Name: the analysis of purkinje cell dendritic morphology in organotypic slice cultures
Uploaded: 2012-03-21T13:00:00
Description: Watch this Scientific Journal Video about The Analysis of Purkinje Cell Dendritic Morphology in Organotypic Slice Cultures at JoVE.com

这项工作得到了巴塞尔大学生物医学系和瑞士国家科学基金会（31003A-116624）。

1。建立器官小脑切片培养出生后小脑切片文化准备从8 P（8）鼠标幼崽使用静态培养方法10。在我们的实验室中，我们使用B6CF1小鼠。在一些实验中也转基因小鼠，用于表达EGFP的浦肯野细胞的选择性。编制片文化需要大约30分钟，每鼠标的小狗，也就是3个小时的一窝6小鼠的幼崽。所有步骤都是在无菌条件下在层流工作台与消毒的外科手术器械进行。 <ol><li> P8鼠标幼?...

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	<li>Bosman, L. W., Hartmann, J., Barski, J. J., Lepier, A., Noll-Hussong, M., Reichardt, L. F., Konnerth, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Requirement+of+TrkB+for+synapse+elimination+in+developing+cerebellar+Purkinje+cells.">Requirement of TrkB for synapse elimination in developing cerebellar Purkinje cells.</a> Brain Cell Biol. 35, 87-101 (2006).</li><li>Boukhtouche, F., Janmaat, S., Vodjdani, G., Gautheron, V., Mallet, J., Dusart, I., Mariani, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Retinoid-related+orphan+receptor+alpha+controls+the+early+steps+of+Purkinje+cell+dendritic+differentiation.">Retinoid-related orphan receptor alpha controls the early steps of Purkinje cell dendritic differentiation.</a> J. Neurosci. 26, 1531-1538 (2006).</li><li>Kapfhammer, J. 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这里介绍的方法允许研究器官小脑切片培养和浦肯野细胞的树突状发展，定量地评价测定树突状树的尺寸和数目的树突状分支点的浦肯野细胞的树突状扩展。当然，一个更广泛，更复杂的定量分析，浦肯野细胞树突是可能的，例如，通过测定总树突长度，执行一个Sholl的分析，确定分形维数的树突状树。对于这种类型的分析，它通常需要手动跟踪整个树的分析程序，例如神经龙力浦肯野细胞树突?...

<table border="1"><tbody><tr><td> 的试剂的名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论（可选） </td></tr><tr><td>组织培养插入</td><td>密理博</td><td> PICM 03050 PICM ORG50 </td><td> PICMORG50插入低的边缘，并允许在显微镜观察活的文化</td></tr><tr><td> MEM </td><td> Gibco公司，Invitrogen公司</td><td> 11012044 </td><td></td></tr><tr><td> Glutamax 1 </td><td> Gibco公司，Invitrogen公司</td><td> 35050038 </td><td></td></tr><tr><td>基底Medum鹰</td><td> Gibco公司，Invitrogen公司</td><td> 41010026 </td><td></td></tr><tr><td>马血清</td><td> Gibco公司，Invitrogen公司</td><td> 26050070 </td><td></td></tr><tr><td>佛12-myrista的特13 - 乙酸酯（PMA） </td><td> TOCRIS </td><td> 1201 </td><td></td></tr><tr><td> （RS）-3,5 - 二羟基-苯基甘氨酸（DHPG） </td><td> TOCRIS </td><td> 0342 </td><td></td></tr><tr><td>兔抗钙结合蛋白D-28K </td><td> Swant </td><td> CB38 </td><td></td></tr><tr><td>反NEUN，克隆A60 </td><td> Chemicon公司，密理博</td><td> MAB377 </td><td></td></tr><tr><td>兔抗GFP </td><td> Abcam公司</td><td> Ab290 </td><td></td></tr></tbody></table>

the analysis of purkinje cell dendritic morphology in organotypic slice cultures

浦肯野细胞是一个有吸引力的模型系统，研究树突状发展，因为他们有一个令人印象深刻的树突状树是严格面向在矢状面和发展主要是在产后期在小型啮齿动物3。此外，几种抗体可用选择性和集中标签包括所有进程的浦肯野细胞，用抗钙结合蛋白Calbindin D28K被最广泛使用的。活细胞的树突观看，都可以通过杰克逊实验室小鼠表达绿色荧光蛋白选择性地在浦肯野细胞11。器官小脑片培养细胞可以轻松的浦肯野细胞树突发育的实验操作，因为大多数的浦肯野细胞树突树的树突状扩张过程中实际发生的培养周期4。我们在座的短，可靠和简单的协议，用于观察和分析浦肯野细胞的树突状形态，生长在organotypiÇ小脑切片文化。出于多种目的，定量评价的浦肯野细胞树突状树是可取的。我们关注两个参数，树突树的大小和分支点的电话号码，可以快速，轻松地确定反小脑钙结合蛋白染色切片文化。这两个参数产生变化的浦肯野细胞树突状树一个可靠的和敏感的措施。使用的例子，治疗与蛋白激酶C（PKC）激活剂PMA和代​​谢型谷氨酸受体1（mGluR1的），我们演示了如何在树突发育的差异可视化和定量评估。的存在下的结合，一个广泛的树突状树，选择性和强烈的免疫染色的方法，覆盖的枝晶生长期间和小鼠模型的浦肯野细胞的特定的EGFP表达，使树突状揭示的机制的浦肯野细胞的强大的模型系统的器官型切片培养发展。

Watch this Scientific Journal Video about The Analysis of Purkinje Cell Dendritic Morphology in Organotypic Slice Cultures at JoVE.com

The Analysis of Purkinje Cell Dendritic Morphology in Organotypic Slice Cultures

我们提出了一个协议，允许查看和定量驴个别浦肯野细胞的树突树的生长在器官小脑切片文化形态。该协议的目的是促进研究的浦肯野细胞树突发育的机制。

浦肯野细胞树突形态的器官切片培养分析

Purkinje cells are an attractive model system for studying dendritic development, because they have an impressive dendritic tree which is strictly ...

Research

JoVE Journal

Neuroscience

The Organotypic Hippocampal Slice Culture Model for Examining Neuronal Injury

海马脑片的器官型检查神经元损伤的文化模式

Organotypic hippocampal slice culture is an in vitro method to examine mechanisms of neuronal injury in which the basic architecture and composition of ...

科研

神经科学

Automated Sholl Analysis of Digitized Neuronal Morphology at Multiple Scales

肖尔自动数字化的神经元形态分析，多尺度

Neuronal morphology plays a significant role in determining how neurons function and communicate1-3.  Specifically, it affects the ability of neurons to ...

Organotypic Hippocampal Slice Cultures

器官型海马切片培养

The hippocampus, a component of the limbic system, plays important roles in long-term memory and spatial navigation 1. Hippocampal neurons can modify the ...

Organotypic Cerebellar Cultures: Apoptotic Challenges and Detection

器官型小脑文化：细胞凋亡的挑战和检测

Organotypic cultures of neuronal tissue were first introduced by Hogue in 1947 1,2 and have constituted a major breakthrough in the field of neuroscience. ...

Whole Cell Recording from an Organotypic Slice Preparation of Neocortex

从大脑皮层的器官型切片制备全细胞记录

We have been studying the expression and functional roles of voltage-gated potassium channels in pyramidal neurons from rat neocortex.  Because of the ...

Analysis of Dendritic Spine Morphology in Cultured CNS Neurons

在培养的中枢神经系统的神经元树突棘形态分析

Dendritic spines are the sites of the majority of excitatory connections within the brain, and form the post-synaptic 
compartment of synapses. These ...

Organotypic Slice Cultures of Embryonic Ventral Midbrain: A System to Study Dopaminergic Neuronal Development in vitro

Organotypic Slice Cultures of Embryonic Ventral Midbrain: A System to Study Dopaminergic Neuronal Development in vitro

胚胎腹侧中脑器官型片文化：一个系统来研究多巴胺能神经元发展在体外</em

The  mouse is an excellent model organism to study mammalian brain development due  to the abundance of molecular and genetic data. However, the ...

Post-embedding Immunogold Labeling of Synaptic Proteins in Hippocampal Slice Cultures

嵌入后突触蛋白的免疫标记海马切片培养

Immunoelectron microscopy is a powerful tool to study biological molecules at the subcellular level. Antibodies coupled to electron-dense markers such as ...

Organotypic Slice Cultures to Study Oligodendrocyte Dynamics and Myelination

器官切片文化研究少突胶质细胞动力学和髓鞘

NG2 expressing cells (polydendrocytes, oligodendrocyte precursor cells) are the fourth major glial cell population in the central nervous system. During ...

The Analysis of Neurovascular Remodeling in Entorhino-hippocampal Organotypic Slice Cultures

神经血管重塑的分析Entorhino，海马脑切片培养

Ischemic brain injury is among the most common and devastating conditions compromising proper brain function and often leads to persisting functional ...