The project was supported by Award AAES 461Hatch/Multistate Grants ALA08-045 and ALA015-1-10026 to N.L.

楽器、刺激ソリューション、およびトコジラミの調製<ol><li> 20ミリリットルボトルに（w / v）の50％KNO 2溶液を調製します。 </li><li>繰り返しにし、溶液からタングステン電極を浸漬することにより、5 VでKNO 2溶液中の2つのタングステン微小電極をシャープ。 <ol><li>大雑把で大幅タングステン線の先端を消費することができる2ディップ/約5分間秒の速度でKNO 2溶液のうちタングステン線の約10mmを浸漬することにより、タングステン線をシャープに。 </li><li>電極の罰金とシャープなポイントになるように繊細に、少なくとも1分間、2ディップ/秒の速度で内溶液のうちワイヤ先端の約1mmを浸漬することによって電極を研ぎます。それはベッドのバグ嗅覚感覚子のクチクラを穿刺するのに十分問題ないはずですが0.2から0.5ミクロンに達するまで頻繁に顕微鏡下で電極先端径を確認してください。 注意：手動で鋭いものの電極稲光、KNO 2溶液にタングステン線の浸漬速度は常に一定ではありません。もっと練習すれば、電極をシャープに比較的一定の速度を維持する方がはるかに簡単です。シャープ時間も電極がどうあるべきか罰金に応じて、不確実です。ここでは、〜0.2ミクロンの直径を有する電極先端部は嗅覚感覚子を通して穿刺のに十分です。 </li></ol></li><li>ストック溶液として1時10 V / Vの初期濃度にニート化合物のジメチルスルホキシド（DMSO）に化学的刺激のそれぞれを希釈。各化学物質のためのストック溶液のそれぞれから、DMSOで再び、実験に必要などのように多くの投与量に応じて、常用希釈系列を作成します。 βピネン及びユーカリプトール - ここでは、10％（+）を使用します。 </li><li>食物を与えられていないか、7日間置き、後送りフォートから（男性または女性のいずれか）大人のベッドのバグ。ディックスコロニー（ケンタッキー大学で博士ヘインズからの贈り物）は、Tで使用します彼はペトリ皿で​​実験します。 注意：ペトリ皿に入れ、ベッドのバグのための正確な数はありません。これは、いくつかまたは多くをすることができます。 </li></ol> 2.トコジラミ触角準備<ol><li>氷上でトコジラミ（2-3分）麻酔し。 </li><li>両面テープで顕微鏡カバースリップ上のアンテナおよび昆虫の体の両方を修正して、細かいハサミで脚を取り外します。 </li><li>着実にテープ上に固執するように静かにアンテナを触れないように、小さなピンを使用してください。 </li><li>操作を容易にし、記録電極（ 図1）のために適切な角度（〜90°）にそれを調整するために歯科用ワックスの小さなボール（〜直径1cm）に対してカバーガラスを置き。 </li><li>一度、ステレオ顕微鏡下ベッドのバグを配置冷光源をオンにし、アンテナが明確に提示されるまで、照明の明るさを調整し、高倍率でトコジラミアンテナの第二の鞭毛に顕微鏡を集中、確保（720X） 。 注意：実験に用いた照明の強度は、実際に実験者の目は照明の強さを感じる方法に依存する、定量化されていません。 </li></ol> 3.シングル感覚子記録<ol><li>信号記録と可視化のためのコンピュータに接続された信号取得制御部、プリアンプ（10倍）を接続します。コンピュータの電源をオンにし、ソフトウェア、 例えば 、AutoSpike32を起動し、メニューバーから「記録」モードをクリックします。そして、波信号の記録を開始するように、「波」を選択します。 注：繰り返しモニターの左から右に実行されている平らな線が表示されるようになるはずです。ここで、記録ウィンドウは40秒続きます。最大波記録は10秒です。選択されたサンプリングレートは96000であり、デジタルサンプリングレートは、オフセット0％無フィルタリング、記録信号の無整流あり240です。必要に応じてソフトウェアでこれらのすべてのパラメータ設定を変更することができます。 </li> &lt;李は&gt;触角感覚子から神経細胞の応答を調色モードを提示するために使用されるプリアンプに接続されたスピーカーをオンにします。 </li><li>安定化されたベッドのバグの腹部に基準電極を挿入します。 注：基準電極を磁気エアテーブルに取り付けられた金属製のスタンドにより開催されました。 </li><li>基準電極は、ベッドのバグの腹部に接続された後、プリアンプに接続し、ベッドのバグのアンテナの後方端に向かって、マイクロマニピュレーターによって操作された記録電極を移動。 </li><li>記録電極は、アンテナの右側先端部と接触しているとき、顕微鏡に切り替えて低倍率で電極を見つけます。 </li><li>電極と触角感覚子の両方が同一平面上にあり、顕微鏡下ではっきりと見えるまで徐々に倍率を増加させながら記録電極を調整します。 注：この時点で、顕微鏡が普通です最高倍率でLY。 </li><li>マイクロマニピュレーターを用いて、感覚子の軸に記録電極を挿入し、バックグラウンドノイズは、活動電位に比べて高い場合、少し深く行きます。 </li><li> βピネン - 一度明確な活動電位を記録感覚子から観察され、10％（+）でマイクロピペットを埋めます。ガラスパスツールピペットの内部に配置された濾紙片（〜3×15 mm）の上にβピネン - 10％（+）の10μlアリコートを堆積させるためにマイクロピペットを使用してください。 <ol><li>刺激コントローラのパルス流管の出口にロードされたピペットを接続し、アンテナを指向管の小さな穴にピペットの先端を配置します。 </li></ol></li><li>すべてのこれらの接続が安定しているときに、連続的な加湿空気流中に刺激の0.5秒のパフ（0.5L /分）を送達するための刺激コントローラのフットスイッチを押し下げます。時footswitc活動電位の記録が同時に開始されますhが押されています。記録処理は、刺激の前に1秒を開始する10秒間の最後になります。 </li><li> 2 500ミリ秒の期間、1の前と刺激後の1のためにオフラインで活動電位をカウントします。前の500ミリ秒の間に記録された自発的な活動から500ミリ秒後に刺激中のスパイク率の変化を引き、2を乗じてスパイク/ sの従来のスケールに数を変換します。 </li></ol> SSRの4刺激交換<ol><li>ベッドのバグのアンテナ上にピペットでβピネンやピペットが除去された後10秒のためのこの特定の匂い物質に対する応答を記録 - フットスイッチがトリガされた後、10％（+）を提供します。 </li><li>テストされる0.001％のユーカリプトールで別の新しいピペットにラベルを付けます。刺激の10μlを新しいピペットに、適用された上に、濾紙の小片を置きます。 </li><li>刺激が完全にvaporiするまで2-5分間待ちますガラスピペットでZED。パルス流管の出口上にピペットを取り付けます。 </li><li>アンテナを志向管の小さな穴にピペットの先端を挿入します。フットスイッチを押し、10秒の記録を開始します。 </li><li>ピペットを外し、0.01％のユーカリプトールで別のピペットを準備します。 </li><li>用量依存性の反応を観察するために触角感覚器のユーカリプトールの投与量（0.001％〜10％）のすべての残りの部分をテストします。ほとんどのテストは、希釈最低用量に希釈します。 </li></ol>

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The authors have nothing to disclose.

単一感覚子記録技術は、広くそのような環境で異なる化学的刺激に対するショウジョウバエ、蚊、ベッドのバグのような昆虫の神経応答を試験する際に使用されています。これらの化学的刺激は、多くの場合、治療の異なる用量を調製するために溶解させ、共通の溶媒で希釈します。しかし、異なる溶媒は、刺激のために非常に異なる放出速度を生成することができます。このようなキイ...

一時的な外部寄生虫のように、彼らの生存、開発、および再生がヒトおよび動物1,2の両方を含むホストからの血液供給源を必要とする意味義務吸血昆虫は、ある共通のベッドのバグトコジラミはL（トコジラミ科半翅目）をlectularius。ウイルス伝播はほとんどによるCに報告されていないが、 lectularius、侵入によって生成された痛烈な迷惑は真剣に物理的にも心理的に3ホストに影響を与えます。導入と化学殺虫剤の普及は、特にDDT、蔓延のリスクを低下させ、1950年代の蔓延の終わりまでに、彼らはもはや深刻な社会問題だったような低レベルでした。しかし、可能な多くの要因は、そのような殺虫剤の使用削減として、世界的にベッドのバグ集団に国民の意識の低下を復活するために導いた、旅行の活動を増加し、殺虫剤4-9に対する抵抗性の発達。&lt;/ P&gt; 環境中の化学手がかりを検出し、このようなアンテナや上顎palpsとして嗅覚器官を通して昆虫によって認識されています。昆虫のアンテナの嗅覚感覚子は、これらの化学手がかりを検出するのに重要な役割を果たしています。化学分子は、キューティクルの表面上の孔を通して触角キューティクルを入力してください。臭気物質は、これらの化学分子に触角リンパバインド中のタンパク質と結合し、嗅覚受容体10の上にそれらを運びます。これらの化学分子が嗅覚受容体11によって認識された後脱分極される神経膜上の非選択的陽イオンチャネルからの嗅覚受容体とそのコレセプター。 単一感覚子記録（SSR）は、化学的又は非化学的刺激のいずれかを適用することによって引き起こされる活動電位の細胞外の変化を検出するために開発されました。感覚子リンパ及び基準電極に記録電極を挿入し虫体（通常複眼または腹部のいずれか）のいくつかの他の部分に、刺激に応答するニューロンの発火率は12を記録することができます。スパイクの数の変化は、特定の刺激に対する昆虫の感度を表します。異なるアイデンティティおよび濃度の化学的刺激は、異なる発射速度および時間的な構造と、異なる神経応答を誘発し、従って特定の化学物質に対する昆虫の符号化処理を調べるために使用することができます。 9溝付きペグCの感覚器、29毛髪様E（E1およびE2）感覚器、および1組Dα、Dβ、Dγ円滑なペグをそれぞれ共通ベッドのバグのために、両方の性的フォームがアンテナに嗅覚感覚子の同じパターンを共有します感覚器13,14。複数のニューロンが感覚子の各タイプにおいて同定されているように、同じ感覚子に収容された別のニューロンからの活動電位を区別することは容易ではないので、この実験の踏太活動電位のL番号は、刺激前後の500ミリ秒の期間にわたってオフラインで計数しました。刺激後の活動電位の数は、刺激の前に活動電位の数から差し引かれ、15秒あたりのスパイクの個々の感覚子で発火率の変化を定量化するために、2つを乗じました。

<table><tbody><tr><td>Tungsten wire</td><td>A-M SYSTEMS</td><td>#716500</td><td>Used for preparing the electrode</td></tr><tr><td>KNO2</td><td>Sigma</td><td>#310484</td><td>Used for sharpening the tungsten wire</td></tr><tr><td>AC Power Supply</td><td>BK Precision</td><td>1653A</td><td>Providing the voltage in sharpening the tungsten wire</td></tr><tr><td>Leica Z6 APO Microscope</td><td>Leica</td><td>10447424</td><td>Used for observing the sensilla on antennae</td></tr><tr><td>Simulus controller</td><td>Syntech</td><td>CS-55</td><td>Used for controlling the stimulus application</td></tr><tr><td>4-Channel USB Acquisition Controller</td><td>Syntech</td><td>IDAC-4</td><td>Real-time on screen display of all signals before and during recording</td></tr><tr><td>Light Source</td><td>SCHOTT</td><td>A20500</td><td>Providing light sources for observation</td></tr><tr><td>Micromanupulator</td><td>Leica</td><td>115378</td><td>Used for minor movement of electrode</td></tr><tr><td>Speaker</td><td>Juster</td><td>95a</td><td>Connected with Acquisition Controller IDAC-4 and providing sound for the signal</td></tr><tr><td>Magnetic stand</td><td>Narishige</td><td>GJ-1</td><td>Used to hold the reference electrode, stablized bed bug and stimulus delivery tube</td></tr><tr><td>TMC Vibration Isolation Table</td><td>TMC</td><td>63-500</td><td>Used for isolating the vibration from the equipments</td></tr><tr><td>Coverslip</td><td>Tedpella</td><td>2225-1</td><td>Used for holding the bed bug</td></tr><tr><td>Double-sided Tape</td><td>3M</td><td>XT6110</td><td>Used for stablizing the bed bug on the coverclip</td></tr><tr><td>Dental Wax</td><td>Dentakit</td><td>DK-R012</td><td>Used for supporting the coverclip where bed bug is stablized&amp;nbsp;</td></tr></tbody></table>

using single sensillum recording to detect olfactory neuron responses of bed bugs to semiochemicals

昆虫嗅覚系は、環境内の情報化学物質を検出するのに重要な役割を果たしています。具体的には、内部の単一または複数の神経細胞を収容触角感覚器は、化学的刺激に応答するの主要な貢献をすると考えられています。直接刺激への暴露後に嗅覚感覚子に活動電位を記録することにより、単一感覚子記録（SSR）技術は、化学的刺激への虫の神経応答を調査するための強力なアプローチを提供します。悪名高い人間の寄生虫であるベッドのバグについては、嗅覚感覚子の複数のタイプが特徴づけられています。本研究では、2つの化学的刺激とSSRの方法を使用してそれらのいずれかの用量依存性応答にベッドのバグ嗅覚感覚子の神経応答を示しました。このアプローチは、develのための貴重な情報を提供するベッドのバグ嗅覚感覚子、上の個々の化学的刺激のための早期のスクリーニングを行うための研究者を可能にします新しいベッドのバグ誘引または忌避剤とメリットベッドバグ制御努力の発。

単一感覚子記録は、昆虫の化学生態学と神経生理学の研究に使用される強力な捜査手法です。別の揮発性化合物に昆虫の神経応答、昆虫の生存と発展に生態学的に関連すると考え特にの調査、昆虫嗅覚プロセスに私たちに貴重な洞察力を与えるだけでなく、潜在的につながる可能性があり、有望な新しい道を開くだけでなく、害虫駆除のために有用な新規試薬の開発?...

Watch this Scientific Journal Video about Using Single Sensillum Recording to Detect Olfactory Neuron Responses of Bed Bugs to Semiochemicals at JoVE.com

Using Single Sensillum Recording to Detect Olfactory Neuron Responses of Bed Bugs to Semiochemicals

Bed bugs rely on olfactory receptor neurons housed in their antennal olfactory sensilla to detect semiochemicals in the environment. Utilizing single sensillum recording, we demonstrate a method to evaluate bed bug response to semiochemicals and explore the coding process involved.

情報化学物質にトコジラミの嗅覚ニューロンの応答を検出するために単一感覚子録音を使用しました

The insect olfactory system plays an important role in detecting semiochemicals in the environment. In particular, the antennal sensilla which house ...

using-single-sensillum-recording-to-detect-olfactory-neuron-responses

Research

JoVE Journal

Neuroscience

9.8K ビュー.  Auburn University. Bed bugs rely on olfactory receptor neurons housed in their antennal olfactory sensilla to detect semiochemicals in the environment. Utilizing single sensillum recording, we demonstrate a method to evaluate bed bug response to semiochemicals and explore the coding process involved.

ビデオ: Using Single Sensillum Recording to Detect Olfactory Neuron Responses of Bed Bugs to Semiochemicals

Single Sensillum Recordings in the Insects Drosophila melanogaster and Anopheles gambiae

The sense of smell is essential for insects to find foods, mates, predators, and oviposition sites3. Insect olfactory sensory neurons (OSNs) are enclosed in sensory hairs called sensilla, which cover the surface of olfactory organs. The surface of each sensillum is covered with tiny pores, through which odorants pass and dissolve in a fluid called sensillum lymph, which bathes the sensory dendrites of the OSNs housed in a given sensillum. The OSN dendrites express odorant receptor (OR) proteins, which in insects function as odor-gated ion channels4, 5. The interaction of odorants with ORs either increases or decreases the basal firing rate of the OSN. This neuronal activity in the form of action potentials embodies the first representation of the quality, intensity, and temporal characteristics of the odorant6, 7.
Given the easy access to these sensory hairs, it is possible to perform extracellular recordings from single OSNs by introducing a recording electrode into the sensillum lymph, while the reference electrode is placed in the lymph of the eye or body of the insect. In Drosophila, sensilla house between one and four OSNs, but each OSN typically displays a characteristic spike amplitude. Spike sorting techniques make it possible to assign spiking responses to individual OSNs. This single sensillum recording (SSR) technique monitors the difference in potential between the sensillum lymph and the reference electrode as electrical spikes that are generated by the receptor activity on OSNs1, 2, 8. Changes in the number of spikes in response to the odorant represent the cellular basis of odor coding in insects. Here, we describe the preparation method currently used in our lab to perform SSR on Drosophila melanogaster and Anopheles gambiae, and show representative traces induced by the odorants in a sensillum-specific manner.

Single Sensillum Recordings in the Insects Drosophila melanogaster and Anopheles gambiae

Electrophysiological responses of olfactory sensory neurons to odorants can be measured in insects using single sensillum recordings. In this video article we will demonstrate how to perform single sensillum recordings in the antennae of the vinegar fly (Drosophila melanogaster) and the maxillary palps of the malaria mosquito (Anopheles gambiae).

昆虫における単一感覚子録音キイロショウジョウバエとハマダラカ</em

The sense of smell is essential for insects to find foods, mates, predators, and oviposition sites3. Insect olfactory sensory neurons (OSNs) are enclosed ...

神経科学

Electrophysiological Measurements from a Moth Olfactory System

Insect olfactory systems provide unique opportunities for recording odorant-induced responses in the forms of electroantennograms (EAG) and single sensillum recordings (SSR), which are summed responses from all odorant receptor neurons (ORNs) located on the antenna and from those housed in individual sensilla, respectively. These approaches have been exploited for getting a better understanding of insect chemical communication. The identified stimuli can then be used as either attractants or repellents in management strategies for insect pests.

Insect olfactory systems provide unique opportunities for recording odorant-induced responses in the forms of electroantennograms (EAG) and single sensillum recordings (SSR), which are summed responses from all odorant receptor neurons (ORNs) located on the antenna and from those housed in individual sensilla, respectively.

蛾嗅覚系の電気生理学的測定

Insect olfactory systems provide unique opportunities for recording odorant-induced responses in the forms of electroantennograms (EAG) and single ...

Electrophysiology of Scorpion Peg Sensilla

We describe a modification of an existing tip-recording technique1,2 for electrophysiologically investigating short, peg-like sensory sensilla3,4. On the mid-ventral surface of all scorpions are two appendages called pectines, which have dense fields of mechano- and chemosensory peg sensilla5,6. One method for assessing chemoresponsiveness of these sensilla uses a tungsten electrode for extracellularly recording neural activity within a sensillum as a volatile odorant is introduced to the sensory field5,7. The limitations of this method include slow data collection and uncontrolled stimulant introduction to, and removal from, the peg field. To overcome these limitations, we developed a new tip-recording technique that uses nonpolar mineral oil as a medium through which to deliver water-based tastants to individual peg sensilla8,9. We have successfully applied this method to obtain sensillar chemoresponses to citric acid, ethanol, and salt. Here we describe the experimental protocol for such a study9. We think this new method may be useful for studying the response properties of other arthropod chemosensory systems, including those of insects10, 11 and crustaceans12.

This article describes an electrophysiological method for isolating chemical stimulation to individual sensilla via extracellular, tip-recordings under mineral oil.

スコーピオンペグ感覚器の電気生理学

We describe a modification of an existing tip-recording technique1,2 for electrophysiologically investigating short, peg-like sensory sensilla3,4. On the ...

Odorant-induced Responses Recorded from Olfactory Receptor Neurons using the Suction Pipette Technique

Animals sample the odorous environment around them through the chemosensory systems located in the nasal cavity. Chemosensory signals affect complex behaviors such as food choice, predator, conspecific and mate recognition and other socially relevant cues. Olfactory receptor neurons (ORNs) are located in the dorsal part of the nasal cavity embedded in the olfactory epithelium. These bipolar neurons send an axon to the olfactory bulb (see Fig. 1, Reisert &amp; Zhao1, originally published in the Journal of General Physiology) and extend a single dendrite to the epithelial border from where cilia radiate into the mucus that covers the olfactory epithelium. The cilia contain the signal transduction machinery that ultimately leads to excitatory current influx through the ciliary transduction channels, a cyclic nucleotide-gated (CNG) channel and a Ca2+-activated Cl- channel (Fig. 1). The ensuing depolarization triggers action potential generation at the cell body2-4.
In this video we describe the use of the "suction pipette technique" to record odorant-induced responses from ORNs. This method was originally developed to record from rod photoreceptors5 and a variant of this method can be found at jove.com modified to record from mouse cone photoreceptors6. The suction pipette technique was later adapted to also record from ORNs7,8. Briefly, following dissociation of the olfactory epithelium and cell isolation, the entire cell body of an ORN is sucked into the tip of a recording pipette. The dendrite and the cilia remain exposed to the bath solution and thus accessible to solution changes to enable e.g. odorant or pharmacological blocker application. In this configuration, no access to the intracellular environment is gained (no whole-cell voltage clamp) and the intracellular voltage remains free to vary. This allows the simultaneous recording of the slow receptor current that originates at the cilia and fast action potentials fired by the cell body9. The difference in kinetics between these two signals allows them to be separated using different filter settings. This technique can be used on any wild type or knockout mouse or to record selectively from ORNs that also express GFP to label specific subsets of ORNs, e.g. expressing a given odorant receptor or ion channel.

Olfactory receptor neurons (ORNs) convert odor signals first into a receptor current that in turn triggers action potentials that are conveyed to second order neurons in the olfactory bulb. Here we describe the suction pipette technique to record simultaneously the odorant-induced receptor current and action potentials from mouse ORNs.

吸引ピペット技術を用いた嗅覚受容ニューロンから記録匂い物質誘発性応答

Animals sample the odorous environment around them through the chemosensory systems located in the nasal cavity. Chemosensory signals affect complex ...

Multi-unit Recording Methods to Characterize Neural Activity in the Locust (Schistocerca Americana) Olfactory Circuits

Detection and interpretation of olfactory cues are critical for the survival of many organisms. Remarkably, species across phyla have strikingly similar olfactory systems suggesting that the biological approach to chemical sensing has been optimized over evolutionary time1. In the insect olfactory system, odorants are transduced by olfactory receptor neurons (ORN) in the antenna, which convert chemical stimuli into trains of action potentials. Sensory input from the ORNs is then relayed to the antennal lobe (AL; a structure analogous to the vertebrate olfactory bulb). In the AL, neural representations for odors take the form of spatiotemporal firing patterns distributed across ensembles of principal neurons (PNs; also referred to as projection neurons)2,3. The AL output is subsequently processed by Kenyon cells (KCs) in the downstream mushroom body (MB), a structure associated with olfactory memory and learning4,5. Here, we present electrophysiological recording techniques to monitor odor-evoked neural responses in these olfactory circuits.
First, we present a single sensillum recording method to study odor-evoked responses at the level of populations of ORNs6,7. We discuss the use of saline filled sharpened glass pipettes as electrodes to extracellularly monitor ORN responses. Next, we present a method to extracellularly monitor PN responses using a commercial 16-channel electrode3. A similar approach using a custom-made 8-channel twisted wire tetrode is demonstrated for Kenyon cell recordings8. We provide details of our experimental setup and present representative recording traces for each of these techniques.

Multi-unit Recording Methods to Characterize Neural Activity in the Locust Schistocerca Americana Olfactory Circuits

We demonstrate variations of the extracellular multi-unit recording technique to characterize odor-evoked responses in the first three stages of the invertebrate olfactory pathway. These techniques can easily be adapted to examine ensemble activity in other neural systems as well.

ローカスの神経活動を特徴づけるためにマルチユニット記録方法（<em&gt; Schistocercaアメリカーナ</em&gt;）嗅覚回路

Detection and interpretation of olfactory cues are critical for the survival  of many organisms. Remarkably, species across phyla have strikingly similar ...

Perforated Patch-clamp Recording of Mouse Olfactory Sensory Neurons in Intact Neuroepithelium: Functional Analysis of Neurons Expressing an Identified Odorant Receptor

Analyzing the physiological responses of olfactory sensory neurons (OSN) when stimulated with specific ligands is critical to understand the basis of olfactory-driven behaviors and their modulation. These coding properties depend heavily on the initial interaction between odor molecules and the olfactory receptor (OR) expressed in the OSNs. The identity, specificity and ligand spectrum of the expressed OR are critical. The probability to find the ligand of the OR expressed in an OSN chosen randomly within the epithelium is very low. To address this challenge, this protocol uses genetically tagged mice expressing the fluorescent protein GFP under the control of the promoter of defined ORs. OSNs are located in a tight and organized epithelium lining the nasal cavity, with neighboring cells influencing their maturation and function. Here we describe a method to isolate an intact olfactory epithelium and record through patch-clamp recordings the properties of OSNs expressing defined odorant receptors. The protocol allows one to characterize OSN membrane properties while keeping the influence of the neighboring tissue. Analysis of patch-clamp results yields&#160;a precise quantification of ligand/OR interactions, transduction pathways and pharmacology, OSNs&#39; coding properties and their modulation at the membrane level.&#160;

Analyzing the physiological properties of olfactory sensory neurons still faces technical limitations. Here we record them through perforated patch-clamp in an intact preparation of the olfactory epithelium in gene-targeted mice. This technique allows the characterization of membrane properties and responses to specific ligands of neurons expressing defined olfactory receptors.

無傷の神経上皮におけるマウス嗅細胞の穿孔パッチクランプ記録：同定された嗅覚受容体を発現するニューロンの機能解析

Analyzing the physiological responses of olfactory sensory neurons (OSN) when stimulated with specific ligands is critical to understand the basis of ...

Using an Adapted Microfluidic Olfactory Chip for the Imaging of Neuronal Activity in Response to Pheromones in Male C. Elegans Head Neurons

The use of calcium indicators has greatly enhanced our understanding of neural dynamics and regulation. The nematode Caenorhabditis elegans, with its completely mapped nervous system and transparent anatomy, presents an ideal model for understanding real-time neural dynamics using calcium indicators. In combination with microfluidic technologies and experimental designs, calcium-imaging studies using these indicators are performed in both free-moving and trapped animals. However, most previous studies utilizing trapping devices, such as the olfactory chip described in Chronis et al., have devices designed for use in the more common hermaphrodite, as the less common male is both morphologically and structurally dissimilar. An adapted olfactory chip was designed and fabricated for increased efficiency in male neuronal imaging with using young adult animals. A turn was incorporated into the worm loading port to rotate the animals and to allow for the separation of the individual neurons within a bilateral pair in 2D imaging. Worms are exposed to a controlled flow of odorant within the microfluidic device, as described in previous hermaphrodite studies. Calcium transients are then analyzed using the open-source software ImageJ. The procedure described herein should allow for an increased amount of male-based C. elegans calcium imaging studies, deepening our understanding of the mechanisms of sex-specific neuronal signaling.

Using an Adapted Microfluidic Olfactory Chip for the Imaging of Neuronal Activity in Response to Pheromones in Male C. Elegans Head Neurons

The use of an adapted &#34;olfactory chip&#34; for the efficient calcium imaging of C. elegans males is described here. Studies of male exposure to glycerol and a pheromone are also shown.

適応マイクロ嗅覚を用いた男性c. の Elegans頭ニューロンのフェロモン応答における神経活動のイメージング

The use of calcium indicators has greatly enhanced our understanding of neural dynamics and regulation. The nematode Caenorhabditis elegans, with its ...

Electrophysiological Recording from Drosophila Trichoid Sensilla in Response to Odorants of Low Volatility

Insects rely on their sense of smell to guide a wide range of behaviors that are critical for their survival, such as food-seeking, predator avoidance, oviposition, and mating. Myriad chemicals of varying volatilities have been identified as natural odorants that activate insect Olfactory Receptor Neurons (ORNs). However, studying the olfactory responses to low-volatility odorants has been hampered by an inability to effectively present such stimuli using conventional odor-delivery methods. Here, we describe a procedure that permits the effective presentation of low-volatility odorants for in vivo Single-Sensillum Recording (SSR). By minimizing the distance between the odor source and the target tissue, this method allows for the application of biologically salient but hitherto inaccessible odorants, including palmitoleic acid, a stimulatory pheromone with a demonstrated effect on ORNs involved in courtship and mating behavior1. Our procedure thus affords a new avenue to assay a host of low-volatility odorants for the study of insect olfaction and pheromone communication.

Electrophysiological Recording from Drosophila Trichoid Sensilla in Response to Odorants of Low Volatility

The overall goal of this protocol is to demonstrate how to present odorants of low volatility for single-sensillum recording from Drosophila olfactory receptor neurons that respond to long-chain cuticular pheromones.

からの電気生理学的記録<em&gt;ショウジョウバエ</em&gt;低揮発性の臭気物質に対するトリコイドセンシラ

Insects rely on their sense of smell to guide a wide range of behaviors that are critical for their survival, such as food-seeking, predator avoidance, ...

Single Sensillum Recordings for Locust Palp Sensilla Basiconica

The palps of locust mouthparts are considered to be conventional gustatory organs that play an important role in a locust&#39;s food selection, especially for the detection of non-volatile chemical cues through sensilla chaetica (previously named terminal sensilla or crested sensilla). There is now increasing evidence that these palps also have an olfactory function. An odorant receptor (LmigOR2) and an odorant-binding protein (LmigOBP1) have been localized in the neurons and accessory cells, respectively, in the sensilla basiconica of the palps. Single sensillum recording (SSR) is used for recording the responses of odorant receptor neurons, which is an effective method for screening active ligands on specific odorant receptors. SSR is used in functional studies of odorant receptors in palp sensilla. The structure of the sensilla basiconica located on the dome of the palps differs somewhat from the structure of those on the antennae. Therefore, when performing an SSR elicited by odorants, some specific advice may be helpful for obtaining optimum results. In this paper, a detailed and highly effective protocol for an SSR from insect palp sensilla basiconica is introduced.

This paper describes a detailed and highly effective protocol for single sensillum recordings from the sensilla basiconica on the palps of insect mouthparts.

イナゴ Palp 触角 Basiconica の単一ゴキブリ録音

The palps of locust mouthparts are considered to be conventional gustatory organs that play an important role in a locust&#39;s food selection, especially ...

昆虫の味覚ニューロン活動の強化された分析のための新しい方法

Base Recording: A Technique for Analyzing Responses of Taste Neurons in Drosophila

Insects taste the external world through taste hairs, or sensilla, that have pores at their tips. When a sensillum comes into contact with a potential food source, compounds from the food source enter through the pore and activate neurons within. For over 50 years, these responses have been recorded using a technique called tip recording. However, this method has major limitations, including the inability to measure neural activity before or after stimulus contact and the requirement for tastants to be soluble in aqueous solutions. We describe here a technique that we call base recording, which overcomes these limitations. Base recording allows the measurement of taste neuron activity before, during, and after the stimulus. Thus, it allows extensive analysis of OFF responses that occur after a taste stimulus. It can be used to study hydrophobic compounds such as long-chain pheromones that have very low solubility in water. In summary, base recording offers the advantages of single-sensillum electrophysiology as a means of measuring neuronal activity - high spatial and temporal resolution, without the need for genetic tools - and overcomes key limitations of the traditional tip recording technique.

著者スポットライト:化学受容の進歩 – 昆虫の匂い受容体からノンコーディングRNAまで

A rarely used method of electrophysiological recording, base recording, allows analysis of features of taste coding that cannot be examined by conventional recording methods. Base recording also allows the analysis of taste responses to hydrophobic stimuli that cannot be studied using traditional electrophysiological methods.

塩基記録:ショウジョウバエの味覚ニューロン応答解析技術

Insects taste the external world through taste hairs, or sensilla, that have pores at their tips. When a sensillum comes into contact with a potential ...

情報化学物質にトコジラミの嗅覚ニューロンの応答を検出するために単一感覚子録音を使用しました

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昆虫における単一感覚子録音キイロショウジョウバエとハマダラカ