Name: quantitative measurement of the immune response and sleep in drosophila
Uploaded: 2012-12-04T13:00:00
Description: Watch this Scientific Journal Video about Quantitative Measurement of the Immune Response and Sleep in Drosophila at JoVE.com

这项工作是由美国国家科学基金会的支持下授予＃IOS-1025627，由美国国立卫生研究院资助项目＃1R21NS078582-01 JAW

该协议使用的设置（ 图1）获得4个不同的读数收集苍蝇的细菌感染。这些输出包括：1）睡眠/唤醒的行为; 2）生存结果; 3）细菌负荷在飞; 4）实时测量体内的的NFκB记者活动。结合，也可在果蝇的遗传工具，这些测量提供机械洞察分子免疫功能和行为之间的联系。 1。测量运动及睡眠苍蝇<ol><li>已安装用于测量运动及睡觉的苍蝇，其中包括果蝇</em...

<ol>
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该协议概述的方法来研究如何行为，特别是睡眠，是与免疫反应参数。这些参数包括细菌量，存活的结果，和NFκB活性在体内荧光素酶报告。这些参数提供了一只苍蝇可以对抗感染的信息。细菌和生存结果是涉及一个简单的测量果蝇的免疫反应参数。 相对E20突变体，缺乏NFκB转录因子，是中央的免疫反应，屈服于迅速细菌感染。遗传性或其他操作的行为也可能影响这些...

<table border="1"><tbody><tr><td> 材料名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论 </td></tr><tr><td></td><td></td><td></td><td> 设备 </td></tr><tr><td>孵化器</td><td> ，波斯富街科学公司</td><td> I30BLLC8 I36VLC8 </td><td>任何孵化器的运行程序化光/温度时间表是适当的。 </td></tr><tr><td> 果蝇酶活性的监视器</td><td> Trikinetics公司，马萨诸塞州沃尔瑟姆</td><td> DAM2 </td><td>如其他地方6，本系统需要计算机接口，软件和其他配件。 </td></tr><tr><td>高硼硅玻璃管</td><td> Trikinetics公司，马萨诸塞州沃尔瑟姆</td><td> PGT-5x65 </td><td></td></tr><tr><td>微孔板闪烁和发光计数器</td><td>Perkin Elmer公司</td><td> TOPCOUNT NXT 12探测器</td><td>可以使用任何能够检测发光酶标仪为这种类型的报告基因分析。 TOPCOUNT包含多个检测器和一个自动化堆垛机，它是能够被编程，以连续读出从多个板。 </td></tr><tr><td>电泳凝胶成像8900 </td><td>阿尔法INNOTECH公司</td><td></td><td>细菌培养成像是可选的，任何一个数字成像系统与可视光的能力是足够的。 </td></tr><tr><td>微管牵引机</td><td> ，Tritech公司研究公司</td><td>成茂PC-10 </td><td></td></tr><tr><td></td><td></td><td></td><td> 耗材 </td></tr><tr><td>高硼硅玻璃毛细管</td><td>世界精密仪器有限公司</td><td> 1B100F-4 </td><td></td></tr><tr><td> 3毫升的注射器</td><td> Fisher Scientific则</td><td> BD 305482 </td><td></td></tr><tr><td>注射器针头</td><td> Fisher Scientific则</td><td> BD 305196 </td><td> 18 G - 切断的针的前端，以防止管子损坏。 </td></tr><tr><td>硅胶管，ID（0.030“）外径（0.065”）壁厚（0.018“） </td><td> VWR </td><td> 60985-706 </td><td>用于安装玻璃毛细管针注射器</td></tr><tr><td> 3通阀</td><td>美国Pharmaseal公司</td><td> K75 </td><td></td></tr><tr><td> Kontes颗粒杵无绳电机</td><td> Fisher Scientific则</td><td> K749540-0000 </td><td></td></tr><tr><td> Kontes颗粒杵</td><td> Fisher Scientific则</td><td> K749521-1590 </td><td></td></tr><tr><td>玻璃球3毫米</td><td> VWR </td><td> 26396-630 </td><td></td></tr><tr><td>微孔板微晶1 + </td><td> Thermo Scientific的</td><td> 7571 </td><td>范围吨96 - 孔板是适当的用于发光的 - 它们必须是不透明的。 </td></tr><tr><td> TopSeal-A :96井的微孔板</td><td>珀金埃尔默</td><td> 6005185 </td><td>微孔板按在粘接密封膜</td></tr><tr><td> D-荧光素钾盐</td><td>金生物技术公司</td><td> LUCNA </td><td></td></tr><tr><td></td><td></td><td></td><td> 软件 </td></tr><tr><td> Insomniac2 </td><td>可根据要求作者</td><td>定制编写的莱斯利阿什莫尔，博士（威斯敏斯特大学） </td><td>基于Matlab软件已被用于常规分析睡眠2,6,11 </td></tr><tr><td> Drosonex </td><td>可根据要求作者</td><td>自定义;写的托马斯·Coradetti（人行道软件） </td><td> A PC MSVC6计划，从原始数据文件，用于生存分析，收集与Trikinetics系统</td></tr><tr><td> Photoshop CS3的</td><td>土砖</td><td></td><td>可用于从数字图像获得号码的菌落形成单位/板（可选） </td></tr></tbody></table>

quantitative measurement of the immune response and sleep in drosophila

甲之间复杂的相互作用的免疫反应和主机行为已经描述了在很宽的范围内的物种。过量的睡眠，特别是，已知作为响应于在哺乳动物中1感染发生，最近也被描述在果蝇 2。人们普遍接受，睡眠在感染的主机是有益的，并且是很重要的一个强有力的免疫系统3,4的维护。然而，支持这一假说的实验证据表明，是有限的4，和多余的睡眠期间的免疫应答的功能尚不清楚。我们使用了多学科的方法来解决这个复杂的问题，果蝇果蝇的简单遗传模型系统，进行研究。我们使用标准的分析测量在果蝇的自发行为，睡眠，并演示了如何使用该法是衡量行为的苍蝇infecteD与细菌的致病性菌株的。该试剂盒也可以用于监测感染的持续时间期间在个别蝇的生存。免疫功能的其他措施包括清除感染，并是在果蝇的天然免疫反应的核心，一个关键的转录因子NFκB的激活能力的苍蝇。生存结果，并在感染细菌清除感染性和耐受性的指标。抵抗是指苍蝇清除感染的能力，而容差定义为主机的能力的限制感染造成的损害，从而尽管有很高水平的系统5内的病原体生存。在感染过程中的实时的NFκB活动的监测提供了洞察生存在感染过程中的分子机制。利用果蝇在这些简单的分析，有利于睡眠的遗传和分子生物学分析和免疫反应，这两个复杂的系统如何往复影响。

<ol><li>感染促进睡眠。在这个例子中，广交会-S（CS）的野生型苍蝇和突变果蝇缺乏NFκB基因，Relish的 （ 相对E20）14，装入两个DAM2活动监视器（每组32为每个基因型）和感染如上所述。苍蝇保持在恒定的光，消除影响生物钟的行为和感染2,7,8。 相对E20的突变体isogenized CS如前所述，11。这两组的苍蝇感染S.粘质沙雷氏菌 ，其结果如图3中所示。在?...

Watch this Scientific Journal Video about Quantitative Measurement of the Immune Response and Sleep in Drosophila at JoVE.com

Quantitative Measurement of the Immune Response and Sleep in Drosophila

要了解免疫反应和行为之间的联系，我们描述了一种方法来测量运动的行为中<em&gt;果蝇</em以及细菌感染的免疫反应能力的苍蝇安装监控求生存，细菌负荷，并实时活动的一个关键的调节先天免疫，NFκB。

定量测定的免疫反应和睡眠模式<em&gt;果蝇</em

A complex interaction between the immune response and  host behavior has been described in a wide range of species. Excess sleep, in  particular, is known ...

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Immunology and Infection

Quantitative Measurement of the Immune Response and Sleep in Drosophila

Multicolor Flow Cytometry Analyses of Cellular Immune Response in Rhesus Macaques

多色流流式细胞仪分析细胞在恒河猴的免疫应答

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科研

免疫与感染

An Introduction to Parasitic Wasps of Drosophila and the Antiparasite Immune Response

An Introduction to Parasitic Wasps of Drosophila and the Antiparasite Immune Response

寄生蜂简介<em&gt;果蝇</em&gt;和Antiparasite的免疫反应的

Most known parasitoid wasp species attack the larval or pupal stages of Drosophila. While Trichopria drosophilae infect the pupal stages of the host (Fig. ...

Quantification of the Respiratory Burst Response as an Indicator of Innate Immune Health in Zebrafish

呼吸爆发的响应为先天免疫健康的斑马鱼中的指标量化

The phagocyte respiratory burst is part of the innate immune response to  pathogen infection and involves the production of reactive oxygen species (ROS). ...

Novel Whole-tissue Quantitative Assay of Nitric Oxide Levels in Drosophila Neuroinflammatory Response

Novel Whole-tissue Quantitative Assay of Nitric Oxide Levels in Drosophila Neuroinflammatory Response

Neuroinflammation is a complex innate immune response vital to the healthy function of the central nervous system (CNS). Under normal conditions, an ...

Using RNA-interference to Investigate the Innate Immune Response in Mouse Macrophages

利用RNA干扰调查的先天免疫反应的小鼠巨噬细胞

Macrophages are key phagocytic innate immune cells.  When macrophages encounter a pathogen, they produce antimicrobial proteins and compounds to kill the ...

The Utilization of Oropharyngeal Intratracheal PAMP Administration and Bronchoalveolar Lavage to Evaluate the Host Immune Response in Mice

口咽部气管内PAMP管理和支气管肺泡灌洗的利用率来评估小鼠宿主的免疫反应

The host immune response to pathogens is a complex biological process. The majority of in vivo studies classically employed to characterize host-pathogen ...

In Vitro Analysis of Myd88-mediated Cellular Immune Response to West Nile Virus Mutant Strain Infection

In Vitro Analysis of Myd88-mediated Cellular Immune Response to West Nile Virus Mutant Strain Infection

在 MyD88的介导的细胞免疫应答西尼罗河病毒变异株感染体外分析

An attenuated West Nile virus (WNV), a nonstructural (NS) 4B-P38G mutant, induced higher innate cytokine and T cell responses than the wild-type WNV in ...

Non-invasive Imaging of the Innate Immune Response in a Zebrafish Larval Model of Streptococcus iniae Infection

Non-invasive Imaging of the Innate Immune Response in a Zebrafish Larval Model of Streptococcus iniae Infection

先天免疫反应中的斑马鱼幼体型号无创成像<em&gt;链球菌iniae</em&gt;感染

The aquatic pathogen, Streptococcus iniae, is responsible for over 100 million dollars in annual losses for the aquaculture industry and is capable of ...

Qualitative and Quantitative Analysis of the Immune Synapse in the Human System Using Imaging Flow Cytometry

影像性流式细胞仪对人体免疫突触的定性和定量分析

The immune synapse is the area of communication between T cells and antigen-presenting cells (APCs). T cells polarize surface receptors and proteins ...

Two-photon Intravital Imaging of Leukocytes During the Immune Response in Lipopolysaccharide-treated Mouse Liver

脂多糖治疗小鼠肝脏免疫应答中白细胞的双光子活体成像

Sepsis is a type of severe infection that can cause organ failure and tissue damage. Although the mortality and morbidity rates associated with sepsis are ...