提供的资金由美国国家科学基金会（MF）。

简介 OMT是一种适应磁性的系绳系统设计纳入一个“虚拟羽模拟器”[1]。以下协议将解释如何正确系绳苍蝇（第一部分），并提供设立的磁体（第二部分）和质量流量调节气味传送系统（第三部分）的战略。这里所描述的信息是这个特殊的系统优化和与其他技术规范的组成部分可能会有所不同。 第1部分：圈养苍蝇这里描述的数据共享的步骤相似，许多以往的研究[2]，但在使用OMT适应[3]。正确的圈养是至关重要的，以确保强大的和可重复的实验试验。小心圈养的重要性不能被夸大了。 <ol><li>收集苍蝇后4-6天羽化，饿死4-6小时。放置kimwipe蘸水在馁，以防止干燥的小瓶。 </li><li>一批的苍蝇从瓶子传送到一个​​小瓶，并插入上Peltier冷却阶段的黄铜块设置在约4摄氏度。适当的温度anesthetizes在大约30秒内的苍蝇。 </li><li>排序冰冷的表面上的冷却阶段的苍蝇。潮湿Kimwipe确保从角质层不断冷却和吸湿排汗。选择圈养的最大的女性。 </li><li>用细刷，推位单形容纳一个野生型雌果蝇的定制的石棺飞。 <img src="/files/ftp_upload/old/1063_2008_11_26_2/1063_Figure-01_thumb.png" alt="" /> 图1。 <a href="https://jove.unicartagenaproxy.elogim.com/files/1063_2008_10_28_1/1063_Figure-01.png">请点击这里看到一个更大的版本的图。</a> </li><li>我们的位置，与装在一个显微一个磁棒一个minutien针（罚款科学工具）。引脚对齐钝端接触，只是后面的两翼之间的头背胸部。头朝向眼睛看时，该引脚应完全垂直。即使少量的“卷”之间的飞行和PIN无法容忍的。侧视角度之间的脚和身体的长轴应在鼻子的角度为30度，相对地平线（图1）。注：在磁铁系绳的优化运行，引脚可能需要修剪长度较短。这将完全取决于悬浮磁铁之间的距离。 </li><li>一旦minutien引脚对齐，使用细钢丝申请的紫外线激活胶（ElectroLite集团）小降minutien针钝端。 </li><li>操作的针，使胶液滴触及头部上方飞的胸部。有了适当的墨滴大小，胶应退火和流量从引脚到胸部。小心不要使用太多胶水。 </li><li>两个20第二阵阵的紫外光固化胶水。 </li><li>将固定飞一边，重复。切成小方块Kimwipe钳给每个系留飞行限制飞行。 </li><li>经过一个小时的恢复，拴苍蝇用于实验的准备。 </li><li>粘合不当可能阻碍一个飞行的能力，以执行在磁铁上。应检查每一个飞行平稳的旋转，在每一个实验，以确保正确粘苍蝇的开始。这可以通过旋转一个足够大的空间时期的条纹图案，引起强烈optomotor反应[4]。如果苍蝇粘在错误的俯仰角，或引脚上的倾斜与辊轴，飞可（i）不旋转，（二）限制在一定范围内的运动，或（iii）在不一致的旋转速度。在上述情况下的飞行应该被丢弃，一个新的取代。如果问题解决了，这是最有可能是不当粘合的情况下。 （如果更换飞不解决问题，见2.4部分） </li></ol>第2部分：校正磁铁正确校正磁铁（稀土magnets.com）是必要的飞行，顺利实现360度范围内的议案。按照下列步骤来正确对齐磁铁。 <ol><li>举行水平较低的环形磁铁是由塑料套管，直接放置在一个明确的真空室顶部（图1，在第3部分进一步描述）。上部杆以上的较低的磁铁，磁铁固定垂直约3 / 4“。 </li><li>粗略对齐上下磁铁视觉直接放在上面的环形磁铁较低的中心点上杆磁铁。它是非常有用的附加罚款规模调整的一个显微（公司锡斯基尤）的磁铁上。 </li><li>环氧V型宝石轴承上部磁铁的底部表面（小部分），以尽量减少旋转摩擦和规范飞行定位。 </li><li>放置在舞台上飞，调整水平和V直到飞上层磁铁的ertical位置可以平稳偏航轴旋转360度。在每次实验开始时，应检查每飞提供平稳的旋转的磁铁，以确保正确对齐和个别minutien引脚不被损坏（见第1.11部分）。 （二）如果磁铁是没有正确对齐飞可（i）不旋转在所有的运动限制在一定范围内，（三）在不一致的速度旋转。如果问题解决，更换一个新的飞，它是最有可能是由于一个粘合的问题（见第1部分）。如果每飞行中存在的这些问题，这是可能的磁铁，将需要进一步调整对齐的问题。 </li><li>警告：与稀土磁铁玩，可好玩了，他们是非常危险的。滥用损坏的设备和/或轻微人身伤害的结果。行使这些诱人的磁铁，但易于被低估极为谨慎。 </li></ol>第3部分：气味交货<ol><li>气味输送系统是基于前面所述之一[5]，是最困难的部分，以优化（图1）。它需要很多的耐心和审判和错误。按照下列步骤来实现稳定的气味追踪。 </li><li>初始设置的气味系统需要运行的窄轨聚四氟乙烯管（小零件）气器（黑貂系统国际）定制小瓶水/气味。这些小瓶的出口直接连接到皮下端口使用相同的特氟隆管的管。 </li><li>定位下一个明确的丙烯酸真空室的舞台，以支持磁铁。我们使用一个安装在真空室相连飞以下开放接近的飞行带来的真空和减少开放的直径4mm的玻璃管。 </li><li>放置到舞台上使用镊子的系留飞行。 </li><li>打开一个有吸引力的气味（苹果醋，效果很好），并直观地验证飞行轨道的气味。如果飞似乎没有气味的偏好，遵循以下三个步骤。这可能需要几个苍蝇，多次试验。 </li><li>调整的气味端口的位置，直至出现飞追踪气味。我们装上micromanipulators气味端口（锡斯基尤公司）和定位4毫米的背和3mm前飞的头。 </li><li>调整质量流率，直至出现飞追踪气味。我们的设置7ml/min（黑貂系统国际MFC - 4）。 </li><li>调整的真空流量，直至出现飞追踪气味。我们将使用流量调节器（科尔帕默）13l/min真空流量。注：大厦真空供电线路可能造成羽状结构的波动，随着时间的推移波动。这可能是有用的流量调节器为恒定的真空流量条件下连接外部的真空泵。 </li><li>一旦设置的端口和流量的位置，定期交换机之间提供的水蒸汽和气味蒸汽使用切换使用气体复用器，直观地验证，苍蝇只能追踪气味的气味时。偶尔残留气味，里面的气味端口可以引出气味追踪的气味时关闭。这可以消除不消除灌装用乙醇皮下管和收集与Kimwipe结束分娩的正确定位的气味端口。使用压缩空气，打击任何剩余的乙醇和恢复实验。 </li></ol>代表性的成果： 当正确地遵循这些步骤，应观察两件事情。首先，不断旋转的视觉刺激下一个拴飞摆在了OMT的旋转顺畅偏航轴360度。 “肉干”圈（称为扫视）是在不断的视觉刺激的情况下自发旋转行为的特点。如果苍蝇显然不是可以顺利地旋转，不当胶合（见第1部分）和偏离磁铁可能是罪魁祸首（见第2部分）。二，打开时所发出的气味，飞头气味的方向。飞的气味时是关闭的，应自发地扫视没有在赛场上的任何单一位置的偏好。如果苍蝇不积极跟踪的气味柱，气味端口的位置，空气流速和/或真空流量可能需要调整（见第3部分）。

<ol>
	<li>Bender, J. A., Dickinson, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Visual+stimulation+of+saccades+in+magnetically+tethered+Drosophila.">Visual stimulation of saccades in magnetically tethered Drosophila.</a> J. Exp. Biol. 209, 3170-3182 (2006).</li><li>Tammero, L. F., Dickinson, M. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Collision-avoidance+and+landing+responses+are+mediated+by+separate+pathways+in+the+fruit+fly,+Drosophila+melanogaster.">Collision-avoidance and landing responses are mediated by separate pathways in the fruit fly, Drosophila melanogaster.</a> J. Exp. Biol. 205, 2785-2798 (2002).</li><li>Duistermars, B. J., Frye, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Crossmodal+visual+input+for+odor+tracking+during+fly+flight.">Crossmodal visual input for odor tracking during fly flight.</a> Curr. Biol. 18, 270-275 (2008).</li><li>Reiser, M. B., Dickinson, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+modular+display+system+for+insect+behavioral+neuroscience.">A modular display system for insect behavioral neuroscience.</a> J. Neurosci. Methods. 167, 127-139 (2008).</li><li>Frye, M. A., Dickinson, M. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Motor+output+reflects+the+linear+superposition+of+visual+and+olfactory+inputs+in+Drosophila.">Motor output reflects the linear superposition of visual and olfactory inputs in Drosophila.</a> J. Exp. Biol. 207, 123-131 (2004).</li></ol>

圈养苍蝇和稳定的气味跟踪争取在这个舞台上是相对比较简单。这需要实践，耐心和反复试验。这个舞台上的许多部件可以进一步优化，或取代在实验的潜力产生变化。例如，较新版本的真空室要小得多，允许更多的空间低于竞技场。此外，稀土永磁材料的强度和类型可以略有不同，并可以添加额外的气味端口，以增加气味的类型和/或在实验中使用浓度。我们注意到，在设备的任何变化将需要量身定制的优化。...

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a magnetic tether system to investigate visual and olfactory mediated flight control in drosophila

它已多年昆虫的视觉线索，以稳定其在风中的标题。许多动物跟踪在风中进行气味。因此，迎风视觉稳定跟踪直接气味追踪艾滋病。但嗅觉信号直接影响视觉跟踪的行为独立于风的线索呢？此外，最近的洪水在果蝇嗅觉神经生理学和神经行为遗传学研究已促使越来越技术上先进和定量的行为分析。在这里，我们修改一个磁性的系绳系统原本是为视觉实验设计配备狭窄的层流气味羽状的舞台。一只苍蝇被粘在一个小钢针，并悬浮在一个磁场，使其能够自由偏航。小直径食物的气味羽状，是直接对果蝇的头部向下，从而引发了一个饥饿的苍蝇稳定跟踪。在这里，我们集中圈养的关键机制，调整磁铁，制订的气味羽，并确认稳定的气味追踪。

Watch this Scientific Journal Video about A Magnetic Tether System to Investigate Visual and Olfactory Mediated Flight Control in Drosophila at JoVE.com

A Magnetic Tether System to Investigate Visual and Olfactory Mediated Flight Control in Drosophila

在这里，我们介绍了如何系绳飞在一个嗅觉磁系绳（OMT）设备。我们描述了如何使稀土永磁材料和气味端口，以及如何为质量流率的刺激传递和真空抽吸，以达到最佳的气味跟踪。

一个磁性的系绳系统，以调查在果蝇的视觉和嗅觉介导的飞行控制

It has been clear for many years that insects use visual cues to stabilize their heading in a wind stream. Many animals track odors carried in the wind. ...

a-magnetic-tether-system-to-investigate-visual-olfactory-mediated

Universidad de Cartagena UDEC

Research

JoVE Journal

Biology

11.2K 次观看.  University of California, Los Angeles. 在这里，我们介绍了如何系绳飞在一个嗅觉磁系绳（OMT）设备。我们描述了如何使稀土永磁材料和气味端口，以及如何为质量流率的刺激传递和真空抽吸，以达到最佳的气味跟踪。

视频: A Magnetic Tether System to Investigate Visual and Olfactory Mediated Flight Control in Drosophila

Early Metamorphic Insertion Technology for Insect Flight Behavior Monitoring

Early Metamorphosis Insertion Technology (EMIT) is a novel methodology for integrating microfabricated neuromuscular recording and actuation platforms on insects during their metamorphic development. Here, the implants are fused within the structure and function of the neuromuscular system as a result of metamorphic tissue remaking. The implants emerge with the insect where the development of tissue around the electronics during pupal development results in a bioelectrically and biomechanically enhanced tissue interface. This relatively more reliable and stable interface would be beneficial for many researchers exploring the neural basis of the insect locomotion with alleviated traumatic effects caused during adult stage insertions. In this article, we implant our electrodes into the indirect flight muscles of Manduca sexta. Located in the dorsal-thorax, these main flight powering dorsoventral and dorsolongitudinal muscles actuate the wings and supply the mechanical power for up and down strokes. Relative contraction of these two muscle groups has been under investigation to explore how the yaw maneuver is neurophysiologically coordinated. To characterize the flight dynamics, insects are often tethered with wires and their flight is recorded with digital cameras. We also developed a novel way to tether Manduca sexta on a magnetically levitating frame where the insect is connected to a commercially available wireless neural amplifier. This set up can be used to limit the degree of freedom to yawing &#8220;only&#8221; while transmitting the related electromyography signals from dorsoventral and dorsolongitudinal muscle groups.

We present a novel surgical procedure to implant electrodes in Manduca sexta during its early metamorphic stages. This technique allows mechanically stable and electrically reliable coupling with the neuromuscular tissue to study flight neurophysiology dynamics. We also present a novel magnetic levitation platform for tethered studies of insect yaw.

早期变质插入技术昆虫飞行行为监控

Early Metamorphosis Insertion Technology (EMIT) is a novel methodology for integrating microfabricated neuromuscular recording and actuation platforms on ...

科研

行为学

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 ...

生物学

Operant Learning of Drosophila at the Torque Meter

For experiments at the torque meter, flies are kept on standard fly medium at 25&deg;C and 60% humidity with a 12hr light/12hr dark regime. A standardized breeding regime assures proper larval density and age-matched cohorts. Cold-anesthetized flies are glued with head and thorax to a triangle-shaped hook the day before the experiment. Attached to the torque meter via a clamp, the fly's intended flight maneuvers are measured as the angular momentum around its vertical body axis. The fly is placed in the center of a cylindrical panorama to accomplish stationary flight. An analog to digital converter card feeds the yaw torque signal into a computer which stores the trace for later analysis. The computer also controls a variety of stimuli which can be brought under the fly's control by closing the feedback loop between these stimuli and the yaw torque trace. Punishment is achieved by applying heat from an adjustable infrared laser.

Measuring the yaw torque of tethered Drosophila with the torque meter allows the neuroscientist exquisite control of the stimulus situation of the experimental animal. Together with the unique genetic tools available in the fruit fly, this paradigm is used for a wide variety of neurobiological research.

果蝇在操作性学习扭力计

For experiments at the torque meter, flies are kept on standard fly medium at 25&deg;C and 60% humidity with a 12hr light/12hr dark regime. A standardized ...

Visually Mediated Odor Tracking During Flight in Drosophila

Flying insects use visual cues to stabilize their heading in a wind stream. Many animals additionally track odors carried in the wind. As such, visual stabilization of upwind tracking directly aids in odor tracking. But do olfactory signals directly influence visual tracking behavior independently from wind cues? Additionally, recent advances in olfactory molecular genetics and neurophysiology have motivated novel quantitative behavioral analyses to assess the behavioral influence of (e.g.) genetically inactivating specific olfactory activation circuits. We modified a magnetic tether system originally devised for vision experiments by equipping the arena with narrow laminar flow odor plumes. Here we focus on experiments that can be performed after a fly is tethered and is able to navigate in the magnetic arena. We show how to acquire video images optimized for measuring body angle, how to judge stable odor tracking, and we illustrate two experiments to examine the influence of visual cues on odor tracking.

Here we describe how to optimize the acquired video image for an olfactory magnetic-tether (OMT) apparatus. We also describe two sample experimental protocols for studying visuo-olfactory fusion.

在果蝇的飞行过程中可视介导的气味追踪

Flying insects use visual cues to stabilize their heading in a wind stream. Many animals additionally track odors carried in the wind. As such, visual ...

Optogenetic Stimulation of Escape Behavior in Drosophila melanogaster

A growing number of genetically encoded tools are becoming available that allow non-invasive manipulation of the neural activity of specific neurons in Drosophila melanogaster1. Chief among these are optogenetic tools, which enable the activation or silencing of specific neurons in the intact and freely moving animal using bright light. Channelrhodopsin (ChR2) is a light-activated cation channel that, when activated by blue light, causes depolarization of neurons that express it. ChR2 has been effective for identifying neurons critical for specific behaviors, such as CO2 avoidance, proboscis extension and giant-fiber mediated startle response2-4. However, as the intense light sources used to stimulate ChR2 also stimulate photoreceptors, these optogenetic techniques have not previously been used in the visual system. Here, we combine an optogenetic approach with a mutation that impairs phototransduction to demonstrate that activation of a cluster of loom-sensitive neurons in the fly's optic lobe, Foma-1 neurons, can drive an escape behavior used to avoid collision. We used a null allele of a critical component of the phototransduction cascade, phospholipase C-&beta;, encoded by the norpA gene, to render the flies blind and also use the Gal4-UAS transcriptional activator system to drive expression of ChR2 in the Foma-1 neurons. Individual flies are placed on a small platform surrounded by blue LEDs. When the LEDs are illuminated, the flies quickly take-off into flight, in a manner similar to visually driven loom-escape behavior. We believe that this technique can be easily adapted to examine other behaviors in freely moving flies.

Optogenetic Stimulation of Escape Behavior in Drosophila melanogaster

Genetically encoded optogenetic tools enable noninvasive manipulation of specific neurons in the Drosophila brain. Such tools can identify neurons whose activation is sufficient to elicit or suppress particular behaviors. Here we present a method for activating Channelrhodopsin2 that is expressed in targeted neurons in freely walking flies.

Optogenetic刺激的逃避行为<em&gt;果蝇</em

A  growing number of genetically encoded tools are becoming available that allow non-invasive  manipulation of the neural activity of specific neurons in ...

神经科学

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 ...

A Behavioral Assay for Mechanosensation of MARCM-based Clones in Drosophila melanogaster

Because of the structural and functional homology to the hair cells of the mammalian inner ear, the neurons that innervate the Drosophila external sense organs provide an excellent model system for the study of mechanosensation. This protocol describes a simple touch behavior in fruit flies which can be used to identify mutations that interfere with mechanosensation. The tactile stimulation of a macrochaete bristle on the thorax of flies elicits a grooming reflex from either the first or third leg. Mutations that interfere with mechanotransduction (such as NOMPC), or with other aspects of the reflex arc, can inhibit the grooming response. A traditional screen of adult behaviors would have missed mutants that have essential roles during development. Instead, this protocol combines the touch screen with mosaic analysis with a repressible cell marker (MARCM) to allow for only limited regions of homozygous mutant cells to be generated and marked by the expression of green fluorescent protein (GFP). By testing MARCM clones for abnormal behavioral responses, it is possible to screen a collection of lethal p-element mutations to search for new genes involved in mechanosensation that would have been missed by more traditional methods.

A Behavioral Assay for Mechanosensation of MARCM-based Clones in Drosophila melanogaster

In order to identify novel mutations affecting mechanosensation, we designed an assay that measures the behavioral response to tactile stimulation of fly bristles in mutant clones generated by the MARCM method. The combination of techniques allows for the identification of mechanosensitive mutations that would otherwise be lethal.

一个行为分析对机械感觉MARCM为基础的克隆<em&gt;果蝇</em

Because of the structural and functional homology to the hair cells of the mammalian inner ear, the neurons that innervate the Drosophila external sense ...

Olfactory Behaviors Assayed by Computer Tracking Of Drosophila in a Four-quadrant Olfactometer

A key challenge in neurobiology is to understand how neural circuits function to guide appropriate animal behaviors. Drosophila melanogaster is an excellent model system for such investigations due to its complex behaviors, powerful genetic techniques, and compact nervous system. Laboratory behavioral assays have long been used with Drosophila to simulate properties of the natural environment and study the neural mechanisms underlying the corresponding behaviors (e.g. phototaxis, chemotaxis, sensory learning and memory)1-3. With the recent availability of large collections of transgenic Drosophila lines that label specific neural subsets, behavioral assays have taken on a prominent role to link neurons with behaviors4-11. Versatile and reproducible paradigms, together with the underlying computational routines for data analysis, are indispensable for rapid tests of candidate fly lines with various genotypes. Particularly useful are setups that are flexible in the number of animals tested, duration of experiments and nature of presented stimuli. The assay of choice should also generate reproducible data that is easy to acquire and analyze. Here, we present a detailed description of a system and protocol for assaying behavioral responses of Drosophila flies in a large four-field arena. The setup is used here to assay responses of flies to a single olfactory stimulus; however, the same setup may be modified to test multiple olfactory, visual or optogenetic stimuli, or a combination of these. The olfactometer setup records the activity of fly populations responding to odors, and computational analytical methods are applied to quantify fly behaviors. The collected data are analyzed to get a quick read-out of an experimental run, which is essential for efficient data collection and the optimization of experimental conditions.

Olfactory Behaviors Assayed by Computer Tracking Of Drosophila in a Four-quadrant Olfactometer

We describe here a behavioral setup and data analysis method for assaying olfactory responses of up to 100 vinegar flies (Drosophila melanogaster). This system may be used with single or multiple olfactory stimuli, and adaptable for optogenetic activation or silencing of neuronal subsets.

通过计算机跟踪检测大鼠嗅觉行为<em&gt;果蝇</em&gt;在四象限嗅

A key challenge in neurobiology is to understand how neural circuits function to guide appropriate animal behaviors. Drosophila melanogaster is an ...

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 ...

Animal Behavior

<h4>Drosophila as a Model Organism</h4>
The common fruit fly, Drosophila melanogaster, is a widely used model organism in biology, though it may be more commonly recognized as a fruit-loving pest. There are multiple reasons that make D. melanogaster an excellent experimental organism: their roughly two-week generation time allows studying multiple generations, they are easily maintained in very small tubes, and their sexual dimorphism enables investigators to easily distinguish between males that have black coloration towards the posterior and females that have banded abdomens.
Thomas Hunt Morgan and his students were among the first to make use of Drosophila, in the early 20th century. In 1933, Morgan received a Nobel Prize for his discovery of the role of chromosomes in heredity, which was founded on his observations of mutant Drosophila. Morgan continued his studies by generating new mutants with radiation. The presence of multiple mutants with discrete effects on the appearance of the fly enabled him to look at patterns of multifactor inheritance. A key aspect of his early work was the discovery of apparent violations of Mendel&#8217;s second law, which states that genes segregate independently of one another. Morgan&#8217;s observations indicated that genes will not segregate independently if they are joined together on the same chromosome. Furthermore, it became possible to determine gene locations on a chromosome relative to each other through analyses of how often they recombined thus triggering gene mapping studies to identify the location of a gene and distances between genes. Consequently, associates of Morgan&#8217;s lab received two additional Nobel Prizes for other studies of Drosophila genetics. Morgan&#8217;s success and the accumulating information about Drosophila inspired others to adopt them as experimental organisms, which added to our knowledge of them and progressively increased their value as a model organism. Other scientists have developed many mutations. Over 27,000 unique Drosophila lines are now maintained as stock cultures at the Drosophila stock centers and are readily available to laboratories that use them as tools for studying diverse aspects of biology, including behavior1.
<h4>Drosophila Ethology</h4>
Animal behavior, or ethology, is the study of how animals interact with each other and their environments. Although it is a relatively young biological field, ethology has important implications since an organism&#8217;s survival depends on how it behaves with regard to possible mates, food sources, and natural obstacles. Studying these organisms helps scientists to understand the contexts of these behaviors and the underlying mechanisms as to how they have evolved. Thus, a goal of the study of animal behavior is to find a genetic basis for the behaviors exhibited by individuals or groups of individuals. For example, kin selection, first proposed by William Hamilton, suggests that individuals will behave altruistically when making a sacrifice for a family member rather than a stranger2. This is because related individuals share more genes than unrelated ones, and the future reproduction by their relatives contributes in a way to their own fitness, or reproductive success. Hence, observing how organisms like Drosophila interact with their environment can allow us to understand how these behaviors are hard-wired. Additionally, these behaviors provide insight into how these organisms have adapted to cope with their environment and survive.
Directional behaviors are commonly studied to understand the basis of how and why an organism moves in its environment. Kinesis and taxis are two forms of directional behaviors. Kinesis involves stimulus-induced movements in random directions, such as the random movement of flies when chased away. On the other hand, taxis involves stimulus-induced movement towards a specific direction, such as moths flying towards a lightbulb. Taxis behaviors are positive if the animal moves towards the stimulus and negative if the animal moves away from the stimulus. Furthermore, taxis behaviors are named based on the specific stimulus that induces them. For example, geotaxis is a response to gravity, where positive geotaxis means that the organism moves with gravity. Phototaxis is movement in response to light, such as positive phototaxis of moths towards a lightbulb. Similarly, chemotaxis is attraction to or avoidance of an airborne chemical cue, such as the negative chemotaxis of many predators, including humans, in response to the smell of skunk spray. Furthermore, there are many other types of taxes that are studied by scientists, including aerotaxis, barotaxis, hydrotaxis, and magnetotaxis, that are movements in response to oxygen, pressure, water, and magnetic fields, respectively.
<h4>Study of Drosophila taxis behavior</h4>
A common and simple method to study Drosophila taxis behavior is to use a choice chamber, which allows the observation of a directional response of the fly to a particular stimulus, such as gravity, light, or a chemical. Drosophila are inserted in the center of the choice chamber and then allowed to wander freely about the chamber, which is designed so that alternative stimuli can be presented at opposing sides. After a period of wandering, the number of flies is counted on either side of the choice chamber. The number of flies on each side of the chamber is then analyzed with the Chi Square test to gauge whether or not there is a preference for a stimulus.
Comparison of directional behavior of mutant and wild-type Drosophila allows researchers to determine the role of the mutated gene in that specific behavior. Moreover, scientists are also trying to understand how animals decide what to do in the presence of combinations of stimuli, including conflicting stimuli3. Together, these studies can help predict how organisms will react to certain stimuli or changes in environmental factors.
<h4>References</h4>
<ol class="references">
	<li>Cook KR, Parks AL, Jacobus LM, Kaufman TC, Matthews K. New research resources at the Bloomington Drosophila Stock Center. Fly. 2010, Vol. 4, 1: 88-91.</li>
	<li>WD, Hamilton. Altruism and related phenomena, mainly in social insects. Ann Rev Ecol Systematics. 1972, Vol. 3, 193-202.</li>
	<li>Gepner R, Skanata MM, Bernat NM, Kaplow M, Gershow M. Computations underlying Drosophila photo-taxis, odor-taxis, and multi-sensory integration. eLife. 2015, Vol. 4, e06229.</li>
</ol>

Animal Behavior: Studying Taxis Behavior of Drosophila - Concept

动物行为

Drosophila as a Model Organism
The common fruit fly, Drosophila melanogaster, is a widely used model organism in biology, though it may be more commonly ...

一个磁性的系绳系统，以调查在果蝇的视觉和嗅觉介导的飞行控制

摘要

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此视频中的章节

早期变质插入技术昆虫飞行行为监控

高分辨率测量的气味驱动在果蝇幼虫的行为

果蝇在操作性学习扭力计

在果蝇的飞行过程中可视介导的气味追踪

Optogenetic刺激的逃避行为

在气味引导行为的高分辨率定量

一个行为分析对机械感觉MARCM为基础的克隆

通过计算机跟踪检测大鼠嗅觉行为

跟踪果蝇幼虫行为对嗅神经元 Optogenetic 刺激的反应

动物行为