一个简单的方法来衡量在改建悬赏寻求行为使用<em>果蝇</em
Summary
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。
Abstract
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。
Introduction
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。
Protocol
Representative Results
Discussion
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…
Disclosures
The authors have nothing to disclose.
Acknowledgements
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。
Materials
| Polystyrene 25 x 95mm Vials | FlyStuff | 32-109 | |
| narrow plastic vials flugs | FlyStuff | 42-102 | |
| Disposable Sterile Needle 18G and 27G | can be acquired by any company | 1.20 X 38mm (18Gx1 1/2") , 0.40 X 13mm (27Gx1/2") | |
| 10x75mm Borosilicate Glass Disposable Culture Tubes | kimble chase | 73500-1075 | |
| calibrated pipets 5ul (microliter) | VWR | 53432-706 | color coded white to contain 5 microliters |
| Mineral Oil | Sigma-Aldrich | M5904 | |
| Sucrose, Molecular Biology Grade | CALBIOCHEM | 573113 | |
| Yeast extract Powder for microbiology | can be acquired by any company | ||
| Ethanol | Sigma-Aldrich | 32221 | |
| standard pipette Tips (micro-pipets) | ThermScientific | T114R-Q | volume- 0.1-20 ul Ultra micro |
| IDENTI-PLUGS (Foam Tube Plugs) | Jaece | L800-A | fits opening 6 to 13mm |
| IDENTI-PLUGS (Foam Tube Plugs) | Jaece | L800-D | fits opening 35 to 45mm |
| virginator fly stock | bloomington drosophila stock center | #24638 | |
| Narrow Vials, Tray Pack (PS) | Genesee Scientific Corporation | # 32-109BR | |
| Drosophila Media Recipes and Methods | Bloomington Drosophila Stock Center | http://flystocks.bio.indiana.edu/Fly_Work/media-recipes/molassesfood.htm | |
| propionic acid | Sigma-Aldrich | P5561 | |
| phosphoric acid | Sigma-Aldrich | W290017 | |
| Methl 4-Hydroxybenzoate | Sigma-Aldrich | H3647 | |
| Agar Agar | can be acquired by any company | ||
| corn meal | can be acquired by any company | ||
| Grandma's molasses | B&G Foods, Inc | not indicated | |
| instant dry yeast | can be acquired by any company | ||
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