Name: measurement of larval activity in the drosophila activity monitor
Uploaded: 2015-04-30T14:00:00.000+00:00
Duration: 9 min 30 s
Description: Watch this Scientific Journal Video about Measurement of Larval Activity in the Drosophila Activity Monitor at JoVE.com

This work was supported by NIH P20GM103643 to I. Meng.

1.准备幼虫<ol><li>为了分析运动或立场偏好所需的幼虫，在标准条件下生长的幼虫到所需的年龄标准使用飞食品11至10测定。 </li><li>请通过拉伸丝印级尼龙网在漏斗网状过滤器。固定网在漏斗颈部橡皮筋。放置在一个烧杯中的漏斗。 </li><li>为了收集幼虫进行分析，舀一铲满包含从培养瓶觅食幼虫的食物，并用RT自来水冲洗过的筛网过滤，收集直接个人幼虫网用画笔10。 </li></ol> 2.准备测定管<ol><li>要准备堵塞试管，精心熬了4％琼脂凝胶倒入培养皿中，以1.5厘米的深度。这将提供一个插头用于管的任一侧时动物被封闭。在4％的溶液是足够致密以防止由对幼虫enetrating插头。 </li><li>确保管是干净，清晰之前，插入幼虫。如果不是，这可能会阻止从被记录的动作。 </li><li>以保证幼虫有足够的水分运动，制备含有少量的热水挤压瓶。倒置的酒瓶，仔细定位走向水槽出口和压榨轻轻地从摄像管排出的水。 </li><li>将瓶子口进入测试管。挤压瓶子提供水汽进入测试管，直到薄层冷凝出现在管的壁。视频1示出了幼虫移动在管与水分的适当的量。 </li><li>以密封幼虫在管，取出从培养皿1.5厘米厚的琼脂凝胶，并放置在一网状表面，以允许气流琼脂下方以使得琼脂塞可插入到管。按该测定管的一端插入的凝胶两次，以插入两个插头凝胶成一端。 </li><li>随着paintb匆忙，将一个幼虫进入管约1.5英寸深，按最近的幼虫年底到琼脂凝胶密封管。所产生的压力将迫使出琼脂凝胶的第二插头从相对端和在管内密封该动物。 </li><li>地方管在MB5多波束果蝇活动监视器（DAM）装置，并调整管&#39;的位置，以使动物不能超越传感器的范围内。 </li><li>为了确保测定管在运输过程中不滑出设备的红外线读取帧时，通过紧固油灰的环每个管在此其接触记录装置周围将灯管保持到位。 </li></ol> 3.测量活动<ol><li>在培养箱中记录活动，以防止可能由于阴影，荧光灯或温度变化在实验室发生的不准确的读数。参见图5为系统的一个建议的布置。 </li><li>设置一个孵化至20℃（ 见图1）。为了避免虚假记载，由于从白炽灯光源的干扰录制过程中，关掉日光灯灯光孵化器，并使用单独的LED光源，同时记录在孵化器。没有任何动物进行试验，以确保光线条件并不异常触发的传感器。应该有这个测试后无记录的移动数据。 </li><li>让幼虫以适应到20℃培养箱设置开始试验前5分钟。 </li><li>要设置DAM系统以所需记录间隔（ 如 1分钟），确保大坝系统录音软件下载到电脑主机12和前打开连接到录音室到PSIU接口DAM系统文件。 </li><li>设置所需的记录频率数据将被保存，选择首选项，然后单击上方或读取间隔选项下方，选择不同的时间框架中的数据将被存储。例如，选择阅读的间隔时间从1秒到1小时。 </li><li>要选择不同的参数记录数据，选择首选项，然后选择相应的框中下输出数据类型（计数，移动，位置和纠缠，见表1）。每个参数提供在管内的幼虫活性的独特的分析。 </li><li>要开始录制，六硼化钙用电话线将显示器连接到电源接口单元（PSIU）。该PSIU连接到电源插座。绿灯会显示相应的连接。 </li><li>通过通用串行总线（USB）电缆到Macintosh或Windows PC进行数据记录连接PSIU。在电脑上打开DAM SystemMB1v6x程序自动开始记录数据。 </li><li>后所需采集时间完成后，选择退出，那么现在退出DAM活动屏幕上;数据将在DAM系统数据自动保存。借此数据文件（ 例如 ，监控1）并拖动<html>到一个单独的文件夹。这将从坝软件存储原始数据，使他们可以在以后进行处理。 </li></ol> 4.准备用于数据处理（DAM FileScan） <ol><li>为了处理原始数据转换成可以理解的格式打开DAM FileScan应用程序并选择选择输入数据的文件夹。然后选择数据文件夹和所需文件，并选择扫描选项。最后选择的bin长到所需的读取周期。这将原始数据组织成由操作者设定的参数，程序将报告收集在该特定范围内的数据。 </li><li>选择输出的数据类型来分析所需要的运动设定（监控计数，移动，住）。下的额外读数菜单选择总和成箱（这是如果仓长度大于所选择的系统的读取间隔）。 </li><li>适当命名的文件并保存。该文件将被存储在相同的位置，从步骤3.9的文件夹。 </li></ol>_title"&gt; 5。访问数据的分析<ol><li>为了收集平均移动，过程从第4步生成为了生成一个电子表格程序表中的原始数据，打开以前保存的文件（步骤4.3），并显示文本导入向导窗口时，选择完成打开在电子表格格式的数据文件。 注:根据分析的数据类型，电子表格将被读取不同。通常情况下测量，研究的时间段将数字记录监视器移动或计数，而每一个人的阅读管会收到一个指定的字母。当在电子表格中查看数据，列KZ表示用于分别测定管1-16的槽。该行表示所收集的数据点。 </li><li>例如，如果收集在1分钟的间隔20分钟的移动，平均的20行来计算移动/分钟等测量的平均数目。 </li></ol>

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The authors declare that they have no competing financial interests.

果蝇幼虫的活性是由多种因素，包括基因型8,13年龄和环境温度2的影响。虽然能够非常详细的分析，强大的录像制品的方法已经开发了那些谁学习运动5，这种详细程度可能是多余的为那些谁希望确定活动的基本参数。这里介绍的方法采用的设备是许多实验室使用，操作方便，产生高重现性的结果，即使是对于那些主要的研究重点是不运动管理。的例子的结果表明，该法可用于检测幼虫经受不同的温度（ 图1）和不同基因型（ 图2）运动性显著变化。 当幼虫，测定在温度范围从5℃ - 35℃，其活性随温度的，除了在20℃至25℃（ 图1）的趋势休息。已经表明通过艾因斯利及其同事认为觅食早期第三龄幼虫喜欢在+/- 2℃的典型25℃的培养温度下。然而，当幼虫进入徘徊中旬第三龄阶段，他们更喜欢有点稍凉2。这一发现是与观察，对于第三龄幼虫活动能力大于在20℃比25℃的一致，这表明动物测定的某些部分是中徘徊的阶段，更积极的更低的温度，而较少在在25℃的正常培养温度。 此方法提供了简洁性，客观性和健壮的吞吐量，但也有局限性。上述的应用程序代表发生在相对短的时间帧测定，这部分是因为目前的设置不公关奥维德幼虫的食物源。保证充足的营养，有必要研究过很长时间的变化，活动或测量昼夜节律在较长时期。在4％琼脂塞可能会限制该室和外部环境，这可能导致幼虫经历低氧条件之间的气体交换。然而，这似乎并不影响一个20分钟测定期间内的活性，因为当每幼虫分钟平均移动期间的每分钟期间进行了分析，幼虫并未以显示活性的任何改变在记录期间（ 图4） 。 由于该设备记录的位置继续它代表捕获更多的运动相比，列举了非自动化的方法有所改进，但有些运动的确逃过检测。非常动物的小运动可能不会触发从该设备的响应，并且幼虫可在圆周的方式中的一个下文将红色光束不破坏相邻光束，导致不准确的低的读数。然而，因为这种类型的错误，预计将发生在所有处理组中，这是不太可能导致错误的结果。虽然第三龄幼虫是这种分析的主要焦点，该装置能够测量所述小得多的第一和第二龄幼虫的运动，以及（ 图3）的。正如预期的那样，每分钟在年轻动物记录移动数比上述更大第三龄动物低。 尽管此设备的全范围的用途尚未被证实，有各种其它修改，可以多样化涉及幼虫研究该装置的用途。例如，该设备允许"停留"测量，其表示时间，在管的确定区域花费。当果蝇各种受雇于这可能会提供有价值的信息幼虫一出租车ssays。通过放置在其一侧的装置，使得管被定向垂直而非水平，人们可以测量幼虫趋地性。为了测量趋光性，光梯度可以建立在管，检测是否幼虫有偏向于光或它的缺席。为了研究趋化作用，一个测试化学物质可以放在琼脂插头和幼虫的位置中的一个可能然后揭示确定它们偏爱或避免化学品。 显示器系统允许各个运动参数，概括在表1通过预实验设置期间选择的所有参数的分析（参见步骤3.6），实验者可以选择在测定后分析哪些参数。然而，如果任何设置未选择，则该数据将不提供对事后分析。应当指出的是，每个时间段选择后​​，数据被冻结在当前计数，并保存到主机。数据收集然后复位至零在此周期之后，再次开始，从而提供了一系列的时间间隔的数据点。必须手动退出，结束数据记录。 涉及该方法未来的研究将集中在使用中的停留参数和它的各种应用。也有可能建立一个协议，该协议将允许研究，以发生在较长的一段时间内，如昼夜研究，通过提供食物和交换的琼脂塞要更多的气体透过性材料14。水分含量将需要控制的好，因为干燥的条件下抑制运动15。目前，该协议提供了一种准确，简单，具有成本效益的方法，在各种实验条件下，以评估幼虫活动的基本参数。

The use of Drosophila as a genetic tool has transformed scientific knowledge of biological systems. Drosophila larvae have been used in a variety of studies including nociception1, development2 and as a model for the study of human disease genes3. Drosophila activity encompasses a range of behaviors that vary under different conditions including temperatures2, exposure to drugs4 and amongst different genotypes. Yet, despite the significant use of the larva as a model organism, a simple, standardized method to analyze larval activity has not been available. Presently, many larval locomotion studies employ sophisticated video analysis software5. While powerful, the complexity of such automated tools may discourage labs that are not already equipped to study locomotion from including analysis of informative activity parameters in their studies. In other current non-automated methods, such as the grid crawling analysis, motion is scored by a human observer, which introduces the possibility of subjectivity and limits throughput to one animal at a time6-7. A similar study used a 5-lane crawling assay, which measured the time it took larvae to travel a certain linear distance8. In such non-automated assays, displacement is measured but this does not account for non-linear travel between the start and end points. As discussed below, the method described here accounts for more of the actual larval movement, is objective, easy to operate, and offers robust throughput.
To easily study larval activity behavior without the compromise of accuracy, efficiency, or cost, this method employs the TriKinetics Drosophila Activity Monitor (DAM), a device often used to study adult activity. Using one device to study both adult and larval activity is cost-effective, and allows direct comparison of motion by animals at these two life stages. The system, featuring the manufacturer&#8217;s highest level of resolution, makes use of 17 infrared detection beams per assay tube, which record larval activity when sensor beams are broken within the 16 individual tubes. The system then automatically saves recorded information to a computer, making it available for manipulation with basic graphing software. The data obtained represents the beams that were broken by individual larvae (which can be converted into a rate), movement when a larva stays within a detector beam and the position of the animals within the assay chambers during a recording period (allowing one to calculate position preference). The system is efficient and relatively simple to operate, and brings highly reproducible basic activity analysis within the reach of any laboratory studying Drosophila larvae.
To demonstrate the power of this assay, data are presented that show its use to verify differences in activity resulting from varying ambient temperatures, as well as through the comparison of a mutant previously described as hypoactive (iav1)9 with a widely analyzed control (w1118).

<table><tbody><tr><td>Drosophila Activity Monitor, Multibeam, 16 tubes, including wires</td><td>TriKinetics Inc.&amp;nbsp;</td><td>MB5</td><td></td></tr><tr><td>Power Supply Interface for Activity Monitor&amp;nbsp;</td><td>TriKinetics Inc.&amp;nbsp;</td><td>PSIU24</td><td></td></tr><tr><td>Glass 80 x 5 mm tubes for Activity Monitor (100)</td><td>TriKinetics Inc.&amp;nbsp;</td><td>PGT 5x80</td><td></td></tr><tr><td>DAMsystemMB1v6x Data Acquisitions Software for Macintonsh OSX (Intel)</td><td>&lt;a href=&quot;http://www.trikinetics.com/&quot;&gt;www.trikinetics.com&lt;/a&gt;</td><td></td><td>free download</td></tr><tr><td>DAMFileScan 108x software for Macintosh</td><td>&lt;a href=&quot;http://www.trikinetics.com/&quot;&gt;www.trikinetics.com&lt;/a&gt;</td><td></td><td>free download</td></tr><tr><td>USB software (PSIUdrivers.zip).</td><td>&lt;a href=&quot;http://www.trikinetics.com/&quot;&gt;www.trikinetics.com&lt;/a&gt;</td><td></td><td>free download</td></tr><tr><td>DAMSystem Notes 308</td><td>&lt;a href=&quot;http://www.trikinetics.com/&quot;&gt;www.trikinetics.com&lt;/a&gt;</td><td></td><td>free download</td></tr><tr><td>Zeiss Stemi 2000C- Stereo Microscope</td><td>Spectra Services</td><td>SP-STEMI2000C-BS</td><td></td></tr><tr><td>Carbon Dioxide</td><td>Maine Oxy</td><td></td><td>anaesthesia</td></tr><tr><td>Fly Pad</td><td>Genesee</td><td>59-114</td><td>surface for sorting anaesthetized flies</td></tr><tr><td>Small paint brush&amp;nbsp;</td><td>Winsor &amp;amp; Newton</td><td>#2 ROUND</td><td>or similar, used for sorting anaesthetized flies</td></tr><tr><td>Silk Screen Printing Mesh (160)</td><td>msj-gallery.com</td><td>SM160W63-3YD</td><td>pore sized used in this protocol was ~ 0.1 mm</td></tr><tr><td>Tegosept</td><td>Genesee</td><td>20-258</td><td>preservative</td></tr><tr><td>Ethanol (190proof)</td><td>Pharmco</td><td>111000190</td><td>used to dissolve Tegosept</td></tr><tr><td>6 oz Square Bottom Bottle (PP)</td><td>Genesee</td><td>32-130</td><td></td></tr><tr><td>&quot;Flugs&quot; for Plastic Fly bottles</td><td>Genesee</td><td>49-100</td><td></td></tr><tr><td>Drosophila Vials, Wide (PS)</td><td>Genesee</td><td>32-117</td><td></td></tr><tr><td>Flugs for wide plastic vials</td><td>Genesee</td><td>49-101</td><td></td></tr><tr><td>Yellow Degerminated Corn Meal</td><td>Gold Medal</td><td></td><td></td></tr><tr><td>Drosophila agar</td><td>LabScientific</td><td>FLY 8020</td><td></td></tr><tr><td>Baker&#039;s Yeast - Red Star</td><td>King Arthur Flour</td><td>1270</td><td></td></tr><tr><td>Granulated Sugar - Extra Fine</td><td>Domino</td><td></td><td></td></tr></tbody></table>

measurement of larval activity in the drosophila activity monitor

Drosophila larvae are used in many behavioral studies, yet a simple device for measuring basic parameters of larval activity has not been available. This protocol repurposes an instrument often used to measure adult activity, the TriKinetics Drosophila activity monitor (MB5 Multi-Beam Activity Monitor) to study larval activity. The instrument can monitor the movements of animals in 16 individual 8 cm glass assay tubes, using 17 infrared detection beams per tube. Logging software automatically saves data to a computer, recording parameters such as number of moves, times sensors were triggered, and animals&#8217; positions within the tubes. The data can then be analyzed to represent overall locomotion and/or position preference as well as other measurements. All data are easily accessible and compatible with basic graphing and data manipulation software. This protocol will discuss how to use the apparatus, how to operate the software and how to run a larval activity assay from start to finish.

图1示出了从控制第三龄幼虫的温度下反应的研究中，瓦特1118的结果，使用的监控装置，以检测在七个不同的温度下在幼虫运动的差异。幼虫洗涤并如上所述放入DAM活动装置，并放置在培养箱设置为所需的温度。该装置然后使其适应环境5分钟记录开始之前。每个幼虫单独为运动在20分钟内和每分钟移动平均数分析，计算为每一个动物，平均为每组32只动物。数据进行分析，并使用电子表格程序作图。幼虫表现出显著增加活性温度从5-35℃，相应增加在5度增量，除了在这一趋势中断在20℃和25℃。 为了验证差异腠LD被控制并检测先前描述的机能减退， 不活动的幼虫（IAV 1）突变体之间检测。数据进行了分析作为运动/分钟对每个32动物，然后计算平均值。 如图2所示 ，分析表明， 不活动的幼虫比的控制移动显著以下。而它们比第三龄幼虫小得多，第一和第二龄幼虫的活性也可测量， 如图3活性在20分钟测定每分钟三龄幼虫被证明保持整个期间相对一致（ 图4）。 <img alt="图1" src="/files/ftp_upload/52684/52684fig1.jpg" /> EAC 35°C -图1. 1118W¯¯幼虫运动被记录在温度5℃的变化。H柱代表32三龄动物，每分钟平均跻身组单独的移动平均运动。 *所有平均数显著彼此不同除了10°C和15°C（p值= 0.116）（不同字母表示一个显著差异，学生t-检验）。 <img alt="图2" src="/files/ftp_upload/52684/52684fig2.jpg" /> 图2.第三龄控制幼虫相比无效 （IAV 1）幼虫在20℃。IAV 1幼虫表现出显著少流动性与对照相比。 <img alt="图3" src="/files/ftp_upload/52684/52684fig3.jpg" /> 图3.测量第一，第二和第三龄幼虫在20分钟的记录间隔的活性在20℃。N = 32每个幼虫阶段。 图4.直方图显示的，在20分钟的记录间隔的每分钟发生移动平均数。32第三龄幼虫进行测定，在20℃。红线代表每分钟移动数平均到整个20分钟时间间隔。 <img alt="图5" src="/files/ftp_upload/52684/52684fig5.jpg" /> 图5.示出了果蝇活动监视器的设置和其连接到PSIU接口单元和桌面计算机。培养箱的内部被示出，但在记录所述培养箱被关闭。 <table border="0" cellpadding="0" cellspacing="0"><colgroup><col span="2" /></colgroup><tbody><tr><td>描述</td><td>参数</td></tr><tr><td>每次数据记录幼虫跨越梁，以及额外的计数被记录，如果幼虫移动，同时单个波束内。 </td><td>计数</td></tr><tr><td>记录数据只有当苍蝇分离的光束之间重新定位，但它不是一个波束内记录运动。 </td><td>移动</td></tr><tr><td>记录每一秒的动物位置在一个记录间隔。此数据表明立场的偏爱时间在每个传感器花了一个试验的过程中的功能。 </td><td>住</td></tr><tr><td>此设置分析动物的一般位置的管内，这表明运动传感器在动物触发。 </td><td>位置</td></tr><tr><td>此设置决定了一组时限内收集的数据被保存到桌面计算机的频率。 </td><td>记录间隔</td></tr></tbody></table> 表1描述measu的rement参数（计数，移动，停留和位置）。 <img alt="视频1" src="/files/ftp_upload/52684/52684vid1.jpg" /> 之前的视频1. DAM系统检测管插入到操作系统的设备。水分被冷凝呼吸提供。这个水平足以维持整个研究期间幼虫运动，而不会导致动物浮动或游泳。 <a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/52684/McParlandlarvamovinginmoistassaytube.mov" target="_blank">请点击此处观看该视频。</a>

Watch this Scientific Journal Video about Measurement of Larval Activity in the Drosophila Activity Monitor at JoVE.com

Measurement of Larval Activity in the Drosophila Activity Monitor

This report describes a method for measuring Drosophila larval activity using the TriKinetics Drosophila Activity Monitor. The device employs infrared beams to detect movements of up to 16 individual animals. Data can be analyzed to represent motion parameters including rates and the positions of the animals within the assay chambers.

幼虫活动的测量<em&gt;果蝇</em&gt;活动监视器

Drosophila larvae are used in many behavioral studies, yet a simple device for measuring basic parameters of larval activity has not been available. This ...

measurement-of-larval-activity-in-the-drosophila-activity-monitor

Nanyang Technological University

Research

JoVE Journal

Biology

Measurement of Larval Activity in the Drosophila Activity Monitor

9.3K 次观看.  University of New England. This report describes a method for measuring Drosophila larval activity using the TriKinetics Drosophila Activity Monitor. The device employs infrared beams to detect movements of up to 16 individual animals. Data can be analyzed to represent motion parameters including rates and the positions of the animals within the assay chambers.

视频: Measurement of Larval Activity in the Drosophila Activity Monitor

Assaying Locomotor Activity to Study Circadian Rhythms and Sleep Parameters in Drosophila

Most life forms exhibit daily rhythms in cellular, physiological and behavioral phenomena that are driven by endogenous circadian (&equiv;24 hr) pacemakers or clocks. Malfunctions in the human circadian system are associated with numerous diseases or disorders. Much progress towards our understanding of the mechanisms underlying circadian rhythms has emerged from genetic screens whereby an easily measured behavioral rhythm is used as a read-out of clock function. Studies using Drosophila have made seminal contributions to our understanding of the cellular and biochemical bases underlying circadian rhythms. The standard circadian behavioral read-out measured in Drosophila is locomotor activity. In general, the monitoring system involves specially designed devices that can measure the locomotor movement of Drosophila. These devices are housed in environmentally controlled incubators located in a darkroom and are based on using the interruption of a beam of infrared light to record the locomotor activity of individual flies contained inside small tubes. When measured over many days, Drosophila exhibit daily cycles of activity and inactivity, a behavioral rhythm that is governed by the animal's endogenous circadian system. The overall procedure has been simplified with the advent of commercially available locomotor activity monitoring devices and the development of software programs for data analysis. We use the system from Trikinetics Inc., which is the procedure described here and is currently the most popular system used worldwide. More recently, the same monitoring devices have been used to study sleep behavior in Drosophila. Because the daily wake-sleep cycles of many flies can be measured simultaneously and only 1 to 2 weeks worth of continuous locomotor activity data is usually sufficient, this system is ideal for large-scale screens to identify Drosophila manifesting altered circadian or sleep properties.

Assaying Locomotor Activity to Study Circadian Rhythms and Sleep Parameters in Drosophila

We describe procedures for recording daily locomotor activity rhythms of Drosophila and subsequent data analysis. Locomotor activity rhythms are a reliable behavioral output of animal circadian clocks and are used as the standard readout of clock function when studying circadian mutants or examining how the environment regulates the circadian system.

含量自发活动，研究昼夜节律和睡眠参数果蝇</em

Most life forms exhibit daily rhythms in cellular, physiological and behavioral phenomena that are driven by endogenous circadian (&equiv;24 hr) ...

科研

神经科学

Measuring Exercise Levels in Drosophila melanogaster Using the Rotating Exercise Quantification System (REQS)

Drosophila melanogaster is a new model organism for studies in exercise biology. To date, two main exercise systems, the Power Tower and the Treadwheel have been described. However, a method to measure the amount of additional animal activity induced through the exercise treatment has been lacking. The Rotating Exercise Quantification System (REQS) fills this need, providing a measure of animal activity for animals experiencing rotational exercise. This protocol details how to use the REQS to assess animal activity during rotational exercise and illustrates the type of data that can be generated. Here, we demonstrate how the REQS is used to measure sex- and strain-specific differences in exercise induced activity. The REQS can also be used to evaluate the impact of various other experimental parameters such as age, diet, or population size on exercise induced activity. In addition, it can be used to compare the efficacy of different exercise training protocols. Importantly, it provides an opportunity to standardize exercise treatments between strains, allowing the researcher to achieve equal amounts of activity between groups if needed. Thus, the REQS is a notable new resource for exercise biologists working with the Drosophila model system and complements existing exercise systems.

Measuring Exercise Levels in Drosophila melanogaster Using the Rotating Exercise Quantification System REQS

The Rotating Exercise Quantification System (REQS) can induce exercise in Drosophila melanogaster through rotation while simultaneously measuring the amount of activity performed by the animals. Here, we present a point-by-point protocol detailing how to measure activity levels of animals experiencing rotational exercise treatments using the REQS.

使用旋转运动量化系统测量果蝇中的运动水平 (满足要求)

Drosophila melanogaster is a new model organism for studies in exercise biology. To date, two main exercise systems, the Power Tower and the Treadwheel ...

遗传学

A Computational Method to Quantify Fly Circadian Activity

In most animals and plants, circadian clocks orchestrate behavioral and molecular processes and synchronize them to the daily light-dark cycle. Fundamental mechanisms that underlie this temporal control are widely studied using the fruit fly Drosophila melanogaster as a model organism. In flies, the clock is typically studied by analyzing multiday locomotor recording. Such a recording shows a complex bimodal pattern with two peaks of activity: a morning peak that happens around dawn, and an evening peak that happens around dusk. These two peaks together form a waveform that is very different from sinusoidal oscillations observed in clock genes, suggesting that mechanisms in addition to the clock have profound effects in producing the observed patterns in behavioral data. Here we provide instructions on using a recently developed computational method that mathematically describes temporal patterns in fly activity. The method fits activity data with a model waveform that consists of four exponential terms and nine independent parameters that fully describe the shape and size of the morning and evening peaks of activity. The extracted parameters can help elucidate the kinetic mechanisms of substrates that underlie the commonly observed bimodal activity patterns in fly locomotor rhythms.

A method to quantify the main temporal features seen in fly circadian locomotor rhythms is presented. The quantification is achieved by fitting fly activity with a multi-parametric model waveform. The model parameters describe the shape and size of the morning and evening peaks of daily activity.

量化飞行昼夜活动的一种计算方法

In most animals and plants, circadian clocks orchestrate behavioral and molecular processes and synchronize them to the daily light-dark cycle. ...

行为学

Quantitative Measurement of the Immune Response and Sleep in Drosophila

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 to occur as a response to infection in mammals 1 and has also recently been described in Drosophila melanogaster2. It is generally accepted that sleep is beneficial to the host during an infection and that it is important for the maintenance of a robust immune system3,4. However, experimental evidence that supports this hypothesis is limited4, and the function of excess sleep during an immune response remains unclear. We have used a multidisciplinary approach to address this complex problem, and have conducted studies in the simple genetic model system, the fruitfly Drosophila melanogaster. We use a standard assay for measuring locomotor behavior and sleep in flies, and demonstrate how this assay is used to measure behavior in flies infected with a pathogenic strain of bacteria. This assay is also useful for monitoring the duration of survival in individual flies during an infection. Additional measures of immune function include the ability of flies to clear an infection and the activation of NF&kappa;B, a key transcription factor that is central to the innate immune response in Drosophila. Both survival outcome and bacterial clearance during infection together are indicators of resistance and tolerance to infection. Resistance refers to the ability of flies to clear an infection, while tolerance is defined as the ability of the host to limit damage from an infection and thereby survive despite high levels of pathogen within the system5. Real-time monitoring of NF&kappa;B activity during infection provides insight into a molecular mechanism of survival during infection. The use of Drosophila in these straightforward assays facilitates the genetic and molecular analyses of sleep and the immune response and how these two complex systems are reciprocally influenced.

Quantitative Measurement of the Immune Response and Sleep in Drosophila

To understand a link between the immune response and behavior, we describe a method to measure locomotor behavior in Drosophila during bacterial infection as well as the ability of flies to mount an immune response by monitoring survival, bacterial load, and real-time activity of a key regulator of innate immunity, NF&kappa;B.

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 ...

免疫与感染

High-Resolution Video Tracking of Locomotion in Adult Drosophila Melanogaster

Flies provide an important model for studying complex behavior due to the plethora of genetic tools available to researchers in this field. Studying locomotor behavior in Drosophila melanogaster relies on the ability to be able to quantify changes in motion during or in response to a given task. For this reason, a high-resolution video tracking system, such as the one we describe in this paper, is a valuable tool for measuring locomotion in real-time. Our protocol involves the use of an initial air pulse to break the flies momentum, followed by a thirty second filming period in a square chamber. A tracking program is then used to calculate the instantaneous speed of each fly within the chamber in 10 msec increments. Analysis software then compiles this data, and outputs a variety of parameters such as average speed, max speed, time spent in motion, acceleration, etc. This protocol will discuss proper feeding and management of flies for behavioral tasks, handling flies without anesthetization or immobilization, setting up a controlled environment, and running the assay from start to finish.

The study of complex locomotor behavior in Drosophila melanogaster is dependent upon the ability to quantify changes in a given fly's movement. This article demonstrates how to do this using a high-resolution tracking system.

高分辨率视频跟踪移动的，在成年果蝇。

Flies provide an important model for studying complex behavior due to the plethora of genetic tools available to researchers in this field.  Studying ...

生物学

Determination of the Spontaneous Locomotor Activity in Drosophila melanogaster

Drosophila melanogaster has been used as an excellent model organism to study environmental and genetic manipulations that affect behavior. One such behavior is spontaneous locomotor activity. Here we describe our protocol that utilizes Drosophila population monitors and a tracking system that allows continuous monitoring of the spontaneous locomotor activity of flies for several days at a time. This method is simple, reliable, and objective and can be used to examine the effects of aging, sex, changes in caloric content of food, addition of drugs, or genetic manipulations that mimic human diseases.

Determination of the Spontaneous Locomotor Activity in Drosophila melanogaster

Drosophila melanogaster are useful in studying genetic or environmental manipulations that affect behaviors such as spontaneous locomotor activity.&#160; Here we describe a protocol that utilizes monitors with infrared beams and data analysis software to quantify spontaneous locomotor activity.&#160;

在自发活动的测定<em&gt;果蝇</em

Drosophila melanogaster has been used as an excellent model organism to study environmental and genetic manipulations that affect behavior. One such ...

A Low-cost Method for Analyzing Seizure-like Activity and Movement in Drosophila

Video tracking systems have been used widely to analyze Drosophila melanogaster movement and detect various abnormalities in locomotive behavior. While these systems can provide a wealth of behavioral information, the cost and complexity of these systems can be prohibitive for many labs. We have developed a low-cost assay for measuring locomotive behavior and seizure movement in D. melanogaster. The system uses a web-cam to capture images that can be processed using a combination of inexpensive and free software to track the distance moved, the average velocity of movement and the duration of movement during a specified time-span. To demonstrate the utility of this system, we examined a group of D. melanogaster mutants, the Bang-sensitive (BS) paralytics, which are 3-10 times more susceptible to seizure-like activity (SLA) than wild type flies. Using this novel system, we were able to detect that the BS mutant bang senseless (bss) exhibits lower levels of exploratory locomotion in a novel environment than wild type flies. In addition, the system was used to identify that the drug metformin, which is commonly used to treat type II diabetes, reduces the intensity of SLA in the BS mutants.

A Low-cost Method for Analyzing Seizure-like Activity and Movement in Drosophila

Using a webcam and a combination of free and inexpensive software programs, locomotive patterns in Drosophila melanogaster can be analyzed to detect differences in the speed, distance, and time of various motor activities.

在分析癫痫样活动和运动的一种低成本方法<em&gt;果蝇</em

Video tracking systems have been used widely to analyze Drosophila melanogaster movement and detect various abnormalities in locomotive behavior. While ...

A Simple Technique to Assay Locomotor Activity in Drosophila

Drosophila melanogaster is an ideal model organism for studying various diseases due to its abundance of advanced genetic manipulation techniques and diverse behavioral features. Identifying behavioral deficiency in animal models is a crucial measure of disease severity, for example, in neurodegenerative diseases where patients often experience impairments in motor function. However, with the availability of various systems to track and assess motor deficits in fly models, such as drug-treated or transgenic individuals, an economical and user-friendly system for precise evaluation from multiple angles is still lacking. A method based on the AnimalTracker application programming interface (API) is developed here, which is compatible with the Fiji image processing program, to systematically evaluate the movement activities of both adult and larval individuals from recorded video, thus allowing for the analysis of their tracking behavior. This method requires only a high-definition camera and a computer peripheral hardware integration to record and analyze behavior, making it an affordable and effective approach for screening fly models with transgenic or environmental behavioral deficiencies. Examples of behavioral tests using pharmacologically treated flies are given to show how the techniques can detect behavioral changes in both adult flies and larvae in a highly repeatable manner.

A Simple Technique to Assay Locomotor Activity in Drosophila

The present protocol assesses the locomotor activity of Drosophila by tracking and analyzing the movement of flies in a hand-made arena using open-source software Fiji, compatible with plugins to segment pixels of each frame based on high-definition video recording to calculate parameters of speed, distance, etc.

一种测定果蝇运动活性的简单技术

Drosophila melanogaster is an ideal model organism for studying various diseases due to its abundance of advanced genetic manipulation techniques and ...

<meta charset="utf8"></head><body>使用 DAM2 系统对 果蝇 进行高通量耐热性测量

Heat Tolerance Assays Using the Drosophila Activity Monitor System: A Guide to an Executable Application for Data Analysis

The study of heat tolerance in&#8239;Drosophila melanogaster has been of particular interest to researchers for decades, with a common approach to assessing heat tolerance being to monitor the time to knockdown (TKD) after exposure to an elevated temperature. Classically, flies are housed in individual vials and placed inside a heated water bath. TKD is then monitored manually by researchers. While very well-established, there remain problems of subjectivity and consistent application of a tangible definition of cessation of all movement, including muscular spasms, when implementing these manual assays. We have developed a high-throughput method for automating heat tolerance assays using the TriKinetics Drosophila Activity Monitors (DAM2). To accompany the DAM2 system, we have written a program and created an easy-use executable to automatically read the last time of movement from the activity data generated. This script then writes to a .csv file the time to heat paralysis (TKD) for each fly. Our data show that this automated DAM2 method is consistent and reliable. Meanwhile, activity profiles created from the activity count data are of interest. These activity profiles can be compiled and have the potential to expand heat tolerance assays to include the relatively unstudied behavioral components of heat tolerance. This protocol will describe in detail how to use the DAM2 system and the HoTDAM! software to estimate heat tolerance in D. melanogaster.

<meta charset="utf8"></head><body>使用 果蝇 活动监测系统进行耐热性测定:数据分析的可执行应用程序指南

This report describes a method for measuring adult Drosophila melanogaster time to knockdown using a Drosophila Activity Monitor (DAM2) in response to an air conduction heat stressor within an incubator chamber. The DAM2 measures activity by recording individual fly movements as they cross an infrared beam. Data analysis is facilitated by a novel executable file created by the authors.

幼虫活动的测量

摘要

探索更多视频

此视频中的章节

含量自发活动，研究昼夜节律和睡眠参数果蝇

使用旋转运动量化系统测量果蝇中的运动水平 (满足要求)

量化飞行昼夜活动的一种计算方法

定量测定的免疫反应和睡眠模式

高分辨率视频跟踪移动的，在成年果蝇。

在自发活动的测定

在分析癫痫样活动和运动的一种低成本方法

一种测定果蝇运动活性的简单技术

使用果蝇活动监测系统进行耐热性测定:数据分析的可执行应用程序指南

使用果蝇活动监测系统进行耐热性测定:数据分析的可执行应用程序指南

幼虫活动的测量

摘要

探索更多视频

此视频中的章节

含量自发活动，研究昼夜节律和睡眠参数果蝇

使用旋转运动量化系统测量果蝇中的运动水平 (满足要求)

量化飞行昼夜活动的一种计算方法

定量测定的免疫反应和睡眠模式

高分辨率视频跟踪移动的，在成年果蝇。

在自发活动的测定

在分析癫痫样活动和运动的一种低成本方法

一种测定果蝇运动活性的简单技术

使用 果蝇 活动监测系统进行耐热性测定:数据分析的可执行应用程序指南

使用 果蝇 活动监测系统进行耐热性测定:数据分析的可执行应用程序指南

使用果蝇活动监测系统进行耐热性测定:数据分析的可执行应用程序指南

使用果蝇活动监测系统进行耐热性测定:数据分析的可执行应用程序指南