Name: a simple way to measure alterations in reward-seeking behavior using drosophila melanogaster
Uploaded: 2016-12-15T13:00:00.000+00:00
Duration: 6 min 57 s
Description: Watch this Scientific Journal Video about A Simple Way to Measure Alterations in Reward-seeking Behavior Using Drosophila melanogaster at JoVE.com

我们感谢U.赫伯利和A. Devineni长期持久的讨论和技术咨询。我们也感谢肖哈特 - 奥弗实验室成员，A. Benzur，L. Kazaz和O.沙洛姆与演示方法的帮助。特别感谢去埃利泽COSTI在实验室建立的飞行系统。这项工作是由以色列科学基金会（14分之384）和玛丽·居里职业集成补助（CIG 631127）的支持。

注:实验设计的总体概述:实验设计包括求偶抑制7-9中雄蝇超过4天的过程中受到奖励和nonrewarding经验，连续3次培训课程的改编协议。 4天 - 在经验阶段结束，苍蝇都在两选择自愿乙醇消费测定法3进行测试。此处的协议包括几个准备步骤，其中的一些可以预先做在一个以上的实验中使用，而另一些应的实验（ 表1）的开始前发生及时。 1.预备步骤<ol><li>准备毛细管馈线系统<ol><li>热量27克针并用它来穿刺沿着小瓶（ 图1）的小孔。 </li><li>切割用刀片在一半（横向）的小瓶插头。标记四个点插塞的表面上，形成一个SQUARe形毛细管持有人的位置之中。使用18G的针通过插头（ 图1b），以使4个孔。 </li><li>用剃刀刀片切micropipets以便它们可以牢固地保持5微升玻璃毛细管，其用作适配器。 </li><li>将4毛细管适配器插入准备好的插头。 </li><li>马克插头上的2乙醇和两个含nonethanol，毛细血管的位置。 注:建议保持2种类型的毛细血管在所有瓶一致的立场。注意，孔的大小，如大孔允许苍蝇的逃逸出来的小瓶。此外，建议使用插塞的光滑的一面（通常是相反的切口侧），为当插入到小瓶中，切割插件创建苍蝇能得到缠结一些小，松散纤维面对向下侧。 </li><li>准备用于毛细管馈线系统大量小瓶和适配器（CAFE）在步骤2.3中描述的实验。 </li></ol></li><li>准备小玻璃小瓶食品<ol><li>收集蝇（步骤1.4）的前一天，安排在微离心管架200的玻璃小瓶中。熔体飞食品（根据玉米面糖蜜配方，见材料的表 ），使用微波炉。倒入2毫升飞食品进玻璃小瓶（尽量不要涂抹食品小瓶墙壁上）。让食物变硬，并用保鲜膜覆盖的小瓶。 注:含有食物的玻璃小瓶可以保持在4℃几周。 </li><li>飞收集的当天，取瓶取出冰箱里，让他们用他们的房子苍蝇之前达到RT。 注意:如果食品小瓶几个星期的时候，请确保食品仍然完好，而不是从小瓶分离，苍蝇可以在食品和玻璃之间的缝隙被困住。 </li></ol></li><li>对于毛细管馈线分析股票制作食品的解决方案<ol><li>预削股票食物溶液（5％蔗糖和5％酵母提取物），溶解5.88克酵母提取物，并在100毫升的蒸馏水5.88克蔗糖。 注:储备溶液是高度易污染。因此，建议将高压釜它（121℃，30分钟，和没有干燥循环），并等分入850微升工作等分试样被保持在4℃直至进一步使用。在原液的成分的浓度为5.88％，并在加入只是其在步骤2.3在使用前15％的乙醇或水（150微升成的预制食品溶液850微升）后会达到5％的最终浓度。 </li></ol></li><li>实验收集苍蝇<ol><li>收集利用CO 2垫200幼稚的雄蝇。 注:使用相关的实验问题的任何实验室应变。 250苍蝇可以从苍蝇五瓶在其羽化挑阶段容易地收集。当收集苍蝇，密切注意山坳那些最近eclosed LECT苍蝇（处女蝇看:鲜艳的苍蝇在他们的腹部胎粪点）。 4日龄在实验中被使用之前 - 实验苍蝇应该单独含食品和〜3岁的小玻璃瓶中保存。 </li><li>用软塞关闭瓶。 注意:在使用口抽吸期间经历相插入女性进入和离开小瓶时相同的软塞将是训练阶段非常有用的。 </li></ol></li><li>收集教练苍蝇 注:所有收集苍蝇应在25℃，70％相对湿度（RH）保持在12小时/ 12小时亮/暗孵化器。在12小时/ 12小时光照/黑暗周期可以改变为一个方便的出发时间。 <ol><li>处女女教练苍蝇<ol><li>收集500处女女教练从WT股票或用作处女接受女性一个virginator股票苍蝇。请的25组每常规尺寸的食品小瓶雌蝇，所有在实验中使用之前4日龄 - 他们OW年龄为3。 </li></ol></li><li>交配雌蝇教练<ol><li>收集从WT股票或股票virginator 300处女雌蝇训练阶段（步骤1.6.1）之前接受交配。 注意:女性苍蝇被关在每个常规尺寸的食品小瓶女10组，年龄以3 - 4日龄在试验后方可使用。 </li></ol></li><li>雄蝇用于生成配接女教练苍蝇<ol><li>收集150雄蝇（不一定是处女）。 注:苍蝇被关在每个常规尺寸的食物和小瓶15岁的男性群体3 - 4日龄与处女女被配对前飞创造交配教练女性。 </li></ol></li></ol></li><li>产生交配雌蝇教练<ol><li>要生成配接雌蝇用作培训师，10个处女女的一组苍蝇与15雄性果蝇通过翻转一种含雄性瓶成女性CONTA进不去小瓶16 -训练阶段前18小时（步骤2.1）10。 注意:这通常应在训练前的晚上放。 1.5:男性之间1的比例飞到雌性果蝇可确保所有女性将被交配。 </li><li>在实验的当天，通过吸男性出用口吸气小瓶的分离从男性交配雌性。每个训练日的开始之前执行这一权利。 </li></ol></li></ol> 2.实验步骤<ol><li>设置了体验阶段<ol><li>开始实验尽可能接近在培养箱的光相位的开始。放置在行为室教练女性（包括处女和配对）。 注:建议以执行一个行为室中的实验，或者任何僻静环境是安静和设定为25℃和60 - 65％RH下。湿度保持50％以上是非常重要的，因为求偶和交配行为小号依赖信息素感应，这是众所周知的是对湿度敏感。 </li></ol></li><li>揭露男性苍蝇奖励和nonrewarding经验<ol><li>含排列在货架离心单雄蝇的玻璃小瓶，定位小瓶沿着机架的空间。 注:这种安排可以让实验者方便地观察对，进行交配的苍蝇没有采取小瓶从机架中。 </li><li>制备（如在步骤2.2.1描述）2基团含有单雄性100玻璃小瓶蝇对接受奖励和nonrewarding交配经历男性单独的机架（ 即拒绝和交配）。 </li><li>吸教练女性，并将其添加到包含雄蝇每个小瓶。设置定时器1小时。与被拒绝的组开始，增加交配雌，每一瓶。然后，移动到要配合男性，并添加一个处女女，每一瓶。 </li><li>密切关注交配遭遇以确保与处女雌性交互的男性的第一个小时之内，并与先前交配雌性交互的雄性不交配成功交配。并监控超过1小时的过程中被拒绝队列中，设定一第二计时器15分钟，并观看男性每15分钟。 </li><li>放电从拒绝队列，托管交配，并从队列交配未在第一堂训练课的结尾结束了交配男性男性。 </li><li>通过吸出先前交配的女性教练成正飞食品小瓶完成第一堂训练课，并让他们在接下来的调理会话。按照此步骤轻轻吸从交配男性组的女性。 </li><li>让苍蝇休息1小时。 </li><li>重复步骤2.2.3 - 2.2.7两次。 </li><li>重复步骤2.2.3训练课 - 2.2.8每天3个多Ð。 </li></ol></li><li>录制消费（CAFE） <ol><li>在最后conditionin的端摹会议（初四），吸出的单身男性，并将它们组合成毛细管馈线瓶。使用软塞以覆盖毛细管馈线小瓶而插入苍蝇，和所有的苍蝇已插入后与现成毛细管馈线插头迅速更换。组6 - 从每个实验组到小瓶的8苍蝇，创建每种条件不低于8小瓶中。 </li><li>轻轻加入5毫升水到插塞的顶部润湿插头。要小心，同时加入了水，以免溅入小瓶。等量水添加到每个小瓶中。 注意:此步骤是为了从毛细管减少蒸发。因此，每天反复进行实验的其余部分。根据不同的相对湿度，随后的几天，可能需要较少的水被添加到该插头。 </li><li>准备加入乙醇或水的股票食物等分以创建的15％的最终浓度两个单独的食品的解决方案。 </li><li>填补了capill与两个食物解白羊，创建5μL的在实验室薄膜条液体食品滴并且允许直到它达到5微升标记溶液填充毛细管。确保使用毛细管，这是接近5μL黑标线的清晰结束。 注意:要一次填充多个毛细血管，就可以连接6 - 8毛细管适配器一起使用它们作为插入保持毛细血管。 </li><li>轻轻拍打在石蜡条毛细血管食物解决方案在黑标定位准确。通过每个毛细管的端浸入矿物油的小滴密封毛细管。这一步最大限度地减少食品解决方案潜在蒸发。 </li><li>插入通过4个适配器/架有两个含有乙醇的毛细管和两个含有nonethanol毛细管在插头直到毛细管的露出侧面勉强的小瓶（从插头面1毫米或更小）的内部偷看。 注意:保持着密切的湿插毛细血管的开放降低了蒸发和可轻松访问的食物，苍蝇通常坐在插头和爬下的毛细管露出来养活。 </li><li>除了包含实验苍蝇瓶，准备不苍蝇的模拟小瓶。这种控制用于评估自然蒸发。 注:尽量保持相比插头毛细管开口的相对相等定位，差异会导致可变蒸发和误导消费数据。在实验过程中保持足够的湿度和毛细管开口从湿插头尽可能接近地定位于湿插头，低湿度和大的距离既可以导致不受控制的蒸发。 </li><li>记录在每个毛细管中的液体的相对位置相对于用尺子以毫米黑色标记线，并且这些值写入下来。 注意:如果解决方案是在黑色L准确定位INE，该值应该等于零，并且如果它是高于或低于，它应该是正的或负的值，分别为。液体的初始水平对于最终O / N消耗值设定的参考点。 </li><li>设置小瓶放回培养箱24小时。在整个实验的恒定时间更换毛细管。 </li><li>从小瓶轻轻取出毛细血管和相对于该黑色标记线测量的液体的水平。 </li><li>仔细检查在模拟小瓶中的液位。 注意:如果该水平下降超过2mm，则表明蒸发高并且消耗水平是不可靠的。 </li><li>用2毫升水轻轻挫伤插头和插入一组新的毛细血管的，如在步骤2.3.3指示 - 2.3.10。设置小瓶放回培养箱24小时。 </li></ol></li><li>确定优先<ol><li>从我减去最终水平计算每个毛细管的总消费量在起点nitial水平。 </li><li> （从含乙醇的毛细血管单独即 nonethanol毛细血管）同时结合了毛细血管的值与同食。 </li><li>减去非乙醇毛细管消耗从含有乙醇的毛细管所消耗的量的量计算乙醇偏好和从同一小瓶的所有毛细管除以总消耗值的值。 </li><li>平均相同的实验组的所有小瓶的优先索引。 <img alt="公式1" src="/files/ftp_upload/54910/54910equation1.jpg" /> 注:Nonethanol 1（NE1），Nonethanol 2（NE2），乙醇1（E1），乙醇2（E2），总nonethanol（共NE）= NE1 + NE2，总乙醇（总E）= E1 + E2，总和消费量（合计金额）=总NE +总E和乙醇偏好指数（PI） </li></ol></li></ol>

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The authors have nothing to disclose.

Here, we illustrate the details of an integrated approach to measure alterations in reward-seeking behavior, based on previous work described by Devineni et al.6 and Shohat-Ophir et al5. The first section of the protocol uses different types of sexual interactions as the experience input, and the second section uses a two-choice feeding assay to assess the effect of experience on the preference to consume ethanol.
As shown by Devineni et al.<sup...

响应于经验行为的修改允许动物来调节他们的环境1其行为的变化。在此过程中，动物与外部环境的不断变化的条件整合其内部的生理状态，随后选择通过另外一个动作，以增加他们的生存和繁殖的机会。奖励制度演变来激励那些通过加强增强即时生存的行为，如吃或喝，或那些确保长期生存，如性行为或照顾子女需要2个人和物种的生存行为。如滥用药物人工化合物也由调解自然奖励2增选神经通路影响奖励制度。 在过去的二十年中，果蝇果蝇已被确立为一个有前途的模型研究molecular和塑造对行为3,4乙醇的影响神经机制。 此前，我们已经确定肽能神经元的果蝇的一个子集（NPF / NPF受体（R）神经元）的夫妻自然奖励，如性经验，获得奖励药物5的动机。 NPF表达双方的性经验和药物的奖励，如乙醇中毒敏感。在NPF表达水平的变化转换为改建乙醇自我管理5，其中高NPF降低和低NPF增加了优先消耗酒精。激活NPF神经元是有益的苍蝇，因为它们显示为具有激活，这也是由减少的乙醇消耗量反射配对的气味强烈偏好。更重要的是，NPF神经元的激活与苍蝇，形成乙醇中毒和气味线索之间的正相关关系的能力干扰。在NPF / R之间的因果关系系统奖励的内存，和乙醇消费量表明，人们可以使用乙醇自我管理作为一项措施在国家奖励5的变化。 在本出版物中，我们证明了窃听到飞自然奖励制度和化验改变奖励状态的综合办法。该方法由两个单独的部分，用于操纵天然奖励相关的经验训练协议，接着两选择毛细管馈线测定（CAFE）评估乙醇自身给药作为在奖励状态的变化的估计。该CAFE测定是类似于在啮齿类动物的研究用于药物自我给药的二瓶的选择测定法和已显示，以反映在苍蝇6成瘾样行为的某些特性。

<table><tbody><tr><td>Polystyrene 25 x 95 mm Vials</td><td>FlyStuff</td><td>32-109</td><td></td></tr><tr><td>narrow plastic vials flugs</td><td>FlyStuff</td><td>42-102</td><td></td></tr><tr><td>Disposable Sterile Needle 18 G and 27 G&amp;nbsp;</td><td>can be acquired by any company</td><td></td><td>1.20 X 38 mm (18 G x 1 1/2&quot;) , 0.40 X 13 mm (27 G x 1/2&quot;)</td></tr><tr><td>10 x 75 mm Borosilicate Glass Disposable Culture Tubes</td><td>kimble chase</td><td>73500-1075</td><td></td></tr><tr><td>calibrated pipets 5 &amp;#956;L</td><td>VWR</td><td>53432-706</td><td>color coded white to contain 5 &amp;#956;L</td></tr><tr><td>Mineral Oil&amp;nbsp;</td><td>Sigma-Aldrich&amp;nbsp;</td><td>M5904</td><td></td></tr><tr><td>Sucrose, Molecular Biology Grade</td><td>CALBIOCHEM</td><td>573113</td><td></td></tr><tr><td>Yeast extract Powder for microbiology</td><td>can be acquired by any company</td><td></td><td></td></tr><tr><td>Ethanol</td><td>Sigma-Aldrich&amp;nbsp;</td><td>32221</td><td></td></tr><tr><td>standard pipette Tips (micro-pipetts)</td><td>ThermScientific</td><td>T114R-Q</td><td>volume: 0.1 - 20 &amp;#956;L (ultramicro)</td></tr><tr><td>IDENTI-PLUGS (Foam Tube Plugs)</td><td>Jaece</td><td>L800-A</td><td>fits opening 6 - 13 mm</td></tr><tr><td>IDENTI-PLUGS (Foam Tube Plugs)</td><td>Jaece</td><td>L800-D</td><td>fits opening 35 - 45 mm</td></tr><tr><td>virginator fly stock&amp;nbsp;</td><td>bloomington drosophila stock center</td><td>#24638</td><td></td></tr><tr><td>Narrow Vials, Tray Pack (PS)</td><td>Genesee Scientific Corporation</td><td>&amp;nbsp;# 32-109BR</td><td></td></tr><tr><td>Drosophila Media Recipes and Methods</td><td>Bloomington Drosophila Stock Center</td><td>http://flystocks.bio.indiana.edu/Fly_Work/media-recipes/molassesfood.htm</td><td></td></tr><tr><td>propionic acid</td><td>Sigma-Aldrich&amp;nbsp;</td><td>P5561</td><td></td></tr><tr><td>phosphoric acid</td><td>Sigma-Aldrich&amp;nbsp;</td><td>W290017</td><td></td></tr><tr><td>Methl 4-Hydroxybenzoate</td><td>Sigma-Aldrich&amp;nbsp;</td><td>H3647</td><td></td></tr><tr><td>Agar Agar</td><td>can be acquired by any company</td><td></td><td></td></tr><tr><td>corn meal</td><td>can be acquired by any company</td><td></td><td></td></tr><tr><td>Grandma&#039;s molasses</td><td>B&amp;amp;G Foods, Inc</td><td>not indicated</td><td></td></tr><tr><td>instant dry yeast</td><td>can be acquired by any company</td><td></td><td></td></tr></tbody></table>

a simple way to measure alterations in reward-seeking behavior using drosophila melanogaster

我们描述了测量在果蝇中乙醇的自我管理（ 果蝇 ）作为在奖励状态的变化代理的协议。我们展示了一个简单的方法来挖掘成飞奖励系统，修改相关的自然奖励的经验，并使用自愿乙醇消耗量作为用于奖励状态的变化的量度。该方法可作为相关的工具来研究发挥内部状态的经验介导的变化作用的神经元和基因。该协议是由两个分立部件:苍蝇暴露于奖励和nonrewarding经验，并测定自愿乙醇消费为动机的量度，以获得药物奖励。这两个部分可以独立使用，以诱导经历调制作为用于进一步的下游分析的初始步骤或作为一个独立的两选择馈送测定法，分别。该协议不需要复杂的设置，因此可以在任何laborator施加y随基本飞行文化工具。

FPreviously，Devineni 等。表明，当果蝇被赋予选择服用含乙醇的食品，它们显示超过食品含有6 nonethanol含乙醇的食品有强烈的偏好。这里显示的是测定天真雄蝇的先天乙醇偏好未经过培训协议时，我们获得了一些有代表性的成果。 朴素广的男蝇成虫羽化后收集，年龄，直到4日龄，并测定其与生俱来的偏爱超过4天?...

Watch this Scientific Journal Video about A Simple Way to Measure Alterations in Reward-seeking Behavior Using Drosophila melanogaster at JoVE.com

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

我们描述诱导奖励和nonrewarding使用乙醇自愿消费在奖励状态变化的措施果蝇（ 果蝇 ）经验的协议。

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 ...

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Research

JoVE Journal

Neuroscience

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

8.3K 次观看.  Bar-Ilan University. 我们描述诱导奖励和nonrewarding使用乙醇自愿消费在奖励状态变化的措施果蝇（ 果蝇 ）经验的协议。 

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

Combining Quantitative Food-intake Assays and Forcibly Activating Neurons to Study Appetite in Drosophila

Food consumption is under the tight control of the brain, which integrates the physiological status, palatability, and nutritional contents of the food, and issues commands to start or stop feeding. Deciphering the processes underlying the decision-making of timely and moderate feeding carries major implications in our understanding of physiological and psychological disorders related to feeding control. Simple, quantitative, and robust methods are required to measure the food ingestion of animals after experimental manipulation, such as forcibly increasing the activities of certain target neurons. Here, we introduced dye-labeling-based feeding assays to facilitate the neurogenetic study of feeding control in adult fruit flies. We review available feeding assays, and then describe our methods step-by-step from setup to analysis, which combine thermogenetic and optogenetic manipulation of neurons controlling feeding motivation with dye-labeled food intake assay. We also discuss the advantages and limitations of our methods, compared with other feeding assays, to help readers choose an appropriate assay.

Combining Quantitative Food-intake Assays and Forcibly Activating Neurons to Study Appetite in Drosophila

Quantitative food-intake assays with dyed food provide a robust and high-throughput means to evaluate feeding motivation. Combining the food consumption assay with thermogenetic and optogenetic screens is a powerful approach to investigate the neural circuits underlying appetite in adult Drosophila melanogaster.

在果蝇中结合定量食物摄入量测定和强迫激活神经元研究食欲

Food consumption is under the tight control of the brain, which integrates the physiological status, palatability, and nutritional contents of the food, ...

科研

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

A Single-fly Assay for Foraging Behavior in Drosophila

For many animals, hunger promotes changes in the olfactory system in a manner that facilitates the search for appropriate food sources. In this video article, we describe an automated assay to measure the effect of hunger or satiety on olfactory dependent food search behavior in the adult fruit fly Drosophila melanogaster. In a light-tight box illuminated by red light that is invisible to fruit flies, a camera linked to custom data acquisition software monitors the position of six flies simultaneously. Each fly is confined to walk in individual arenas containing a food odor at the center. The testing arenas rest on a porous floor that functions to prevent odor accumulation. Latency to locate the odor source, a metric that reflects olfactory sensitivity under different physiological states, is determined by software analysis. Here, we discuss the critical mechanics of running this behavioral paradigm and cover specific issues regarding fly loading, odor contamination, assay temperature, data quality, and statistical analysis.

A Single-fly Assay for Foraging Behavior in Drosophila

In this video article, we describe an automated assay to measure the effect of hunger or satiety on olfactory dependent food search behavior in the adult fruit fly Drosophila melanogaster.

单苍蝇检测方法的觅食行为的<em&gt;果蝇</em

For many animals, hunger promotes changes in the olfactory system in a manner that facilitates the search for appropriate food sources. In this video ...

糖诱导的果蝇觅食行为分析

Tracking Sugar-Elicited Local Searching Behavior in Drosophila

Foraging behavior is essential for the survival of organisms as it enables them to locate and acquire essential food resources. In Drosophila, hunger triggers a distinct search behavior following the consumption of small quantities of a sugar solution. This report presents a simple experimental setup to study sugar-elicited search behavior with the aim of uncovering the underlying mechanisms. Minute quantities of concentrated sugar solution elicit sustained searching behavior in flies. The involvement of path integration in this behavior has been established, as flies utilize their trajectory to return to the sugar location. The most recent findings provide evidence of temporal modulation in the initiation and intensity of the search behavior after sugar intake. We have also used this setup for artificial activation of specific taste-receptor neurons in the pharynx, which elicits the search behavior. The Drosophila neurogenetic toolkit offers a diverse array of tools and techniques that can be combined with the sugar-elicited search behavior paradigm to study the neural and genetic mechanisms underlying foraging. Understanding the neural basis of hunger-driven searching behavior in flies contributes to the field of neurobiology as a whole, offering insights into the regulatory mechanisms that govern feeding behaviors not only in other organisms but also in humans.

作者聚焦:通过觅食和交流中的跨物种洞察探索行为通路

This protocol describes a behavioral assay for recording sugar-elicited search behavior using Drosophila melanogaster. The assay can be utilized to study feeding and foraging-related behaviors, as well as the underlying neuronal mechanisms.

追踪果蝇中糖诱导的局部搜索行为

Foraging behavior is essential for the survival of organisms as it enables them to locate and acquire essential food resources. In Drosophila, hunger ...

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 ...

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. ...

The CApillary FEeder Assay Measures Food Intake in Drosophila melanogaster

For most animals, feeding is an essential behavior for securing survival, and it influences development, locomotion, health and reproduction. Ingestion of the right type and quantity of food therefore has a major influence on quality of life. Research on feeding behavior focuses on the underlying processes that ensure actual feeding and unravels the role of factors regulating internal energy homeostasis and the neuronal bases of decision-making. The model organism Drosophila melanogaster, with its great variety of genetically traceable tools for labeling and manipulating single neurons, allows mapping of neuronal networks and identification of molecular signaling cascades involved in the regulation of food intake. This report demonstrates the CApillary FEeder assay (CAFE) and shows how to measure food intake in a group of flies for time spans ranging from hours to days. This easy-to-use assay consists of glass capillaries filled with liquid food that flies can freely access and feed on. Food consumption in the assay is accurately determined using simple measurement tools. Herein we describe step-by-step the method from setup to successful execution of the CAFE assay, and provide practical examples to analyze the food intake of a group of flies under controlled conditions. The reader is guided through possible limitations of the assay, and advantages and disadvantages of the method compared to other feeding assays in D. melanogaster are evaluated.

The CApillary FEeder Assay Measures Food Intake in Drosophila melanogaster

The CApillary FEeder (CAFE) assay is a simple, budget-friendly, highly reliable method for investigating mechanisms underlying food intake. Used with the highly versatile genetic model organism Drosophila melanogaster, it provides a powerful means of gaining new insights into regulatory mechanisms of food intake.

毛细管馈线分析测量的摄食<em&gt;果蝇</em

For most animals, feeding is an essential behavior for securing survival, and it influences development, locomotion, health and reproduction. Ingestion of ...

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.

一个简单的方法来衡量在改建悬赏寻求行为使用

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此视频中的章节

在果蝇中结合定量食物摄入量测定和强迫激活神经元研究食欲

欲求联想嗅觉学习

单苍蝇检测方法的觅食行为的

追踪果蝇中糖诱导的局部搜索行为

追踪果蝇中糖诱导的局部搜索行为

追踪果蝇中糖诱导的局部搜索行为

在气味引导行为的高分辨率定量

高通量检测，以检查个体产蛋首

毛细管馈线分析测量的摄食

测量并改变配套驱动马累