Name: paradigms for pharmacological characterization of c. elegans synaptic transmission mutants
Uploaded: 2008-08-18T17:08:00
Description: Watch this Scientific Journal Video about Paradigms for Pharmacological Characterization of C. elegans Synaptic Transmission Mutants at JoVE.com

我们要感谢考德威尔实验室所有成员的合作精神。 March of Dimes的职业生涯从美国国家科学基金会奖，以海关总署从一个罗勒奥康纳学者奖，以及来自霍华德休斯医学研究所的阿拉巴马大学本科生科研的科学项目资助，资助的神经元的兴奋和癫痫研究中的考德威尔实验室。

涕灭威曝光范式<ol><li>在第一天，确保至少30个年轻的成年阶段各基因型和每个复制的蠕虫将于第二天，涕灭威的检测。这是最好的，如果实验者选择了50多个L4阶段到新鲜NGM板蠕虫（最好没有制霉菌素），其中含有大肠杆菌作为食物来源的大肠杆菌 （最好OP50），并成长为12-24小时，他们在一个一致的和宽松的温度（20 ° C至22 ° C是最好的，虽然25℃是好的）。 </li><li>第二天...

<ol>
	<li>Mahoney, T. R., Luo, S., Nonet, M. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analysis+of+synaptic+transmission+in+Caenorhabditis+elegans+using+an+aldicarb-sensitivity+assay.">Analysis of synaptic transmission in Caenorhabditis elegans using an aldicarb-sensitivity assay.</a> Nat. Protoc. 1, 1772-1777 (2006).</li><li>Williams, S. N., Locke, C. J., Braden, A. L., Caldwell, K. A., Caldwell, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epileptic-like+convulsions+associated+with+LIS-1+in+the+cytoskeletal+control+of+neurotransmitter+signaling+in+Caenorhabditis+elegans.">Epileptic-like convulsions associated with LIS-1 in the cytoskeletal control of neurotransmitter signaling in Caenorhabditis elegans.</a> Hum. Mol. Genet.. 13, 2043-2059 (2004).</li></ol>

当前的协议与 C涕灭威曝光线虫不允许实验者区分与特定的赤字乙酰胆碱在突触传递中的广义赤字的传输和突变体的突变体，突变体的两个类表现出耐涕灭威。同样，涕灭威不能用来确定突变GABA的传输或广义故障的具体赤字负调节乙酰胆碱的传输，因为这两个类的突变体表现出过敏到涕灭威。我们云台暴露实验，涕灭威暴露实验结果相结合，结果使研究人员能够更好地表征突触传递的突变体。 <p...

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<td>Aldicarb</td>
<td>Reagent</td>
<td>Supelco, Inc.</td>
<td>PS734</td>
<td>Purchasable from Sigma</td>
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<td>Pentylenetetrazole</td>
<td>Reagent</td>
<td>Sigma</td>
<td>P6500</td>
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paradigms for pharmacological characterization of c. elegans synaptic transmission mutants

线虫，线虫，已成为研究神经递质的权宜之计模型。线虫在动物模型中是独一无二的，解剖学和神经系统的连接已经从电子显微镜确定和完善了药理实验。在这段视频中，我们描述了如何两个互补的神经兴奋剂，涕灭威，乙酰胆碱酯酶抑制剂，和伽马氨基丁酸（GABA）受体拮抗剂，戊四氮（PTZ），可受聘具体特征线虫神经肌肉接头的信号（NMJs），方便我们对立的神经回路的了解。 302线虫的神经元，十九电机D型GABA能神经元支配体壁肌肉（BWMs），而GABA能神经元，称为RMEs，头部肌肉的支配。相反，三十九个运动神经元表达的兴奋性神经递质，乙酰胆碱（ACH），拮抗GABA的传输在BWMs协调运动。 GABA能和胆碱能运动神经元在体壁NMJs的对立性质最初由激光烧蚀和后来挟着涕灭威曝光。涕灭威在野生型蠕虫当然时间或剂量反应瘫痪急性暴露的结果。然而，兴奋胆碱传输损失抗性涕灭威，蠕虫NMJs少乙酰胆碱的积累，导致少BWMs刺激。与乙酰胆碱的特定或一般的突触功能的突变体，可观察到涕灭威的电阻。与对立的传输γ-氨基丁酸和乙酰胆碱，GABA的传输损失，或失败负调节乙酰胆碱的释放一致，赋予过敏涕灭威。虽然涕灭威曝光导致的许多蠕虫病毒的神经递质基因同源的隔离，涕灭威曝光单独不能有效地判断当时的角色线虫在特定的运动神经元的基因和途径。为此，我们引入了一个互补的实验方法，它使用云台。 神经传递突变体显示清晰的表型，有别于涕灭威，引起瘫痪，到PTZ。野生型蠕虫病毒，以及与乙酰胆碱的释放或接收的具体无能的突变体，并没有显示明显的敏感性云台。然而，氨基丁酸的突变体，以及一般的突触功能的突变体，显示在时间的课程或剂量反应的方式，前抽搐。到BWMs突变体不能负调控一般神经递质的释放，因此，分泌过量的乙酰胆碱，成为云台瘫痪。离散突变类的PTZ诱导的表型表明，涕灭威和线虫的PTZ曝光范式互补的做法，可能会加速我们的神经传递的理解。此外，视频演示了如何进行药理实验，应建立对线虫的研究相一致方法。

Watch this Scientific Journal Video about Paradigms for Pharmacological Characterization of C. elegans Synaptic Transmission Mutants at JoVE.com

Paradigms for Pharmacological Characterization of C. elegans Synaptic Transmission Mutants

本视频演示了如何利用互补的方法，研究线虫，线虫突触功能的神经兴奋剂，涕灭威和戊四氮（PTZ），。这种互补的方法也可能被用来阐明调节神经元的同步进化上保守的机制和癫痫和癫痫发作的影响。

为线虫突触传递的突变体的药理特性的范式

The nematode, Caenorhabditis elegans, has become an expedient model for studying neurotransmission. C. elegans is unique among animal models, as the ...

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