这项研究是由普通医学科学研究所（GM099883和GM074675）的赠款，以EAK的资助者在研究设计，数据收集和分析，发布决定，或准备的手稿没有作用的支持。我们感谢的人从我们在哈佛医学院加工车间的行为设备的设计（联系人<a href="http://mikesmachine.com">:http://mikesmachine.com）。</a> 作者的贡献:ST，BC设计的行为装置。 ST设计和优化实验方案。 ST和BC进行分析实验。 ST，EAK和BC写文章。

注:以下是实验性的协议，我们用它来 ​​允许可靠触发侵略对雄性果蝇之间（果蝇）的一步一步的说明。这个过程导致的"失败者"效应的形成。使用该协议，一个短期的"胜利者"效果也已被最近报道5,6-，图1示出了行为实验的时间线。 <img alt="图1" src="/files/ftp_upload/53395/53395fig1.jpg" /> 图1.时间线侵略的实验。第1天，后期雄蜂蛹（当翅膀变成黑色）被隔离在新配制的隔离小瓶。在新兴的成人后的5当日4日龄雄性果蝇麻醉与CO 2和涂上不同的颜色标识。第7天，食品杯和战斗室是准备设立的行为实验。两个6-7日龄成年男性被装入室，并能互动。 <a href="https://www.jove.com/files/ftp_upload/53395/53395fig1large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> 1.第一天:蛹的社会隔离<ol><li>提高，在25℃，50％相对湿度保持苍蝇股环境控制12小时光照/ 12小时黑暗周期的孵化器。 </li><li>准备隔离瓶:热鲜飞食品，直到融化。使用巴斯德吸管，转移大约的熔化飞食品1.5 ml的成单空玻璃试管（1.6×10厘米）。让食物凝固。 </li><li>隔离蛹:用细的画笔，轻轻地从股票瓶中取出后期雄蛹（当翅膀变成黑色），并把它放在一个新鲜烹制的食品含有试管的一侧。关闭隔离小瓶棉（ 图2）。 </li><lI&gt;含有蛹到12小时光照/ 12小时黑暗培养箱中在25℃和相对湿度50％放置隔离小瓶中。 </li></ol><img alt="图2" src="/files/ftp_upload/53395/53395fig2.jpg" /> 图2.社会隔离，一个后期男性蛹被放置在侧上新鲜制备的隔离管中。含蛹隔离瓶放置在12小时光照/ 12小时黑暗的孵化器在25℃，相对湿度50％。 <a href="https://www.jove.com/files/ftp_upload/53395/53395fig2large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> 2.第5天:绘画蝇的个人识别目的<ol><li>用4日龄雄性苍蝇和现有的行为实验2天执行此步骤在第5天，以减少的 CO 2麻醉的负面影响。 </li><li>麻醉蝇用CO 2。 </l我&gt; <li>用细牙签，放置一个小点上胸部丙烯酸涂料。避免将油漆的头，翅膀，腹部或腿部（ 图3）。 </li><li>让油漆干燥大约10秒钟，然后轻轻地传回苍蝇到原来孤立的小瓶。 </li><li>返回隔离小瓶于12小时光照/ 12小时黑暗培养箱中在25℃和相对湿度50％。 </li></ol><img alt="图3" src="/files/ftp_upload/53395/53395fig3.jpg" /> 图3.个体识别。麻醉带有CO 2，一小滴丙烯酸漆飞后涂抹在胸部。对于个体识别的目的，不同的颜色时（白色和蓝色在这里）。请参阅以下有关使用二氧化碳在这一步的讨论。 <a href="https://www.jove.com/files/ftp_upload/53395/53395fig3large.jpg" target="_blank">请点击此处查看本figu的放大版本。</a>回覆。 3，第7天:准备设备<ol><li>准备食物杯: <ol><li>热火新鲜飞食品，直到它融化。 </li><li>使用巴斯德吸管，转移大约0.8 ml的食物到螺旋盖小瓶（1.5厘米直径，1厘米高）的盖子，并允许冷却。避免气泡。 </li><li>通过混合干酵母几滴之水鲜酵母膏。当厚，应用一小滴酵母膏到食品杯表面的用牙签（图4）的中心。 </li></ol></li></ol><img alt="图4" src="/files/ftp_upload/53395/53395fig4.jpg" /> 图4.准备食物的杯子。一种食品杯新鲜飞食品准备。食品凝固后，酵母膏点放置在食用杯面的中心。 <a href="https://www.jove.com/files/ftp_upload/53395/53395fig4large.jpg" target="_blank">请速到v</a>IEW这个图的放大版本。 <ol start="2"><li>准备个人战斗钱伯斯: <ol><li>使用前用水每次彻底清洗装置和个人战斗腔室。干燥，使用前纸巾。 </li><li>将食物杯在每个战斗室的中心和地方解决了一滴腻子。 </li><li>覆盖战斗腔室与该装置的盖子（图5）。 </li><li>放置装置成高湿度环境（60％），在25℃。 </li><li>放置装置之上的光源以照亮战斗腔室，并设置一台摄像机用于从上方的记录。 </li></ol></li></ol><img alt="图5" src="/files/ftp_upload/53395/53395fig5.jpg" /> 图5.准备个别战斗室中。将新鲜制备的食品杯用酵母膏的表面上的点被固定到洗涤个人音响ghting室。钱伯斯被关闭的两片盖允许插入一个不透明的塑料分隔成单个的战斗室。 <a href="https://www.jove.com/files/ftp_upload/53395/53395fig5large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> 4，第7天:行为实验<ol><li>执行行为实验用6-7日龄雄蝇。 </li><li>将塑料分隔成各个作战室。 </li><li>从隔离管中取出棉塞和定位下方开放孔的管在装置的一侧。 </li><li>让飞进入战斗室由负趋地性，通常在几秒钟之内。 </li><li>关闭滑动壁飞已经离开所述管并进入该腔室后。 </li><li>重复此过程，介绍第二蝇进入战斗室的设备的另一端。 </li><li>放置装置背面被照明videotapiNG的位置。 </li><li>开始录像并取下塑料分隔，让苍蝇进行互动。 <ol><li>侵略实验: <ol><li>在这些实验中，记录打架至少20分钟，以确保强支配关系的形成。 </li><li>后所需的时间，停止录像。 </li><li>通过反转上述步骤卸下苍蝇战斗室。 </li><li>彻底用温和的清洁剂和水每次使用和清洗后清洗行为装置。 </li></ol></li><li>对于"失败者"的试验研究: 注意:在这些实验中，第一打架被认为是调节阶段和第二打斗作为测试阶段。休息之间的战斗的时间可能有所不同，但10分钟可形成强大的"失败者"的效果。 <ol><li>按照上述执行的第一次战斗的过程。 </li><li>关键的一步:为了诱导的"失败者"的心态，E的​​形成nsure首先打架至少有20分钟长，以建立强大的统治地位的关系。 20分钟后，重新插入塑料分隔成战斗室的苍蝇分离。 </li><li>经过10分钟休息时间，轻轻地撕下塑料分隔，让苍蝇进行交互的第二战。 </li><li>在第二个20分钟战斗结束，停止录像。 </li><li>删除苍蝇作为上述行为室。 </li><li>再次，彻底清洁行为装置。 </li></ol></li></ol></li></ol> 5.行为分析<ol><li>传输影片<ol><li>连接照相机到计算机。 </li><li>转换苍蝇.MTS为.mov格式的​​苍蝇，并加入电影剪辑在一起，使用的视频转换软件，并选择"加入所选的剪辑"。 </li><li>观察和分析每部电影与视频阅读器软件。 </li></ol></li><li>计分行为 注:这可以手动执行，我们荷兰国际集团的行为分析程序软件（如果可用）。下图描述了一个"失败者"的效果可能的人工分析。 <ol><li>得分苍蝇上的食物杯之间的第一次遭遇的时间（遭遇是对苍蝇至少持续2秒之间的相互作用）和外观第一刺的时间（一种行为模式，其中一个飞升至高它的后腿和捕捉下来，并试图抓住对手）。随后，比分延迟弓步（首先遇到的与第一弓步之间的间隔）。 </li><li>在战斗中遭遇数和箭步蹲的总数。 注意:如果需要，也可以得分等行为模式，如翼的威胁，拳击，击剑，撤退或遇到的第一个冲刺前的数字，例如。 </li><li>分数建立霸主地位（第一次遇到，当一只苍蝇务虚连续三次断食杯前后的时间之间的时间的时间呃接收弓步）。注意这飞是赢家，哪些战斗的失败者。 </li></ol></li><li>计算一个"失败者"的影响<ol><li>划分战斗在前面的失败者失去第二次战斗（失败者的损失:LL）的数量由打架的总数，由100所获得的比例代表"失败者"的效果倍增: [（＃苍蝇LL）/（＃总拼）]×100 </li><li>使用卡方检验来比较"失败者"效应是否是从50％的预期值统计学差异。 </li></ol></li></ol>

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<li>Greenspan, R. J. <a target="_blank" href="http://scholar.google.com/scholar?hl=en&safe=off&q=author%3aGreenspan%2C+RJ+%22Fly+Pushing%3A+The+theory+and+Practice+of+Drosophila+Genetics%22">Fly Pushing: The theory and Practice of Drosophila Genetics</a>. , Cold Spring Harbor Laboratory Press. Cold Spring Harbor. (1997).</li>
<li>Cabral, L. G., Foley, B. R., Nuzhdin, S. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Does+sex+trade+with+violence+among+genotypes+in+Drosophila+melanogaster%3F%5BTitle%5D)+AND+%22PLoS+One%22%5BJournal%5D)">Does sex trade with violence among genotypes in Drosophila melanogaster?</a> PLoS One. 3 (4), e1986(2008).</li>
</ol>

The authors have nothing to disclose.

利用果蝇以其强大的遗传方式作为模式生物的侵略研究的新时代开始了十几年前在可靠的战斗行为，可以单对苍蝇1,7所获得的引进新的实验舞台上， 8,17,19。这些领域包括所需的资源（食物杯，潜在配偶）和失败者的空间飞行，以退为进。当用这些领域获得的结果，加入到约果蝇拼行为以前的知识， 果蝇确立为用于检查像侵略一个复杂的行为非常理想的模型系统，用...

侵略在动物系统是紧密联系在一起的采集和如食物，领地和配偶的资源控股关联。鉴于这种起着个人健身的重要作用，这是不奇怪的侵略整个动物王国中发展而来的。作为一个自适应的特点是直接受益的个体，较强的学习和记忆组件与侵略有关 。在对社会地位的竞争，以往的战斗体验影响了后来的比赛结果。在一般情况下，以前输球阅历的增加，​​并赢得经验减少，失去以后的比赛的可能性（被称为"输家"和"赢家"的影响）。 "失败者"的影响已经在广泛的物种中观察到，一些报告表明，这些都可以持续数天，而"赢家"的影响通常是持续时间较短2,10,11的。 攻击行为的第一次报告在果蝇（D. ampelophila）是由斯特蒂文特于1915年在涉及性别认同和性别选择12纸。半个世纪后的今天，男性的水果更完整的检查飞行侵略所做，所描述的大多数在果蝇打架观察到的行为模式。这些实验主要是进行了小团体的男性和女性的苍蝇观察到小时，13-15。近来，通过加入强大的遗传工具，二进单一性别的战斗的设置和观测，D。更短的时间果蝇已成为侵略1,16的生物学研究的重要模式系统。同性对雄性和雌性果蝇的打架分析表明，侵略涉及刻板的行为模式，从一个到另一个在统计学上可靠的方式1,17过渡。一些观察到的行为模式是性别特异性，而其他人在战斗观察S在男女双方。男打架去更高强度级别比女性打架，结果有明确的"赢家"和"输家"的主导地位关系的形成。目前，许多实验室已经开始调查了生化18，神经7,19-21，和遗传22基础侵略。不幸的是，一元分析的研究深入了解拼动力学和的形成和维持支配关系是有问题的，由于使用的实验程序在不同实验室的多重的。基本上所有的文献中所描述的技术涉及处理苍蝇和操纵，导入到舞台19,23,24和行为实验3,4中转移出来的苍蝇的竞技场。轻柔抽吸是操纵苍蝇3,4,23-25 ​​最常见的方式，但冷或CO 2麻醉也使用9,26，即使previous研究已经报告说，这些程序都有飞的行为27,28有害影响。持续至少24小时的使用任何麻醉剂后，建议尽量减少其对行为的影响29,30。 苍蝇从以前的战斗经验和修改他们的行为模式的使用在新的情况下，这意味着学习和记忆陪伴，并激动遭遇的后果。在这个意义上，飞打架就像在其中飞得知一个战略工作，然后在随后的遭遇越来越多的时候使用它操作条件反射的学习情况。苍蝇改变自己的战斗策略后主导地位的关系都在打架成立至今，凭借赢家冲刺越来越多，失败者越来越少。经过一段时间的分离的，以往的失败者显示更顺从的行为和非常可能失去2与幼稚FL成对第二战斗IES或往届获奖者3,4。然而，由于缺乏一个实验的过程，不包括装卸苍蝇的取得了"失败者"的效果很难详尽的研究。在最近的研究中，我们比较常规用于实验室（吸入和冷麻醉）引入飞进行为室新方法，消除了处理两个实验程序。结果表明，冷麻醉有更大的负面影响侵略不是愿望，但即使抽吸苍蝇减少攻击性的水平。吸入没有，但是，造成伴随侵略的学习和记忆非常显著的影​​响。继相同的协议有两个实验程序（愿望，也没有处理），当苍蝇的处理是从实验过程5,6消除只观察到一个强大的"失败者"的效果。 理想情况下， 果蝇的侵略在Labo研究ratories应包括苍蝇通常在野外遇到（对资源的争夺，领土保卫和空间来逃脱）环境状况。此外，实验条件应进行优化，以可靠地诱导，观察和解释所检查的行为（尽量减少动物的处理，限制使用二氧化碳作为麻醉剂，规范的实验程序）。在试图解决大多数或所有这些问题，我们设计了消除处理苍蝇前，中，向他们介绍行为赛场后，新的行为装置。与这些设备中，蝇使用其固有负趋地性，以被移动，并转移进和移出的斗争​​腔室。通过消除处理苍蝇，该协议的目的是:（1）降低个体之间的变异性的行为; （二）减少所需的时间苍蝇相互作用并产生清晰显性关系（男性）; （三）可靠地诱发足够强大behavio拉尔发生变化，允许组建"失败者"的效果。 本协议描述一个新的实验设备和一步一步的过程来分析的侵略，并允许在D.强烈的"失败者"效应的形成果蝇。我们期望这一行为装置，可以很容易地适用于通过果蝇表现出其他社会行为的研究。

<table><tbody><tr><td>Drosophila melanogaster pupa</td><td>Bloomington Stock Center</td><td></td><td></td></tr><tr><td>Standard fly food</td><td></td><td></td><td>Fabricated in-house.&amp;nbsp;</td></tr><tr><td>Behavioral chambers</td><td>Mike machine</td><td></td><td>Fabricated in-house. Directly&amp;nbsp; contact the compagny for more informations.</td></tr><tr><td>Borosilicate glass vials</td><td>VWR International&amp;nbsp;</td><td>47729-576</td><td>16 x 10 mm</td></tr><tr><td>Cotton</td><td>Fisherbrand</td><td>22-456-881</td><td>Any brand can be used</td></tr><tr><td>Pasteur pipettes</td><td>VWR International&amp;nbsp;</td><td>53300-567</td><td></td></tr><tr><td>Paintbrush&amp;nbsp;</td><td>Blick Art Material</td><td>06157-7030</td><td>Round , Size 3/0</td></tr><tr><td>Toothpick</td><td>N/A</td><td>N/A</td><td>Any brand can be used</td></tr><tr><td>Acrylic paint (blue/white)</td><td>Blick Art Material</td><td>01637-5172/01637-1022</td><td>Any brand can be used</td></tr><tr><td>Dry active yeast</td><td>Sigma</td><td>YSC2-500G</td><td></td></tr><tr><td>Srew cap tube</td><td>VWR International&amp;nbsp;</td><td>10011-394&amp;nbsp;</td><td>15 mm diameter, 10 mm height</td></tr><tr><td>Eppendorf</td><td>VWR International&amp;nbsp;</td><td>22363212</td><td></td></tr><tr><td>Tape</td><td>N/A</td><td>N/A</td><td>Any brand can be used</td></tr><tr><td>Plastic slices</td><td>Electron Microscopy Sciences</td><td>70329-40</td><td>22 x 40 x 0.25 mm Thickness</td></tr><tr><td>Light source (bulb)</td><td>VWR International&amp;nbsp;</td><td>500003-418</td><td>Any brand can be used</td></tr><tr><td>Timer</td><td>VWR International&amp;nbsp;</td><td>62344-641</td><td>Any brand can be used</td></tr><tr><td>Incubator</td><td>Percival</td><td></td><td>Directly contact the constructor for more informations.</td></tr><tr><td>Carbon Dioxide Dry (CO&lt;sub&gt;2&lt;/sub&gt;)</td><td>Medical-Technical Gases, Inc</td><td>14H31</td><td></td></tr><tr><td>Binocular</td><td>Nikon</td><td>SMZ-745</td><td>Any brand can be used</td></tr><tr><td>Camera (SONY Handycam&amp;nbsp; HDR-CX330)</td><td>B&amp;amp;H</td><td>SOHDRCX330B</td><td>Any brand can be used</td></tr><tr><td>Computer</td><td></td><td></td><td>With a minimum of 1.4 Ghz Processor, running Microsoft Windows or Machintosh HD</td></tr><tr><td>ClipWrap</td><td>Download online</td><td></td><td>Any importing software can be used</td></tr><tr><td>QuickTime Player</td><td>Download online</td><td></td><td>Any reading software can be used</td></tr><tr><td>GraphPad Software</td><td>Online</td><td></td><td>Any statistical software can be used</td></tr></tbody></table>

a new approach that eliminates handling for studying aggression and the "loser" effect in drosophila melanogaster

果蝇攻击行为是由刻板行为模式的顺序表达（分析参见图1）。这种复杂的行为是由遗传，荷尔蒙和环境因素的影响。正如在许多生物，以往的战斗体验影响果蝇的战斗策略和以后比赛的结果:失去的斗争增加了失去以后的比赛的可能性，揭示了"失败者"效应可能涉及学习和记忆2-4。伴随像侵略复杂的社会行为表达的学习和记忆，是敏感的前期试验动物5,6的处理。许多实验程序被用在不同的实验室研究侵略7-9，然而，没有常规使用的协议，排除处理苍蝇当前可用。在这里，我们报告说，省去了处理果蝇的一个新的行为装置，使用insteað他们与生俱来的负面geotactic响应移动的动物进入或退出战斗室。在这个协议中，含有食品杯小圆形斗争领域被引入之前苍蝇分成相等的两半通过一个可移动的塑料滑块。苍蝇通过推拉房门及趋地进入他们的家庭隔离小瓶室。在去除塑料滑块，苍蝇可以自由交互。经过指定的时间段，苍蝇是通过滑块进行后续实验再次分离。所有这一切都没有办理个人苍蝇容易。该设备提供了一种新的方法来研究的侵略和相关的学习和记忆，其中包括在飞行战斗"失败者"效应的形成。此外，这种新的通用行为装置可以被用来研究苍蝇等社会行为和应，在一般情况下，是用于调查在基本行为过程经验相关的变化的兴趣。

本节介绍行为腔室的设计和一组典型的按照以上测定侵略和的"失败者"效应的形成描述的协议行为实验的分析。还示出的其他行为的样品，可以使用该装置来测量。 图6示出了新装置的示意图。该装置由三个圆筒形孔（尺寸:22.86毫米直径17毫米的高度），这些战斗的腔室。两片盖关闭顶部的战斗室和允许插入的薄不透明的塑料分频器，分割室分成相等的两部分。在...

Watch this Scientific Journal Video about A New Approach that Eliminates Handling for Studying Aggression and the "Loser" Effect in Drosophila melanogaster at JoVE.com

A New Approach that Eliminates Handling for Studying Aggression and the "Loser" Effect in Drosophila melanogaster

在果蝇打架的行为模式观察，争取动态，以及相关的学习和记忆都是通过实验条件的影响。这里介绍的协议描述了一种新的程序，完全消除了在实验操作苍蝇。这提高了战斗的动态，并允许形成强大的"失败者"的效果。

一种新的方法，省去了处理的研究侵略和"失败者"的影响<em&gt;果蝇</em

Aggressive behavior in Drosophila melanogaster is composed of the sequential expression of stereotypical behavioral patterns (for analysis see 1). This ...

a-new-approach-that-eliminates-handling-for-studying-aggression-loser

Research

JoVE Journal

Neuroscience

A New Approach that Eliminates Handling for Studying Aggression and the "Loser" Effect in Drosophila melanogaster

9.6K 次观看.  Harvard Medical School. 在果蝇打架的行为模式观察，争取动态，以及相关的学习和记忆都是通过实验条件的影响。这里介绍的协议描述了一种新的程序，完全消除了在实验操作苍蝇。这提高了战斗的动态，并允许形成强大的"失败者"的效果。 

视频: A New Approach that Eliminates Handling for Studying Aggression and the "Loser" Effect in Drosophila melanogaster

Studying Aggression in Drosophila (fruit flies)

Aggression is an innate behavior that evolved in the framework of defending or obtaining resources. This complex social behavior is influenced by genetic, hormonal and environmental factors. In many organisms, aggression is critical to survival but controlling and suppressing aggression in distinct contexts also has become increasingly important. In recent years, invertebrates have become increasingly useful as model systems for investigating the genetic and systems biological basis of complex social behavior. This is in part due to the diverse repertoire of behaviors exhibited by these organisms. In the accompanying video, we outline a method for analyzing aggression in Drosophila whose design encompasses important eco-ethological constraints. Details include steps for: making a fighting chamber; isolating and painting flies; adding flies to the fight chamber; and video taping fights. This approach is currently being used to identify candidate genes important in aggression and in elaborating the neuronal circuitry that underlies the output of aggression and other social behaviors.

Studying Aggression in Drosophila fruit flies

学习在果蝇的侵略（果蝇）

Aggression is an innate behavior that evolved in the framework of defending or obtaining resources.  This complex social behavior is influenced by ...

科研

生物学

A Method to Test the Effect of Environmental Cues on Mating Behavior in Drosophila melanogaster

An individual's sexual drive is influenced by genotype, experience and environmental conditions. How these factors interact to modulate sexual behaviors remains poorly understood. In Drosophila melanogaster, environmental cues, such as food availability, affect mating activity offering a tractable system to investigate the mechanisms modulating sexual behavior. In D. melanogaster, environmental cues are often sensed via the chemosensory gustatory and olfactory systems. Here, we present a method to test the effect of environmental chemical cues on mating behavior. The assay consists of a small mating arena containing food medium and a mating couple. The mating frequency for each couple is continuously monitored for 24 h. Here we present the applicability of this assay to test environmental compounds from an external source through a pressurized air system as well as manipulation of the environmental components directly in the mating arena. The use of a pressurized air system is especially useful to test the effect of very volatile compounds, while manipulating components directly in the mating arena can be of value to ascertain a compound's presence. This assay can be adapted to answer questions about the influence of genetic and environmental cues on mating behavior and fecundity as well as other male and female reproductive behaviors.

A Method to Test the Effect of Environmental Cues on Mating Behavior in Drosophila melanogaster

We demonstrate an assay to analyze the environmental and genetic cues that influence mating behavior in the fruit fly Drosophila melanogaster.

测试环境线索对交配行为的影响的方法<em&gt;黑腹果蝇</em

An individual's sexual drive is influenced by genotype, experience and environmental conditions. How these factors interact to modulate sexual behaviors ...

行为学

Appetitive Associative Olfactory Learning in Drosophila Larvae

In the following we describe the methodological details of appetitive associative olfactory learning in Drosophila larvae. The setup, in combination with genetic interference, provides a handle to analyze the neuronal and molecular fundamentals of specifically associative learning in a simple larval brain.
Organisms can use past experience to adjust present behavior. Such acquisition of behavioral potential can be defined as learning, and the physical bases of these potentials as memory traces1-4. Neuroscientists try to understand how these processes are organized in terms of molecular and neuronal changes in the brain by using a variety of methods in model organisms ranging from insects to vertebrates5,6. For such endeavors it is helpful to use model systems that are simple and experimentally accessible. The Drosophila larva has turned out to satisfy these demands based on the availability of robust behavioral assays, the existence of a variety of transgenic techniques and the elementary organization of the nervous system comprising only about 10,000 neurons (albeit with some concessions: cognitive limitations, few behavioral options, and richness of experience questionable)7-10.
Drosophila larvae can form associations between odors and appetitive gustatory reinforcement like sugar11-14. In a standard assay, established in the lab of B. Gerber, animals receive a two-odor reciprocal training: A first group of larvae is exposed to an odor A together with a gustatory reinforcer (sugar reward) and is subsequently exposed to an odor B without reinforcement 9. Meanwhile a second group of larvae receives reciprocal training while experiencing odor A without reinforcement and subsequently being exposed to odor B with reinforcement (sugar reward). In the following both groups are tested for their preference between the two odors. Relatively higher preferences for the rewarded odor reflect associative learning - presented as a performance index (PI). The conclusion regarding the associative nature of the performance index is compelling, because apart from the contingency between odors and tastants, other parameters, such as odor and reward exposure, passage of time and handling do not differ between the two groups9.

Appetitive Associative Olfactory Learning in Drosophila Larvae

Drosophila larvae are able to associate odor stimuli with gustatory reward. Here we describe a simple behavioral paradigm that allows the analysis of appetitive associative olfactory learning.

欲求联想嗅觉学习<em&gt;果蝇</em&gt;幼虫

In the following we describe the methodological  details of appetitive associative olfactory learning in Drosophila larvae. The setup, in combination with ...

神经科学

Drosophila Passive Avoidance Behavior as a New Paradigm to Study Associative Aversive Learning

This protocol describes a new paradigm for analyzing aversive associative learning in adult flies (Drosophila melanogaster). The paradigm is analogous to passive avoidance behavior in laboratory rodents in which animals learn to avoid a compartment where they have previously received an electric shock. The assay takes advantage of negative geotaxis in flies, which manifests as an urge to climb up when they are placed on a vertical surface. The setup consists of vertically oriented upper and lower compartments. On the first trial, a fly is placed into a lower compartment from where it usually exits&#160;within 3-15 s, and steps into the upper compartment where it receives an electric shock. During the second trial, 24 h later, the latency is significantly increased. At the same time, the number of shocks is decreased compared to the first trial, indicating that flies formed long-term memory about the upper compartment. The recordings of latencies and number of shocks could be performed with a tally counter and a stopwatch or with an Arduino-based simple device. To illustrate how the assay can be used, the passive avoidance behavior of D. melanogaster and D. simulans male and female were characterized here. Comparison of latencies and number of shocks revealed that both D. melanogaster and D. simulans flies efficiently learned the passive avoidance behavior. No statistical differences were observed between male and female flies. However, males were a little faster while entering the upper compartment on the first trial, while females received a slightly higher number of shocks in every retention trial. The Western diet (WD) significantly impaired learning and memory in male flies while flight exercise counterbalanced this effect. Taken together, the passive avoidance behavior in flies offers a simple and reproducible assay that could be used for studying basic mechanisms of learning and memory.

Drosophila Passive Avoidance Behavior as a New Paradigm to Study Associative Aversive Learning

This work describes a simple behavioral paradigm that allows the analysis of aversive associative learning in adult fruit flies. The method is based on suppressing the innate negative geotaxis behavior due to the association formed between a specific environmental context and an electric shock.

果蝇 被动回避行为作为研究联想性厌恶学习的新范式

This protocol describes a new paradigm for analyzing aversive associative learning in adult flies (Drosophila melanogaster). The paradigm is analogous to ...

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 ...

High Throughput Assay to Examine Egg-Laying Preferences of Individual Drosophila melanogaster

Recently, egg-laying preference of Drosophila has emerged as a genetically tractable model to study the neural basis of simple decision-making processes. When selecting sites to deposit their eggs, female flies are capable of ranking the relative attractiveness of their options and choosing the &#34;greater of two goods.&#34; However, most egg-laying preference assays are not practical if one wants to take a systematic genetic screening approach to search for the circuit basis underlying this simple decision-making process, as they are population-based and laborious to set up. To increase the throughput of studying of egg-laying preferences of single females, we developed custom chambers that each can simultaneously assay egg-laying preferences of up to thirty individual flies as well as a protocol that ensures each female has a high egg-laying rate (so that their preference is readily discernable and more convincing). Our approach is simple to execute and produces very consistent results. Additionally, these chambers can be equipped with different attachments to allow video recording the egg-laying animals and to deliver light for optogenetics studies. This article provides the blueprints for fabricating these chambers and the procedure for preparing the flies to be assayed in these chambers.

High Throughput Assay to Examine Egg-Laying Preferences of Individual Drosophila melanogaster

This protocol describes a high throughput assay for testing egg-laying preferences of Drosophila melanogaster at single-animal resolution. This assay provides a simple, efficient, and scalable platform to identify genes and circuit components that control a simple decision-making process.

高通量检测，以检查个体产蛋首<em&gt;果蝇</em

Recently, egg-laying preference of Drosophila has emerged as a genetically tractable model to study the neural basis of simple decision-making processes. ...

A Simple Way to Measure Alterations in Reward-seeking Behavior Using Drosophila melanogaster

We describe a protocol for measuring ethanol self-administration in fruit flies (Drosophila melanogaster) as a proxy for changes in reward states. We demonstrate a simple way to tap into the fly reward system, modify experiences related to natural reward, and use voluntary ethanol consumption as a measure for changes in reward states. The approach serves as a relevant tool to study the neurons and genes that play a role in experience-mediated changes of internal state. The protocol is composed of two discrete parts: exposing the flies to rewarding and nonrewarding experiences, and assaying voluntary ethanol consumption as a measure of the motivation to obtain a drug reward. The two parts can be used independently to induce the modulation of experience as an initial step for further downstream assays or as an independent two-choice feeding assay, respectively. The protocol does not require a complicated setup and can therefore be applied in any laboratory with basic fly culture tools.

A Simple Way to Measure Alterations in Reward-seeking Behavior Using Drosophila melanogaster

We describe a protocol for inducing rewarding and nonrewarding experiences in fruit flies (Drosophila melanogaster) using voluntary ethanol consumption as a measure for changes in reward states.

一个简单的方法来衡量在改建悬赏寻求行为使用<em&gt;果蝇</em

We describe a protocol for measuring ethanol self-administration in fruit flies (Drosophila melanogaster) as a proxy for changes in reward states. We ...

Measuring and Altering Mating Drive in Male Drosophila melanogaster

Despite decades of investigation, the neuronal and molecular bases of motivational states remain mysterious. We have recently developed a novel, reductionist, and scalable system for in-depth investigation of motivation using the mating drive of male Drosophila&#160;melanogaster (Drosophila), the methods for which we detail here. The behavioral paradigm centers on the finding that male mating drive decreases alongside fertility over the course of repeated copulations and recovers over ~3 d. In this system, the powerful neurogenetic tools available in the fly converge with the genetic accessibility and putative wiring diagram available for sexual behavior. This convergence allows rapid isolation and interrogation of small neuronal populations with specific motivational functions. Here we detail the design and execution of the satiety assay that is used to measure and alter courtship motivation in the male fly. Using this assay, we also demonstrate that low male mating drive can be overcome by stimulating dopaminergic neurons. The satiety assay is simple, affordable, and robust to influences of genetic background. We expect the satiety assay to generate many new insights into the neurobiology of motivational states.

Measuring and Altering Mating Drive in Male Drosophila melanogaster

This article describes a behavioral assay that uses male mating drive in Drosophila melanogaster to study motivation. Using this method, researchers can utilize advanced fly neurogenetic techniques to uncover the genetic, molecular, and cellular mechanisms that underlie this motivation.

测量并改变配套驱动马累<em&gt;果蝇</em

Despite decades of investigation, the neuronal and molecular bases of motivational states remain mysterious. We have recently developed a novel, ...

Drosophila Courtship Conditioning As a Measure of Learning and Memory

Many insights into the molecular mechanisms underlying learning and memory have been elucidated through the use of simple behavioral assays in model organisms such as the fruit fly, Drosophila melanogaster. Drosophila is useful for understanding the basic neurobiology underlying cognitive deficits resulting from mutations in genes associated with human cognitive disorders, such as intellectual disability (ID) and autism. This work describes a methodology for testing learning and memory using a classic paradigm in Drosophila known as courtship conditioning. Male flies court females using a distinct pattern of easily recognizable behaviors. Premated females are not receptive to mating and will reject the male&#39;s copulation attempts. In response to this rejection, male flies reduce their courtship behavior. This learned reduction in courtship behavior is measured over time, serving as an indicator of learning and memory. The basic numerical output of this assay is the courtship index (CI), which is defined as the percentage of time that a male spends courting during a 10 min interval. The learning index (LI) is the relative reduction of CI in flies that have been exposed to a premated female compared to na&#239;ve flies with no previous social encounters. For the statistical comparison of LIs between genotypes, a randomization test with bootstrapping is used. To illustrate how the assay can be used to address the role of a gene relating to learning and memory, the pan-neuronal knockdown of Dihydroxyacetone phosphate acyltransferase (Dhap-at) was characterized here. The human ortholog of Dhap-at, glyceronephosphate O-acyltransferase (GNPT), is involved in rhizomelic chondrodysplasia punctata type 2, an autosomal-recessive syndrome characterized by severe ID. Using the courtship conditioning assay, it was determined that Dhap-at is required for long-term memory, but not for short-term memory. This result serves as a basis for further investigation of the underlying molecular mechanisms.

Drosophila Courtship Conditioning As a Measure of Learning and Memory

This protocol describes a Drosophila learning and memory assay called courtship conditioning. This classic assay is based on a reduction of male courtship behavior after sexual rejection by a non-receptive premated female. This natural form of behavioral plasticity can be used to test learning, short-term memory, and long-term memory.

果蝇调理作为学习和记忆的措施

Many insights into the molecular mechanisms underlying learning and memory have been elucidated through the use of simple behavioral assays in model ...

果蝇 自闭症谱系障碍的遗传和行为特征模型

Paradigms for Behavioral Assessment in Drosophila Model of Autism Spectrum Disorder

Autism Spectrum Disorder (ASD) encompasses a heterogeneous group of neurodevelopmental disorders with common behavioral symptoms including deficits in social interaction and ability for communication, enhanced restricted or repetitive behaviors, and also, in some cases, learning disability and motor deficit. Drosophila has served as an unparalleled model organism for modeling a great number of human diseases. As many genes have been implicated in ASD, fruit flies have emerged as a powerful and efficient way to test the genes putatively involved with the disorder. As hundreds of genes, with varied functional roles, are implicated in ASD, a single genetic fly model of ASD is unfeasible; instead, individual genetic mutants, gene knockdowns, or overexpression-based studies of the fly homologs of ASD-associated genes are the common means for gaining insight regarding molecular pathways underlying these gene products. A host of behavioral techniques are available in Drosophila which provide easy readout of deficits in specific behavioral components. Social space assay and aggression and courtship assays in flies have been shown to be useful in assessing defects in social interaction or communication. Grooming behavior in flies is an excellent readout of repetitive behavior. Habituation assay is used in flies to estimate the ability for habituation learning, which is found to be affected in some ASD patients. A combination of these behavioral paradigms can be utilized to make a thorough assessment of the human ASD-like disease state in flies. Using Fmr1 mutant flies, recapitulating Fragile-X syndrome in humans, and POGZ-homolog row knockdown in fly neurons, we have shown quantifiable deficits in social spacing, aggression, courtship behavior, grooming behavior, and habituation. These behavioral paradigms are demonstrated here in their simplest and straightforward forms with an assumption that it would facilitate their widespread use for research on ASD and other neurodevelopmental disorders in fly models.

作者聚焦：探索果蝇的自闭症谱系障碍症状 — 遗传模型和行为测试

Autism Spectrum Disorder (ASD) is associated with impaired social and communicative behavior and the emergence of repetitive behavior. For studying the interrelation between ASD genes and behavioral deficits in the Drosophila model, five behavioral paradigms are described in this paper for assaying social spacing, aggression, courtship, grooming, and habituation behavior.

自闭症谱系障碍果 蝇 模型的行为评估范式

Autism Spectrum Disorder (ASD) encompasses a heterogeneous group of neurodevelopmental disorders with common behavioral symptoms including deficits in ...

一种新的方法，省去了处理的研究侵略和"失败者"的影响

摘要

探索更多视频

学习在果蝇的侵略（果蝇）

测试环境线索对交配行为的影响的方法

欲求联想嗅觉学习

果蝇被动回避行为作为研究联想性厌恶学习的新范式

简单的检测方法的社交回避的量化

高通量检测，以检查个体产蛋首

一个简单的方法来衡量在改建悬赏寻求行为使用

测量并改变配套驱动马累

果蝇调理作为学习和记忆的措施

自闭症谱系障碍果蝇模型的行为评估范式

一种新的方法，省去了处理的研究侵略和"失败者"的影响

摘要

探索更多视频

学习在果蝇的侵略（果蝇）

测试环境线索对交配行为的影响的方法

欲求联想嗅觉学习

果蝇 被动回避行为作为研究联想性厌恶学习的新范式

简单的检测方法的社交回避的量化

高通量检测，以检查个体产蛋首

一个简单的方法来衡量在改建悬赏寻求行为使用

测量并改变配套驱动马累

果蝇调理作为学习和记忆的措施

自闭症谱系障碍果 蝇 模型的行为评估范式

果蝇被动回避行为作为研究联想性厌恶学习的新范式

自闭症谱系障碍果蝇模型的行为评估范式