我々は、これらの実験技術の開発中に技術的なインプットを提供するための博士ダングリーンに感謝。この作品は、NIHグラントR01 - EY016849Aとスミスファミリー新奨励賞によってサポートされていました。

パートI：タングステンインガラス電極の作製<ol><li>グラファイトリングは、硝酸ナトリウム（0.71g/mL）と水1に溶解した水酸化カリウム（0.34g/mL）の濃縮液で満たされたビーカーに入れられ、ビーカーが変更されたミシンの縫製腕の下に置かれている。 </li><li>それは部分的に浸漬されるようにタングステン線（0.5ミリメートルの直径、6センチメートル長さ）は溶液中で（1〜2センチメートル）ミシンアームにテープで固定されています。 </li><li> 2Vは、グラファイトリングに縫製アームと負のリード線にプラス側のリード線を取り付けることによりタングステンワイヤを介して適用されます。 </li><li>ミシンがオンになっているとタングステンワイヤが電解鋭いポイントに金属をエッチングする2分、のための〜4Hzの速度で溶液中に繰り返し浸漬する。 </li><li>エッチングされたワイヤは、マニュアルマニピュレータ及びガラスピペット（0.25ミリメートルOD、0.175ミリメートルID）ガラスの引き手と1mmの先端に引っ張らのシャフトを持つ微細な指導（40倍の倍率）の下に整列点に配置されます。電線挿入中に動かないようにするため、ガラスは、一時的にパテを顕微鏡ステージに固定されている。 </li><li>ワイヤはシャフトの下、ガラスのunpulled最後に慎重に進んでおり、1μmの開口部を通してガラスの小片は、ガラス中のタングステンのタイトなフィットを示し、ビデオの先端を絶つために見られている。ワイヤーは、ラット光ファイバー録音用先端を越えて5〜10ミリメートルの周りに拡張する必要があります。 </li><li>シアノアクリレートは、ガラスのunpulled最後に適用され、硬化後の電気インピーダンスを測定して、1MΩの電極は、一般的に最も効果的場所。 </li></ol>パートII：定位セットアップとスパイクトレインレコーディング<ol><li>刺激の表示システムは、脳に電極を配置し、低下させるための定位固定装置とマイクロドライブと一緒に、ビデオに示されている。 </li><li>網膜スパイク列の記録を実行するためにラットを麻酔し、後に麻痺です。標準的な手順は、（図示せず）薬物送達および麻痺の間に機械的な換気のために気管（Y -チューブ）カニューレのための大腿静脈にカニューレの外科的挿入のために使用されています。実験は、動物用の端子です。 </li><li>準備手続の後、動物は加熱毛布の上に配置され、頭部は耳と歯のバーで、としてラット脳アトラス2で定義されて、定位位置に固定されています。 </li><li>直腸プローブは37℃体温を維持する恒温制御ユニット℃に介して加熱毛布を制御する、挿入される</li><li>金属針は、心電図やモニターの心拍数を測定するためにリード線として使用されます。 </li><li>皮膚は、頭蓋骨の上に開かれ、頭蓋骨のプレートの交点（ブレグマ）が公開されています。カスタム設計されたクロスバーのフレームを所定の位置に設定されている、円はクロスバーの穴に通しbegma周り頭蓋骨に記されています、とクロスバーは、一時的に削除されます。 </li><li>定位システムはブレグマ上のガイドの針を配置することによって較正されています。 5 mmの穴が著しい場所で頭蓋骨にドリルされ、骨が削除され、脳を露出させる。パテの細いリングが所定の位置にリセットされたクロスバー、の穴周りのシールを形成するために頭蓋骨の開口部の周りに置かれます。 </li><li>動物は、しばらく麻酔の安定した平面になっている場合、それはデータ収集時に目の動きを防ぐために、麻痺を注射されています。呼吸が停止した後、動物は機械的に換気される。カスタムプログラムでは、実験を通して換気と生理学的パラメータを追跡し、任意のパラメータが正常なレベルの外に行く必要の補償作用が必要な場合は実験者に知らせます。 </li><li>ガイド針はタングステン電極とbackloadedと脳に低下する。脳の浸透後にクロスバーの穴は、寒天（図示せず）が充填されている、とマイクロドライブのアセンブリは、機械的安定性のためのクロスバーにクランプされます。先端が視交叉が配置されている頭蓋骨、下は約8〜9ミリメートルになるまで、タングステン電極は、ガイド針アウト（2μm/sec）その後徐々に高度です。 </li><li>網膜神経節細胞に属するスパイクは、視覚入力への応答によって識別されます。単一の光ファイバの分離時に記録された細胞の受容野は、視覚的な空間にマッピングされ、その応答特性は、漂流格子や関心のある他の視覚的なパターンで特徴付けられる。システムの安定性は、数時間の3つのユニットの録音が可能になります。 </li></ol>材料成人ブラウンノルウェーラット（250〜400グラム）が商用ベンダーから購入し、規制（12/12）明/暗サイクル下で飼育した。麻酔は塩酸ケタミンとキシラジン（70、2 mg / kgを、それぞれ、ヘンリーシャイン社）の腹腔内注射で誘導とexperの期間中維持されたカテーテルを通して静脈内ブドウ糖、生理食塩水と混合ケタミンとキシラジンの投与（30及び1 mg / kg / hrの）、およびガラミンのtriethiodide（40 mg / kg体重/時、フィッシャー社）（0.13ミリメートルOD、スモールパーツ社とiment右大腿静脈で）。ガラミンは、目の動きを麻痺させる混合物に含まれています。心拍数と血圧の変動性によって評価される麻酔の安定した平面を維持するために必要なポンプ（WPI社）の注入速度を調節した。麻痺した後、動物は機械的にインラインで測定した呼気終末CO 2を維持するために、必要に応じて調整率で、60から80呼吸/分（2ccの体積）で気管カニューレを介して（ハーバード装置、モデル683）換気された30％でカプノメーター（ノバメトリックス社、モデル710Sp）。体温は恒温毛布の制御システム（ハーバード装置株式会社）を介して調節した。体温、呼吸終期のCO 2、心拍数、血圧を連続的にLabVIEWのプログラムで実験を通してモニターした。視覚刺激は、800x600ピクセルの解像度で100Hzで動作してソニーのマルチスキャン17E CRTモニター（30 cd / m 2の輝度を意味する）に発表されました。データ収集とモニター出力は、ビデオイメージプロセッサ（ケンブリッジリサーチシステム株式会社、ビット+ +）と心理物理学ツールボックス4との組み合わせでMATLABやLabVIEWで記述されたカスタムソフトウェアで制御されていました。動物は、コンタクトレンズ（眼インスツルメンツ社）を通じて、16.5センチメートルの距離で刺激ディスプレイ（40.4 X 30.2センチメートル）表示、および点眼液は眼の潤いを保つために、実験中に定期的に適用されました。刺激へのアクセス可能な視野は、フローティングテーブルの表面（TMC社）上記の定位固定装置（CoをStoelting）上昇14センチメートルで動物をマウントし、一緒にテーブルの上に移動し、カスタム設計のそりの上にモニターを設定し、逆により最大化されていた鼻は約± 100 °の円弧。これは60度を超えると35度目の高さ以下と± 42度横方向にモニタの中心からの延長repositionable表示フィールドを提供する。タングステンelectodesは、ガラスチップを形成するために使用された小型部品株式会社とフリードリヒとディモック社標準micropippeteプーラー（サッタインスツルメンツ、モデルP - 97）から得られたカスタムホウケイ酸ガラスから得られる線で作製した。電極インピーダンスのテスターは、朴エレクトロニクスから購入した。電極は、電動マイクロドライブシステム（ニューポート社、StepperMike）で進めていた。心電図および視神経の信号は、高入力インピーダンスの多電極アンプ（FHC社、X -セル× 4）で増幅し、ろ過した。ハートビートは、ウィンドウのディスクリミネータ（WPI社、モデル121）で検出した。神経の活動電位は、デジタルスパイク弁別（FHC社、APM）で0.1msの分解能で検出し、タイムスタンプが付けられた。スパイクディスクリミネータは、スパイク時間が刺激にロックされていたようにビデオ画像プロセッサからのトリガ信号によってゲートだった。

<ol>
	<li>Levick, W. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Another+tungsten+microelectrode.">Another tungsten microelectrode.</a> Med Biol Eng. 10 (4), 510-515 (1972).</li><li>Paxinos, G., Watson, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The rat brain in stereotaxic coordinates. , (1998).</li><li>Freeman, D. K., Heine, W. F., Passaglia, C. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+maintained+discharge+of+rat+retinal+ganglion+cells.">The maintained discharge of rat retinal ganglion cells.</a> Vis Neurosci. 25 (4), 535-548 (2008).</li><li>Brainard, D. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+psychophysics+toolbox.">The psychophysics toolbox.</a> Spat Vis. 10, 433-436 (1997).</li></ol>

すべての実験手順は、ボストン大学の動物実験使用の委員会によって承認された。

光ファイバーの録音は、そのまま目を必要とする網膜情報の符号化と伝送に関する実験的な質問に対処するための魅力的なアプローチです。電極が交差し、非クロス視神経線維の活動が単一の電極への浸透で記録することができます視交叉や消化管に配置されている場合、また、両眼のシグナル伝達特性は、ほぼ同じ生理的な状態で検討することができます。光ファイバーの録音は、げっ歯類のようなビジョンの研究で使用されている他の人気の動物モデルでそれらの小さいサイズに多分により猫に共通のものですが、。我々は、ラットにおける単繊維の活動を拾いに成功した、どれも同じようなインピーダンスと材料の商業電極、様々な試みている、私たちの電極のようないくつかの時間ができるためにそれを記録するどころか。これは、典型的な商用のものの広い丸い先端とは対照的に長く、鋭い先端を持っている私たちの電極、、の特定の幾何学は神経節細胞軸索スパイクの信頼性と安定した単離のために重要であることを意味します。これらの電極を作製する方法を示すに加えて、我々は、 生体の視覚神経生理学的研究のために私たちのカスタム設計の定位システムを示しています。システムは、環境振動から、電磁ノイズからの高インピーダンスの微小電極を保護するために構築されます。これは、特に近くに位置するコンピュータのモニタで、長期間の軸索（のような細胞体に比べて）によって生成された小さな活動電位を記録するために重要です。それは数十マイクロボルトのオーダーの時間以上と騒音レベルの代表的な記録時間で、優れた安定性と信号対雑音比でそうする。これらの機能は、網膜や他の脳領域の神経線維から生体内で録音することを目的とビジョンの研究者のためのセットアップは特に有用です。

single-unit in vivo recordings from the optic chiasm of rat

視覚的な世界に関する情報は、視神経を構成する網膜神経節細胞の軸索における活動電位のシーケンスで脳に伝達される。<em&gt; in vivoで</emいくつかの動物モデルにおける神経節細胞のスパイク列の&gt;録音はどのように初期視覚システムのプロセスについて知られていることの多くを明らかにし、視覚的情報をエンコードしている。しかし、そのような録音は、最も一般的な動物モデルの一つ、ラットで小径軸索によってトリガされるスパイクを検出することが困難である可能性によりまれてきた。ラットを含む多くの網膜疾患のモデルは、眼を持つかのように、目を乱すことなく、神経節細胞の機能的性質を特徴づけるための必要性を動機<em&gt; in vitroで</em&gt;レコーディング。ここで、我々は麻酔ラットの視交叉から神経節細胞スパイク列を記録するための方法を示しています。まず、ラットにおける単神経節細胞の軸索からの電気的活動を拾うことができるタングステンインガラス電極を作製する方法を示します。電極は、我々がしようとしたこと商業目的のものを上回っています。私たちは、その後のために私たちのカスタム設計の定位システムを示しています<em&gt; in vivoで</em&gt;視覚神経生理学の実験や動物の準備と視交叉における信頼性と安定した電極配置のための私たちの手続き。

Watch this Scientific Journal Video about Single-unit In vivo Recordings from the Optic Chiasm of Rat at JoVE.com

Single-unit In vivo Recordings from the Optic Chiasm of Rat

網膜神経節細胞は、眼からの活動電位の配列との脳に視覚情報を送信する。ここで、我々は、単一の神経節細胞の活動電位を記録する方法を示しています<em&gt; in vivoで</em&gt;麻酔ラットから。

シングルユニット in vivoで</emラットの視交叉から>録音

Information about the visual world is transmitted to the brain in sequences of action potentials in retinal ganglion cell axons that make up the optic ...

single-unit-in-vivo-recordings-from-the-optic-chiasm-of-rat

Nanyang Technological University

Research

JoVE Journal

Neuroscience

Single-unit In vivo Recordings from the Optic Chiasm of Rat

11.6K ビュー.  Boston University. 網膜神経節細胞は、眼からの活動電位の配列との脳に視覚情報を送信する。ここで、我々は、単一の神経節細胞の活動電位を記録する方法を示しています in vivoで麻酔ラットから。

ビデオ: Single-unit In vivo Recordings from the Optic Chiasm of Rat

Electrophysiological Recordings from the Giant Fiber Pathway of D. melanogaster

When startled adult D. melanogaster react by jumping into the air and flying away. In many invertebrate species, including D. melanogaster, the "escape" (or "startle") response during the adult stage is mediated by the multi-component neuronal circuit called the Giant Fiber System (GFS). The comparative large size of the neurons, their distinctive morphology and simple connectivity make the GFS an attractive model system for studying neuronal circuitry. The GFS pathway is composed of two bilaterally symmetrical Giant Fiber (GF) interneurons whose axons descend from the brain along the midline into the thoracic ganglion via the cervical connective. In the mesothoracic neuromere (T2) of the ventral ganglia the GFs form electro-chemical synapses with 1) the large medial dendrite of the ipsilateral motorneuron (TTMn) which drives the tergotrochanteral muscle (TTM), the main extensor for the mesothoracic femur/leg, and 2) the contralateral peripherally synapsing interneuron (PSI) which in turn forms chemical (cholinergic) synapses with the motorneurons (DLMns) of the dorsal longitudinal muscles (DLMs), the wing depressors. The neuronal pathway(s) to the dorsovental muscles (DVMs), the wing elevators, has not yet been worked out (the DLMs and DVMs are known jointly as indirect flight muscles - they are not attached directly to the wings, but rather move the wings indirectly by distorting the nearby thoracic cuticle) (King and Wyman, 1980; Allen et al., 2006). The di-synaptic activation of the DLMs (via PSI) causes a small but important delay in the timing of the contraction of these muscles relative to the monosynaptic activation of TTM (~0.5 ms) allowing the TTMs to first extend the femur and propel the fly off the ground. The TTMs simultaneously stretch-activate the DLMs which in turn mutually stretch-activate the DVMs for the duration of the flight. The GF pathway can be activated either indirectly by applying a sensory (e.g."air-puff" or "lights-off") stimulus, or directly by a supra-threshold electrical stimulus to the brain (described here). In both cases, an action potential reaches the TTMs and DLMs solely via the GFs, PSIs, and TTM/DLM motoneurons, although the TTMns and DLMns do have other, as yet unidentified, sensory inputs. Measuring "latency response" (the time between the stimulation and muscle depolarization) and the "following to high frequency stimulation" (the number of successful responses to a certain number of high frequency stimuli) provides a way to reproducibly and quantitatively assess the functional status of the GFS components, including both central synapses (GF-TTMn, GF-PSI, PSI-DLMn) and the chemical (glutamatergic) neuromuscular junctions (TTMn-TTM and DLMn-DLM). It has been used to identify genes involved in central synapse formation and to assess CNS function.

Electrophysiological Recordings from the Giant Fiber Pathway of D. melanogaster

The Giant Fiber System is a simple neuronal circuit of adult Drosophila melanogaster containing the largest neurons in the fly. We describe the protocol for monitoring synaptic transmission through this pathway by recording post synaptic potentials in dorsal longitudinal (DLM) and tergotrochanteral (TTM) muscles following direct stimulation of the Giant Fiber interneurons.

の巨大線維経路からの電気生理学的記録 D.キイロ</em

When startled adult D. melanogaster react by jumping into the air and flying away. In many invertebrate species, including D. melanogaster, the "escape" ...

神経科学

Retrograde Fluorescent Labeling Allows for Targeted Extracellular Single-unit Recording from Identified Neurons In vivo

The overall goal of this method is to record single-unit responses from an identified population of neurons. In vivo electrophysiological recordings from individual neurons are critical for understanding how neural circuits function under natural conditions. Traditionally, these recordings have been performed 'blind', meaning the identity of the recorded cell is unknown at the start of the recording. Cellular identity can be subsequently determined via intracellular1, juxtacellular2 or loose-patch3 iontophoresis of dye, but these recordings cannot be pre-targeted to specific neurons in regions with functionally heterogeneous cell types. Fluorescent proteins can be expressed in a cell-type specific manner permitting visually-guided single-cell electrophysiology4-6. However, there are many model systems for which these genetic tools are not available. Even in genetically accessible model systems, the desired promoter may be unknown or genetically homogenous neurons may have varying projection patterns. Similarly, viral vectors have been used to label specific subgroups of projection neurons7, but use of this method is limited by toxicity and lack of trans-synaptic specificity. Thus, additional techniques that offer specific pre-visualization to record from identified single neurons in vivo are needed. Pre-visualization of the target neuron is particularly useful for challenging recording conditions, for which classical single-cell recordings are often prohibitively difficult8-11. The novel technique described in this paper uses retrograde transport of a fluorescent dye applied using tungsten needles to rapidly and selectively label a specific subset of cells within a particular brain region based on their unique axonal projections, thereby providing a visual cue to obtain targeted electrophysiological recordings from identified neurons in an intact circuit within a vertebrate CNS.
The most significant novel advancement of our method is the use of fluorescent labeling to target specific cell types in a non-genetically accessible model system. Weakly electric fish are an excellent model system for studying neural circuits in awake, behaving animals12. We utilized this technique to study sensory processing by "small cells" in the anterior exterolateral nucleus (ELa) of weakly electric mormyrid fish. "Small cells" are hypothesized to be time comparator neurons important for detecting submillisecond differences in the arrival times of presynaptic spikes13. However, anatomical features such as dense myelin, engulfing synapses, and small cell bodies have made it extremely difficult to record from these cells using traditional methods11, 14. Here we demonstrate that our novel method selectively labels these cells in 28% of preparations, allowing for reliable, robust recordings and characterization of responses to electrosensory stimulation.

Retrograde Fluorescent Labeling Allows for Targeted Extracellular Single-unit Recording from Identified Neurons In vivo

Retrograde transport of fluorescent dye labels a sub-population of neurons based on anatomical projection. Labeled axons can be visually targeted in vivo, permitting extracellular recording from identified axons. This technique facilitates recording when neurons cannot be labeled through genetic manipulation or are difficult to isolate using 'blind' in vivo approaches.

逆行性蛍光標識は、特定された神経細胞から標的細胞外シングルユニット記録を可能にします<em生体内で&gt;</em

The overall goal of this method is to record single-unit responses from an identified population of neurons. In vivo electrophysiological recordings from ...

A Procedure for Implanting Organized Arrays of Microwires for Single-unit Recordings in Awake, Behaving Animals

In vivo electrophysiological recordings in the awake, behaving animal provide a powerful method for understanding neural signaling at the single-cell level. The technique allows experimenters to examine temporally and regionally specific firing patterns in order to correlate recorded action potentials with ongoing behavior. Moreover, single-unit recordings can be combined with a plethora of other techniques in order to produce comprehensive explanations of neural function. In this article, we describe the anesthesia and preparation for microwire implantation. Subsequently, we enumerate the necessary equipment and surgical steps to accurately insert a microwire array into a target structure. Lastly, we briefly describe the equipment used to record from each individual electrode in the array. The fixed microwire arrays described are well-suited for chronic implantation and allow for longitudinal recordings of neural data in almost any behavioral preparation. We discuss tracing electrode tracks to triangulate microwire positions as well as ways to combine microwire implantation with immunohistochemical techniques in order to increase the anatomical specificity of recorded results.

Implanting organized arrays of microwires for use in single-unit electrophysiological recordings presents a number of technical challenges.&#160;Methods for performing this technique and the equipment necessary are described.&#160;Also, the beneficial use of organized microwire arrays to record from distinct neural subregions with high spatial selectivity is discussed.

動物を振る舞う、覚醒中のシングルユニット記録のためにマイクロワイヤの組織化された配列を移植するための手順

In vivo electrophysiological recordings in the awake, behaving animal provide a powerful method for understanding neural signaling at the single-cell ...

A Guide to In vivo Single-unit Recording from Optogenetically Identified Cortical Inhibitory Interneurons

A major challenge in neurophysiology has been to characterize the response properties and function of the numerous inhibitory cell types in the cerebral cortex. We here share our strategy for obtaining stable, well-isolated single-unit recordings from identified inhibitory interneurons in the anesthetized mouse cortex using a method developed by Lima and colleagues1. Recordings are performed in mice expressing Channelrhodopsin-2 (ChR2) in specific neuronal subpopulations. Members of the population are identified by their response to a brief flash of blue light. This technique &#8211; termed &#8220;PINP&#8221;, or Photostimulation-assisted Identification of Neuronal Populations &#8211; can be implemented with standard extracellular recording equipment. It can serve as an inexpensive and accessible alternative to calcium imaging or visually-guided patching, for the purpose of targeting extracellular recordings to genetically-identified cells. Here we provide a set of guidelines for optimizing the method in everyday practice. We refined our strategy specifically for targeting parvalbumin-positive (PV+) cells, but have found that it works for other interneuron types as well, such as somatostatin-expressing (SOM+) and calretinin-expressing (CR+) interneurons.

A Guide to In vivo Single-unit Recording from Optogenetically Identified Cortical Inhibitory Interneurons

Here we describe our strategy for obtaining stable, well-isolated single-unit recordings from identified inhibitory interneurons in the anesthetized mouse cortex. Neurons expressing ChR2 are identified by their response to blue light. The method uses standard extracellular recording equipment, and serves as an inexpensive alternative to calcium imaging or visually-guided patching.

へのガイド<em&gt;インビボ</emOptogenetically同定された皮質抑制性介在から&gt;シングル単位記録

A major challenge in neurophysiology has been to characterize the response properties and function of the numerous inhibitory cell types in the cerebral ...

Minimally-invasive Technique for Injection into Rat Optic Nerve

The rat optic nerve is a useful model for stem cell regeneration research. Direct injection into the rat optic nerve allows delivery into the central nervous system in a minimally-invasive surgery without bone removal. This technique describes an approach to visualization and direct injection of the optic nerve following minor fascial dissection from the orbital ridge, using a conjunctival traction suture to gently pull the eye down and out. Representative examples of an injected optic nerve show successful injection of dyed beads.

Direct injection into the rat optic nerve is useful for regenerative research. We demonstrate a minimally-invasive technique for direct injection into a rat optic nerve that does not involve opening the skull. Using this method, surgical complications are minimized and recovery is more rapid.

ラット視神経への注入のための低侵襲テクニック

The rat optic nerve is a useful model for stem cell regeneration research. Direct injection into the rat optic nerve allows delivery into the central ...

Visual Evoked Potential Recording in a Rat Model of Experimental Optic Nerve Demyelination

The visual evoked potential (VEP) recording is widely used in clinical practice to assess the severity of optic neuritis in its acute phase, and to monitor the disease course in the follow-up period. Changes in the VEP parameters closely correlate with pathological damage in the optic nerve. This protocol provides a detailed description about the rodent model of optic nerve microinjection, in which a partial demyelination lesion is produced in the optic nerve. VEP recording techniques are also discussed. Using skull implanted electrodes, we are able to acquire reproducible intra-session and between-session VEP traces. VEPs can be recorded on individual animals over a period of time to assess the functional changes in the optic nerve longitudinally. The optic nerve demyelination model, in conjunction with the VEP recording protocol, provides a tool to investigate the disease processes associated with demyelination and remyelination, and can potentially be employed to evaluate the effects of new remyelinating drugs or neuroprotective therapies.

Focal demyelination is induced in the optic nerve using lysolecithin microinjection. Visual evoked potentials are recorded via skull electrodes implanted over the visual cortex to examine the signal conduction along the visual pathway in vivo. This protocol details the surgical procedures underlying electrode implantation and optic nerve microinjection.

実験視神経の脱髄のラットモデルにおける視覚誘発電位の記録

The visual evoked potential (VEP) recording is widely used in clinical practice to assess the severity of optic neuritis in its acute phase, and to ...

Determination of Photoreceptor Cell Spectral Sensitivity in an Insect Model from In Vivo Intracellular Recordings

Intracellular recording is a powerful technique used to determine how a single cell may respond to a given stimulus. In vision research, intracellular recording has historically been a common technique used to study sensitivities of individual photoreceptor cells to different light stimuli that is still being used today. However, there remains a dearth of detailed methodology in the literature for researchers wishing to replicate intracellular recording experiments in the eye. Here we present the insect as a model for examining eye physiology more generally. Insect photoreceptor cells are located near the surface of the eye and are therefore easy to reach, and many of the mechanisms involved in vision are conserved across animal phyla. We describe the basic procedure for in vivo intracellular recording of photoreceptor cells in the eye of a butterfly, with the goal of making this technique more accessible to researchers with little prior experience in electrophysiology. We introduce the basic equipment needed, how to prepare a live butterfly for recording, how to insert a glass microelectrode into a single cell, and finally the recording procedure itself. We also explain the basic analysis of raw response data for determining spectral sensitivity of individual cell types. Although our protocol focuses on determining spectral sensitivity, other stimuli (e.g., polarized light) and variations of the method are applicable to this setup.

Determination of Photoreceptor Cell Spectral Sensitivity in an Insect Model from In Vivo Intracellular Recordings

The electrophysiological technique of intracellular recording is demonstrated and used to determine spectral sensitivities of single photoreceptor cells in the compound eye of a butterfly.

昆虫モデルにおける視細胞分光感度の決意から<em&gt;インビボ</em&gt;細胞内記録

Intracellular recording is a powerful technique used to determine how a single cell may respond to a given stimulus. In vision research, intracellular ...

Acute In Vivo Electrophysiological Recordings of Local Field Potentials and Multi-unit Activity from the Hyperdirect Pathway in Anesthetized Rats

Converging evidence shows that many neuropsychiatric diseases should be understood as disorders of large-scale neuronal networks. To better understand the pathophysiological basis of these diseases, it is necessary to precisely characterize in which way the processing of information is disturbed between the different neuronal parts of the circuit. Using extracellular in vivo electrophysiological recordings, it is possible to accurately delineate neuronal activity within a neuronal network. The application of this method has several advantages over alternative techniques, e.g., functional magnetic resonance imaging and calcium imaging, as it allows a unique temporal and spatial resolution and does not rely on genetically engineered organisms. However, the use of extracellular in vivo recordings is limited since it is an invasive technique that cannot be universally applied. In this article, a simple and easy to use method is presented with which it is possible to simultaneously record extracellular potentials such as local field potentials and multiunit activity at multiple sites of a network. It is detailed how a precise targeting of subcortical nuclei can be achieved using a combination of stereotactic surgery and online analysis of multi-unit recordings. Thus, it is demonstrated, how a complete network such as the hyperdirect cortico-basal ganglia loop can be studied in anesthetized animals in vivo.

Acute In Vivo Electrophysiological Recordings of Local Field Potentials and Multi-unit Activity from the Hyperdirect Pathway in Anesthetized Rats

In this study, the methodology is presented on how to perform multi-site in vivo electrophysiological recordings from the hyperdirect pathway under urethane anesthesia.

急性<em&gt;インビボ</em&gt;麻酔ラットにおけるHyperdirect経路からの局所場ポテンシャルとマルチユニット活性の電気生理学的記録

Converging evidence shows that many neuropsychiatric diseases should be understood as disorders of large-scale neuronal networks. To better understand the ...

Simultaneous Recordings of Cortical Local Field Potentials, Electrocardiogram, Electromyogram, and Breathing Rhythm from a Freely Moving Rat

Monitoring the physiological dynamics of the brain and peripheral tissues is necessary for addressing a number of questions about how the brain controls body functions and internal organ rhythms when animals are exposed to emotional challenges and changes in their living environments. In general experiments, signals from different organs, such as the brain and the heart, are recorded by independent recording systems that require multiple recording devices and different procedures for processing the data files. This study describes a new method that can simultaneously monitor electrical biosignals, including tens of local field potentials in multiple brain regions, electrocardiograms that represent the cardiac rhythm, electromyograms that represent awake/sleep-related muscle contraction, and breathing signals, in a freely moving rat. The recording configuration of this method is based on a conventional micro-drive array for cortical local field potential recordings in which tens of electrodes are accommodated, and the signals obtained from these electrodes are integrated into a single electrical board mounted on the animal's head. Here, this recording system was improved so that signals from the peripheral organs are also transferred to an electrical interface board. In a single surgery, electrodes are first separately implanted into the appropriate body parts and the target brain areas. The open ends of all of these electrodes are then soldered to individual channels of the electrical board above the animal's head so that all of the signals can be integrated into the single electrical board. Connecting this board to a recording device allows for the collection of all of the signals into a single device, which reduces experimental costs and simplifies data processing, because all data can be handled in the same data file. This technique will aid the understanding of the neurophysiological correlates of the associations between central and peripheral organs.

This study introduces a method for the simultaneous recording of local field potentials in the brain, electrocardiograms, electromyograms, and breathing signals of a freely moving rat. This technique, which reduces experimental costs and simplifies data analysis, will contribute to the understanding of the interactions between the brain and peripheral organs.

皮質局所電界電位、心電図、筋電図、呼吸リズム自由行動ラットからの同時録音

Monitoring the physiological dynamics of the brain and peripheral tissues is necessary for addressing a number of questions about how the brain controls ...

Investigating Object Representations in the Macaque Dorsal Visual Stream Using Single-unit Recordings

Previous studies have shown that neurons in parieto-frontal areas of the macaque brain can be highly selective for real-world objects, disparity-defined curved surfaces, and images of real-world objects (with and without disparity) in a similar manner as described in the ventral visual stream. In addition, parieto-frontal areas are believed to convert visual object information into appropriate motor outputs, such as the pre-shaping of the hand during grasping. To better characterize object selectivity in the cortical network involved in visuomotor transformations, we provide a battery of tests intended to analyze the visual object selectivity of neurons in parieto-frontal regions.

A detailed protocol to analyze object selectivity of parieto-frontal neurons involved in visuomotor transformations is presented.

単一ユニットの録音を使用してサルの背側視覚ストリームのオブジェクト表現を調査

Previous studies have shown that neurons in parieto-frontal areas of the macaque brain can be highly selective for real-world objects, disparity-defined ...

シングルユニット in vivoで録音