We would like to thank members of the Tessier lab for critical reading of this manuscript and helpful suggestions during the preparation of this protocol.

1.饥饿<ol><li>制备通过在标飞小瓶的底部饱和18.2兆欧水棉球飞饥饿小瓶中。另外，同样在药瓶内的角度饱和滤纸的18.2MΩ水的地方一小片。 </li><li>收集到的苍蝇套〜100只的二氧化碳垫，然后苍蝇添加到小瓶的准备。 注意:最好的结果是用动物是少于5天获得。然而，动物的确切年龄可被控制作为实验变量，以确定随时间的口味偏好的变化。 </li><li>使用棉球或泡沫塞子以固定小瓶封闭。放在自己身边瓶在环境控制孵化器。保持在25℃的温度，并在70％以上的湿度。离开瓶触及24小时。 </li></ol> 2.口味偏好分析<ol><li>制备用于测定所有促味剂在同一天一小号测试。 注意:要使用将根据实验的问题各不相同的确切促味剂被要求。以下是在本协议所使用的例子促味剂。请参阅优化第4节。 <ol><li>结合10微升的100毫米蔗糖溶液，13微升的红色食用色素和977微升18.2MΩ水准备控制呈味物质（1毫米蔗糖）。 </li><li>结合50微升100毫米蔗糖溶液，10微升蓝色食用色素，和940微升18.2MΩ水准备实验呈味物质（5毫米蔗糖）。 </li></ol></li><li>使用以下述方式制备的标准的100毫米×15毫米的塑料陪替氏培养皿使测定室: <ol><li>放置在12点和最靠近所述板的边缘控制促味剂的3个10微升滴在6点钟另一个3滴。确保液滴之间的间隔是相似的。 </li><li>放置最靠近所述板的边缘实验促味剂的3个10微升滴在3点和一个其他3滴在9点钟。确保液滴之间的间隔是相似的。 </li><li>根据需要重复尽可能多的重复步骤2.2.1和2.2.2。 </li></ol></li><li>空1瓶〜100饿死的苍蝇到的 CO 2垫或许足够用来麻醉所有动物（约10秒）。刷动物进入事先准备好的测定室的中间，并与菜盖上盖子。 应避免较长的CO 2的曝光时间，以改善恢复时间和限制干扰取食行为:注意。暴露于冰（〜5分钟）可以用于麻醉，以避免可能由即使是有限的曝光产生的 CO 2的行为的效果。 </li><li>放置测定室在一个不透明的纸板箱。请务必使用条件和基因型被检测到标签的包装盒外面。 </li><li>将整个设置程序（从步骤2.4所含纸箱内测定室）到25℃的培养箱具有至少70％的湿度为2小时。 </li&gt; <li>重复步骤2.3到2.5的所有重复。 </li><li> 2小时后，放置在测定室中，仍含有纸箱内，直接放入-20℃的冰箱，直到准备定量。 </li></ol> 3.口味偏好量化分析<ol><li>允许单个测定室温至室温（约5分钟）。 </li><li>在解剖镜下，用刷子或钳子的基础上，其腹部的颜色组动物:红色，蓝色，紫色或清除（ 图1）。 </li><li>记录动物在每个分组的数目。考虑没有参加试验明确动物，因此不包括他们在任何计算。 </li><li>根据下列方程之一计算优先指标: <ol><li>如果感兴趣的实验促味剂被添加到红色染料，然后用（N- 红色 + 0.5N 紫色 ）/（N 红色 + N 蓝色 + N purplE）。 </li><li>如果实验促味剂被添加到蓝色染料，然后调整公式（N- 蓝 + 0.5N 紫色 ）/（N 蓝色 + N 红色 + N 紫色 ）。 </li></ol></li><li>重复计算所有的实验条件和重复。 </li></ol> 4.口味偏好分析的优化<ol><li>经验确定的食品着色指标浓度被使用，因此食品着色不影响味道试验的结果，如下所示: <ol><li>使用如步骤2.1所示相同的基化合物（ 例如 5毫米蔗糖）准备4促味，但省略了食用色素。 </li><li>加1.3％的红色食品着色的促味剂的一种。使剩余的3促味剂在每个管不同浓度的蓝色食用色素（ 例如 ，0.6％，1％和1.3％）。 </li><li>完整的协议步骤2.2至3.4为每对呈味物质:1.3％红与0.6％的蓝色; 1.3％的红色与1％蓝色和1.3％的红色与1.3％的蓝色。 </li><li>重复步骤4.1.1-4.1.3与蓝色食用色素不同百分比直到偏好指数平均值为0（ 图2）的值。 注:作为一个起点，再加上1％的蓝色食用色素1.3％的红色食用色素通常会产生良好的效果。如果蓝色食用色素没有令人满意的浓度可以匹配到1.3％的染料，则步骤4.1.1通过4.1.3可以与不同浓度的红色着色和蓝色食品着色恒定浓度的重复。 </li><li>分析用相同的优化食用色素浓度待测试的所有条件。 </li></ol></li></ol>

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The authors declare that they have no competing financial interests.

我们已经描述了一个简单而有效的协议，用于确定在果蝇味道偏好。在实验中经常使用这个实验版本，以确定感知不同品质（苦，甜，酸，咸和鲜味）呈味物质的味觉受体（GRS）的贡献。 果蝇基因组包含大约60个基因其通过选择性剪接8,9-编码68鉴定味觉受体。然而，其他蛋白质，如离子型谷氨酸受体和TRP通道也被证明在味道10-13中发挥作用。因此，在昆虫味道辨别能?...

动物用chemosensation从不利条件，除了区分了有利条件。这种看法可能是这样的事情作为确定最好的食物来源，避免有毒物质或确定最佳的交配伙伴1关键。 Chemosensation通常分为两种感官部分:嗅觉和味觉的感官。这些感官的主要鉴别特征是，嗅觉（气味）被使用，而味觉（味道）需要与非易失性基板的身体接触来样周围的气态化学环境。这两种形式的感官刺激而被处理，并在大脑中产生相应的吸引力或排斥行为2解码神经系统的反应。这些感官因此对动物生存至关重要。 果蝇果蝇是不断增长的普及在了解使用模式生物ING昆虫如何感知气味和滋味。果蝇由于可用于分子，细胞，和行为途径的解剖遗传工具的丰富提供优于其他模型系统的巨大的优势。在过去15年的工作一直在表征特定细胞的身份，神经元受体和信号参与了嗅觉和味觉机制特别的工具。现在， 果蝇遗传学的功率被用来进一步阐明这些过程是如何在单个神经元和单电路级3-6编码。因此，它们提供了测定轻松打进改建感觉通路的读数是对这些字段的持续进步至关重要。 而大量的已知如何嗅觉信号进行编码并在大脑中处理的，要少得多理解有关在味觉通路类似的机制。我们在这里描述了可用于确定味道preferen一个协议行政长官在果蝇 ， 果蝇，像哺乳动物，一般喜欢甜味的化合物，而不是苦味化合物。这些食物源的任何组合可以在本实验设计来确定已知的遗传改变如何影响口味的选择可以利用。此外，药理学干预策略可以类似地评估其对动物的口味偏好效应。该测定的便利性和灵活性使得它有用的范例为理解在果蝇味觉感知的性质。

<table><tbody><tr><td>Blue Food Coloring (Water, Propylene Glycol, FD&amp;amp;C Blue 1 and Red 40, Propylparaben)</td><td>McCormick</td><td>N/A</td><td></td></tr><tr><td>Cryo/Freezer Boxes w/o Dividers</td><td>Fisher</td><td>03-395-455</td><td></td></tr><tr><td>Dumont #5 Forceps</td><td>Fine Science Tools</td><td>11251-20</td><td></td></tr><tr><td>Glacial Acetic Acid</td><td>Fisher</td><td>BP2401-500</td><td></td></tr><tr><td>Leica S6 E Stereozoom 0.63X-4.0X microscope</td><td>W. Nuhsbaum, Inc.</td><td>10446294</td><td></td></tr><tr><td>Petri dish (100 mm x 15 mm)</td><td>BD Falcon</td><td>351029</td><td>Reuseable if thoroughly washed and dried</td></tr><tr><td>Quick-Snap Microtubes</td><td>Alkali Scientific Inc.</td><td>C3017</td><td></td></tr><tr><td>Red Food Coloring (Water, Propylene Glycol, FD&amp;amp;C Reds 40 and 3, Propylparaben)</td><td>McCormick</td><td>N/A</td><td></td></tr><tr><td>Sucrose</td><td>IBI Scientific</td><td>IB37160</td><td></td></tr></tbody></table>

taste preference assay for adult drosophila

Olfactory and gustatory perception of the environment is vital for animal survival. The most obvious application of these chemosenses is to be able to distinguish good food sources from potentially dangerous food sources. Gustation requires physical contact with a chemical compound which is able to signal through taste receptors that are expressed on the surface of neurons. In insects, these gustatory neurons can be located across the animal's body allowing taste to play an important role in many different behaviors. Insects typically prefer compounds containing sugars, while compounds that are considered bitter tasting are avoided. Given the basic biological importance of taste, there is intense interest in understanding the molecular mechanisms underlying this sensory modality. We describe an adult Drosophila taste assay which reflects the preference of the animals for a given tastant compound. This assay may be applied to animals of any genetic background to examine the taste preference for a desired soluble compound.

从口味偏好分析一些典型结果如下所示。在大多数实验中腹部着色强度的一些变化可以看出（ 图1）。在腹部是否强烈或弱任何色素被认为是一个积极的摄入。因此，建议研究者将比分动物盲目的实验条件，同时，以限制任何潜在的偏见。 同样重要的是要选择食品染料不影响优先测定的结果的浓度。的使用各种食?...

Watch this Scientific Journal Video about Taste Preference Assay for Adult Drosophila at JoVE.com

Taste Preference Assay for Adult Drosophila

Taste is an important sensory process which facilitates attraction to beneficial substances and avoidance of toxic substances. This protocol describes a simple ingestion assay for determining Drosophila gustatory preference for a given chemical compound.

口味偏好测定成人<em&gt;果蝇</em

Olfactory and gustatory perception of the environment is vital for animal survival.  The most obvious application of these chemosenses is to be able to ...

taste-preference-assay-for-adult-drosophila

Research

JoVE Journal

Neuroscience

Taste Preference Assay for Adult Drosophila

9.6K 次观看.  Indiana University School of Medicine-South Bend. Taste is an important sensory process which facilitates attraction to beneficial substances and avoidance of toxic substances. This protocol describes a simple ingestion assay for determining Drosophila gustatory preference for a given chemical compound.

视频: Taste Preference Assay for Adult Drosophila

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 ...

科研

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 ...

Testing Drosophila Olfaction with a Y-maze Assay

Detecting signals from the environment is essential for animals to ensure their survival. To this aim, they use environmental cues such as vision, mechanoreception, hearing, and chemoperception through taste, via direct contact or through olfaction, which represents the response to a volatile molecule acting at longer range. Volatile chemical molecules are very important signals for most animals in the detection of danger, a source of food, or to communicate between individuals. Drosophila melanogaster is one of the most common biological models for scientists to explore the cellular and molecular basis of olfaction. In order to highlight olfactory abilities of this small insect, we describe a modified choice protocol based on the Y-maze test classically used with mice. Data obtained with Y-mazes give valuable information to better understand how animals deal with their perpetually changing environment. We introduce a step-by-step protocol to study the impact of odorants on fly exploratory response using this Y-maze assay.

Testing Drosophila Olfaction with a Y-maze Assay

We present a choice test to reveal the influence of odorants on Drosophila behavior using a Y-maze assay.

测试<em&gt;果蝇</em&gt;用Y型迷宫测定嗅觉

Detecting signals from the environment is essential for animals to ensure their survival. To this aim, they use environmental cues such as vision, ...

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. ...

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

We describe a protocol for measuring ethanol self-administration in fruit flies (Drosophila melanogaster) as a proxy for changes in reward states. We demonstrate a simple way to tap into the fly reward system, modify experiences related to natural reward, and use voluntary ethanol consumption as a measure for changes in reward states. The approach serves as a relevant tool to study the neurons and genes that play a role in experience-mediated changes of internal state. The protocol is composed of two discrete parts: exposing the flies to rewarding and nonrewarding experiences, and assaying voluntary ethanol consumption as a measure of the motivation to obtain a drug reward. The two parts can be used independently to induce the modulation of experience as an initial step for further downstream assays or as an independent two-choice feeding assay, respectively. The protocol does not require a complicated setup and can therefore be applied in any laboratory with basic fly culture tools.

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

We describe a protocol for inducing rewarding and nonrewarding experiences in fruit flies (Drosophila melanogaster) using voluntary ethanol consumption as a measure for changes in reward states.

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 ...

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 ...

A Rapid Food-Preference Assay in Drosophila

To select food with nutritional value while avoiding the consumption of harmful agents, animals need a sophisticated and robust taste system to evaluate their food environment. The fruit fly, Drosophila melanogaster, is a genetically tractable model organism that is widely used to decipher the molecular, cellular, and neural underpinnings of food preference. To analyze fly food preference, a robust feeding method is needed. Described here is a two-choice feeding assay, which is rigorous, cost-saving, and fast. The assay is Petri-dish-based and involves the addition of two different foods supplemented with blue or red dye to the two halves of the dish. Then, ~70 prestarved, 2-4-day-old flies are placed in the dish and&#160;allowed to choose between blue and red foods in the dark for about 90 min. Examination of the abdomen of each fly is followed by the calculation of the preference index. In contrast to multiwell plates, each Petri dish takes only ~20 s to fill and saves time and effort. This feeding assay can be employed to quickly determine whether flies like or dislike a particular food.

A Rapid Food-Preference Assay in Drosophila

We present a protocol for a two-choice feeding assay for flies. This feeding assay is fast and easy to run and is suitable not only for small-scale laboratory research, but also for high-throughput behavioral screens in flies.

果蝇的快速食物偏好分析

To select food with nutritional value while avoiding the consumption of harmful agents, animals need a sophisticated and robust taste system to evaluate ...

Feeding Test: Assessment of Feeding Preferences in Drosophila melanogaster

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).
This video describes a feeding assay for fruit flies in which colored food is used to monitor feeding and the effect added compounds may have on feeding activity. The example protocol tests the effects of several endocrine disrupting chemicals (EDC).

喂养测试:黑腹果蝇喂养偏好的评估

Source: Bovier, T. F., et al. Methods to Test Endocrine Disruption in Drosophila melanogaster. J. Vis. Exp. (2019).
This video describes a feeding assay ...

JoVE Encyclopedia of Experiments

生物学

黑腹果蝇（果蝇）

成虫

Temperature Gradient Assay: A Method to Test Temperature Preference in Drosophila Larvae

Source: Liu, J., et al. <a href="https://www.jove.com/video/57963/" target="_blank">A Temperature Gradient Assay to Determine Thermal Preferences of Drosophila Larvae</a>. J. Vis. Exp. (2018).
Drosophila fruit flies can distinguish small changes in temperature and therefore select environments with favorable thermal conditions. This video describes a behavioral assay that tests the temperature preference of Drosophila larvae on a linear thermal gradient.

温度梯度测定:一种测试果蝇幼虫温度偏好的方法

Source: Liu, J., et al. A Temperature Gradient Assay to Determine Thermal Preferences of Drosophila Larvae. J. Vis. Exp. (2018).
Drosophila fruit flies ...

幼虫

Taste Preference Assay: A Method for Measuring Feeding Behavior in Drosophila

Source: Bantel, A. P. and Tessier, C. R. <a href="https://www.jove.com/video/54403/" target="_blank">Taste Preference Assay for Adult Drosophila</a>. J. Vis. Exp. (2016).
This video describes the taste preference assay, a behavioral method used to measure attraction or avoidance towards colored solutions that taste differently by assessing the fly&#39;s abdominal coloration after ingestion of the preferred substance. The featured protocol demonstrates the procedure used to measure flies&#39; preference towards solutions of varying sucrose concentrations.

味觉偏好测定:一种测量果蝇摄食行为的方法

Source: Bantel, A. P. and Tessier, C. R. Taste Preference Assay for Adult Drosophila. J. Vis. Exp. (2016).
This video describes the taste preference ...

口味偏好测定成人

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欲求联想嗅觉学习

单苍蝇检测方法的觅食行为的

测试

高通量检测，以检查个体产蛋首

一个简单的方法来衡量在改建悬赏寻求行为使用

毛细管馈线分析测量的摄食

果蝇的快速食物偏好分析

喂养测试:黑腹果蝇喂养偏好的评估

温度梯度测定:一种测试果蝇幼虫温度偏好的方法

味觉偏好测定:一种测量果蝇摄食行为的方法