和神经胶质细胞之间的直接膜接触信号。此外，大量的神经元和神经胶质细胞，在传统的共培养体系缺乏必要的分辨率，观察到一个单一的轴突和神经胶质细胞之间的相互作用。 在这项研究中，我们描述了一种新的轴突和神经胶质细胞共培养系统与使用平台（MCP）的微流体文化。在此共同培养系统中，神经元和神经胶质细胞培养在两个单独的腔室，通过连接多个中央通道。在这个微文化平台，神经元的过程（特别是轴突）可以通过中央通道进入的胶质。结合强大的荧光蛋白标记，该系统可以直接检查/树突状轴突和胶质细胞相互作用的信号转导通路之间，这样的小号轴突神经胶质细胞，神经胶质细胞介导的​​受体贩运神经元的终端，和神经胶质细胞介导的​​轴突生长介导的转录调控。的窄直径的腔室，也显着到胶质室禁止神经元的富集培养基的流，促进轴突/树突和胶质表面之间的直接膜 - 蛋白质相互作用的探测。

轴突诱导GLT1的启动子活性在星形胶质细胞延时成像分析隔间的神经元和星形胶质细胞的共培养系统允许的神经元的过程，尤其是在轴突，以选择性地与星形胶质细胞相互作用。不同类型的轴突神经胶质细胞相互作用的成功建立在组装MCP的轴突和星形胶质细胞（或其他神经胶质细胞）共培养后，可以研究，如轴突诱导活化的星形胶质细胞的基因启动子活性，星形胶质细胞诱导的?...

Watch this Scientific Journal Video about Imaging Analysis of Neuron to Glia Interaction in Microfluidic Culture Platform MCP-based Neuronal Axon and Glia Co-culture System at JoVE.com

Imaging Analysis of Neuron to Glia Interaction in Microfluidic Culture Platform MCP-based Neuronal Axon and Glia Co-culture System

本研究介绍了一种新的神经元轴突和神经胶质细胞共培养（ASTRO）平台的程序。在此共培养体系中的一个单一的轴突（及单一的神经胶质细胞）之间的直接相互作用，操纵变得可行，允许机械分析的相互神经胶质信令。

神经元对神经胶质细胞在微流控文化互动平台（MCP）的神经元轴突和神经胶质细胞共培养系统的影像学分析

Proper  neuron to glia interaction is critical to physiological function of the central  nervous system (CNS). This bidirectional communication is ...

Research

JoVE Journal

Neuroscience

Imaging Analysis of Neuron to Glia Interaction in Microfluidic Culture Platform (MCP)-based Neuronal Axon and Glia Co-culture System

Bilaminar Co-culture of Primary Rat Cortical Neurons and Glia

二层共培养小学的大鼠皮层神经元和神经胶质

This video will guide you through the process of culturing rat cortical neurons in the presence of a glial feeder layer, a system known as a bilaminar or ...

科研

神经科学

Analysis of Developing Tooth Germ Innervation Using Microfluidic Co-culture Devices

微流控采用共培养开发设备牙胚支配分析

Innervation plays a key role in the development, homeostasis and regeneration of organs and tissues. However, the mechanisms underlying these phenomena ...

Utilizing Combined Methodologies to Define the Role of Plasma Membrane Delivery During Axon Branching and Neuronal Morphogenesis

采用组合运用各种方法，定义质膜交付在轴突分支和神经元形态发生中的作用

During neural development, growing axons extend to multiple synaptic partners by elaborating axonal branches. Axon branching is promoted by extracellular ...

The Indirect Neuron-astrocyte Coculture Assay: An In Vitro Set-up for the Detailed Investigation of Neuron-glia Interactions

The Indirect Neuron-astrocyte Coculture Assay: An In Vitro Set-up for the Detailed Investigation of Neuron-glia Interactions

间接神经元的星形胶质细胞共培养试验：一个<em&gt;体外</em&gt;设置 - 为神经元的神经胶质细胞相互作用的详细调查

Proper neuronal development and function is the prerequisite of the developing and the adult brain. However, the mechanisms underlying the highly ...

M&#252;ller Glia Cell Activation in a Laser-induced Retinal Degeneration and Regeneration Model in Zebrafish

激光诱导的斑马鱼视网膜变性及再生模型中的 m ü ller 胶质细胞活化

A fascinating difference between teleost and mammals is the lifelong potential of the teleost retina for retinal neurogenesis and regeneration after ...

Axonal Transport of Organelles in Motor Neuron Cultures using Microfluidic Chambers System

使用微流体腔系统在运动神经元培养物中有机细胞的斧头迁移

Motor neurons (MNs) are highly polarized cells with very long axons. Axonal transport is a crucial mechanism for MN health, contributing to neuronal ...

Isolation and Culture of Primary Neurons and Glia from Adult Rat Urinary Bladder

来自成人大鼠尿膀胱的原发神经元和胶质的隔离与培养

The lower urinary tract has two main functions, namely, periodic urine storage and micturition; these functions are mediated through central and ...

Targeted Neuronal Injury for the Non-Invasive Disconnection of Brain Circuitry

脑回路非侵入性断开的靶向神经元损伤

Surgical intervention can be quite effective for treating certain types of medically intractable neurological diseases. This approach is particularly ...

Dissection and Isolation of Murine Glia from Multiple Central Nervous System Regions

The methods presented here demonstrate laboratory procedures for the dissection of four different regions of the central nervous system (CNS) from murine ...

A Cell Culture Model for Studying the Role of Neuron-Glia Interactions in Ischemia

研究神经-胶质相互作用在缺血中的作用的细胞培养模型

Ischemic stroke is a clinical condition characterized by hypoperfusion of brain tissue, leading to oxygen and glucose deprivation, and the consequent ...