The authors acknowledge continued funding the Natural Sciences and Engineering Council of Canada (NSERC) to MBS.

1.准备葡萄板和控股瓶的幼虫采集<ol><li>为了使持有瓶，切孔的6盎司飞培养瓶，大到足以适合供气苍蝇小瓶插头（ 图1D）的一面。 </li><li>使葡萄板，通过煮沸琼脂，葡萄汁制备250毫升葡萄汁培养基（1.8％琼脂，45％葡萄汁，2.5％乙酸，2.5％的乙醇）和大部分的水，降温至70℃（搅拌，同时冷却），然后添加乙酸和乙醇，并把最多体积水。 </li><li>使用注射器（没有针）来填充35×10毫米的培养皿的盖子，直至葡萄汁介质的表面，使盖（ 图1A）的唇部上方的圆顶。 </li><li>在潮湿和气密容器商店葡萄板在4ºC直到使用（最多2周）。 </li></ol> 2.食品板的制备<ol><li>标准的准备飞食品的配方。 注意:给T在路径长度EST饲养食品品质的影响，我们使用低营养飞食品:10％蔗糖，1.3％琼脂，0.8％酒石酸钠钾，0.1％磷酸二氢钾，0.05％氯化钠，0.05％氯化钙，0.05％氯化镁， 0.05％硫酸铁，5％酵母和0.5％的丙酸水溶液;和高营养飞食品:1.5％蔗糖，1.4％琼脂，3％葡萄糖，1.5％玉米粉，1％小麦胚芽，1％大豆粉，3％糖蜜，3.5％酵母，0.5％的丙酸，0.2％Tegosept和1 ％乙醇在水中。对于这两种食品中添加丙酸，Tegosept和乙醇前高压灭菌的解决方案，并冷却至50℃。 </li><li>倒加入40ml飞食品为100×15毫米的培养皿。 </li><li>在潮湿和密闭容器储存食品板块在4ºC直到使用（最多2周）。 </li></ol> 3.准备测试板<ol><li>制备58×25厘米，厚6毫米的黑色有机玻璃板10，1mm深的圆形的孔，直径10厘米和布置在2×5天ESIGN（图1C）中测试幼虫上。 注:最多30幼虫可以一次如果3个或更多的上述树脂玻璃板的可进行测试。可替代地，幼虫可在培养皿中，在10厘米直径的测试。该选项是更费力的，并允许幼虫较低的数字进行测试。 </li><li>得到黑色39×8厘米，厚6毫米的有机玻璃吊具上的有机玻璃试验板扩频酵母膏。 </li><li>制备100×15毫米的培养皿盖子用于通过预先分离塔底罐盖记录幼虫路长度。 </li></ol> 4.设置家长人群<ol><li>保持饲养条件在整个实验常数（ 例如 ，25ºC，60％湿度和12:12光:对标飞食品暗周期）。 </li><li>提高相同的饲养条件的家长群体。 </li><li>收集100 - 150名女性和50 - 75男性，年龄为他们5±1天为最佳的生育能力。 </li><li>允许˚Females和男性在一个6盎司苍蝇培养瓶与标准的飞食品交配过夜。 </li><li>转飞入控股瓶和安全葡萄板（在葡萄板少量鲜酵母膏将促进产蛋;图1B）在用胶带（ 图1E）的顶部。 </li><li>离开瓶底部倒立（图1F），与葡萄板以允许成年人的葡萄板产卵。 </li><li>弃去前葡萄板后24小时，并放置一个新的葡萄板在保持瓶子。这将是从幼虫收集第二天葡萄板。 注:葡萄板块将有正在组建，成立了板，使他们能够早日第二天被清除后，幼虫22小时的清除（ 例如 ，在12日下午成立了板，并在10清楚次日上午）。明智的做法是放弃的第一个葡萄板，以避免女性可能已经隐瞒和Tha任何鸡蛋ŧ可能在发展中前进。 </li></ol> 5.葡萄板收集L1（一龄）幼虫<ol><li>在一个60×15毫米的培养皿取下瓶子，他们建立了22个小时后发生葡萄板。 </li><li>将与控股瓶鲜酵母膏新的葡萄板。 </li><li>清除并丢弃使用解剖探头种子葡萄板上的所有幼虫。 </li><li>清除孵育葡萄板在标准条件下4小时。 </li><li> 4小时后，挑每株100的L1幼虫从葡萄板和放置在食品板。 <ol><li>广场上轻轻食物的幼虫（不穿孔食品和埋葬的幼虫），并通过分发整个食品板块幼虫避免拥挤。 </li></ol></li><li>孵育食品板块，直到实验动物是流浪开始前约10小时。 注: - 孵化后96小时在25ºC，通常徘徊10小时之前，对应于72。该EXAC孵育时间t用量应通过收集一组初始的幼虫对每个菌株和记录的时间数，直到徘徊的开始开始实验之前确定（游荡幼虫将食品搬出到食品的侧面和盖板）。 注意:如果需要的话，同样的父母可用于收集3幼虫 - 通过重复步骤4.7至5.5的测试4天。改变在其中菌株建立和测试跨天，以避免在路径长度的测试顺序效应的顺序。测试相同的"控制权"株/株测试的每一天，以控制天的影响。 </li><li>如果许多菌株被测试，错开幼虫的设置和收集时间，以允许各菌株在适当的时间点测试以后。 </li></ol> 6.幼虫路径长度测试<ol><li>溶于水干活性面包酵母以1:2的比例（重量比体积）。成品酵母膏应该比巴薄ncake面糊，在起重勺子出糊剂时的一种方式，它滴落和叶迅速融化的糊的表面上带的后面。 注意:酵母膏的稠度是重要的，应是在整个实验常数。确切的酵母:水的比例可能会根据不同的品牌和批次的酵母进行调整。 </li><li>轻轻一挑第一株幼虫从使用画笔食品板块进行测试。 </li><li>收集幼虫在培养皿中，用水轻轻洗涤以除去所有的食品。 <ol><li>快速洗幼虫用1 - 2毫升水，轻轻地轻拍幼虫干燥用实验室擦拭避免将在测试板上的幼虫时传输任何水。为了避免精神紧张，保持湿润的幼虫，但水不沉。 </li></ol></li><li>排列并排三个测试板侧，并设置一个5分钟计时器为每个测试板。 </li><li>应用酵母膏到测试板的顶部，并使用吊具，扩散薄捻在每个测试板，使得所有的孔中有酵母的偶数层的酵母膏的呃。 </li><li>轻轻将一个幼虫在每个的中心孔，将第一幼虫上每个板之后开始的5分钟的定时器。注:三个板可在同一时间内完成，得到30 /应变/测试一天中的样本大小，最多共500幼虫。 </li><li>放置在每个顶部培养皿盖好。 </li><li>当计时器用完，无需从板取出幼虫，开始使用永久性标记追查酵母留下的路径长度。在相同的顺序幼虫迹路长度被放置在测试板上。 <ol><li>执行以相同的速度跟踪作为幼虫放置在平板上。在以后的数据分析的一致性，用于所有路径长度相同的标记。建议永久性标记。 </li></ol></li><li>重复步骤6.2至6.8.1对于每个要测试的菌株。立即进行测试，以避免对PA饥饿效应前拾取从食物中每一株TH-长度。 </li></ol> 7.食品剥夺<ol><li>如果幼虫是在试验前禁食，如在步骤6.2至6.3中所述收集幼虫，但食物剥夺时所需量的早期（ 例如 ，如果幼虫要禁食4小时，收集幼虫4小时前测试时间）。 </li><li>划分洗涤幼虫分为两组，并放置食物剥夺组中的60×15毫米的培养皿用5毫升1％琼脂和对照组在培养皿用5毫升标准食物。 </li><li>将重量上的培养皿倒盖（否则幼虫都能够为食搜索时爬出来）的顶部。 </li><li>孵育先前建立的食物匮乏时期和步骤6.2到6.9描述的测试继续进行。 </li><li>如果许多菌株进行测试，错开剥夺食物开始时间，以允许在适当的时间点每种菌株的检测。 </li></ol> 8.数据分析注意:本节描述了使用ImageJ 7或斐济8对于路径长度的处理的数据分析的方法，尽管其他软件可以被使用。 <ol><li>使用一个光表来从培养皿盖所有跟踪路长度转移到白纸（盖可以用乙醇洗涤，并为进一步路径长度记录重复使用）。 </li><li>跟踪在纸张的片材之一50mm的直参考线。 </li><li>扫描路径长度在300 dpi的分辨率，并保存为位图或TIFF图像文件。 </li><li>打开图像文件，选择"进程"选项卡&gt;"二进制"&gt;"使二进制"。那么"进程"选项卡下选择"二进制"&gt;"缩略"。这产生的无论迹线的厚度的一个像素宽度线。 </li><li>确保每个路径的长度显示为无交叉连续一行。 </li><li>单独使用"直立任何交集NE工具棒"（跟踪）工具"和着色线白色（路径的连续性可以与被检查"时，整个路径应当强调黄色（ 图2）。 </li><li>选择"分析"选项卡&gt;设置测量，然后选择"雷池"和"显示标签"。 </li><li>选择"分析"选项卡&gt;"设置规模"，选择"清除水垢"。 </li><li>在图像，只能选择参考线，选择"分析"选项卡&gt;"分析粒子"，并选择"显示:大纲"，"显示结果"和"清除成绩表"。注意:输出将显示对应于50毫米标线的像素数。 </li><li>选择"分析"标签&gt;"设置规模"，并在"以像素为单位的距离"中的像素数和在"已知距离"对应的距离（50毫米）填充。 </li><li>最后，选择都是一个基因型的路径长度图纸TA的时候，选择"分析"选项卡&gt;"分析粒子"，并选择"显示:大纲"，"显示结果"和"清除成绩表"。 <ol><li>通过选择区域并填充它白色移除所选区域内的任何书面标签或明显外来标记（它们将被识别为路径长度）。 </li></ol></li><li>目测"纲要"的输出文件，以确保周边​​被正确确定。输出将显示一个路径长度在毫米为在选择的每个路径长度图。这很容易被导出到电子表格，并与任何统计软件进行分析。 </li></ol>

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The authors have nothing to disclose.

这里列出的路径长度的测定提供了果蝇幼虫觅食行为一个强大的和简单的措施。协议遵循Sokolowski 2中描述的一般方法，但一直以来在问候效率和实验对照得到改善。据我们所知，这方法是测量幼虫路径长度的唯一可用的方法。原始版本的路径长度协议2，3，15上的培养皿16测试幼虫酵母膏的薄层用刷子施加，后来的玻璃扩散杆。更近的所述协议10的版本中?...

由于白基因在托马斯亨特摩根的实验室在1910年发现，果蝇， 黑腹果蝇 （ 果蝇 ），已被用作各种生物过程的分子和生理基础的研究的模型。 D的普及果蝇很大程度上从遗传工具的相当数量和品种茎。 果蝇的体积小，相对易于处理和短代时间使其为遗传学研究的理想模型。同样重要的是果蝇表现出许多由更复杂的生物，包括哺乳动物表达的表型的能力。这包括复杂的表型，如行为站在生物体和其环境之间的接口。由于对果蝇这样，行为学研究作出了重大贡献我们的基因和环境如何调解行为的理解1。 其中D的第一研究果蝇幼虫的行为通过测量幼虫2路长度而喂养调查幼虫取食策略个体差异。路径长度被定义为通过在酵母单个幼虫行进的总距离，五分钟内。在多伦多这两个实验室株和苍蝇从人口自然在他们的觅食行为的变化，并有遗传因素在路径长度的个体差异。两个幼虫取食摇身一变，从定量路径长度分布描述他们被称为月球车和保姆。流动站表现出更长的路径长度而穿过较大的区域，而不是保姆食品基物上。使用此路径长度分析，百丽德等 3所映射的觅食（对于）基因基本以分散的区域，这些个体行为差异对chromosome- 2（24A3-24C5）。 D.在米elanogaster 用于基因稍后克隆4和显露是一个的cGMP依赖性蛋白激酶5，在果蝇生理和行为和其他生物6的调节剂。 在这里，我们列出了最初在Sokolowski 2开发的幼虫路径长度分析当前协议。虽然该试验的某些方面已经改变多年来，设计背后的概念还没有。我们还提供数据来说明分析的潜力，以评估在果蝇幼虫觅食行为的个体差异的遗传和环境的贡献。幼虫路径长度测定法是简单，有效，但健壮。单人可以测试高达500幼虫容易在四小时的结果可以具有高水平的再现性来获得。最初开发本地化为 ，它可以在遗传筛选，数量性状基因座的映射被使用，并且在研究基因逐环境（GXE）的相互作用。此外，它的简单性和可重复性使其成为本科教学的重要资源。

<table><tbody><tr><td>6 oz&amp;nbsp;fly culture bottles&amp;nbsp;</td><td>Fisher Scientific&amp;nbsp;</td><td>AS355&amp;nbsp;</td><td></td></tr><tr><td>Fly vial plugs</td><td>Droso-Plugs</td><td>59-201</td><td></td></tr><tr><td>35 x 10 mm Petri dishes&amp;nbsp;</td><td>Falcon</td><td>351008</td><td></td></tr><tr><td>100 x 15 mm Petri dishes&amp;nbsp;</td><td>Fisher</td><td>875712</td><td></td></tr><tr><td>60 x 15 mm Petri dishes</td><td>VWR</td><td>25384-168&amp;nbsp;</td><td></td></tr><tr><td>Dissecting probes</td><td>Almedic</td><td>2325-58-5300&amp;nbsp;</td><td></td></tr><tr><td>Yeast</td><td>Lab Scientific</td><td>FLY-8040-20F</td><td></td></tr></tbody></table>

foraging path-length protocol for drosophila melanogaster larvae

果蝇幼虫路径长度的表型是用来研究行为变化的遗传和环境的贡献建立措施。幼虫路径长度法的开发是为了衡量后来被链接到觅食基因觅食行为的个体差异。幼虫路径长度是易攻入性状便于大样本的收集，以最低的成本，为遗传筛选。在这里，我们提供当前协议的用于第一使用Sokolowski幼虫路径长度测定的详细描述。该协议详细介绍了如何处理重复性试验动物，进行行为测定和分析数据。的测定法可如何用于测量响应于环境变化行为可塑性，通过操纵进给环境之前，执行测定的例子中，还提供。最后，合适的测试设计以及环境因素，可以修改幼虫路径长度如食品质量，发育年龄和一天的效果进行了讨论。

<html>在流动站和保姆为菌株和食物剥夺对路径长度的影响的路径长度的差异在图中示出。 3.数据收集测试连续三天呈现显著应变效应（F（1,421）= 351.89，P &lt;2.20×10 -16;图3A），与流浪旅行比保姆更远。除了 ​​应变的效果，也有一个显著食物治疗效果（F（1，421）= 461.62，P &lt;2.20×10 -16;图3...

Watch this Scientific Journal Video about Foraging Path-length Protocol for Drosophila melanogaster Larvae at JoVE.com

Foraging Path-length Protocol for Drosophila melanogaster Larvae

觅食路径长度协议<em&gt;果蝇</em&gt;幼虫

We provide a detailed protocol for a Drosophila melanogaster foraging path-length assay. We discuss the preparation and handling of test animals, how to perform the assay and analyze the data.

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

foraging-path-length-protocol-for-drosophila-melanogaster-larvae

Research

JoVE Journal

Neuroscience

Foraging Path-length Protocol for Drosophila melanogaster Larvae

9.3K 次观看.  University of Toronto. We provide a detailed protocol for a Drosophila melanogaster foraging path-length assay. We discuss the preparation and handling of test animals, how to perform the assay and analyze the data.

视频: Foraging Path-length Protocol for Drosophila melanogaster Larvae

High-resolution Measurement of Odor-Driven Behavior in Drosophila Larvae

Olfactory responses in Drosophila larvae have been traditionally studied in Petri dishes comprising a single peripheral odor source. In this behavioral paradigm, the experimenter usually assumes that the rapid diffusion of odorant molecules from the source leads to the creation of a stable gradient in the dish. To establish a quantitative correlation between sensory inputs and behavioral responses, it is necessary to achieve a more thorough characterization of the odorant stimulus conditions. In this video article, we describe a new method allowing the construction of odorant gradients with stable and controllable geometries. We briefly illustrate how these gradients can be used to screen for olfactory defects (full and partial anosmia) and to study more subtle features of chemotaxis behavior.

In this video article, we describe a new method allowing the construction of odorant gradients with stable and controllable geometries. We briefly illustrate how these gradients can be used to screen for olfactory defects (full and partial anosmia) and to study more subtle features of chemotaxis behavior.

高分辨率测量的气味驱动在果蝇幼虫的行为

Olfactory responses in Drosophila larvae have been traditionally studied in Petri dishes comprising a single peripheral odor source. In this behavioral ...

科研

生物学

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 ...

Functional Analysis of the Larval Feeding Circuit in Drosophila

The serotonergic feeding circuit in Drosophila melanogaster larvae can be used to investigate neuronal substrates of critical importance during the development of the circuit. Using the functional output of the circuit, feeding, changes in the neuronal architecture of the stomatogastric system can be visualized. Feeding behavior can be recorded by observing the rate of retraction of the mouth hooks, which receive innervation from the brain. Locomotor behavior is used as a physiological control for feeding, since larvae use their mouth hooks to traverse across an agar substrate. Changes in feeding behavior can be correlated with the axonal architecture of the neurites innervating the gut. Using immunohistochemistry it is possible to visualize and quantitate these changes. Improper handling of the larvae during behavior paradigms can alter data as they are very sensitive to manipulations. Proper imaging of the neurite architecture innervating the gut is critical for precise quantitation of number and size of varicosities as well as the extent of branch nodes. Analysis of most circuits allow only for visualization of neurite architecture or behavioral effects; however, this model allows one to correlate the functional output of the circuit with the impairments in neuronal architecture.

Functional Analysis of the Larval Feeding Circuit in Drosophila

The feeding circuit in Drosophila melanogaster larvae serves a simple yet powerful model that allows changes in feeding rate to be correlated with alterations in the stomatogastric neural circuitry. This circuit is composed of central serotonergic neurons that send projections to the mouth hooks as well as the foregut.

在幼虫馈电电路的功能分析果蝇</i

The serotonergic  feeding circuit in Drosophila  melanogaster larvae can be used to investigate neuronal substrates of  critical importance during the ...

An Affordable and Efficient "Homemade" Platform for Drosophila Behavioral Studies, and an Accompanying Protocol for Larval Mitochondrial Respirometry

The usefulness of Drosophila as a model organism for the study of human diseases, behaviors and basic biology is unquestionable. Although practical, Drosophila research lacks popularity in developing countries, possibly due to the misinformed idea that establishing a lab and performing relevant experiments with such tiny insects is difficult and requires expensive, specialized apparatuses. Here, we describe how to build an affordable flylab to quantitatively analyze a myriad of behavioral parameters in D. melanogaster, by 3D-printing many of the necessary pieces of equipment. We provide protocols to build in-house vial racks, courtship arenas, apparatuses for locomotor assays, etc., to be used for general fly maintenance and to perform behavioral experiments using adult flies and larvae. We also provide protocols on how to use more sophisticated systems, such as a high resolution oxygraph, to measure mitochondrial oxygen consumption in larval samples, and show its association with behavioral changes in the larvae upon the xenotopic expression of the mitochondrial alternative oxidase (AOX). AOX increases larval activity and mitochondrial leak respiration, and accelerates development at low temperatures, which is consistent with a thermogenic role for the enzyme. We hope these protocols will inspire researchers, especially from developing countries, to use Drosophila to easily combine behavior and mitochondrial metabolism data, which may lead to information on genes and/or environmental conditions that may also regulate human physiology and disease states.

<meta charset="utf8"></head><body>一个经济实惠且高效的“自制”果蝇行为研究平台，以及幼虫线粒体呼吸测定的配套方案

We provide protocols for anyone with a "maker culture" mind to start building a flylab for quantitative analysis of a myriad of behavioral parameters in Drosophila melanogaster, by 3D-printing many of the necessary pieces of equipment. We also describe a high resolution respirometry protocol using larvae to combine behavioral and mitochondrial metabolism data.

一个经济实惠且高效的“自制”果 蝇 行为研究平台，以及幼虫线粒体呼吸测定的配套方案

The usefulness of Drosophila as a model organism for the study of human diseases, behaviors and basic biology is unquestionable. Although practical, ...

行为学

Using Linear Agarose Channels to Study Drosophila Larval Crawling Behavior

Drosophila larval crawling is emerging as a powerful model to study neural control of sensorimotor behavior. However, larval crawling behavior on flat open surfaces is complex,&#160;including: pausing, turning, and meandering. This complexity in the repertoire of movement hinders detailed analysis of the events occurring during a single crawl stride cycle. To overcome this obstacle, linear agarose channels were made that constrain larval behavior to straight, sustained, rhythmic crawling. In principle, because agarose channels and the Drosophila larval body are both optically clear, the movement of larval structures labeled by genetically-encoded fluorescent probes can be monitored in intact, freely-moving larvae. In the past, larvae were placed in linear channels and crawling at the level of whole organism, segment, and muscle were analyzed1. In the future, larvae crawling in channels can be used for calcium imaging to monitor neuronal activity. Moreover, these methods can be used with larvae of any genotype and with any researcher-designed channel. Thus the protocol presented below is widely applicable for studies using the Drosophila larva as a model to understand motor control.

Using Linear Agarose Channels to Study Drosophila Larval Crawling Behavior

The Drosophila larva is a powerful model system to study neural control of behavior. This publication describes the use of linear agarose channels to elicit sustained bouts of linear crawling and methods to quantify the dynamics of larval structures during repetitive crawling behavior.

用线性琼脂糖渠道研究<em&gt;果蝇</em&gt;幼虫爬行行为

Drosophila larval crawling is emerging as a powerful model to study neural control of sensorimotor behavior. However, larval crawling behavior on flat ...

Tracking Drosophila Larval Behavior in Response to Optogenetic Stimulation of Olfactory Neurons

The ability of insects to navigate toward odor sources is based on the activities of their first-order olfactory receptor neurons (ORNs). While a considerable amount of information has been generated regarding ORN responses to odorants, the role of specific ORNs in driving behavioral responses remains poorly understood. Complications in behavior analyses arise due to different volatilities of odorants that activate individual ORNs, multiple ORNs activated by single odorants, and the difficulty in replicating naturally observed temporal variations in olfactory stimuli using conventional odor-delivery methods in the laboratory. Here, we describe a protocol that analyzes Drosophila larval behavior in response to simultaneous optogenetic stimulation of its ORNs. The optogenetic technology used here allows for specificity of ORN activation and precise control of temporal patterns of ORN activation. Corresponding larval movement is tracked, digitally recorded, and analyzed using custom written software. By replacing odor stimuli with light stimuli, this method allows for a more precise control of individual ORN activation in order to study its impact on larval behavior. Our method could be further extended to study the impact of second-order projection neurons (PNs) as well as local neurons (LNs) on larval behavior. This method will thus enable a comprehensive dissection of olfactory circuit function and complement studies on how olfactory neuron activities translate in to behavior responses.

Tracking Drosophila Larval Behavior in Response to Optogenetic Stimulation of Olfactory Neurons

This protocol analyzes navigational behavior of Drosophila larva in response to simultaneous optogenetic stimulation of its olfactory neurons. Light of 630 nm wavelength is used to activate individual olfactory neurons expressing a red-shifted channel rhodopsin. Larval movement is simultaneously tracked, digitally recorded, and analyzed using custom-written software.

跟踪果蝇幼虫行为对嗅神经元 Optogenetic 刺激的反应

The ability of insects to navigate toward odor sources is based on the activities of their first-order olfactory receptor neurons (ORNs). While a ...

Light Spot-Based Assay for Analysis of Drosophila Larval Phototaxis

The larvae of Drosophila melanogaster show obvious light-avoiding behavior during the foraging stage. Drosophila larval phototaxis can be used as a model to study animal avoidance behavior. This protocol introduces a light-spot assay to investigate larval phototactic behavior. The experimental set-up includes two main parts: a visual stimulation system that generates the light spot, and an infrared light-based imaging system that records the process of larval light avoidance. This assay allows tracking of the behavior of larva before entering, during encountering, and after leaving the light spot. Details of larval movement including deceleration, pause, head casting, and turning can be captured and analyzed using this method.

Light Spot-Based Assay for Analysis of Drosophila Larval Phototaxis

This protocol introduces a light-spot assay to investigate Drosophila larval phototactic behavior. In this assay, a light spot is generated as light stimulation, and the process of larval light avoidance is recorded by an infrared light-based imaging system.

光斑分析分析果蝇拉瓦尔光轴

The larvae of Drosophila melanogaster show obvious light-avoiding behavior during the foraging stage. Drosophila larval phototaxis can be used as a model ...

Lifespan Protocol for Drosophila melanogaster: Generating Survival Curves to Identify Differences in Fly Longevity

Source: Bovier, T. F., et al. <a href="https://www.jove.com/video/59535/" target="_blank">Methods to Test Endocrine Disruption in Drosophila melanogaster</a>. J. Vis. Exp. (2019).
Scientists use the Drosophila lifespan protocol to measure how different experimental conditions affect flies&#39; longevity and generate survival curves in the process to quantify the experimental outcomes. The sample protocol features the assay being applied to study the effects on longevity from exposure to varying concentrations of endocrine disruptor chemicals, or EDCs, such as 17-&#945;-ethinylestradiol (EE2).

黑腹果蝇的寿命方案:生成生存曲线以确定果蝇寿命的差异

Source: Bovier, T. F., et al. Methods to Test Endocrine Disruption in Drosophila melanogaster. J. Vis. Exp. (2019).
Scientists use the Drosophila lifespan ...

觅食路径长度协议

摘要

探索更多视频

高分辨率测量的气味驱动在果蝇幼虫的行为

欲求联想嗅觉学习

单苍蝇检测方法的觅食行为的

在幼虫馈电电路的功能分析果蝇

一个经济实惠且高效的“自制”果蝇行为研究平台，以及幼虫线粒体呼吸测定的配套方案

一个经济实惠且高效的“自制”果蝇行为研究平台，以及幼虫线粒体呼吸测定的配套方案

用线性琼脂糖渠道研究

跟踪果蝇幼虫行为对嗅神经元 Optogenetic 刺激的反应

光斑分析分析果蝇拉瓦尔光轴

黑腹果蝇的寿命方案:生成生存曲线以确定果蝇寿命的差异

觅食路径长度协议

摘要

探索更多视频

高分辨率测量的气味驱动在果蝇幼虫的行为

欲求联想嗅觉学习

单苍蝇检测方法的觅食行为的

在幼虫馈电电路的功能分析果蝇

一个经济实惠且高效的“自制”果 蝇 行为研究平台，以及幼虫线粒体呼吸测定的配套方案

一个经济实惠且高效的“自制”果 蝇 行为研究平台，以及幼虫线粒体呼吸测定的配套方案

用线性琼脂糖渠道研究

跟踪果蝇幼虫行为对嗅神经元 Optogenetic 刺激的反应

光斑分析分析果蝇拉瓦尔光轴

黑腹果蝇的寿命方案:生成生存曲线以确定果蝇寿命的差异

一个经济实惠且高效的“自制”果蝇行为研究平台，以及幼虫线粒体呼吸测定的配套方案

一个经济实惠且高效的“自制”果蝇行为研究平台，以及幼虫线粒体呼吸测定的配套方案