We wish to thank Mr Wabbel for preparing the wires and Mr Tietsch for constructing the plugs and cages according to our plans. This work was supported by the Deutsche Forschungsgemeinschaft (Sonderforschungsbereich 688). Felix Fluri holds a fellowship of the Interdisziplin&#228;re Zentrum f&#252;r Klinische Forschung (IZKF), University Clinics W&#252;rzburg, Germany.

動物実験は、ヴュルツブルク大学と法的状態当局（ウンターフランケン、承認番号：54-2531.01-102 / 13）によって承認された、実験ストロークでの研究のための推奨に従って行っ17を研究し、現在の動物実験：の報告in vivo実験ガイドライン（http://www.nc3rs.org.uk/arrive-guidelines）。 1.麻酔<ol><li>手続きの期間中、供給ガス（酸素）およびイソフルランの適切な量を確保するための麻酔システムをチェックしてください。定位固定装置の切歯バーをノーズコーンを接続し、-3.3ミリメートルに切歯バーを置きます。 </li><li>供給ガス（2 L /分）をオンにします。ボックスにラットを置き、上部を密封します。 3.5％にイソフルラン気化器の電源をオンにします。 </li><li>ラットが横臥のとき麻酔ガスは、切歯バーに固定されたノーズコーンに流れるように、システムを切り替えます。 </li><li> RAを削除ボックス室からトンと耳と目の間の領域を剃ります。 Jodosept PVPを浸した綿棒を使用して、任意の緩い毛を取り除くために剃った領域を綿棒。 </li><li>ポジションノーズコーンにおけるラット（ 図1）とO 2（1リットル/分）でイソフルラン2.5％で麻酔を続けます。インターデジタルの領域をつまんで麻酔のレベルを確認してください。ラットを十分に麻酔されている場合は、守備の反射は（足のすなわち 、撤退を）廃止されています。 </li><li>手順の間刺激に呼吸し、応答を監視し、必要に応じて気化器を調整します。 </li><li>麻酔下ながら乾燥を防ぐために、目に獣医軟膏を適用します。監視およびフィードバック制御加熱方式により37±0.5℃の体温を維持します。 </li></ol> 2.手術<ol><li>全体の手術中に無菌手術野を保管してください。 surgeon&#39;s手は無菌であり、動作フィールドは、滅菌すると、のみcarefuを移動LLYと無菌状態を壊さないように覚えています。これも一つの楽器を下に設定することができるの無菌領域（ すなわち、滅菌防水ドレープ）を有する含まれています。 </li><li>剃っエリアの中心にメピバカイン皮下0.2ミリリットルを注入。メピバカインは、最大3時間の作用持続時間を有する局所麻酔薬です。さらに、手術領域を麻酔ます。 </li><li>メスを用いて、正中切開は耳の間に開始し、2センチメートルに向かって延在作ります。骨膜（皮膚の下に光沢のある膜）も切開されていることを確認します。 4クランプ（ 図2）で頭蓋骨を公開します。 </li><li>綿棒を使用して、冠状および矢状縫合が露出するまで静かに骨膜を取り除きます。その後、綿ウールで血を止めます。 </li><li>プローブホルダに固定された針を用いてブレグマの座標を決定し、黒のサインペンで針の先端をマーク。前方/後方（AP）を使用して、正中線/横（ML）と背腹（DV）の駆動ネジは、直接ブレグマの上に針の先端を置きます。 </li><li> APとMLバーニアスケールの測定値を取る：APの読みから3.6ミリメートルを引き、右STNに電極注入のための読書のMLから2.5ミリメートル、または左STNに電極注入の2.5ミリメートルを追加します。この位置は、頭蓋骨の表面に針の先端を下げた後、フェルトペンの色素によってマークされます。 </li><li>定位固定装置の大型プローブホルダーに歯科用ドリルを固定します。 すなわち、頭蓋骨上の印を付けた点-計算された領域に歯科用ドリルを移動します。硬膜が表示されるまで、顕微鏡で見ると、（頭蓋骨は約1mmの厚さである）頭蓋骨の貫通孔（直径約1mm）を開けます。マイクロ解剖鉗子または滅菌針を用いて硬膜を撤回。硬膜は、電極の先端を破壊するのに十分なタフです。 </li><li>各前頭鱗であり、int型に歯科用ドリルで穴を開け電極穴にerparietal鱗の反対。定位固定装置からのプローブホルダを外します。静脈血管が頭蓋骨の下に縫合糸をたどるよう頭蓋骨縫合糸をドリルしないでください。 </li><li> 5つの穴のそれぞれに骨ねじをねじ込みます。深すぎるでネジをねじ込むことは避けてください。ステンレス製ねじ（M1.6）は、ねじの2-3ターンは十分に脳に圧力をかけることなく、ねじを保持します。巻き数は、ねじのピッチに依存します。マイクロマニピュレーターで、電極（ 図3）とプローブホルダーを固定します。 </li><li>先端がほぼブレグマに触れられるまで、AP、MLとDVドライブネジを使用して、電極とプローブホルダを移動します。ブレグマでのAP、MLとDVバーニアスケールの測定値に注意してください。朗読が行われた場合には、移動中に頭蓋骨をこするから電極を防ぐために、電極を数ミリメートルを上げます。電極に挿入する必要がある位置の座標を決定するために穴にMLの読み取りには、APの読み取りに3.6ミリメートルを追加し、追加（または減算）2.5ミリメートル。 </li><li> APとMLドライブネジを使用して、計算された位置に電極を移動します。この時点で、電極先端が掘削電極穴の真上に位置する必要があります。その後、顕微鏡を通して見ることにより、硬膜（ 図4）のレベルに電極を下げます。このレベルは、DV方向のゼロレベルとして機能します。その後、静かに顕微鏡で見ることによって、脳への電極の先端を挿入します。 </li><li>記録システムのコネクタに電極ピンを接続します。定位固定装置（ 図5）で、ラットの上にファラデーケージを入れて（またはアルミホイルでそれを置き換えてください）。で働いていたされている部屋のカウンターポイズと定位固定装置を接地してください。 </li><li>記録システムを起動します。可能な場合、またadvancin時は、1単位の放電/軟膏の音響信号を得るためにスピーカーを使用G電極。 </li><li>ゆっくり電極を進める中の電気活動を記録することにより、脳内に電極を挿入します。硬膜から7.5の間と8.1ミリメートルの深さで、STNの特定の電気的活動は、（ 図6）は、通常、検出可能です。 18：STNのニューロンの典型的な活動が不規則な発火パターンと高発火率によって特徴付けられる（40.9±12.9 Hzの周波数を意味します）。 </li><li>録音中に、（0.8から1.0パーセントに、 例えば ）できるだけ多くの麻酔を減らします。低麻酔動物は明確に電気脳の活性を示します。 </li><li>電極を下げたときに、頭蓋骨の表面で移動された任意の血液や脳脊髄液を離れて綿棒。 </li><li>歯科用セメントの少量を混合し、電極の周りに、小さなへら（ 図7）を使用して、5本のネジの4の周りにそれを適用します。第五のネジは、プラグのアース線を固定するために使用されます。 </li><li>歯科用セメントが固定されたときに記録システムの電極ホルダーとコネクタから電極ピンを外します。 </li><li>歯科用セメントで固定されませんでしたネジを外します。電極ピンのプラグを入れてください。第五ネジ（ 図8）とプラグのアース線を固定してください。 </li><li>歯科用セメントを混合し、プラグの周りにそれを適用します。セメントが厚くなるように、キャップを形成するために、プラグの周りにそれを成形。動物に害を与えると（ 図9Aおよび B）硬化中にそれらを除去することができる歯科用セメントのシャープなエッジを避けてください。 </li><li>傷のエッジを創面切除し、前面にキャップの後ろに縫合糸でそれらを閉じます。傷のエッジを消毒します。 </li><li>スイベルに固定されたワイヤにヘッドプラグを接続します。定位固定装置からラットを削除します。 </li><li> 2〜3日間毎日一回、その後の介入の最後にトラマドール（12.5ミリグラム/キログラム、腹腔内）を適用し、。 THERMAと清潔なケージにラットを置き、リットルのサポートは、このケージ（ 図10）にスイベルを修正し、1時間慎重に検査します。 </li><li>それは胸骨横臥位を維持するのに十分な意識を取り戻したまで無人の動物を放置しないでください。完全に回復するまで、他の動物の会社に手術を受けた動物を返さないでください。 </li></ol> 3.刺激<ol><li>インピーダンスメータを用いて、刺激の前の動物の抵抗を決定します。 </li><li>ワイヤと電流出力を有するワイヤの他端のプラグと刺激装置のアース線用の出力とスイベルのプラグを接続します。刺激をプログラムするために、コンピュータと刺激を接続します。 </li><li>プログラム内の刺激のパラメータを選択してください。例えば、パーキンソン病に使用されるパラメータは、パルス長さ：60μ秒;周波数：130 Hzの。ジスキネジアが認識されるまで増加する電流振幅を有するラットを刺激します。目を削減ジスキネジアを誘発強度以下の10〜20％で、または神経学的徴候が消え、動物が快適になるまで電子電界強度。単相矩形パルスをこの研究で使用しました。 </li><li>実験を完了した後、イソフルランで動物を安楽死さ：5％以上にisofurane流量や濃度を調整します。停止を呼吸した後、1分までイソフルラン暴露を続けます。 </li></ol>

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The authors declare that they have no competing financial interests.

<html>本研究では、ラットのSTNにモノポーラ慢性電極を移植するための命令のステップバイステップのセットを提供します。低インピーダンスのタングステン電極は、多くの場合、DBS 18,19のために使用されているが、白金/イリジウム白金（Pt / Ir）から製の単極電極は、約1MΩのインピーダンスを持っていたことを採用しました。 Pt / Irの電極はまた、それらの好ましい特性のパーキンソ...

4または腹側中間視床5 -脳深部刺激（DBS）は、内部の淡蒼球1、視床下核（STN）2のような特定の脳構造に埋め込 ​​まれた電極を介した電気インパルスの配信に基づいて治療法です。過去20年間では、この治療は、パーキンソン病1のための強力な治療ツールとして確立されている- 4、ジストニア6と震え7、および慢性疼痛7を変調するためにも使用され、精神障害（ すなわち 、強迫性障害8、大うつ病9）または難治性てんかん10,11。また、DBSは、将来的には、耐火動脈高血圧12や起立性低血圧13のための治療選択肢になるかもしれません。 効果の基礎となる生理学的メカニズムDBSのよくわかっていないままです。麻酔をかけたげっ歯類での研究は、臨床的にDBS 14適用模倣高頻度刺激に神経応答への洞察を提供してきました。しかしながら、これらの研究は、DBS効果の行動の裏付けを欠くだけでなく、14を適用した刺激パラメータに応じてかなりのばらつきが生じます。 より簡潔に意識げっ歯類におけるDBSの行動への影響と根底にある機構を調べるために、刺激のセットアップには、特定の要件を満たすことが必要です。 DBSは、主に、長期療法として使用されている（例えば、パーキンソン病、慢性の痛み）。ユニットはプラグと同様に、外部刺激へのプラグからのワイヤと電極で構成されるようにこのように、げっ歯類における刺激セットアップを設計する必要があります。頭蓋骨に固定されたときや、本機は、軽量で割れないでなければなりません。また、移動の自由はstimula中のラットのために不可欠です長期にわたってる。 DBSの標的構造は小さいです。例えば、ラットにおけるSTNは1.2mmの長さおよび0.8 mmの3,15の体積を有します。したがって、電極は、核が挿入中に損傷しないように設計されており、正確にニーズを標的としなければなりません。げっ歯類で行わ最もDBS研究は、標的構 ​​造に対して電極のランドマークベースの定位挿入を使用しているように、エラーレートがPaxinosとWatsonの16に従って座標を用いた場合であっても、比較的高いとすることができます。これは、統計的に意味のある結果に到達するために必要な動物の多数を生じます。 本研究では、電極注入法は、電極を進めながらmicrorecording系を用いて高精度にSTNを標的とする、導入されます。また、刺激システムは、刺激された動物のための移動性の高度を可能にするだけでなく、連続stimulatiを保証しない提示されていますラットの頭の上に（ステンレス製スプリングで保護されている）刺激線の確実な固定を介した上で。

<table><tbody><tr><td>Pt/Ir electrode</td><td>FHC Inc.</td><td>UE</td><td>Custom-made: Specification: UEPSEGSECN1M</td></tr><tr><td>Plugs</td><td>GT Labortechnik (Arnstein/Germany)</td><td></td><td>Custom-made</td></tr><tr><td>Pin header</td><td>DISTRELEC</td><td>143-95-324</td><td>single-row, 90&amp;deg; 1x3 datamate, Type M80-8420342</td></tr><tr><td>Socket</td><td>DISTRELEC</td><td>143-95-621</td><td>single-row,straight 2 mm pole no.1 x 3 datamate, Type M80-8400342</td></tr><tr><td>Stainless steel spring</td><td>Plastics ONE</td><td>SS0102</td><td>Part-#: .120 X .156 Spring ID (mm): 3.0&amp;nbsp; Spring OD (mm): 4.0</td></tr><tr><td>Dental cement/Paladur</td><td>Heraeus Kulzer</td><td>64707938</td><td>Liquid, 500 ml</td></tr><tr><td>Dental cement/Paladur</td><td>Heraeus Kulzer</td><td>64707954</td><td>Powder, rose, 500 g</td></tr><tr><td>Head screw</td><td>Hummer &amp;amp; Reiss</td><td>V2ADIN84 M1.6x3</td><td></td></tr><tr><td>Jodosept PVP</td><td>Vetoquinol</td><td></td><td>435678/E04</td></tr><tr><td>Mepivacain 1%</td><td>AstraZeneca</td><td></td><td>PZN03338515</td></tr><tr><td>Epinephrine</td><td>Sanofi-Aventis</td><td></td><td>PZN00176118</td></tr><tr><td>Tramadolhydrochloride</td><td>Rotexmedica</td><td></td><td>38449.00.00</td></tr></tbody></table>

microelectrode guided implantation of electrodes into the subthalamic nucleus of rats for long-term deep brain stimulation

脳深部刺激（DBS）は、特発性パーキンソン病、ジストニアまたは振戦などのいくつかの神経障害のために広く使用され、効果的な治療です。 DBSは、中枢神経系の特定の深い解剖学的構造への電気刺激の配信に基づいています。しかし、DBSの効果のメカニズムは謎のままです。これは、特にラットに、動物モデルにおいてDBSの影響を調査の関心につながっています。 DBSは、長期的な治療であるため、研究はDBS後数週間を発生する神経回路の分子遺伝的変化に焦点を置くべきです。ラットは所定の位置に刺激に対する動物の頭からの導線を維持するのに問題が発生し、そのケージに動き回るため、ラットの長期DBSは困難です。また、ラットの脳内の刺激のための標的構造は、小型であり、したがって、電極は、容易に必要な位置に配置することができません。このように、長期的なstimulaためのセットアップ約1MΩのインピーダンスと白金/イリジウム電極を用いたラットの化は、この研究のために開発されました。これらの仕様と電極は、適切な刺激だけでなく、DBSのためのターゲット領域を識別するために、脳深部構造の記録だけでなくすることができます。我々のセットアップでは、ワイヤ用のプラグと電極を頭蓋骨に固定4本のネジで固定する歯科用セメントに埋め込まれました。刺激へのプラグからワイヤーをステンレス製のスプリングにより保護されていました。スイベルは、もつれたならワイヤを防止するために回路に接続しました。全体的に、この刺激のセットアップは、ラットのための自由移動性の高い学位を提供し、長期的な強度を保持するために、ヘッドプラグだけでなく、プラグと刺激間の配線の接続を可能にします。

記録システムを用いてラットのSTNに電極を移植する - ここに提示されるようなことは - 動物あたり約1時間かかりますDBSための効果的かつ正確な手順です。このモデルは、かなりマイナーな手順です：手術を施したラット10匹のうち、すべての介入を生き延びました。介入後の二十四時間は、各ラットの状態をモニターし、何の動物は、重大度コードに応じて1以上3点を達成していません。連?...

Watch this Scientific Journal Video about Microelectrode Guided Implantation of Electrodes into the Subthalamic Nucleus of Rats for Long-term Deep Brain Stimulation at JoVE.com

Microelectrode Guided Implantation of Electrodes into the Subthalamic Nucleus of Rats for Long-term Deep Brain Stimulation

A method for implanting electrodes into the subthalamic nucleus (STN) of rats is described. Better localization of the STN was achieved by using a microrecording system. Furthermore, a stimulation set-up is presented that is characterized by long-lasting connections between the head of the animal and the stimulator.

微小電極は、長期的な脳深部刺激のためのラットの視床下核に電極の移植ガイド付き

Deep brain stimulation (DBS) is a widely used and effective therapy for several neurologic disorders, such as idiopathic Parkinson&#8217;s disease, ...

microelectrode-guided-implantation-electrodes-into-subthalamic

Research

JoVE Journal

Neuroscience

19.6K ビュー.  University Hospital Wuerzburg. A method for implanting electrodes into the subthalamic nucleus (STN) of rats is described. Better localization of the STN was achieved by using a microrecording system. Furthermore, a stimulation set-up is presented that is characterized by long-lasting connections between the head of the animal and the stimulator.

ビデオ: Microelectrode Guided Implantation of Electrodes into the Subthalamic Nucleus of Rats for Long-term Deep Brain Stimulation

Intra-Operative Behavioral Tasks in Awake Humans Undergoing Deep Brain Stimulation Surgery

Deep brain stimulation (DBS) is a surgical procedure that directs chronic, high frequency electrical stimulation to specific targets in the brain through implanted electrodes. Deep brain stimulation was first implemented as a therapeutic modality by Benabid et al. in the late 1980s, when he used this technique to stimulate the ventral intermediate nucleus of the thalamus for the treatment of tremor 1. Currently, the procedure is used to treat patients who fail to respond adequately to medical management for diseases such as Parkinson's, dystonia, and essential tremor. The efficacy of this procedure for the treatment of Parkinson's disease has been demonstrated in well-powered, randomized controlled trials 2. Presently, the U.S. Food and Drug Administration has approved DBS as a treatment for patients with medically refractory essential tremor, Parkinson's disease, and dystonia. Additionally, DBS is currently being evaluated for the treatment of other psychiatric and neurological disorders, such as obsessive compulsive disorder, major depressive disorder, and epilepsy. 
DBS has not only been shown to help people by improving their quality of life, it also provides researchers with the unique opportunity to study and understand the human brain. Microelectrode recordings are routinely performed during DBS surgery in order to enhance the precision of anatomical targeting. Firing patterns of individual neurons can therefore be recorded while the subject performs a behavioral task. Early studies using these data focused on descriptive aspects, including firing and burst rates, and frequency modulation 3. More recent studies have focused on cognitive aspects of behavior in relation to neuronal activity 4,5. This article will provide a description of the intra-operative methods used to perform behavioral tasks and record neuronal data with awake patients during DBS cases. Our exposition of the process of acquiring electrophysiological data will illuminate the current scope and limitations of intra-operative human experiments.

Deep brain stimulation surgery offers a unique opportunity to examine information encoding in the awake human brain. This article will describe intra-operative methods used to perform cognitive and behavioral tasks while simultaneously acquiring physiological data such as EMG, single-unit neuronal activity and/or local field potentials.

脳深部刺激術を受けた覚醒ヒトにおける術中行動作業

Deep brain stimulation (DBS) is a surgical procedure that directs chronic, high frequency electrical stimulation to specific targets in the brain through ...

医学

A General Method for Evaluating Deep Brain Stimulation Effects on Intravenous Methamphetamine Self-Administration

Substance use disorders, particularly to methamphetamine, are devastating, relapsing diseases that disproportionally affect young people. There is a need for novel, effective and practical treatment strategies that are validated in animal models. Neuromodulation, including deep brain stimulation (DBS) therapy, refers to the use of electricity to influence pathological neuronal activity and has shown promise for psychiatric disorders, including drug dependence. DBS in clinical practice involves the continuous delivery of stimulation into brain structures using an implantable pacemaker-like system that is programmed externally by a physician to alleviate symptoms. This treatment will be limited in methamphetamine users due to challenging psychosocial situations. Electrical treatments that can be delivered intermittently, non-invasively and remotely from the drug-use setting will be more realistic. This article describes the delivery of intracranial electrical stimulation that is temporally and spatially separate from the drug-use environment for the treatment of IV methamphetamine dependence. Methamphetamine dependence is rapidly developed in rodents using an operant paradigm of intravenous (IV) self-administration that incorporates a period of extended access to drug and demonstrates both escalation of use and high motivation to obtain drug.

This article describes the delivery of intracranial electrical stimulation that is temporally and spatially separate from the drug-use environment for the treatment of IV methamphetamine dependence.

静脈内メタンフェタミン自己管理に脳深部刺激効果を評価するための一般的な方法

Substance use disorders, particularly to methamphetamine, are devastating, relapsing diseases that disproportionally affect young people.  There is a need ...

行動学

Surgical Implantation of Chronic Neural Electrodes for Recording Single Unit Activity and Electrocorticographic Signals

The success of long-term electrophysiological recordings often depends on the quality of the implantation surgery. Here we provide useful information for surgeons who are learning the process of implanting electrode systems. We demonstrate the implantation procedure of both a penetrating and a surface electrode. The surgical process is described from start to finish, including detailed descriptions of each step throughout the procedure. It should also be noted that this video guide is focused towards procedures conducted in rodent models and other small animal models. Modifications of the described procedures are feasible for other animal models.

We provide useful information for surgeons who are learning the process of implanting chronic neural recording electrodes. Techniques for both penetrating and surface electrode systems are described in a rodent animal model.

単一ユニット活動とElectrocorticographic信号を記録するための慢性神経電極の外科的移植

The success of long-term electrophysiological recordings often depends on the quality of the implantation surgery.  Here we provide useful information for ...

神経科学

A Wireless, Bidirectional Interface for In Vivo Recording and Stimulation of Neural Activity in Freely Behaving Rats

In vivo electrophysiology is a powerful technique to investigate the relationship between brain activity and behavior at a millisecond and micrometer scale. However, current methods mostly rely on tethered cable recordings or only use unidirectional systems, allowing either recording or stimulation of neural activity, but not at the same time or same target. Here, a new wireless, bidirectional device for simultaneous multichannel recording and stimulation of neural activity in freely behaving rats is described. The system operates through a single portable head stage that both transmits recorded activity and can be targeted in real-time for brain stimulation using a telemetry-based multichannel software. The head stage is equipped with a preamplifier and a rechargeable battery, allowing stable long-term recordings or stimulation for up to 1 h. Importantly, the head stage is compact, weighs 12 g (including battery) and thus has minimal impact on the animal&#180;s behavioral repertoire, making the method applicable to a broad set of behavioral tasks. Moreover, the method has the major advantage that the effect of brain stimulation on neural activity and behavior can be measured simultaneously, providing a tool to assess the causal relationships between specific brain activation patterns and behavior. This feature makes the method particularly valuable for the field of deep brain stimulation, allowing precise assessment, monitoring, and adjustment of stimulation parameters during long-term behavioral experiments. The applicability of the system has been validated using the inferior colliculus as a model structure.

A Wireless, Bidirectional Interface for In Vivo Recording and Stimulation of Neural Activity in Freely Behaving Rats

A wireless, bidirectional system for multi-channel neural recordings and stimulation in freely behaving rats is introduced. The system is light and compact, thus having minimal impact on the animal&#180;s behavioral repertoire. Moreover, this bidirectional system provides a sophisticated tool to assess causal relationships between brain activation patterns and behavior.

ワイヤレス、 In Vivo記録そして自由行動下ラットにおける神経活動の刺激のための双方向インタ フェース

In vivo electrophysiology is a powerful technique to investigate the relationship between brain activity and behavior at a millisecond and micrometer ...

Deep Brain Stimulation with Simultaneous fMRI in Rodents

In order to visualize the global and downstream neuronal responses to deep brain stimulation (DBS) at various targets, we have developed a protocol for using blood oxygen level dependent (BOLD) functional magnetic resonance imaging (fMRI) to image rodents with simultaneous DBS. DBS fMRI presents a number of technical challenges, including accuracy of electrode implantation, MR artifacts created by the electrode, choice of anesthesia and paralytic to minimize any neuronal effects while simultaneously eliminating animal motion, and maintenance of physiological parameters, deviation from which can confound the BOLD signal.&#160;Our laboratory has developed a set of procedures that are capable of overcoming most of these possible issues. For electrical stimulation, a homemade tungsten bipolar microelectrode is used, inserted stereotactically at the stimulation site in the anesthetized subject. In preparation for imaging, rodents are fixed on a plastic headpiece and transferred to the magnet bore. For sedation and paralysis during scanning, a cocktail of dexmedetomidine and pancuronium is continuously infused, along with a minimal dose of isoflurane; this preparation minimizes the BOLD ceiling effect of volatile anesthetics. In this example experiment, stimulation of the subthalamic nucleus (STN) produces BOLD responses which are observed primarily in ipsilateral cortical regions, centered in motor cortex. Simultaneous DBS and fMRI allows the unambiguous modulation of neural circuits dependent on stimulation location and stimulation parameters, and permits observation of neuronal modulations free of regional bias. This technique may be used to explore the downstream effects of modulating neural circuitry at nearly any brain region, with implications for both experimental and clinical DBS.

This protocol describes a standard method for simultaneous functional magnetic resonance imaging and deep brain stimulation in the rodent. The combined use of these experimental tools allows for the exploration of global downstream activity in response to electrical stimulation at virtually any brain target.

げっ歯類での同時のfMRIと脳深部刺激

In order to visualize the global and downstream neuronal responses to deep brain stimulation (DBS) at various targets, we have developed a protocol for ...

Analysis of Gene Expression Changes in the Rat Hippocampus After Deep Brain Stimulation of the Anterior Thalamic Nucleus

Deep brain stimulation (DBS) surgery, targeting various regions of the brain such as the basal ganglia, thalamus, and subthalamic regions, is an effective treatment for several movement disorders that have failed to respond to medication. Recent progress in the field of DBS surgery has begun to extend the application of this surgical technique to other conditions as diverse as morbid obesity, depression and obsessive compulsive disorder. Despite these expanding indications, little is known about the underlying physiological mechanisms that facilitate the beneficial effects of DBS surgery. One approach to this question is to perform gene expression analysis in neurons that receive the electrical stimulation. Previous studies have shown that neurogenesis in the rat dentate gyrus is elicited in DBS targeting of the anterior nucleus of the thalamus1. DBS surgery targeting the ATN is used widely for treatment refractory epilepsy. It is thus of much interest for us to explore the transcriptional changes induced by electrically stimulating the ATN. In this manuscript, we describe our methodologies for stereotactically-guided DBS surgery targeting the ATN in adult male Wistar rats. We also discuss the subsequent steps for tissue dissection, RNA isolation, cDNA preparation and quantitative RT-PCR for measuring gene expression changes. This method could be applied and modified for stimulating the basal ganglia and other regions of the brain commonly clinically targeted. The gene expression study described here assumes a candidate target gene approach for discovering molecular players that could be directing the mechanism for DBS.

The mechanism underlying the therapeutic effects of Deep Brain Stimulation (DBS) surgery needs investigation. The methods presented in this manuscript describe an experimental approach to examine the cellular events triggered by DBS by analyzing the gene expression profile of candidate genes that can facilitate neurogenesis post DBS surgery.

視床前核の脳深部刺激後のラット海馬における遺伝子発現の変化の解析

Deep brain stimulation (DBS) surgery, targeting various regions of the brain such as the basal ganglia, thalamus, and subthalamic regions, is an effective ...

Chronic Transcranial Electrical Stimulation and Intracortical Recording in Rats

Transcranial electrical stimulation (TES) is a powerful and relatively simple approach to diffusely influence brain activity either randomly or in a closed-loop event-triggered manner. Although many studies are focusing on the possible benefits and side-effects of TES in healthy and pathologic brains, there are still many fundamental open questions regarding the mechanism of action of the stimulation. Therefore, there is a clear need for a robust and reproducible method to test the acute and the chronic effects of TES in rodents. TES can be combined with regular behavioral, electrophysiological, and imaging techniques to investigate neuronal networks in vivo. The implantation of transcranial stimulation electrodes does not impose extra constraints on the experimental design while it offers a versatile, flexible tool to manipulate brain activity. Here we provide a detailed, step-by-step protocol to fabricate and implant transcranial stimulation electrodes to influence brain activity in a temporally constrained manner for months.

This detailed protocol describes transcranial stimulation electrode placement on the temporal bone in order to investigate the short- and long-term effects of transcranial electrical stimulation in freely moving rats.

慢性的な経頭蓋電気刺激およびラットの皮質内の記録

Transcranial electrical stimulation (TES) is a powerful and relatively simple approach to diffusely influence brain activity either randomly or in a ...

An Invasive Method for the Activation of the Mouse Dentate Gyrus by High-frequency Stimulation

Electrical high-frequency stimulation (HFS), using implanted electrodes targeting various brain regions, has been proven as an effective treatment for various neurological and psychiatric disorders. HFS in the deep region of the brain, also named deep-brain stimulation (DBS), is becoming increasingly important in clinical trials. Recent progress in the field of high-frequency DBS (HF-DBS) surgery has begun to spread the possibility of utilizing this invasive technique to other situations, such as treatment for major depression disorder (MDD), obsessive-compulsive disorder (OCD), and so on. 
Despite these expanding indications, the underlying mechanisms of the beneficial effects of HF-DBS remain enigmatic. To address this question, one approach is to use implanted electrodes that sparsely activate distributed subpopulations of neurons by HFS. It has been reported that HFS in the anterior nucleus of the thalamus could be used for the treatment of refractory epilepsy in the clinic. The underlying mechanisms might be related to the increased neurogenesis and altered neuronal activity. Therefore, we are interested in exploring the physiological alterations by the detection of neuronal activity as well as neurogenesis in the mouse dentate gyrus (DG) before and after HFS treatment. 
In this manuscript, we describe methodologies for HFS to target the activation of the DG in mice, directly or indirectly and in an acute or chronic manner. In addition, we describe a detailed protocol for the preparation of brain slices for c-fos and Notch1 immunofluorescent staining to monitor the neuronal activity and signaling activation and for bromodeoxyuridine (BrdU) labeling to determine the neurogenesis after the HF-DBS induction. The activation of the neuronal activity and neurogenesis after the HF-DBS treatment provides direct neurobiological evidence and potential therapeutic benefits. Particularly, this methodology can be modified and applied to target other interested brain regions such as the basal ganglia and subthalamic regions for specific brain disorders in the clinic.

This protocol shows how to set up a reliable HFS method in mice. Neurons throughout the hippocampal dentate gyrus are electrically stimulated by HFS directly and indirectly in vivo. Neuronal activity and molecular signaling are examined by c-fos and Notch1 immunofluorescent staining, respectively; neurogenesis is quantified by bromodeoxyuridine labeling assay.

高周波刺激によるマウスの海馬歯状回の活性化のための侵襲的な方法

Electrical high-frequency stimulation (HFS), using implanted electrodes targeting various brain regions, has been proven as an effective treatment for ...

Subdural Soft Electrocorticography (ECoG) Array Implantation and Long-Term Cortical Recording in Minipigs

Neurological impairments and diseases can be diagnosed or treated using electrocorticography (ECoG) arrays. In drug-resistant epilepsy, these help delineate the epileptic region to resect. In long-term applications such as brain-computer interfaces, these epicortical electrodes are used to record the movement intention of the brain, to control the robotic limbs of paralyzed patients. However, current stiff electrode grids do not answer the need for high-resolution brain recordings and long-term biointegration. Recently, conformable electrode arrays have been proposed to achieve long-term implant stability with high performance. However, preclinical studies for these new implant technologies are needed to validate their long-term functionality and safety profile for their translation to human patients. In this context, porcine models are routinely employed in developing medical devices due to their large organ sizes and easy animal handling. However, only a few brain applications are described in the literature, mostly due to surgery limitations and integration of the implant system on a living animal.
Here, we report the method for long-term implantation (6 months) and evaluation of soft ECoG arrays in the minipig model. The study first presents the implant system, consisting of a soft microfabricated electrode array integrated with a magnetic resonance imaging (MRI)-compatible polymeric transdermal port that houses instrumentation connectors for electrophysiology recordings. Then, the study describes the surgical procedure, from subdural implantation to animal recovery. We focus on the auditory cortex as an example target area where evoked potentials are induced by acoustic stimulation. We finally describe a data acquisition sequence that includes MRI of the whole brain, implant electrochemical characterization, intraoperative and freely moving electrophysiology, and immunohistochemistry staining of the extracted brains.
This model can be used to investigate the safety and function of novel design of cortical prostheses; mandatory preclinical study to envision translation to human patients.

Subdural Soft Electrocorticography ECoG Array Implantation and Long-Term Cortical Recording in Minipigs

Here, we present a method for the long-term performance and safety assessment of soft subdural electrode arrays in a minipig model, describing surgical method and tools, postoperative magnetic resonance imaging, electrophysiology of the auditory cortex, electrochemical properties of the implant, and postmortem immunochemistry.

ミニブタにおける硬膜下軟性皮質電図(ECoG)アレイ移植と長期皮質記録

Neurological impairments and diseases can be diagnosed or treated using electrocorticography (ECoG) arrays. In drug-resistant epilepsy, these help ...

生物工学

Implanting a Wireless Bidirectional Microelectrode System in a Rat Brain

Source: Melo-Thomas, L., et al., <a href="https://app.jove.com/v/56299">A Wireless, Bidirectional Interface for In Vivo Recording and Stimulation of Neural Activity in Freely Behaving Rats.</a> J. Vis. Exp. (2017)
The video demonstrates a procedure for implanting a wireless bidirectional multichannel electrode unit into a rat brain to record and stimulate multichannel neural networks.

ラット脳への無線双方向微小電極システムの移植

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. ...

微小電極は、長期的な脳深部刺激のためのラットの視床下核に電極の移植ガイド付き

要約

さらに動画を探す

この動画の章

脳深部刺激術を受けた覚醒ヒトにおける術中行動作業

静脈内メタンフェタミン自己管理に脳深部刺激効果を評価するための一般的な方法

単一ユニット活動とElectrocorticographic信号を記録するための慢性神経電極の外科的移植

ワイヤレス、 In Vivo記録そして自由行動下ラットにおける神経活動の刺激のための双方向インタフェース

げっ歯類での同時のfMRIと脳深部刺激

視床前核の脳深部刺激後のラット海馬における遺伝子発現の変化の解析

慢性的な経頭蓋電気刺激およびラットの皮質内の記録

高周波刺激によるマウスの海馬歯状回の活性化のための侵襲的な方法

ミニブタにおける硬膜下軟性皮質電図(ECoG)アレイ移植と長期皮質記録

ラット脳への無線双方向微小電極システムの移植

微小電極は、長期的な脳深部刺激のためのラットの視床下核に電極の移植ガイド付き

要約

さらに動画を探す

この動画の章

脳深部刺激術を受けた覚醒ヒトにおける術中行動作業

静脈内メタンフェタミン自己管理に脳深部刺激効果を評価するための一般的な方法

単一ユニット活動とElectrocorticographic信号を記録するための慢性神経電極の外科的移植

ワイヤレス、 In Vivo記録そして自由行動下ラットにおける神経活動の刺激のための双方向インタ フェース

げっ歯類での同時のfMRIと脳深部刺激

視床前核の脳深部刺激後のラット海馬における遺伝子発現の変化の解析

慢性的な経頭蓋電気刺激およびラットの皮質内の記録

高周波刺激によるマウスの海馬歯状回の活性化のための侵襲的な方法

ミニブタにおける硬膜下軟性皮質電図(ECoG)アレイ移植と長期皮質記録

ラット脳への無線双方向微小電極システムの移植

ワイヤレス、 In Vivo記録そして自由行動下ラットにおける神経活動の刺激のための双方向インタフェース