Name: inducing dendritic growth in cultured sympathetic neurons
Uploaded: 2012-03-21T12:00:00
Description: Watch this Scientific Journal Video about Inducing Dendritic Growth in Cultured Sympathetic Neurons at JoVE.com

这项工作是从美国国立卫生研究院（赠款R21的NS45037和以R01 ES014901）的资金支持。

1。培养基的制备（C2的介质） <ol><li>中列出的顺序，添加以下，无菌的，一次性的500毫升三角瓶： <table border="1"><tbody><tr><td> 量 </td><td> 元件 </td><td> 终浓度 </td></tr><tr><td>第190笔资料毫升</td><td> DMEM培养基（低糖） </td><td> N / A </td></tr><tr><td> 10毫升</td><td>脂肪酸免费BSA（DMEM培养液20mg/mL时） </td><td> 50微克/毫升</td></tr><tr><td> 2.8毫升</td><td> L-谷氨酰胺</td><td> 1.4?...

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I., Higgins, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glial+cells+promote+dendritic+development+in+rat+sympathetic+neurons+in+vitro.">Glial cells promote dendritic development in rat sympathetic neurons in vitro.</a> Glia. 1, 380-392 (1988).</li><li>Lein, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effects+of+extracellular+matrix+and+osteogenic+protein-1+on+the+morphological+differentiation+of+rat+sympathetic+neurons.">The effects of extracellular matrix and osteogenic protein-1 on the morphological differentiation of rat sympathetic neurons.</a> Int. J. Dev. 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I., Banker, G., Goslin, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Culturing Nerve Cells. , 177-295 (1991).</li><li>Beck, H. N., Drahushuk, K., Jacoby, D. B., Higgins, D., Lein, P. 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这种模式的优点包括：（一）文化是一个没有其他细胞类型17,23交感神经元的同质人口组成;（二）交感神经神经生长因子的要求建立的，它允许使用工作定义和界定媒体的各种媒介，与基板增长和维持交感神经元23（三）在这些文化中的神经元统一回应基底膜，BMP-7和民进党的其他骨形态发生蛋白的树突促进活动60A亚科17,18,20,21;及（d）基底膜或BMP诱导的树突形成细胞的存?...

<table border="1"><tbody><tr><td> 试剂名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论 </td></tr><tr><td>盖玻片</td><td> bellco </td><td> 1943-10012 </td><td> 12毫米的圆形盖玻片＃1厚度，德国玻璃</td></tr><tr><td>弯曲的尖滴管</td><td> bellco </td><td> 1273-50003 </td><td></td></tr><tr><td>聚-D-赖氨酸</td><td>西格玛</td><td> P0899-100MG </td><td> 1毫克/毫升原液</td></tr><tr><td>蒸馏水</td><td> Gibco公司</td><td> 15230 </td><td></td></tr><tr><td>精细镊子杜蒙没有。 4和5 </td><td>的罚款科学工具（FST）的</td><td> 11252-30 11241-30 </td><td></td></tr><tr><td> 20表针</td><td>碧迪</td><td> 305176 </td><td></td></tr><tr><td>莱博维茨的L-15培养基</td><td> Invitrogen公司</tD&gt; <td> 11415 </td><td></td></tr><tr><td>胶原酶</td><td>沃辛顿</td><td> 4176 </td><td> 1毫克/毫升</td></tr><tr><td> Dispase酶</td><td>罗氏公司</td><td> 04942078001 </td><td> 5毫克/毫升</td></tr><tr><td>汉克的平衡盐溶液（钙，镁） </td><td> Invitrogen公司</td><td> 14185-052 </td><td></td></tr><tr><td> DMEM培养液低血糖</td><td> Invitrogen公司</td><td> 11885 </td><td></td></tr><tr><td> L-谷氨酰胺</td><td> Invitrogen公司</td><td> 25030 </td><td> 20毫米</td></tr><tr><td>胰岛素/转/硒</td><td> Invitrogen公司</td><td> 51500-056 </td><td> 100X </td></tr><tr><td>脂肪酸的免费牛血清白蛋白</td><td> Calbiochem公司</td><td> 126609 </td><td></td></tr><tr><td>神经生长因子</td><td>哈伦生物制品</td><td> 5017 </td><td></td></tr><tr><td>火腿的F-12营养培养基（架F-12） </td><td> Invitrogen公司</td><td> 11765 </td><tD&gt; </td></tr><tr><td>阿糖胞苷</td><td>西格玛</td><td> C1768 </td><td></td></tr><tr><td>基底膜</td><td> BD公司</td><td> 356234 </td><td>避免反复冻融</td></tr><tr><td> BMP-7 </td><td> R＆D系统Curis公司</td><td> 345-BP BMP7的-07H </td><td>骨形态发生蛋白4，5和6的工作同样以及BMP-7，有时更容易获得商业</td></tr><tr><td>多聚甲醛</td><td>西格玛</td><td> P6148 </td><td></td></tr><tr><td>的Triton X-100 </td><td>费舍尔</td><td> BP151-500 </td><td></td></tr><tr><td>小鼠抗大鼠MAP2的抗体</td><td>伯格单克隆抗体公司</td><td> SMI52 </td><td> 1/5000 </td></tr><tr><td>的Alexa Fluor 546山羊抗鼠</td><td> Invitrogen公司</td><td> A11003 </td><td> 1/10000 </td></tr><tr><td>安装媒体</td><td>延长抗淬灭分子探针套件</td> &lt;TD&gt; P7481 </td><td></td></tr></tbody></table>

inducing dendritic growth in cultured sympathetic neurons

树突状乔木的形状决定的总突触输入一个神经细胞可以接收1-3，并影响这些输入4-6的类型和分布。树突生长和可塑性的改变模式与7个实验模型中的神经行为功能受损，被认为有助于神经紊乱8-10和11-13的神经退行性疾病中观察到的临床症状。这种意见强调精确调节树突形态功能的重要性，并建议确定枝晶生长的机制，控制，不仅促进神经元的连接是如何在正常发展的规管的认识，但还可以提供各种神经系统疾病的新型治疗策略的洞察力。 枝晶生长的机理研究，将极大地促进了可用性Ø发模型系统，使神经元实验从一个国家，他们在不延长树突之一，在他们阐述树突状乔木与体内对口切换。从颈上神经节（SCG）围产期啮齿类动物中分离交感神经元的原代培养提供了这样一种模式。定义培养基中的血清和神经节神经胶质细胞的情况下培养时，交感神经元延长一个单一的过程，这是轴突，这个单极状态持续几周文化14,15个月。然而，无论是骨形态发生蛋白-7（BMP-7），16,17或18基底膜培养基除了引发这些神经元的延长，树突的形态，生化和功能的标准，满足多个进程。交感神经元脱离围产期啮齿动物的上海建工，并在特定条件下生长的同质人口的神经元19一致响应基底膜，BMP-7和其他骨形态发生蛋白decapentaplegic（民进党）和60A亚科17,18,20,21枝促进活动。重要的是，基底膜和BMP诱导的树突的形成发生在细胞的存活或轴突生长17,18变化的情况下。 在这里，我们描述了如何设置从上海建工围产期大鼠的交感神经元的分离培养，使他们顺应选择性枝促进基底膜或BMP的活动。

Watch this Scientific Journal Video about Inducing Dendritic Growth in Cultured Sympathetic Neurons at JoVE.com

Inducing Dendritic Growth in Cultured Sympathetic Neurons

我们描述了使用骨形态发生蛋白-7（BMP-7）或基底膜的协议，主要从围产期大鼠颈上神经节（SCG）分离交感神经元选择性诱导的树突状增长。

诱导培养的交感神经元的树突状增长

The shape of the dendritic arbor determines the total synaptic input a neuron can receive 1-3, and influences the types and distribution of these inputs ...

Research

JoVE Journal

Neuroscience

Fluorescence Recovery After Photobleaching (FRAP) of Fluorescence Tagged Proteins in Dendritic Spines of Cultured Hippocampal Neurons

Fluorescence Recovery After Photobleaching FRAP of Fluorescence Tagged Proteins in Dendritic Spines of Cultured Hippocampal Neurons

漂白后荧光标记蛋白的荧光恢复（FRAP）在培养海马神经元的树突棘

FRAP has been used to quantify the mobility of GFP-tagged proteins. Using a strong excitation laser, the fluorescence of a GFP-tagged protein is bleached ...

科研

神经科学

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 ...

Quantitative Analysis of Synaptic Vesicle Pool Replenishment in Cultured Cerebellar Granule Neurons using FM Dyes

使用调频染料培养的小脑颗粒神经突触小泡池补充的定量分析

After neurotransmitter release in central nerve terminals, SVs are rapidly retrieved by endocytosis.  Retrieved SVs are then refilled with ...

Genetic Study of Axon Regeneration with Cultured Adult Dorsal Root Ganglion Neurons

培养成年的背根神经节神经元轴突再生的遗传研究

It is well known that mature neurons in the central nervous system (CNS) cannot regenerate their axons after injuries due to diminished intrinsic ability ...

Voltage-sensitive Dye Recording from Axons, Dendrites and Dendritic Spines of Individual Neurons in Brain Slices

电压敏感染料记录单个神经元在大脑切片的轴突，树突和树突棘

Understanding the biophysical properties and functional organization of single neurons and how they process information is fundamental for understanding ...

Imaging pHluorin-tagged Receptor Insertion to the Plasma Membrane in Primary Cultured Mouse Neurons

成像pHluorin标记插入到质膜受体原代培养的小鼠神经元

A better understanding of the mechanisms governing receptor trafficking between the plasma membrane (PM) and intracellular compartments requires an ...

Fast Micro-iontophoresis of Glutamate and GABA: A Useful Tool to Investigate Synaptic Integration

谷氨酸和GABA快速微离子导入：一个有用​​的工具，调查突触整合

One of the fundamental interests in neuroscience is to understand the integration of excitatory and inhibitory inputs along the very complex structure of ...

Imaging Dendritic Spines of Rat Primary Hippocampal Neurons using Structured Illumination Microscopy

成像大鼠原代海马神经元树突棘使用结构照明显微镜

Dendritic spines are protrusions emerging from the dendrite of a neuron and represent the primary postsynaptic targets of excitatory inputs in the brain. ...

Quantification of Endosome and Lysosome Motilities in Cultured Neurons Using Fluorescent Probes

使用荧光探针定量培养的神经元中的内体和溶酶体运动

In the brain, membrane trafficking systems play important roles in regulating neuronal functions, such as neuronal morphology, synaptic plasticity, ...

Measuring Synaptic Vesicle Endocytosis in Cultured Hippocampal Neurons

海马神经元中突触囊泡吞的测量

During endocytosis, fused synaptic vesicles are retrieved at nerve terminals, allowing for vesicle recycling and thus the maintenance of synaptic ...