Name: a new approach that eliminates handling for studying aggression and the "loser" effect in drosophila melanogaster
Uploaded: 2015-12-30T15:00:00
Description: 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

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....

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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 border="0" cellpadding="0" cellspacing="0" width="665">
	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Drosophila melanogaster pupa</td>
			<td style="width:163px;">Bloomington Stock Center</td>
			<td style="width:119px;"></td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Standard fly food</td>
			<td style="width:163px;"></td>
			<td style="width:119px;"></td>
			<td style="width:167px;">Fabricated in-house.&#160;</td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:216px;">Behavioral chambers</td>
			<td style="width:163px;">Mike machine</td>
			<td style="width:119px;"></td>
			<td style="width:167px;">Fabricated in-house. Directly&#160; contact the compagny for more informations.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Borosilicate glass vials</td>
			<td style="width:163px;">VWR International&#160;</td>
			<td style="width:119px;">47729-576</td>
			<td style="width:167px;">16 x 10 mm</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Cotton</td>
			<td style="width:163px;">Fisherbrand</td>
			<td style="width:119px;">22-456-881</td>
			<td style="width:167px;">Any brand can be used</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Pasteur pipettes</td>
			<td style="width:163px;">VWR International&#160;</td>
			<td style="width:119px;">53300-567</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Paintbrush&#160;</td>
			<td style="width:163px;">Blick Art Material</td>
			<td style="width:119px;">06157-7030</td>
			<td style="width:167px;">Round , Size 3/0</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Toothpick</td>
			<td style="width:163px;">N/A</td>
			<td style="width:119px;">N/A</td>
			<td style="width:167px;">Any brand can be used</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Acrylic paint (blue/white)</td>
			<td style="width:163px;">Blick Art Material</td>
			<td style="width:119px;">01637-5172/01637-1022</td>
			<td style="width:167px;">Any brand can be used</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Dry active yeast</td>
			<td style="width:163px;">Sigma</td>
			<td style="width:119px;">YSC2-500G</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Srew cap tube</td>
			<td style="width:163px;">VWR International&#160;</td>
			<td style="width:119px;">10011-394&#160;</td>
			<td style="width:167px;">15 mm diameter, 10 mm height</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Eppendorf</td>
			<td style="width:163px;">VWR International&#160;</td>
			<td style="width:119px;">22363212</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Tape</td>
			<td style="width:163px;">N/A</td>
			<td style="width:119px;">N/A</td>
			<td style="width:167px;">Any brand can be used</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Plastic slices</td>
			<td style="width:163px;">Electron Microscopy Sciences</td>
			<td style="width:119px;">70329-40</td>
			<td style="width:167px;">22 x 40 x 0.25 mm Thickness</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Light source (bulb)</td>
			<td style="width:163px;">VWR International&#160;</td>
			<td style="width:119px;">500003-418</td>
			<td style="width:167px;">Any brand can be used</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Timer</td>
			<td style="width:163px;">VWR International&#160;</td>
			<td style="width:119px;">62344-641</td>
			<td style="width:167px;">Any brand can be used</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Incubator</td>
			<td style="width:163px;">Percival</td>
			<td style="width:119px;"></td>
			<td style="width:167px;">Directly contact the constructor for more informations.</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Carbon Dioxide Dry (CO2)</td>
			<td style="width:163px;">Medical-Technical Gases, Inc</td>
			<td style="width:119px;">14H31</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Binocular</td>
			<td style="width:163px;">Nikon</td>
			<td style="width:119px;">SMZ-745</td>
			<td style="width:167px;">Any brand can be used</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Camera (SONY Handycam&#160; HDR-CX330)</td>
			<td style="width:163px;">B&#38;H</td>
			<td style="width:119px;">SOHDRCX330B</td>
			<td style="width:167px;">Any brand can be used</td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:216px;">Computer</td>
			<td style="width:163px;"></td>
			<td style="width:119px;"></td>
			<td style="width:167px;">With a minimum of 1.4 Ghz Processor, running Microsoft Windows or Machintosh HD</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">ClipWrap</td>
			<td style="width:163px;">Download online</td>
			<td style="width:119px;"></td>
			<td style="width:167px;">Any importing software can be used</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">QuickTime Player</td>
			<td style="width:163px;">Download online</td>
			<td style="width:119px;"></td>
			<td style="width:167px;">Any reading software can be used</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">GraphPad Software</td>
			<td style="width:163px;">Online</td>
			<td style="width:119px;"></td>
			<td style="width:167px;">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...

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 ...

Research

JoVE Journal

Neuroscience

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

Single Sensillum Recordings in the Insects Drosophila melanogaster and Anopheles gambiae

Single Sensillum Recordings in the Insects Drosophila melanogaster and 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

Paired Nanoinjection and Electrophysiology Assay to Screen for Bioactivity of Compounds using the Drosophila melanogaster Giant Fiber System

Paired Nanoinjection and Electrophysiology Assay to Screen for Bioactivity of Compounds using the Drosophila melanogaster Giant Fiber System

电检测屏幕的配对Nanoinjection和生物活性化合物的使用果蝇巨人纤维系统

Screening compounds for in vivo activity can be used as a first step to identify candidates that may be developed into pharmacological agents1,2. We ...

Visualization of Proprioceptors in Drosophila Larvae and Pupae

Visualization of Proprioceptors in Drosophila Larvae and Pupae

可视化本体感受器<em&gt;果蝇</em&gt;幼虫和蛹

Proprioception is the ability to sense the motion, or position, of body parts by responding to stimuli arising within the body. In fruitflies and other ...

Determination of the Spontaneous Locomotor Activity in Drosophila melanogaster

Determination of the Spontaneous Locomotor Activity in Drosophila melanogaster

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 ...

Straightforward Assay for Quantification of Social Avoidance in Drosophila melanogaster

Straightforward Assay for Quantification of Social Avoidance 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

Conditions Affecting Social Space in Drosophila melanogaster

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

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

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

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

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 ...

Foraging Path-length Protocol for Drosophila melanogaster Larvae

Foraging Path-length Protocol for Drosophila melanogaster Larvae

觅食路径长度协议<em&gt;果蝇</em&gt;幼虫

The Drosophila melanogaster larval path-length phenotype is an established measure used to study the genetic and environmental contributions to behavioral ...

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

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

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 ...