The authors want to thank Dr. Tadmiri Venkatesh for his encouragement to prepare this manuscript, Dova Brenman and Selwyn Chui for their constructive comments, Elyssa Burg and Wayne Rasband for designing the macro in ImageJ for all interfly distances and for nearest neighbor distances. The authors also want to thank the reviewers for their constructive comments.
A.A.A, M.C.C. and A.F.S were responsible for research design; S.N.J., A.A.A, M.N., Z.R., and A.J.M. performed the experiments. A.A.A, M.N., A.J.M. and M.C.C and A.F.S. analyzed the data; A.R.M. and A.F.S. wrote the manuscript.
This work was supported by PSC-CUNY research awards, jointly funded by The Professional Staff Congress and The City University of New York to A.F.S.; by internal funding from Western University to A.F.S.; by a Mathematics, Biology, Chemistry, and Geology majors scholarship for teachers-in-training and by a Louis Stokes Alliance for Minority Participation scholarship to A.A.A.

1.设备和试剂创造了内部（材料为他人见列表） <ol><li>制备果蝇冷麻醉装置如前所述8。 </li><li>准备一个飞吸气如前所述13。 </li><li>准备社会空间室和持有人。 <ol><li>命令或使玻璃板和丙烯酸垫片，以创造社会空间室。每个社会空间室由两个方形玻璃板（17.6厘米×17.6厘米，厚度为0.3厘米），两个直角三角形丙烯酸间隔物（16.5厘米，8.9厘米的碱基的高度，厚度为0.3厘米），并且两个矩形的空间（9厘米×1.5厘米，厚度为0.3厘米）（ 图1A）。 </li><li>装配玻璃和丙烯酸类社会空间室，使得其与 ​​图1B中的腔室。为此，通过将在玻璃上的一个方形窗格顶部的一个三角形垫片平面，使得启动直角三角形垫片的对准的平方窗格的一个角。下一步，放置在广场上窗格顶部的第二个三角形隔板持平，反映的第一个三角垫片。 注意:请一定要戴上手套，以免污染与你的手油和香料的作品。 </li><li>将两个小矩形隔板平放在方形面板的顶部，以及未涉及窗格的一面。广场上飞机前的丙烯酸垫片，现在应该形成一个三角形的舞台。 </li><li>放置玻璃的第二方形窗格上的丙烯酸系衬垫使得其上面低于玻璃原窗格对齐的顶部。 </li><li>使用四种长尾夹来固定窗格和隔板一起以形成社会空间室。将一个剪辑邻近的拐角处，在每个三角间隔物的长边，将它们固定在玻璃板内部。在这些相同侧，放置一个夹在相邻的拐角处，在每个矩形隔板，将它们固定在另一边的玻璃板。 </li><li>装配一个支座和一个烧瓶钳使得钳将保持在直立位置的社会空间室，作为社会空间室搁置在其上的工作表面的上边缘。 </li><li>对于实验中的每个重复，组建一个社会空间室，并支持立场。 </li></ol></li><li>准备攀登试验反应装置检查苍蝇测试的爬坡能力。 <ol><li>使用逆流装置，如前所述14,15。 </li></ol></li><li>通过执行实验上覆盖着白色的长椅盖在白色背景前一个作业面确保均匀的照明条件。 </li></ol> 2.准备蝇实验前<ol><li>维护苍蝇含标准果蝇的瓶子飞的食物。它们保存在一个孵育室在25℃在12小时光照/黑暗循环。 </li><li>一到两天之前，电子商务xperiment，收集并在冷麻醉性苍蝇，如前所述8。 <ol><li>寒意冷麻醉机在-4℃。将飞入50ml塑料管。完全淹没塑料管含在保温冰桶苍蝇成冰，等待至少5分钟，苍蝇变得动弹不得。 </li><li>将苍蝇在冰冷的麻醉设备的聚乙烯薄膜。在RT，使用立体显微镜来的男性从女性分开。保持苍蝇成群，含标准果蝇瓶飞的食物，不超过40苍蝇每小瓶。 </li></ol></li><li>在实验开始前两小时，确保在试验时将执行的房间的温度为24-25℃，湿度大约为50％。 </li><li>转移苍蝇到含有食品新的小瓶，并将它们放置2小时在哪里的实验将要执行的工作表面上。 注:对于这些experimenTS，所使用的蝇株果蝇品系:广-特殊或广-S（CS）， 瓦特1118铯10（或W - 瓦特1118异型杂交的10倍到广-S），俄勒冈州和萨马尔罕苍蝇都从我们的实验室库存16;埃尔伍德苍蝇被收集在2011年秋季在亨廷顿的埃尔伍德附近，长岛，纽约，美国8。除了当指定，否则，给出的结果，得到与广东-S蝇（在红色色调）。 </li></ol> 3.执行实验<ol><li> （ - 以小时为单位的时间光发病后 - 时间授时ZT 4〜7）进行实验12点和下午3点之间。 </li><li>准备的社会空间，室内苍蝇的转移。 <ol><li>放置一个社会空间室平放在工作台上，用含有最接近你的身体矩形隔板一侧。 </li><li>删除ØNE夹最近你的身体，并向外滑动一个长方形的间隔，产生的矩形隔板之间大约为1厘米的差距。 </li><li>使用磁带和标记，标号社会空间室与性别，应变和重复次数的上角。注意确保磁带不包括在内三角舞台的一部分。 </li></ol></li><li>转移苍蝇进入社会空间室。 <ol><li>转让航班从含有食物进入一个新的空药瓶的瓶。吸苍蝇从空药瓶，并将其转移到社会空间室。 <ol><li>吸入吸引苍蝇入抽吸器的尖端。头放入的社会空间室的矩形隔板之间1厘米的差距，并在呼气一致的速度，迫使苍蝇进入内三角的舞台。 </li><li>立即矩形隔板滑回原位，关闭内三角竞技场的基部，然后将粘合剂剪辑重新打开。 </li></ol></l我&gt; </ol></li><li>上的冲击垫，它位于不同长凳比在该实验工作发生时，保持社会空间室直立，使得矩形隔板是在底侧。英镑三次，以确保所有的苍蝇已经下降到了舞台的底部。 注:由于亚克力间隔伸出最小室尺寸（ 图1D）中，砰的肘部在工作表面上，同时保持一个安全的扶住设备，以确保苍蝇已经跌到了谷底。 </li><li>启动计时器。 </li><li>放置在工作表面上的社会空间室和使用的支持立场，瓶夹，以保持直立。将尺子或已知长度的平贴在社会空间室的贴纸，但不涉及任何部位的内三角舞台。 <ol><li>对于水平放置的实验中，不要放在支撑支架上的社会空间室;而不是躺在工作台上的社会空间室持平。 </li></o升&gt; </li><li>当苍蝇纷纷落户，一般经过大约30分钟，以社会空间室的照片。确保帧包含整个内三角舞台上，统治者和标签。 </li><li>重复实验每个基因型和条件（最好3个内部重复和3个独立的重复）。 </li></ol> 4.分析的社会空间数据<ol><li>在导入ImageJ的17以下宏<ol><li> ImageJ的可以在这里找到:http://rsbweb.nih.gov/ij/。 </li><li>您可以通过保存在下面的宏在中的ImageJ / plugins文件夹命名为"Measure_Distances.txt"文件，在宏子文件夹中，并使用帮助&gt;更新菜单命令创建的插件菜单中"测量距离"命令。" </li><li>在一个.txt复制文件中提供补充数据，并最初发表在3宏。 </li></ol></li><li>上传照片到电脑，并开立一个图像重新使用ImageJ。 <ol><li>在图片中的标尺上画线从0厘米〜1厘米的标记创建一个规模，并分析选项卡下，选择设置缩放功能可以设置一个比例为1厘米的距离（同样的方法进行的贴纸已知长度）。选择应用规模的图片前，在全局选项。 </li><li>使用图像选项卡，使得它包括所有的苍蝇下的裁剪功能裁剪图片，同时消除尽可能多的画面尽可能的休息。 </li><li>使画面的黑白从图片选项卡下的类型选项中选择8位。 </li><li>在图像选项卡的调节选项下将阈值特征取下图像中的所有的背景噪音。拖动滑块以增强或删除的对比度，使得每个蝇的本体可以清楚地看到，而不在图像中的任何其他标​​记。如果有斑纹是不是苍蝇，捕捉它们与矩形工具，并将其删除。 </li><li>摆好测量通过分析选项卡下的设置测量工具。选择面积，质量，质心的中心，并显示标签。接下来，选择分析颗粒分析选项卡下的功能来创建所有的黑点代表苍蝇编号列表。 <ol><li>当使用分析颗粒;集大小0.01-0.1（不选择像素单元）;圆0.00-1.00;显示轮廓;显示结果，并加入到经理。 </li><li>确保每个黑点（每个粒子）是在原始图片，通过编号的列表进行比较，以原始图象的苍蝇的精确表示。 注意:如果一些苍蝇非常接近一起被算作一次粒子，手动绘制白线分离两个蝇二进制图像上。 </li></ol></li><li>虽然列表中选择，使用最近邻 - 列出的距离宏插件标签第二个宏选项下创建一个包含距离一个新的列表，在厘米，从EACH飞到离他们最近的邻居。 </li></ol></li><li>从复制ImageJ的最近邻距离并粘贴到一个电子表格程序的一列。 <ol><li>编译所有从每个重复数据到同一个电子表格，并组织由基因型和条件。 </li></ol></li><li>使用统计软件进行数据分析。在这个实验中，图垫棱镜（6版本的MacOSX，格拉夫派得软件圣地亚哥加利福尼亚美国，www.graphpad.com）被用来进行单向ANOVA和秩和检验测试，Tukey的和邓尼特的事后检验。 </li><li>的距离的分布遵循非参数分布1，和数据被表示为框和Tukey的晶须（消除异常值）。 </li></ol> 5.登山分析（控制行为） <ol><li>使用登山法作为控制测试蝇攀爬能力14,15,18。 <ol><li>使用逆流装置，使得三个试验Ç一个可并行运行。 </li></ol></li><li>转移50到100天真果蝇（果蝇尚未前测试）在三个不同的测试小瓶，并且它们卡入第 1次，3次和最后时隙在逆流装置的底部，以在相反的位置新鲜空小瓶中。 </li><li>放置在空槽新鲜空小瓶在装置的底部。 </li><li>挖掘下来的装置三次，以使得所有的苍蝇都在每个管的底部，以启动试验。 </li><li>启动定时器，持续15秒（时间足以为3-7天老广-S〜100％，达到了顶级小瓶-见图3F）。 </li><li>滑动逆流装置的上部，由一个槽来置换顶小瓶中。 </li><li>通过点击设备下来，把所有的苍蝇在底部瓶收集苍蝇。 <ol><li> 1次和第2次管（或3次和第4次， 第 5次</sUp&gt;和6 次 ），分别对应于没有达到顶部苍蝇和爬到山顶小瓶苍蝇。 </li></ol></li><li>计算性能指数（PI）。 <ol><li>算蝇每个小瓶的数目。 </li><li>该PI是苍蝇能攀上上管的百分比。 </li><li>在PI的装置遵循正态分布1，和数据被表示为平均值加或减标准误差的平均值（SEM）的柱形图。 注意:可替换地，在PI可被计算为苍蝇能达到较低小瓶（老年人苍蝇的上部的百分比，5秒的足够的时间以量化的差在感染与非爬坡能力感染12个天龄广-S从静止图像量化:看到的结果）。而其他人使用的惊吓引起的负趋地表现不同的措施，同时也是适当的量化爬坡能力19-22。 </li></ol></li></ol>

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P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Systemic+bacterial+infection+and+immune+defense+phenotypes+in+Drosophila+melanogaster.">Systemic bacterial infection and immune defense phenotypes in Drosophila melanogaster.</a> JoVE. , (2015).</li><li>Fernandez, R. W., Akinleye, A. A., Nurilov, M., Rouzyi, Z., Simon, A. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> , (2013).</li><li>Suh, G. S. B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+single+population+of+olfactory+sensory+neurons+mediates+an+innate+avoidance+behaviour+in+Drosophila.">A single population of olfactory sensory neurons mediates an innate avoidance behaviour in Drosophila.</a> Nature. 431, 854-859 (2004).</li><li>Helfrich-Fo&#776;rster, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differential+control+of+morning+and+evening+components+in+the+activity+rhythm+of+Drosophila+melanogaster.--Sex-specific+differences+suggest+a+different+quality+of+activity.">Differential control of morning and evening components in the activity rhythm of Drosophila melanogaster.--Sex-specific differences suggest a different quality of activity.</a> J. Biol. Rhythms. 15, 135-154 (2000).</li><li>Fujii, S., Krishnan, P., Hardin, P. E., Amrein, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nocturnal+male+sex+drive+in+Drosophila.">Nocturnal male sex drive in Drosophila.</a> Curr Biol. 17, 244-251 (2007).</li><li>Pellegrino, A. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Weather+forecasting+by+insects:+modified+sexual+behaviour+in+response+to+atmospheric+pressure+changes.">Weather forecasting by insects: modified sexual behaviour in response to atmospheric pressure changes.</a> PLoS One. 8, e75004 (2013).</li></ol>

The authors declare that they have no competing financial interests.

在这个协议中，对社会空间的量化的详细过程进行了说明。一些关键步骤，以确保实验成功的是:1）清理和设置设备，以保持自己的油脂和香料关闭设备的内室，2）确保苍蝇被收集之前至少有一天，当一直使用的手套实验，以减少冷麻醉的任何潜在影响，3）在实验前2小​​时，提供一种含有新鲜食物，以确保苍蝇不饿死，并已清洁自己新小瓶中，4）在这些2小时，让苍蝇使其恢复到其新的环境，...

社会互动是至关重要的一个组内的整体正常发展的个人和健康，并能在众多的品种可以观察到，从人类（ 智人 ），以简单的有机体，例如果蝇（ 果蝇 ）5,6。单个果蝇或人共享共同方式处理过程中这些互动的感觉信息，它是否是:听觉，视觉，嗅觉，触觉，味觉或。我们和其他人推测可能有是一种潜在的共享neurocircuitry底层行为反应社会互动和神经元细胞和参与可能是进化保守的基因7。一旦初始相互作用已经发生，相互作用的个人之间的社会空间要么增加（社交回避8）或减小（组形成/聚合5）。更复杂的相互作用，如侵略或求爱，就可以成行。 果蝇的稳定基团，如在以下的研究1-4,9使用。社会空间是指一种苍蝇和其最接近的邻居10之间的距离的量度。社会空间是D的一个特定人群一致当实验条件将被保留（平均约1-2个体的长度），并且改变相对于所述蝇社会经验，如果个体已经被保持在隔离1增加果蝇 。正确的目标是要维持正常的社会距离，而不是传统的添味剂或CVA感知1。因此，衡量社会空间可作为诊断工具来分析社会交往和量化D.社会行为黑1，我们在这里描述的细节如何执行此量化，并共同实验变量的影响程度如何这种行为。 我们表明，在该试验进行腔室，以及蝇的数目的取向 - 在一定程度上 - 做影响社会的空间。这是以前表明腔的几何形状影响的苍蝇11,12自发探索性运动，这种现象可能会最终影响他们决定定居。然而，只要在蝇密度（飞/厘米2）和腔室方向保持相同，苍蝇的社会空间也保持不变。该测定的鲁棒性的事实，使用不同的腔室的尺寸，形状独立实验室，和取向可以复制由WH的突变体所显示的结果示出ITE基因（影响眼部色素沉着），这是一个增加社会空间（1个垂直三角形或横向循环，用气流在3水平平方）。 我们的结果还表明，保持在该社会空间实验进行时间是至关重要的结果的一致性，因为我们表明，男性，但不是雌性，进一步分开，在晚上。然而，看到白天和晚上的时间之间的差异不是由于苍蝇的活动不同，和大家讨论的论点，表示不与社会空间相关的活动水平。 最后，有遗传基础的社会空间的测定，由白突变体所指示已经描述的1,3，和苍蝇，我们在这里介绍的各种近交系和捕获的野生株之间的差异。 因此，这个实验使一个很好的工具˚F或研究的遗传效应以及环境因素。

<table><tbody><tr><td>Stereo Zoom Microscope&amp;nbsp;</td><td>Nikon&amp;nbsp;&amp;nbsp;</td><td>SMZ-645</td><td>Any other standard scope for fly handling would work</td></tr><tr><td>Small paint brushes&amp;nbsp;</td><td></td><td></td><td>for pushing flies</td></tr><tr><td>Jazz-mix</td><td>Fisher</td><td>33545</td><td>any other standard drosophila food would work</td></tr><tr><td></td><td></td><td></td><td></td></tr><tr><td>Mini-Alarm Timer/Stopwatch</td><td></td><td></td><td></td></tr><tr><td>&amp;nbsp;Sharpie pens</td><td></td><td></td><td></td></tr><tr><td>Adhesive Tape</td><td></td><td></td><td></td></tr><tr><td>Medium size binder clips</td><td>Staples</td><td></td><td>to hold the chambers together: 1-1/4&quot; (32mm) medium clips with 5/8&quot; capacity</td></tr><tr><td>small SupportStands</td><td>Carolina&amp;nbsp;</td><td>707161</td><td>to hold the chambers in a vertical orientation</td></tr><tr><td>Buret clamps</td><td>Carolina</td><td>707362</td><td></td></tr><tr><td>Digital Camera</td><td>Nikon</td><td>&amp;nbsp;Coolpix S8000&amp;nbsp;</td><td>to take the still pictures</td></tr><tr><td>small ruler</td><td></td><td></td><td>to be able to scale the picture</td></tr><tr><td>trifold board and white bench cover</td><td></td><td></td><td>to provide a white background, and a homogeneous light.</td></tr><tr><td>pounding pad</td><td></td><td></td><td>any mouse pad works.</td></tr><tr><td></td><td></td><td></td><td></td></tr><tr><td>Prism 6</td><td>GraphPad Software Inc.</td><td>Prism 6 for Mac OS X</td><td>Any statistical analysis software with t-test, one-way and two ANOVA would work</td></tr></tbody></table>

conditions affecting social space in drosophila melanogaster

此处所描述的社会空间测定可用于定量的果蝇melanogasteř社会互动-或其它小昆虫-以直接的方式。如我们以前表明图1，在一个二维室，我们首先迫使苍蝇形成紧密基，随后使它们采取彼此其优选距离。之后苍蝇纷纷落户，我们测量到最近的邻居（或社会空间）的距离，处理静态图片，免费在线软件（ImageJ的）。到最近的邻居的距离的分析，使研究人员能够确定遗传和环境因素对社会互动的效果，同时控制了潜在的混杂因素。不同的因素，例如爬坡能力，天，性别的时间，和苍蝇的数目，可以修改蝇社会间距。因此，我们提出了一系列的实验控制，以减轻这些混杂影响。此法可用于至少两个目的。首先，研究人员能够确定自己喜欢的环境转变（如隔离，温度，压力或毒素）会影响社会的间距1,2。其次，研究人员可以剖析社会行为1,3这种基本形式的遗传和神经基础。具体而言，我们用它作为诊断工具，研究认为参与在其他生物，如候选基因孤独症在人中4社会行为直系同源基因的作用。

社会空间室可以作为一种工具，以量化果蝇的社会行为。丙烯酸垫片和玻璃板被夹在一起以形成内三角竞技场，提供了一个在其中苍蝇能形成稳定的基团没有许多潜在的混杂线索的存在二维区域。当苍蝇被转移到垂直领域，它们是由被窃听下来吓了一跳，他们通过逃逸行为作出回应:负趋地。他们爬到直立的三角形的顶点，强行纳入一个紧密的组，然后提供时间，以进一步将形成彼此，并探...

Watch this Scientific Journal Video about Conditions Affecting Social Space in Drosophila melanogaster at JoVE.com

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 ...

conditions-affecting-social-space-in-drosophila-melanogaster

Research

JoVE Journal

Neuroscience

Conditions Affecting Social Space in Drosophila melanogaster

12.1K 次观看.  University of Western Ontario. 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.

视频: Conditions Affecting Social Space in Drosophila melanogaster

Rearing the Fruit Fly Drosophila melanogaster Under Axenic and Gnotobiotic Conditions

The influence of microbes on myriad animal traits and behaviors has been increasingly recognized in recent years. The fruit fly Drosophila melanogaster is a model for understanding microbial interactions with animal hosts, facilitated by approaches to rear large sample sizes of Drosophila under microorganism-free (axenic) conditions, or with defined microbial communities (gnotobiotic). This work outlines a method for collection of Drosophila embryos, hypochlorite dechorionation and sterilization, and transfer to sterile diet. Sterilized embryos are transferred to sterile diet in 50 ml centrifuge tubes, and developing larvae and adults remain free of any exogenous microbes until the vials are opened. Alternatively, flies with a defined microbiota can be reared by inoculating sterile diet and embryos with microbial species of interest. We describe the introduction of 4 bacterial species to establish a representative gnotobiotic microbiota in Drosophila. Finally, we describe approaches for confirming bacterial community composition, including testing if axenic Drosophila remain bacteria-free into adulthood.

Rearing the Fruit Fly Drosophila melanogaster Under Axenic and Gnotobiotic Conditions

A method for rearing Drosophila melanogaster under axenic and gnotobiotic conditions is presented. Fly embryos are dechorionated in sodium hypochlorite, transferred aseptically to sterile diet, and reared in closed containers. Inoculating diet and embryos with bacteria leads to gnotobiotic associations, and bacterial presence is confirmed by plating whole-body Drosophila homogenates.

饲养果蝇<em&gt;果蝇</em&gt;在无菌和悉生条件

The influence of microbes on myriad animal traits and behaviors has been increasingly recognized in recent years. The fruit fly Drosophila melanogaster is ...

科研

发育生物学

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

Methods to Assay Drosophila Behavior

Drosophila melanogaster, the fruit fly, has been used to study molecular mechanisms of a wide range of human diseases such as cancer, cardiovascular disease and various neurological diseases1. We have optimized simple and robust behavioral assays for determining larval locomotion, adult climbing ability (RING assay), and courtship behaviors of Drosophila. These behavioral assays are widely applicable for studying the role of genetic and environmental factors on fly behavior. Larval crawling ability can be reliably used for determining early stage changes in the crawling abilities of Drosophila larvae and also for examining effect of drugs or human disease genes (in transgenic flies) on their locomotion. The larval crawling assay becomes more applicable if expression or abolition of a gene causes lethality in pupal or adult stages, as these flies do not survive to adulthood where they otherwise could be assessed. This basic assay can also be used in conjunction with bright light or stress to examine additional behavioral responses in Drosophila larvae. Courtship behavior has been widely used to investigate genetic basis of sexual behavior, and can also be used to examine activity and coordination, as well as learning and memory. Drosophila courtship behavior involves the exchange of various sensory stimuli including visual, auditory, and chemosensory signals between males and females that lead to a complex series of well characterized motor behaviors culminating in successful copulation. Traditional adult climbing assays (negative geotaxis) are tedious, labor intensive, and time consuming, with significant variation between different trials2-4. The rapid iterative negative geotaxis (RING) assay5 has many advantages over more widely employed protocols, providing a reproducible, sensitive, and high throughput approach to quantify adult locomotor and negative geotaxis behaviors. In the RING assay, several genotypes or drug treatments can be tested simultaneously using large number of animals, with the high-throughput approach making it more amenable for screening experiments.

Methods to Assay Drosophila Behavior

Drosophila melanogaster is a genetically and behaviorally tractable model system that has been used to understand the molecular and cellular basis of many important biological processes for over a century 1. Drosophila has been well exploited to gain insights into the genetic basis of fly behavior.

对检测方法果蝇行为

Drosophila melanogaster, the fruit fly, has been used to study molecular mechanisms of a wide range of human diseases such as cancer, cardiovascular ...

神经科学

An Automated Method to Determine the Performance of Drosophila in Response to Temperature Changes in Space and Time

Temperature is a ubiquitous environmental factor that affects how species distribute and behave. Different species of Drosophila fruit flies have specific responses to changing temperatures according to their physiological tolerance and adaptability. Drosophila flies also possess a temperature sensing system that has become fundamental to understanding the neural basis of temperature processing in ectotherms. We present here a temperature-controlled arena that permits fast and precise temperature changes with temporal and spatial control to explore the response of individual flies to changing temperatures. Individual flies are placed in the arena and exposed to pre-programmed temperature challenges, such as uniform gradual increases in temperature to determine reaction norms or spatially distributed temperatures at the same time to determine preferences. Individuals are automatically tracked, allowing the quantification of speed or location preference. This method can be used to rapidly quantify the response over a large range of temperatures to determine temperature performance curves in Drosophila or other insects of similar size. In addition, it can be used for genetic studies to quantify temperature preferences and reactions of mutants or wild-type flies. This method can help uncover the basis of thermal speciation and adaptation, as well as the neural mechanisms behind temperature processing.

An Automated Method to Determine the Performance of Drosophila in Response to Temperature Changes in Space and Time

Here we present a protocol to automatically determine the locomotor performance of Drosophila at changing temperatures using a programmable temperature-controlled arena that produces fast and accurate temperature changes in time and space.

一种自动测定果蝇在空间和时间温度变化中的性能的方法

Temperature is a ubiquitous environmental factor that affects how species distribute and behave. Different species of Drosophila fruit flies have specific ...

Flypub To Study Ethanol Induced Behavioral Disinhibition and Sensitization

Alcohol use disorder (AUD) remains a serious problem in our society. To develop effective interventions for addiction, it is important to understand the underlying neurobiological mechanisms, for which diverse experimental approaches and model systems are needed. The main ingredient of alcoholic beverages is ethanol, which causes adaptive changes in the central nervous system and behavior upon chronic intake. Behavioral sensitization (i.e., escalated responses) in particular represents a key adaptive change underlying addiction. Most ethanol-induced behavioral sensitization studies in animal models have been conducted on the locomotor activating effect of ethanol. A prominent effect of ethanol is behavioral disinhibition. Behavioral sensitization to the disinhibition effect of ethanol, however, is underrepresented. To address this issue, we developed the Flypub assay that allows measuring the escalated increase in disinhibited courtship activities upon recurring ethanol exposure in Drosophila melanogaster.&#160;Here, we report the step-by-step Flypub assay including assembly of ethanol exposure chambers, setup of the assay station, criteria for fly care and collection, ethanol delivery, quantification of disinhibited courtship activities, data processing and statistical analysis. Also provided are how to troubleshoot critical steps, overcome limitations and expand its utility to assess additional ethanol-induced behaviors. The Flypub assay in combination with powerful genetic tools in Drosophila melanogaster will facilitate the task of discovering the mechanism underlying ethanol-induced behavioral sensitization.

The Flypub assay measures the behaviors that the fruit fly Drosophila melanogaster displays under the influence of ethanol. The assay can be readily mastered by experimenters at all levels and applied to various vaporized stimuli, facilitating substance abuse and addiction studies.

飞行酒吧研究乙醇诱导的行为抑制和敏感性

Alcohol use disorder (AUD) remains a serious problem in our society. To develop effective interventions for addiction, it is important to understand the ...

Determination of the Spontaneous Locomotor Activity in Drosophila melanogaster

Drosophila melanogaster has been used as an excellent model organism to study environmental and genetic manipulations that affect behavior. One such behavior is spontaneous locomotor activity. Here we describe our protocol that utilizes Drosophila population monitors and a tracking system that allows continuous monitoring of the spontaneous locomotor activity of flies for several days at a time. This method is simple, reliable, and objective and can be used to examine the effects of aging, sex, changes in caloric content of food, addition of drugs, or genetic manipulations that mimic human diseases.

Determination of the Spontaneous Locomotor Activity in Drosophila melanogaster

Drosophila melanogaster are useful in studying genetic or environmental manipulations that affect behaviors such as spontaneous locomotor activity.&#160; Here we describe a protocol that utilizes monitors with infrared beams and data analysis software to quantify spontaneous locomotor activity.&#160;

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

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 ...

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

Aggressive behavior in Drosophila melanogaster is composed of the sequential expression of stereotypical behavioral patterns (for analysis see 1). This complex behavior is influenced by genetic, hormonal and environmental factors. As in many organisms, previous fighting experience influences the fighting strategy of flies and the outcome of later contests: losing a fight increases the probability of losing later contests, revealing &#34;loser&#34; effects that likely involve learning and memory 2-4. The learning and memory that accompanies expression of complex social behaviors like aggression, is sensitive to pre-test handling of animals 5,6. Many experimental procedures are used in different laboratories to study aggression 7-9, however, no routinely used protocol that excludes handling of flies is currently available. Here, we report a new behavioral apparatus that eliminates handling of flies, using instead their innate negative geotactic responses to move animals into or out of fighting chambers. In this protocol, small circular fight arenas containing a food cup are divided into two equal halves by a removable plastic slider prior to introduction of flies. Flies enter chambers from their home isolation vials via sliding chamber doors and geotaxis. Upon removal of plastic sliders, flies are free to interact. After specified time periods, flies are separated again by sliders for subsequent experimentation. All of this is done easily without handling of individual flies. This apparatus offers a novel approach to study aggression and the associated learning and memory, including the formation of &#34;loser&#34; effects in fly fights. In addition, this new general-purpose behavioral apparatus can be employed to study other social behaviors of flies and should, in general, be of interest for investigating experience-related changes in fundamental behavioral processes.

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

During fruit fly fights, the behavioral patterns observed, fight dynamics, and associated learning and memory are influenced by experimental conditions. The protocol presented here describes a novel procedure that entirely eliminates handling of flies during experiments. This improves fight dynamics and allows formation of strong &#34;loser&#34; effects.

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 ...

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 ...

在条件影响社会空间

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饲养果蝇

测试环境线索对交配行为的影响的方法

对检测方法果蝇行为

一种自动测定果蝇在空间和时间温度变化中的性能的方法

飞行酒吧研究乙醇诱导的行为抑制和敏感性

在自发活动的测定

简单的检测方法的社交回避的量化

一种新的方法，省去了处理的研究侵略和"失败者"的影响

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