国立科学財団（MF）で提供される資金。

はじめ OMTは、&quot;仮想プルームシミュレーター&quot;を組み込むために設計された[1]磁気テザーシステムの適応です。次のプロトコルは、どのように適切にテザーのハエ（パート1）を説明し、磁石（パート2）とマスフロー規制臭気デリバリーシステム（その3）を設定するための戦略を提供します。ここで説明する情報は、この特定のシステム用に最適化されており、他の技術仕様を持つコンポーネントによって異なる場合があります。 パート1：テザリングのハエここで説明するテザリングをするための手順は、多くの先行研究と類似している[2]しかし、OMT法での使用に適合[3]。適切なテザリングは、堅牢で再現性の実験的試行を確保することが重要です。慎重にテザリングの重要性は誇張することはできません。 <ol><li> 4-6日の後の羽化を飛ぶ収集し、4〜6時間のためにそれらを飢えさせる。キムワイプで乾燥を防ぐために、食べ物のないバイアルに水で湿らせた置きます。 </li><li>小さなバイアルに瓶からハエのバッチを転送し、摂氏約4度で設定されたペルチェ冷却ステージ上に真鍮のブロックに挿入します。適切な温度は約30秒以内にハエをanesthetizes。 </li><li>冷却ステージの冷たい表面上のハエをソートします。湿らせたキムワイプはキューティクルから一定の冷却と水分の上がりを確実にします。テザリングのための最大の女性を選択してください。 </li><li>細かいブラシを使用して、単一の野生型雌ミバエを収容する形特注の石棺の位置に単一のフライを押してください。 <img src="/files/ftp_upload/old/1063_2008_11_26_2/1063_Figure-01_thumb.png" alt="" /> 図1。 <a href="https://jove.unicartagenaproxy.elogim.com/files/1063_2008_10_28_1/1063_Figure-01.png">図の拡大バージョンをご覧いただくにはこちらをクリックしてください。</a> </li><li>我々は、マイクロマニピュレータに取り付けられた磁気ロッドとminutienピン（罰金科学のツール）を置きます。ピンは、平滑末端がちょうど2つの翼の間に頭の後ろに背胸部に触れるように配置されることです。目に向かって正面から見たときにピンが完全に垂直でなければなりません。フライとピンの間に&quot;ロール&quot;のであっても少量を許容することはできません。ピンと体の長軸の間にサイドビューの角度は水平線（図1）に対して相対的に30度の機首アップの角度になっているはず。注：磁石のテザーで最適な動作では、ピンはケーブル長を短くするためにトリミングする必要があります。これにより、サスペンションの磁石間の距離に完全に依存します。 </li><li> minutienのピンが整列されると、minutienピンの平滑末端にUV -活性化接着剤（ElectroLiteコーポレーション）の小滴を適用するために細い鋼線を使用してください。 </li><li>接着剤の液滴がちょうど頭の上にハエの胸部に触れるようにピンを操作します。適切なドロップサイズで、接着剤は、アニールすべきと胸部の上にピンから流れます。あまりにも多くの接着剤を使用しないように注意してください。 </li><li> UV光の2つ20秒のバーストで接着剤を治す。 </li><li>固定フライを脇に置き、繰り返し。キムワイプの小さな四角をスライスし、鉗子で飛行を抑制するために、各係留飛行するものを与える。 </li><li>回復の一時間後に、テザーハエは、実験の準備が整いました。 </li><li>不適切な接着には、磁石の分野で実行するハエの能力を妨げる可能性があります。すべてのハエは、ハエが適切に接着されるように、すべての実験の開始時に滑らかな回転のためにチェックする必要があります。これは、強力な微細運動の応答を引き出すために十分に大きな空間的な周期で縞模様を回転させることによって達成することができます[4]。ハエが間違っピッチ角に接着またはロール軸を基準にしてピンに傾いていると、ハエ5月は、（i）の動きの制限された範囲を持っている、または（iii）一貫性のないで回転（ii）全く回転しない速度。上記のケースではハエを破棄し、新しいものに置き換えられます。この問題が解決した場合、それはおそらく不適切な接着のケースだった。 （フライを交換しても問題が解決しない場合は、第2.4参照） </li></ol>パート2：磁石の位置を揃える磁石（希土類- magnets.com）を揃えることが適切動きのスムーズな360度の範囲を達成するためにフライが必要である。適切に磁石を整列させるには、次の手順に従います。 <ol><li>下のリング磁石は、プラスチックスリーブで水平に保持し、明確な真空チャンバーの上に直接配置されます（図1、パート3で詳しく説明）。上部の棒磁石を下側磁石の上に垂直に約3 / 4&quot;固定されています。 </li><li>ほぼ直接下部リング磁石の中心点の上に上部の棒磁石を置くことによって視覚的に上下の磁石の位置を合わせます。それは、微細調整のためにマイクロマニピュレータ（シスキユー社）に、上部磁石を添付すると便利です。 </li><li>エポキシ回転摩擦を最小限にし、フライの位置を標準化するために、上部磁石の底面上にV -宝石軸受（スモールパーツ）。 </li><li>アリーナでフライを配置し、水平方向とvを調整するフライまで、上部磁石のertical位置は、円滑かつ着実にヨー軸に360度回転させることができます。すべてのハエは、磁石が適切に整列されており、個々のminutienピンは（パート1.11参照）損傷していないことを保証するためにすべての実験の開始時に滑らかな回転のためにチェックする必要があります。磁石が適切にフライを整列されていない場合5月は、（i）すべての（ⅱ）で回転しない動きの制限された範囲を（iii）の一貫性のない速度で回転する必要があります。問題は新しいものとフライを交換することによって解決されている場合、それは（パート1を参照）最も可能性の高い接着の問題によるものであった。これらの問題は、すべてのハエのために存在する場合、それは可能性が高い磁石とさらに調整が必要とされるのアラインメントの問題です。 </li><li>警告：楽しいことができる希土類磁石で遊んでいる間、彼らは非常に危険です。破損した機器及び/またはマイナー身体的危害の結果を悪用。これらの魅力的な、まだ簡単に過小評価して磁石では細心の注意を払ってください。 </li></ol>パート3：臭配信<ol><li>臭気の配信システムは、前述のいずれかに基づいている[5]および最適化するために過酷な部分（図1）である。それは忍耐と試行錯誤の多くを必要とします。安定した臭いの追跡を実現するためには、次の手順に従います。 </li><li>臭気のシステムの初期セットアップは、カスタム構築された水/臭バイアルへのガスマルチプレクサ（セーブルシステムズインターナショナル）から狭いゲージテフロンチューブ（スモールパーツ）を実行している必要があります。これらのバイアルの出口は、同じテフロンチューブを使用して皮下ポートのチューブに直接接続します。 </li><li>透明なアクリルの真空チャンバは、磁石をサポートするために、アリーナの下に配置されている。私たちは、ハエに近い開き真空を持参し、開口部の直径を減らすために、真空チャンバに接続されているフライの下にマウントさ4ミリメートルのガラス管を使用してください。 </li><li>鉗子を使用して舞台にテザーフライを置きます。 </li><li>魅力的な臭気（リンゴ酢がうまく機能）をオンにし、視覚的にハエが匂いを追跡することを確認してください。フライは、匂いに対する嗜好を持っていないように見える場合は、次の3つの手順に従ってください。これは、いくつかのハエ、そしていくつかの臨床試験が必要になる場合があります。 </li><li>フライは、匂いを追跡するために表示されるまで、臭気のポートの位置を調整します。我々は、マイクロマニピュレータ上に臭気のポートをマウント（シスキユー社）とハエの頭に4ミリメートル背側および3mm前方に配置されます。 </li><li>フライは、匂いを追跡するために表示されるまで、質量流量を調整します。私たちの設定が7ml/min（セーブルシステムズインターナショナルMFC - 4）です。 </li><li>フライは、匂いを追跡するために表示されるまで、真空の流量を調整します。我々は、フローレギュレータ（コールパーマー）を使用して13l/minで真空流量を設定する。注：建物の真空供給ラインは、プルームの構造の変動を引き起こして時間の経過とともに変動する可能性があります。それは一定の真空流量条件のフローレギュレータに外付けの真空ポンプを接続するために役に立つかもしれません。 </li><li>一度ポートの位置と流量が設定されているが、定期的にガスのマルチプレクサを使用して切替を使用し、視覚的にその場だけ臭いがオンになって臭いを追跡することを確認する水蒸気や臭気の蒸気を提供切り替える。時折、臭気のポート内部の残臭は臭いがオフになっている匂いの追跡を引き出すことができます。これは、エタノールで皮下注射管を充填し、キムワイプで配信終わりにそれを収集することにより適切な位置臭気のポートを削除せずに除去することができます。圧縮空気を使用して、残りのエタノールを吹き消すと実験を再開する。 </li></ol>代表的な結果： これらの手順が正しく実行されると二つのことが観察されるべきである。最初に、一定の回転視覚刺激下OMTに配置されたテザーフライは、スムーズにヨー軸に360度回転する必要があります。 &quot;ジャーキー&quot;ターンは、（サッカードと呼ばれる）を一定の視覚刺激の非存在下での自発的な転換動作の特徴です。ハエが明らかにスムーズに回転できない場合、不適切な接着（パート1を参照）とずれ磁石は（パート2を参照）せいにするかもしれない。第二に、臭気がオンになっているときに、ハエは臭いの方向に向かうはずです。匂いをオフにすると、フライは競技場内の任意の単一の位置のための希望なしに自発的にサッカードがあります。ハエが積極的に臭羽毛を追跡しない場合は、臭気のポートの位置、空気の流量および/または真空の流量は、（パート3を参照）調整が必要な場合があります。

<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>

ハエをテザリングとこの分野で安定的な臭気の追跡を誘発することは比較的簡単です。それは練習、忍耐と試行錯誤が必要です。この分野の多くの部分は、さらに最適化や実験的な電位の変動をもたらすために代替できる場合もあります。例えば、真空チャンバ内の新しいバージョンは、はるかに小さいですし、舞台下に多くのスペースを許可する。また、希土類磁石の強さや種類が若干変化することがで?...

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

それは虫が風の流れでその見出しを安定させるために視覚的な手がかりを使用することを長年にわたって明らかになっている。多くの動物は、風で運ばれた匂いを追跡。このように、風上の視覚的な安定は、臭気の追跡に直接補助追跡。しかし、嗅覚の信号は、直接風の手がかりから独立して視覚的な追跡の動作に影響を与えるのですか？また、ショウジョウバエの神経生理学と嗅覚の神経行動遺伝学に関する研究の最近の洪水では、これまで以上に技術的に洗練され、定量的な行動アッセイを動機としています。ここで、我々は元々狭い層流臭プルームでアリーナを搭載することにより、ビジョンの実験のために考案された磁気テザーシステムを変更。フライは小さなスチールピンに接着し、自由にヨーを可能にする磁場中に懸濁されている。小径食品の匂いのプルームは、空腹フライで安定したトラッキングを引き出す、ハエの頭の上に下向きにしている。ここでは、磁石を合わせ臭羽毛を考案し、安定的な臭気の追跡を確認、テザリングの重要なメカニズムに焦点を当てる。

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

ここでは、どのようにテザー嗅覚磁気テザー（OMT）装置でフライをして説明します。我々は、希土類磁石と臭気のポート、およびどのように刺激の配信と最適な臭気の追跡を実現するために真空吸引の両方の質量流量を設定することを整列する方法について説明します。

ショウジョウバエの視覚と嗅覚を介したフライトコントロールを調査するために磁気テザーシステム

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 ビュー.  University of California, Los Angeles. ここでは、どのようにテザー嗅覚磁気テザー（OMT）装置でフライをして説明します。我々は、希土類磁石と臭気のポート、およびどのように刺激の配信と最適な臭気の追跡を実現するために真空吸引の両方の質量流量を設定することを整列する方法について説明します。

ビデオ: 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.

昆虫飛翔挙動監視のための早期変成挿入技術

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

行動学

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.

ショウジョウバエの幼虫の匂い駆動型の動作の高分解能測定

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.

トルクメーターでのショウジョウバエのオペラント学習

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.

ショウジョウバエのフライトの間に視覚的に仲介臭気の追跡

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.

で逃避行動のOptogenetic刺激<em&gt;キイロショウジョウバエ</em

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

神経科学

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.

臭気誘導挙動の高分解能定量<em&gt;キイロショウジョウバエ</em&gt;使用<em&gt; Flywalk</em&gt;パラダイム

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.

でMARCMベースのクローンのMechanosensationための行動アッセイ<em&gt;キイロショウジョウバエ</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.

のコンピュータ追跡することによってアッセイ嗅覚行動<em&gt;ショウジョウバエ</em&gt; 4象限オルファクトメーターで

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.

嗅覚ニューロンの光刺激に対する応答のショウジョウバエ幼虫の行動を追跡

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

動物の行動

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 ...

ショウジョウバエの視覚と嗅覚を介したフライトコントロールを調査するために磁気テザーシステム

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昆虫飛翔挙動監視のための早期変成挿入技術

ショウジョウバエの幼虫の匂い駆動型の動作の高分解能測定

トルクメーターでのショウジョウバエのオペラント学習

ショウジョウバエのフライトの間に視覚的に仲介臭気の追跡

で逃避行動のOptogenetic刺激

臭気誘導挙動の高分解能定量

でMARCMベースのクローンのMechanosensationための行動アッセイ

のコンピュータ追跡することによってアッセイ嗅覚行動

嗅覚ニューロンの光刺激に対する応答のショウジョウバエ幼虫の行動を追跡

動物の行動