Name: preparation of neuronal cultures from midgastrula stage drosophila embryos
Uploaded: 2007-07-04T00:00:00
Description: Watch this Scientific Journal Video about Preparation of Neuronal Cultures from Midgastrula Stage Drosophila Embryos at JoVE.com

这项工作是由美国国立卫生研究院授予NS27501 DKOD支持。这从霍华德休斯医学研究所教授格兰特DKOD工作的额外支持。

一，果蝇胚胎收集<ol><li>准备鸡蛋收集板<ol><li>煮沸200毫升与8克琼脂DH 2 O </li><li>冷却至50 ° C </li><li>加入2 ml乙醇和2毫升乙酸</li><li>倒入35 mm塑料培养皿中，上衣</li><li>酷存储在密封容器中，并在4 ° C </li><li>使大约80板</li></ol></li><li>准备酵母膏<ol><li>溶解于水Fleishmans积极烘烤酵母的形式粘贴</li><li> 20-30秒杀死酵母微波（小心沸腾了） </li><li>商店在10 CC（无针头注射器，在4 ° C的2个星期） <...

<ol>
	<li>Rohrbough, J., O'Dowd, D. K., Baines, R. A., Broadie, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cellular+bases+of+behavioral+plasticity:+Establishing+and+modifying+synaptic+circuits+in+the+Drosophila+Genetic+System.">Cellular bases of behavioral plasticity: Establishing and modifying synaptic circuits in the Drosophila Genetic System.</a> J. Neurobiol. 54, 254-271 (2003).</li><li>O'Dowd, D. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Voltage-gated+currents+and+firing+properties+of+embryonic+Drosophila+neurons+grown+in+a+chemically+defined+medium.">Voltage-gated currents and firing properties of embryonic Drosophila neurons grown in a chemically defined medium.</a> J. Neurobiol. 27, 113-126 (1995).</li><li>Lee, D., O'Dowd, D. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fast+excitatory+synaptic+transmission+mediated+by+nAChR+in+cultured+Drosophila+neurons.">Fast excitatory synaptic transmission mediated by nAChR in cultured Drosophila neurons.</a> J. Neurosci. 19, 5311-5321 (1999).</li><li>Lee, D., O'Dowd, D. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cyclic-AMP-dependent+plasticity+at+excitatory+cholinergic+synapses+in+Drosophila+neurons:+Alterations+in+the+memory+mutant+dunce.">Cyclic-AMP-dependent plasticity at excitatory cholinergic synapses in Drosophila neurons: Alterations in the memory mutant dunce.</a> J. Neurosci. 20, 2104-2111 (2000).</li><li>Hodges, D. D., Lee, D., Boswell, K., Preston, C. F., Hall, L. M., O'Dowd, D. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=tipE+regulates+sodium-dependent+repetitive+firing+in+Drosophila+neurons.">tipE regulates sodium-dependent repetitive firing in Drosophila neurons.</a> Mol. Cell Neurosci. 19, 402-416 (2002).</li><li>Lee, D., Su, H., Dowd, O., K, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=GABA+receptors+containing+Rdl+subunits+mediate+fast+inhibitory+synaptic+transmission+in+Drosophila+neurons.">GABA receptors containing Rdl subunits mediate fast inhibitory synaptic transmission in Drosophila neurons.</a> J. Neurosci. 23, 4625-4634 (2003).</li></ol>

在果蝇的文化准备midgastrula阶段的胚胎的神经元产生的，其中许多鸿沟之前，在体外分化成神经细胞前体。因此，该系统提供了一个独特的机会，在神经元发育的早期阶段（Rohrbaugh等，2003）中的重要探索遗传和环境因素。我们已经表明，生长在一个简单的定义介质的神经元分化成神经元电兴奋，也形成功能性突触连接（直径多德，1995年李和O多德，1999年）。使用分析突变体和/或药理操作，在这个模型系统可用于...

在这个媒体所作的文化，必须保持在5％CO2培养箱约22-24 ° C。我们使用一个标准的哺乳动物组织培养孵化器放置在寒冷的房间，并加热到23 ° C总是用灭菌过滤后的水，并在媒体和补充只保留容器。不要将pH计或搅拌媒体其他目的使用的栏。

preparation of neuronal cultures from midgastrula stage drosophila embryos

该视频演示了midgastrula阶段果蝇胚胎的主要神经文化的过程。收集胚胎和dechorionation使用漂白水的方法证明。我们说明了一嘴吸痰管使用玻璃吸管，从单一胚胎细胞清除。分散成小（5升）下降中等未涂层的玻璃盖玻片上的每一个embyro细胞的方法是证明。一个电镀后1小时，通过显微镜说明首选的细胞密度。大多数细胞的生存时，在定义中等增长是神经母细胞分裂前12-24小时延长神经炎进程的一个或多个次文化。通过显微镜说明突起生长的水平，预计在2天的体外分支在一个健康的文化。的文化是生长在一个简单的碳酸氢钠为主的定义介质在5％二氧化碳培养箱中培养，在22-24 ° C。神经炎进程继续阐述文化的第一个星期，当他们从邻近的细胞，他们往往形成功能性突触连接的突起联系。在这些文化中的神经元表达电压门控钠，钙，钾通道，电兴奋。这种文化系统是用于研究分子遗传和环境因素，调节神经细胞的分化，兴奋性突触的形成/功能。

Watch this Scientific Journal Video about Preparation of Neuronal Cultures from Midgastrula Stage Drosophila Embryos at JoVE.com

Preparation of Neuronal Cultures from Midgastrula Stage Drosophila Embryos

这个视频演示midgastrula阶段果蝇胚胎的主要神经文化的准备。生活文化的意见表明电镀和后2天在碳酸氢盐为基础的定义中等的增长有区别的神经元细胞1小时后。神经元电兴奋，并形成突触连接。

从Midgastrula舞台果蝇胚胎神经文化的制备

This video illustrates the procedure for making primary neuronal cultures from midgastrula stage Drosophila embryos. The methods for collecting embryos ...

Research

JoVE Journal

Biology

Primary Neuronal Cultures from the Brains of Late Stage Drosophila Pupae

从晚期果蝇蛹的大脑原发性神经文化

In this video, we demonstrate the preparation of primary neuronal cultures from the brains of late stage Drosophila pupae. The procedure begins with the ...

科研

生物学

Placing Growth Factor-Coated Beads on Early Stage Chicken Embryos

配售早期鸡胚生长因子包被的磁珠

The neural tube expresses many proteins in specific spatiotemporal patterns during development.   These proteins have been shown to be critical for cell ...

In Ovo Electroporations of HH Stage 10 Chicken Embryos

在OVO的HH阶段10个鸡胚Electroporations

Large size and external development of the chicken embryo have long made it a valuable tool in the study of developmental biology.  With the advent of ...

Neuronal Cell Cultures from Aplysia for High-Resolution Imaging of Growth Cones

从海兔的神经细胞培养的高分辨率成像生长锥

Neuronal growth cones are the highly motile structures at the tip of axons that can detect guidance cues in the environment and transduce this information ...

The Preparation of Drosophila Embryos for Live-Imaging Using the Hanging Drop Protocol

The Preparation of Drosophila Embryos for Live-Imaging Using the Hanging Drop Protocol

编制果蝇实时成像的胚胎使用悬滴协议

Green fluorescent protein (GFP)-based timelapse live-imaging is a powerful technique for studying the genetic regulation of dynamic processes such as ...

Preparation of Aplysia Sensory-motor Neuronal Cell Cultures

Preparation of Aplysia Sensory-motor Neuronal Cell Cultures

制备海兔感觉运动神经元细胞培养

The nervous system of the marine mollusk Aplysia californica is relatively simple, consisting of approximately 20,000 neurons.  The neurons are large (up ...

Preparation of embryos for Electron Microscopy of the Drosophila embryonic heart tube

Preparation of embryos for Electron Microscopy of the Drosophila embryonic heart tube

电子显微镜对胚胎的制备果蝇胚胎心管

The morphogenesis of the Drosophila  embryonic heart tube has emerged as a valuable model system for studying cell migration, cell-cell adhesion and cell ...

In-vivo Centrifugation of Drosophila Embryos

In-vivo Centrifugation of Drosophila Embryos

果蝇胚胎的体内离心

A major strategy for purifying and isolating different types of intracellular organelles is to separate them from each other based on differences in ...

Upright Imaging of Drosophila Embryos

Upright Imaging of Drosophila Embryos

直立成像果蝇胚胎

Several well-known morphogenetic gradients and cellular movements occur along the dorsal/ventral axis of the Drosophila embryo. However, the current ...

Primary Cell Cultures from Drosophila Gastrula Embryos

Primary Cell Cultures from Drosophila Gastrula Embryos

原代细胞培养从果蝇原肠胚

Here we describe a method for preparing and culturing primary cells dissociated from Drosophila gastrula embryos.   In brief, a large amount of staged ...