这项工作是由美国国立卫生研究院（AG023088为BR）的资助。

注：广-S品系是从布卢明顿库存中心获得的标准野生型背景线。 1，食品制作和配方1000毫升食物注意：本节介绍的协议准备食物。大型金属罐是用来在同一时间准备大约18L的食物。这里所描述的协议的小型化，并使用千毫升H 2 O的食品灭菌两次。 <ol><li>混合113克蔗糖和28克啤酒酵母643毫升的水。留下的热板集配料，在25℃下用搅拌棒混合在整个15分钟。 </li><li> 20分钟高压灭菌食品的解决方案。 </li><li>混合49克玉米面和8.1克琼脂268毫升水中，并添加到在步骤1.2中所述的高压灭菌的食品混合物。用大汤匙或打蛋器拌匀。 </li><li>高压釜食品混合物另外20分钟。 </li><li>将食物放在盘子里，让降温持续混合与搅拌棒。我F额外的解决方案应该被添加到食品中，如米非司酮（RU486），保持食物的热板设置在60℃，并加入解决方案当食物达到所需要的温度。 </li><li>溶解2.4克tegosept在10.7毫升100％EtOH中，并保持在一个冷板有搅拌器，使其完全溶解并混合约15分钟。 </li><li>添加tegosept解决食物时，食物的温度为60°C并充分混合。 </li><li>使用泵或食品分配器倒约10ml的食品，用于广泛的小瓶中。通过使用食品贩卖机可以在同一时间同时倒入食物转化为100宽，塑料瓶（1盘）。 </li><li>盖上的Kimwipes和奶酪布的小瓶，把食物在室温下放置12-24小时冷却下来。保持食物在4℃和3-4周内使用。热身食物室温用于飞工作之前。 </li></ol> 2，准备玻璃瓶的<ol><li>根据步骤1中列出的协议准备食物。 </li&gt; <li>等分5毫升食物进入每个窄，玻璃小瓶，这是正确的大小为人口显示器。这个量的食物应足够低，以低于人口显示器的最低环。 </li><li>食后冷却至室温，覆盖海绵塞的小瓶中，并保持在4℃长达2周。因为食物在一个小瓶的量是相当低的，最好是用食物一两个星期内，以避免任何干燥。 </li><li>热身小瓶至室温后方可使用。 </li></ol>家长蝇3。维护<ol><li>成长的苍蝇在广塑小瓶用标准的实验室食物，并保持小瓶在湿润，温度控制环境室在25°C在12小时亮/暗周期。日光周期开始于6:00 AM在这个实验室。 </li><li>在上午的明确成蝇从该家长苍蝇会被收集瓶。 </li><li>收集新eclosed FLIEs和他们分开了性别上的内8小时的CO 2垫后羽化，以确保女性的苍蝇是处女。苍蝇开始羽化后交配8小时。 </li><li>当处女男和女苍蝇是5和10日龄之间，把男10例，女10苍蝇与标准食品和活性酵母在上面几个晶粒的小瓶。 注意：通过使用蝇数相同，保证它们在小瓶两天控制幼虫的密度。除了活性酵母的促进产蛋。 </li><li>保持苍蝇交配和产卵的温度控制的环境室在25℃，12小时的光/暗周期为2天。设置5-10小瓶父母苍蝇。 </li><li>通过苍蝇到一个新的塑料瓶每隔一天，时刻保持与鸡蛋的小瓶在孵化器在25°C。 </li></ol> 4，收集实验蝇<ol><li> 9天后苍蝇会从小瓶到E关闭那里的parenta升苍蝇产下的卵（在步骤3.6中所述）。清除并丢弃在第一天的eclosed苍蝇和返回小瓶孵化器。大多数eclosed第1天的苍蝇是女性。苍蝇更同步的人口将E关闭第2天。 </li><li>在24小时内到位新eclosed苍蝇对CO 2垫，并收集25名男性和25雌性果蝇每小瓶用画笔或金属汤匙。保持果蝇对CO 2垫很短的时间周期，以减少CO 2的任何影响。写在小瓶羽化的日子。聚集在至少5瓶重复实验和对照组。 </li><li>保持在温控环境箱小瓶在25℃与12小时的光/暗周期。 </li><li>用漏斗通过苍蝇到一个新的塑料瓶每隔一天。 </li><li>年龄苍蝇，直到实验所需的年龄为止。 </li></ol> 5，设置了移动显示器<ol><li>将在一个温度控制的恒温箱人口监视。 </li><li> 通过 5路分离器（多线），它可以连接多达5个单独的显示器一开口PSIU连接每台显示器配备4芯电话线连接到电源接口单元（PSIU）。请参阅图1A和2B。 </li><li>该PSIU连接线电源插座（100-240V）。将电源输出插头插入2交配PSIU的一个插口。相邻的绿色指示灯亮起绿色的正确连接时。 </li><li>该PSIU连接到通用串行总线（USB）的硬件。连接USB电缆USB硬件与Macintosh或Windows PC之间进行数据记录。这将是最好有一个既能用于数据采集专用电脑，因为集合在同一时间运行数天。 </li><li>下载的USB软件（PSIUdrivers.zip）。 USB驱动程序软件所使用的电源接口，需要下载一次。它合成一个该计算机程序和PSIU /活动监视器之间的数据链路。对于PC使用的COM端口和Macintosh上使用一个简单的串行端口。 </li><li>按照制造商注308.pdf提供的说明下载的计算机程序适用于Macintosh OSX（英特尔）或Windows PC上（XP/Vista/7的）方案。 </li><li>启动计算机，程序和设置程序通过点击首选项，灯光或显示器。该程序将运行，直到用户选择了“退出”以停止该程序。如果计算机程序或计算机关闭显示器将继续计数束中断，但计数不会被记录，直到程序重新启动。在这种情况下，该第一读取将包括自上次的PSIU发送的数据到计算机中的所有计数。 </li><li>选择首选项选项卡，然后选择串口，PSIU为Macintosh和COM的PC。 </li><li>选择的读取间隔，从秒不等，分钟或一个小时。 </li> &lt;LI&gt;选择显示器：每个显示器都有唯一的编号，是由制造商提供。选择显示器范围对应于由制造商给定的监视器的数量。 </li><li>灯框：确保所有的显示器已正确连接，它的特点是旁边上的软件监视器号码开了绿灯。红灯表示连接已丢失，一个黑盒子表示系统处于关闭或设置不正确了。 </li></ol> 6，设置实验<ol><li>从4℃取出装有食物的玻璃小瓶，让他们温暖至室温。 </li><li>单独的男性和女性的苍蝇一样的年龄对CO 2垫。对于老龄化的研究有可能早在3日龄开始流动的研究。 </li><li>把10名男或女10进苍蝇食物含有每个玻璃小瓶。使用至少三个小瓶中的每个实验组和对照线的苍蝇和每个性别。 </li><li>保持六站在他们一边ALS，直到苍蝇的CO 2回收，以保证苍蝇不被卡住的食物。独立的苍蝇在大约上午8:00，离开他们约2小时，在室温下由CO 2回收。 </li><li>将安置在孵化人口显示器内部的小瓶。 </li><li>弃去最初的24小时内收集的数据苍蝇放入孵化器后，让他们适应新的环境。 </li><li>通过后3天或4天的苍蝇到新的小瓶，以避免食物的干燥。如果苍蝇很容易死亡或者40日龄以上的老年人，收集后2天2天，并利用数据传递苍蝇。此外，使用每组三个以上的小瓶，以确保有足够的重复。从小瓶用死苍蝇的数据应被忽略，不包括在分析。 </li></ol> 7，运行活动监视器和计算的总自发活动<ol><li>选择首选项 - 间隔进行数据采集&lt;。BR /&gt;注：该计算机程序允许收集数据的时间间隔从1秒到60分钟。 10和30分钟周期已经被发现，而无需时间点的绝大多数提供有关流动性充足的信息。在选定的时间段，该程序将发送当前的总计数为每台监视器的计算机，并开始从零重新计数。该计算机程序存储在由计算机数据系统中创建一个新文件夹中的数据。收集在每个显示器中的数据被分别存储，和单独的文本文件的每个小瓶中创建的。只要在程序操作的数据被连续收集。 </li><li>在实验结束时，扫描使用FileScan110X用于Macintosh OSX（英特尔）或SystemMB108用于Windows的PC（XP/Vista/7的）程序的数据。 注：扫描程序消除了重复的读数，并确保该录音是完整的。 </li><li>保存一个特定的时间和p中收集的数据eriod天。选择一个实验名称，并从分析计算机数据文件夹复制文件。 注意：在这个时候，活动的时间间隔是可以改变的，并转换成不同的。原来的数据将保持存储在计算机的数据文件夹，只要他们没有被删除，可以检索到。 </li></ol> 8，数据分析<ol><li>收集在文本文件中的数据复制到Excel电子表格中的列进行数据分析。通过该软件收集的数据是在列，其中包含表示在一段由研究者选择的时间在单个显示器总活性号码。 注：收集每个监视器的数据是在单独的文本文件。有32列每个监视器。前六个栏是空的，只包含0;接下来的三个包含在底部环收集的数据，在中间，并且在顶环。的信道的其余部分可以被删除，因为它们不包含任何数据。每个环输出每次单个值。看截屏图2中的原始数据。 </li><li>计算总活性在期望时间内为每个代表活性的三种不同的红外光束的高度，收集的总和显示器。 注意：该时间段的范围可以从几个小时，24小时或数天。 </li><li>确定平均运动活性和3个屏幕表示3次生物学重复的标准偏差。 注意：这些数据可以用于统计显着性通过使用一定数量的测试分析。双尾学生t-检验，单向方差分析（ANOVA）和杜克HSD事后检验可用于确定在24小时前自发活动16的一些环境或遗传操作的影响。还有一些可以使用的和先前已发表​​17的其他方案。 </li></ol>

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P., Allada, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Processing circadian data collected from the Drosophila Activity Monitoring (DAM) System. Protoc. 11, (2010).</li><li>Ardekani, R., Tavar&#233;, S., Tower, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessing+senescence+in+Drosophila+using+video+tracking.">Assessing senescence in Drosophila using video tracking.</a> Methods Mol. Biol. 965, 501-516 (2013).</li></ol>

我们什么都没有透露。

苍蝇的自发活动是由许多因素，如年龄，遗传背景，性别和影响2,13,18,19。此外，环境因素，诸如食物，温度环境，除了不同的药物，和白天/黑夜光周期的卡路里含量可影响蝇的活性。例如，雄性苍蝇同龄具有更高的自发运动量相比，女性（ 图1）。因此，苍蝇相同年龄和性别的，应相互比较。当检验遗传操作上飞活性，如过表达或特定基因的功能丧失的效果，实验组和对照?...

果蝇， 果蝇 ，已被用来作为一种有价值的模式生物来研究相关 ​​复杂的行为，如学习和记忆，社会交往，侵略，滥用药物，睡眠，感觉功能，求偶，交配和1,2机制。已通过多种协议研究的一个行为是自发活动。负趋地性是用于测量果蝇活动开发了第一种方法，而这种协议包括测量苍蝇的那个到达小瓶的一定高度后的苍蝇都摇动到容器1,3的底部的百分比。此方法具有简单，廉价的优点，并且由于它不要求它能够在任何实验室中进行任何特殊的设备。它已被用来作为一种有价值的筛查工具，研究苍蝇的流动性3种不同的基因操作的效果。但是，它是时间和劳动力密集型的一ND有偏见的，由于小瓶和人类录音变量晃动的可能性。 在由快速迭代负趋地性（环）方法4,5，这需要飞瓶以下的苍蝇晃动底部照片的发展负趋地性方法进行了改进。本协议的优点是它的灵敏度和测试大量蝇小瓶在同一时间的可能性。然而，该协议仍具有人为错误的可能性，并只测量负趋地性。其他实验室使用了简单的观察在培养瓶，以确定自主活动6。 最近已经开发了几种视频记录系统，用于测量苍蝇的运动行为。一个视频监控协议提供时间记录7之前的调整。通过Slawson 等人描述的方法。还使用一个空气脉冲停止movemen吨，直到记录的开始，这有可能是应激对动物7。这种方法提供的平均速度，最大速度，时间花费在运动等。另一个三维跟踪系统测量个体的苍蝇在约0.2秒内自由飞行起飞8的最大速度信息。一种三维视频监控协议使用果蝇表达GFP和装有过滤器，允许荧光的检测来确定飞流动性9多台摄像机。在这个协议苍蝇往往表现出由于果蝇文化的形状圆柱形飞行模式，这是潜在的小瓶10。这种方法是通过使用一个圆顶，使测量两只苍蝇11的自发运动改善。使用相机来自动监控和量化果蝇的个人和社会行为的高通量方法一直还描述12。 Zou 等人开发，它使用两个电脑辅助相机记录一生的行为和动作，如静止，移动，飞行，吃，喝，或个别实蝇果实蝇死亡13一个行为监控系统（BMS）。几个其它视频系统已被开发并监控蝇行为活动14,15。 在这里，我们描述了量化果蝇的活动，利用人口监测的方法。这些监视器都装在温度和湿度控制孵化器在25°C下12小时昼夜光周期。每个群体都有监视器放置在环定位在3个不同高度的红外光束。每次一飞跨环移动它中断了红外线光束，其上记录由一个微处理器独立地记录和计数小瓶内蝇的活性。微处理器上传小瓶内的总活动的电脑用户定义intervaLS，可以从1秒至60分钟。这里介绍的方法提供充足的时间过得来适应新的环境，并允许苍蝇多达120人口的自发活动的同时测量。此外，我们描述准备食物，飞维护，设置在温度控制恒温箱的流动性人口的显示器，以及可能影响业绩的潜在因素。该方法可用于研究不同环境或遗传修改如何影响蝇自发活动。

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Sucrose FCC Food Grade 100 LB,</td>
			<td>Fisher Scientific MP Biomedicals</td>
			<td>ICN90471380</td>
			<td></td>
		</tr>
		<tr>
			<td>Brewer&#8217;s Yeast</td>
			<td>Fisher Scientific MP Biomedicals</td>
			<td>ICN90331280</td>
			<td></td>
		</tr>
		<tr>
			<td>Drosophila Agar Fine</td>
			<td>SciMart</td>
			<td>DR-820-25F</td>
			<td></td>
		</tr>
		<tr>
			<td>Cornmeal</td>
			<td>Fisher Scientific MP Biomedicals</td>
			<td>ICN90141125</td>
			<td></td>
		</tr>
		<tr>
			<td>Methyl4-hydroxybenzoate, tegosept</td>
			<td>Sigma</td>
			<td>H5501-5KG</td>
			<td></td>
		</tr>
		<tr>
			<td>EtOH</td>
			<td>Pharmco-AAPER</td>
			<td>111000200</td>
			<td></td>
		</tr>
		<tr>
			<td>Active Dry Yeast</td>
			<td>Fisher Scientific</td>
			<td>ICN10140001</td>
			<td></td>
		</tr>
		<tr>
			<td>Fly CO2 pad</td>
			<td>LabScientific</td>
			<td>BGSU-7</td>
			<td></td>
		</tr>
		<tr>
			<td>Stereo Microscope</td>
			<td>Olympus</td>
			<td>SZ40</td>
			<td></td>
		</tr>
		<tr>
			<td>Drosophila carbon dioxide (CO2) tank</td>
			<td>Airgas</td>
			<td>UN1013</td>
			<td></td>
		</tr>
		<tr>
			<td>Small paint brush for pushing the flies</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Shell vial wide</td>
			<td>Fischer Scientific</td>
			<td>AS519</td>
			<td></td>
		</tr>
		<tr>
			<td>Buzzplugs for wide plastic vials</td>
			<td>Fischer Scientific</td>
			<td>AS275</td>
			<td></td>
		</tr>
		<tr>
			<td>Glass vials (25x95mm)</td>
			<td>Fischer Scientific Kimble 60931-8</td>
			<td>AS-574</td>
			<td></td>
		</tr>
		<tr>
			<td>Sponge plugs for glass vials</td>
			<td>SciMart</td>
			<td>DR-750</td>
			<td></td>
		</tr>
		<tr>
			<td>Drosophila Food Dispenser</td>
			<td>Applied Scientific (Fischer Scientific)</td>
			<td>AS780Q</td>
			<td></td>
		</tr>
		<tr>
			<td>DPM Drosophila Population Monitor</td>
			<td>Trikinetics Inc.</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>DC Power Supply with line cord</td>
			<td>Trikinetics Inc.</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>PSIU9 The Power Supply Interface Unit</td>
			<td>Trikinetics Inc.</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Telephone cables and 5 way splitters</td>
			<td>Trikinetics Inc.</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Universal Serial Bus (USB) hardware</td>
			<td>Trikinetics Inc.</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Macintosh or Windows PC with UCB port</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>DAMSystem308X Data Acquisition Software for Macintoch OSX (Intel)</td>
			<td>www.trikinetics.com</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>DAMSystem308 Data Acquisition Software for Windows PC (XP/Vista/7)</td>
			<td>www.trikinetics.com</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>DAMFileScan108X software for Macintosh</td>
			<td>www.trikinetics.com</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>DAMFileScan108X software for Windows PC (XP/Vista/7)</td>
			<td>www.trikinetics.com</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>USB software (PSIUdrivers.zip)</td>
			<td>www.trikinetics.com</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>DAMSystem Notes 308</td>
			<td><a href="http://www.trikinetics.com/Downloads/DAMSystem%20Notes%20308.pdf">(http://www.trikinetics.com/Downloads/DAMSystem%20Notes%20308.pdf</a></td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

determination of the spontaneous locomotor activity in drosophila melanogaster

果蝇已被用来作为一个极好的模型有机体，以研究影响行为环境和遗传操作。一个这样的行为是自发活动。在这里，我们描述了我们的协议，它利用果蝇种群监测和跟踪系统，可连续监测苍蝇的自发活动的好几天的时间。该方法简单，可靠，客观和可以用来检查老化，性别，改变食物的卡路里含量，另外的药物，或模仿人类疾病的遗传操作的效果。

<html>在果蝇的自发活动依赖于飞性别（ 图3A），食品（ 图3B）的卡路里含量和光/暗周期。一旦灯被关闭飞活性显着降低。 图3A示出24小时的男性和女性的苍蝇自发活动记录。在x轴以星号标记当光被关掉的时间和过渡到暗循环。 图3B示出收集在三个群体监视器男性的平均自发活动之间的标准偏差拥有3日龄玉米食品。收集在24小时内自发运动?...

Watch this Scientific Journal Video about Determination of the Spontaneous Locomotor Activity in Drosophila melanogaster at JoVE.com

Determination of the Spontaneous Locomotor Activity in Drosophila melanogaster

果蝇是研究影响的行为，如自发活动的遗传或环境的操作非常有用。在这里，我们描述了一个协定，它利用监测与红外光束和数据分析软件来量化自发活动。

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 ...

determination-spontaneous-locomotor-activity-drosophila

Research

JoVE Journal

Neuroscience

Determination of the Spontaneous Locomotor Activity in Drosophila melanogaster

12.6K 次观看.  University of Connecticut Health Center.  果蝇是研究影响的行为，如自发活动的遗传或环境的操作非常有用。在这里，我们描述了一个协定，它利用监测与红外光束和数据分析软件来量化自发活动。 

视频: Determination of the Spontaneous Locomotor Activity in Drosophila melanogaster

Single Sensillum Recordings in the Insects Drosophila melanogaster and Anopheles gambiae

The sense of smell is essential for insects to find foods, mates, predators, and oviposition sites3. Insect olfactory sensory neurons (OSNs) are enclosed in sensory hairs called sensilla, which cover the surface of olfactory organs. The surface of each sensillum is covered with tiny pores, through which odorants pass and dissolve in a fluid called sensillum lymph, which bathes the sensory dendrites of the OSNs housed in a given sensillum. The OSN dendrites express odorant receptor (OR) proteins, which in insects function as odor-gated ion channels4, 5. The interaction of odorants with ORs either increases or decreases the basal firing rate of the OSN. This neuronal activity in the form of action potentials embodies the first representation of the quality, intensity, and temporal characteristics of the odorant6, 7.
Given the easy access to these sensory hairs, it is possible to perform extracellular recordings from single OSNs by introducing a recording electrode into the sensillum lymph, while the reference electrode is placed in the lymph of the eye or body of the insect. In Drosophila, sensilla house between one and four OSNs, but each OSN typically displays a characteristic spike amplitude. Spike sorting techniques make it possible to assign spiking responses to individual OSNs. This single sensillum recording (SSR) technique monitors the difference in potential between the sensillum lymph and the reference electrode as electrical spikes that are generated by the receptor activity on OSNs1, 2, 8. Changes in the number of spikes in response to the odorant represent the cellular basis of odor coding in insects. Here, we describe the preparation method currently used in our lab to perform SSR on Drosophila melanogaster and Anopheles gambiae, and show representative traces induced by the odorants in a sensillum-specific manner.

Single Sensillum Recordings in the Insects Drosophila melanogaster and Anopheles gambiae

Electrophysiological responses of olfactory sensory neurons to odorants can be measured in insects using single sensillum recordings. In this video article we will demonstrate how to perform single sensillum recordings in the antennae of the vinegar fly (Drosophila melanogaster) and the maxillary palps of the malaria mosquito (Anopheles gambiae).

在昆虫单感器记录果蝇按蚊</em

The sense of smell is essential for insects to find foods, mates, predators, and oviposition sites3. Insect olfactory sensory neurons (OSNs) are enclosed ...

科研

神经科学

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) ...

Assaying Locomotor, Learning, and Memory Deficits in Drosophila Models of Neurodegeneration

Advances in genetic methods have enabled the study of genes involved in human neurodegenerative diseases using Drosophila as a model system1. Most of these diseases, including Alzheimer's, Parkinson's and Huntington's disease are characterized by age-dependent deterioration in learning and memory functions and movement coordination2. Here we use behavioral assays, including the negative geotaxis assay3 and the aversive phototaxic suppression assay (APS assay)4,5, to show that some of the behavior characteristics associated with human neurodegeneration can be recapitulated in flies. In the negative geotaxis assay, the natural tendency of flies to move against gravity when agitated is utilized to study genes or conditions that may hinder locomotor capacities. In the APS assay, the learning and memory functions are tested in positively-phototactic flies trained to associate light with aversive bitter taste and hence avoid this otherwise natural tendency to move toward light. Testing these trained flies 6 hours post-training is used to assess memory functions. Using these assays, the contribution of any genetic or environmental factors toward developing neurodegeneration can be easily studied in flies.

Assaying Locomotor, Learning, and Memory Deficits in Drosophila Models of Neurodegeneration

Behavior assays for measuring locomotor functions, learning, and memory abilities in Drosophila.

含量的运动能力，学习和记忆的赤字果蝇</em神经退行性疾病>模型

Advances in genetic methods have enabled the study of genes involved in human neurodegenerative diseases using Drosophila as a model system1. Most of ...

Labeling of Single Cells in the Central Nervous System of Drosophila melanogaster

In this article we describe how to individually label neurons in the embryonic CNS of Drosophila melanogaster by juxtacellular injection of the lipophilic fluorescent membrane marker DiI. This method allows the visualization of neuronal cell morphology in great detail. It is possible to label any cell in the CNS: cell bodies of target neurons are visualized under DIC optics or by expression of a fluorescent genetic marker such as GFP. After labeling, the DiI can be transformed into a permanent brown stain by photoconversion to allow visualization of cell morphology with transmitted light and DIC optics. Alternatively, the DiI-labeled cells can be observed directly with confocal microscopy, enabling genetically introduced fluorescent reporter proteins to be colocalised. The technique can be used in any animal, irrespective of genotype, making it possible to analyze mutant phenotypes at single cell resolution.

Labeling of Single Cells in the Central Nervous System of Drosophila melanogaster

We present a technique for labeling single neurons in the central nervous system (CNS) of Drosophila embryos, which allows the analysis of neuronal morphology by either transmitted light or confocal microscopy.

标记的单细胞在中枢神经系统<em&gt;果蝇</em

In this  article we describe how to individually label neurons in the embryonic CNS of Drosophila melanogaster by juxtacellular injection of the ...

Optogenetic Stimulation of Escape Behavior in Drosophila melanogaster

A growing number of genetically encoded tools are becoming available that allow non-invasive manipulation of the neural activity of specific neurons in Drosophila melanogaster1. Chief among these are optogenetic tools, which enable the activation or silencing of specific neurons in the intact and freely moving animal using bright light. Channelrhodopsin (ChR2) is a light-activated cation channel that, when activated by blue light, causes depolarization of neurons that express it. ChR2 has been effective for identifying neurons critical for specific behaviors, such as CO2 avoidance, proboscis extension and giant-fiber mediated startle response2-4. However, as the intense light sources used to stimulate ChR2 also stimulate photoreceptors, these optogenetic techniques have not previously been used in the visual system. Here, we combine an optogenetic approach with a mutation that impairs phototransduction to demonstrate that activation of a cluster of loom-sensitive neurons in the fly's optic lobe, Foma-1 neurons, can drive an escape behavior used to avoid collision. We used a null allele of a critical component of the phototransduction cascade, phospholipase C-&beta;, encoded by the norpA gene, to render the flies blind and also use the Gal4-UAS transcriptional activator system to drive expression of ChR2 in the Foma-1 neurons. Individual flies are placed on a small platform surrounded by blue LEDs. When the LEDs are illuminated, the flies quickly take-off into flight, in a manner similar to visually driven loom-escape behavior. We believe that this technique can be easily adapted to examine other behaviors in freely moving flies.

Optogenetic Stimulation of Escape Behavior in Drosophila melanogaster

Genetically encoded optogenetic tools enable noninvasive manipulation of specific neurons in the Drosophila brain. Such tools can identify neurons whose activation is sufficient to elicit or suppress particular behaviors. Here we present a method for activating Channelrhodopsin2 that is expressed in targeted neurons in freely walking flies.

Optogenetic刺激的逃避行为<em&gt;果蝇</em

A  growing number of genetically encoded tools are becoming available that allow non-invasive  manipulation of the neural activity of specific neurons in ...

Optogenetic Perturbation of Neural Activity with Laser Illumination in Semi-intact Drosophila Larvae in Motion

Drosophila larval locomotion is a splendid model system in developmental and physiological neuroscience, by virtue of the genetic accessibility of the underlying neuronal components in the circuits1-6. Application of optogenetics7,8 in the larval neural circuit allows us to manipulate neuronal activity in spatially and temporally patterned ways9-13. Typically, specimens are broadly illuminated with a mercury lamp or LED, so specificity of the target neurons is controlled by binary gene expression systems such as the Gal4-UAS system14,15. In this work, to improve the spatial resolution to "sub-genetic resolution", we locally illuminated a subset of neurons in the ventral nerve cord using lasers implemented in a conventional confocal microscope. While monitoring the motion of the body wall of the semi-intact larvae, we interactively activated or inhibited neural activity with channelrhodopsin16,17 or halorhodopsin18-20, respectively. By spatially and temporally restricted illumination of the neural tissue, we can manipulate the activity of specific neurons in the circuit at a specific phase of behavior. This method is useful for studying the relationship between the activities of a local neural assembly in the ventral nerve cord and the spatiotemporal pattern of motor output.

Optogenetic Perturbation of Neural Activity with Laser Illumination in Semi-intact Drosophila Larvae in Motion

Here we describe a protocol for optogenetic manipulation of motoneuronal activity while monitoring changes in motor output (muscle contraction) in semi-intact Drosophila larvae using lasers within a conventional confocal microscope. This technique enables researchers to achieve local perturbation of neural activity in a few neuromeres to elucidate the dynamics of motor circuits.

光遗传学的扰动激光照射半完好的神经活动<em&gt;果蝇</em&gt;幼虫运动

Drosophila larval locomotion is a splendid model system in developmental and physiological neuroscience, by virtue of the genetic accessibility of the ...

Straightforward Assay for Quantification of Social Avoidance in Drosophila melanogaster

Drosophila melanogaster is an emerging model to study different aspects of social interactions. For example, flies avoid areas previously occupied by stressed conspecifics due to an odorant released during stress known as the Drosophila stress odorant (dSO). Through the use of the T-maze apparatus, one can quantify the avoidance of the dSO by responder flies in a very affordable and robust assay. Conditions necessary to obtain a strong performance are presented here. A stressful experience is necessary for the flies to emit dSO, as well as enough emitter flies to cause a robust avoidance response to the presence of dSO. Genetic background, but not their group size, strongly altered the avoidance of the dSO by the responder flies. Canton-S and Elwood display a higher performance in avoiding the dSO than Oregon and Samarkand strains. This behavioral assay will allow identification of mechanisms underlying this social behavior, and the assessment of the influence of genes and environmental conditions on both emission and avoidance of the dSO. Such an assay can be included in batteries of simple diagnostic tests used to identify social deficiencies of mutants or environmental conditions of interest.

Straightforward Assay for Quantification of Social Avoidance in Drosophila melanogaster

Here, we present a protocol to quantify the avoidance of stressed individuals. This paradigm is powerful yet user-friendly and can be used to assess the influence of genes and environment on one kind of social interaction in Drosophila melanogaster.

简单的检测方法的社交回避的量化<em&gt;果蝇</em

Drosophila melanogaster is an emerging model to study different aspects of social interactions. For example, flies avoid areas previously occupied by ...

Conditions Affecting Social Space in Drosophila melanogaster

The social space assay described here can be used to quantify social interactions of Drosophila melanogaster &#8212; or other small insects &#8212; in a straightforward manner. As we previously demonstrated 1, in a two-dimensional chamber, we first force the flies to form a tight group, subsequently allowing them to take their preferred distance from each other. After the flies have settled, we measure the distance to the closest neighbor (or social space), processing a static picture with free online software (ImageJ). The analysis of the distance to the closest neighbor allows researchers to determine the effects of genetic and environmental factors on social interaction, while controlling for potential confounding factors. Diverse factors such as climbing ability, time of day, sex, and number of flies, can modify social spacing of flies. We thus propose a series of experimental controls to mitigate these confounding effects. This assay can be used for at least two purposes. First, researchers can determine how their favorite environmental shift (such as isolation, temperature, stress or toxins) will impact social spacing 1,2. Second, researchers can dissect the genetic and neural underpinnings of this basic form of social behavior 1,3. Specifically, we used it as a diagnostic tool to study the role of orthologous genes thought to be involved in social behavior in other organisms, such as candidate genes for autism in humans 4.

Conditions Affecting Social Space in Drosophila melanogaster

The effect of genes and environment on social space of Drosophila melanogaster can be quantified through a powerful but straightforward analytical paradigm. We show here different factors that can affect this social space, and thus need to be taken into consideration when designing experiments in this paradigm.

在条件影响社会空间<em&gt;果蝇</em

The social space assay described here can be used to quantify social interactions of Drosophila melanogaster &#8212; or other small insects &#8212; in a ...

High-resolution Quantification of Odor-guided Behavior in Drosophila melanogaster Using the Flywalk Paradigm

In their natural environment, insects such as the vinegar fly Drosophila melanogaster are bombarded with a huge amount of chemically distinct odorants. To complicate matters even further, the odors detected by the insect nervous system usually are not single compounds but mixtures whose composition and concentration ratios vary. This leads to an almost infinite amount of different olfactory stimuli which have to be evaluated by the nervous system.
To understand which aspects of an odor stimulus determine its evaluation by the fly, it is therefore desirable to efficiently examine odor-guided behavior towards many odorants and odor mixtures. To directly correlate behavior to neuronal activity, behavior should be quantified in a comparable time frame and under identical stimulus conditions as in neurophysiological experiments. However, many currently used olfactory bioassays in Drosophila neuroethology are rather specialized either towards efficiency or towards resolution.
Flywalk, an automated odor delivery and tracking system, bridges the gap between efficiency and resolution. It allows the determination of exactly when an odor packet stimulated a freely walking fly, and to determine the animal&#180;s dynamic behavioral reaction.

High-resolution Quantification of Odor-guided Behavior in Drosophila melanogaster Using the Flywalk Paradigm

The automated tracking system Flywalk is used for high-resolution quantification of odor-guided behavior in Drosophila melanogaster.

在气味引导行为的高分辨率定量<em&gt;果蝇</em&gt;使用<em&gt; Flywalk</em&gt;范式

In their natural environment, insects such as the vinegar fly Drosophila melanogaster are bombarded with a huge amount of chemically distinct odorants. To ...

A Behavioral Assay for Mechanosensation of MARCM-based Clones in Drosophila melanogaster

Because of the structural and functional homology to the hair cells of the mammalian inner ear, the neurons that innervate the Drosophila external sense organs provide an excellent model system for the study of mechanosensation. This protocol describes a simple touch behavior in fruit flies which can be used to identify mutations that interfere with mechanosensation. The tactile stimulation of a macrochaete bristle on the thorax of flies elicits a grooming reflex from either the first or third leg. Mutations that interfere with mechanotransduction (such as NOMPC), or with other aspects of the reflex arc, can inhibit the grooming response. A traditional screen of adult behaviors would have missed mutants that have essential roles during development. Instead, this protocol combines the touch screen with mosaic analysis with a repressible cell marker (MARCM) to allow for only limited regions of homozygous mutant cells to be generated and marked by the expression of green fluorescent protein (GFP). By testing MARCM clones for abnormal behavioral responses, it is possible to screen a collection of lethal p-element mutations to search for new genes involved in mechanosensation that would have been missed by more traditional methods.

A Behavioral Assay for Mechanosensation of MARCM-based Clones in Drosophila melanogaster

In order to identify novel mutations affecting mechanosensation, we designed an assay that measures the behavioral response to tactile stimulation of fly bristles in mutant clones generated by the MARCM method. The combination of techniques allows for the identification of mechanosensitive mutations that would otherwise be lethal.

一个行为分析对机械感觉MARCM为基础的克隆<em&gt;果蝇</em

Because of the structural and functional homology to the hair cells of the mammalian inner ear, the neurons that innervate the Drosophila external sense ...