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。

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			<td>Pt/Ir electrode</td>
			<td>FHC Inc.</td>
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			<td>Custom-made: Specification: UEPSEGSECN1M</td>
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			<td>Plugs</td>
			<td>GT Labortechnik (Arnstein/Germany)</td>
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			<td>Pin header</td>
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			<td>Part-#: .120 X .156 Spring ID (mm): 3.0&#160; Spring OD (mm): 4.0</td>
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			<td>Epinephrine</td>
			<td>Sanofi-Aventis</td>
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			<td>Tramadolhydrochloride</td>
			<td>Rotexmedica</td>
			<td></td>
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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

Universidad de Cartagena UDEC

Research

JoVE Journal

Neuroscience

19.5K 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

TMS: Using the Theta-Burst Protocol to Explore Mechanism of Plasticity in Individuals with Fragile X Syndrome and Autism

Fragile X Syndrome (FXS), also known as Martin-Bell Syndrome, is a genetic abnormality found on the X chromosome.1,2 Individuals suffering from FXS display abnormalities in the expression of FMR1 - a protein required for typical, healthy neural development.3 Recent data has suggested that the loss of this protein can cause the cortex to be hyperexcitable thereby affecting overall patterns of neural plasticity.4,5
In addition, Fragile X shows a strong comorbidity with autism: in fact, 30% of children with FXS are diagnosed with autism, and 2 - 5% of autistic children suffer from FXS.6
Transcranial Magnetic Stimulation (a non-invasive neurostimulatory and neuromodulatory technique that can transiently or lastingly modulate cortical excitability via the application of localized magnetic field pulses 7,8) represents a unique method of exploring plasticity and the manifestations of FXS within affected individuals. More specifically, Theta-Burst Stimulation (TBS), a specific stimulatory protocol shown to modulate cortical plasticity for a duration up to 30 minutes after stimulation cessation in healthy populations, has already proven an efficacious tool in the exploration of abnormal plasticity.9,10 
Recent studies have shown the effects of TBS last considerably longer in individuals on the autistic spectrum - up to 90 minutes.11 This extended effect-duration suggests an underlying abnormality in the brain's natural plasticity state in autistic individuals - similar to the hyperexcitability induced by Fragile X Syndrome.
In this experiment, utilizing single-pulse motor-evoked potentials (MEPs) as our benchmark, we will explore the effects of both intermittent and continuous TBS on cortical plasticity in individuals suffering from FXS and individuals on the Autistic Spectrum.

In this article, we examine the effects of Theta-Burst TMS stimulation on cortical plasticity in individuals suffering from Fragile X syndrome and individuals on the autistic spectrum.

TMS：脆弱X症候群と自閉症の可塑性のMechasnismを探検するシータバーストプロトコルを使用する

Fragile X Syndrome (FXS), also known as Martin-Bell Syndrome, is a genetic abnormality found on the X chromosome.1,2 Individuals suffering from FXS ...

神経科学

The NeuroStar TMS Device: Conducting the FDA Approved Protocol for Treatment of Depression

The Neuronetics NeuroStar Transcranial Magnetic Stimulation (TMS) System is a class II medical device that produces brief duration, pulsed magnetic fields. These rapidly alternating fields induce electrical currents within localized, targeted regions of the cortex which are associated with various physiological and functional brain changes.1,2,3
In 2007, O'Reardon et al., utilizing the NeuroStar device, published the results of an industry-sponsored, multisite, randomized, sham-stimulation controlled clinical trial in which 301 patients with major depression, who had previously failed to respond to at least one adequate antidepressant treatment trial, underwent either active or sham TMS over the left dorsolateral prefrontal cortex (DLPFC). The patients, who were medication-free at the time of the study, received TMS five times per week over 4-6 weeks.4
The results demonstrated that a sub-population of patients (those who were relatively less resistant to medication, having failed not more than two good pharmacologic trials) showed a statistically significant improvement on the Montgomery-Asberg Depression Scale (MADRS), the Hamilton Depression Rating Scale (HAMD), and various other outcome measures. In October 2008, supported by these and other similar results5,6,7, Neuronetics obtained the first and only Food and Drug Administration (FDA) approval for the clinical treatment of a specific form of medication-refractory depression using a TMS Therapy device (FDA approval K061053).
In this paper, we will explore the specified FDA approved NeuroStar depression treatment protocol (to be administered only under prescription and by a licensed medical profession in either an in- or outpatient setting).

In this article, we examine the methodology and considerations relevant to the FDA approved depression treatment protocol using the Neuronetics NeuroStar TMS device.

NeuroStar TMSデバイス：うつ病の治療薬としてFDA承認されたプロトコルを実施

The Neuronetics NeuroStar Transcranial Magnetic Stimulation (TMS) System is a class II medical device that produces brief duration, pulsed magnetic ...

A Polished and Reinforced Thinned-skull Window for Long-term Imaging of the Mouse Brain

In vivo imaging of cortical function requires optical access to the brain without disruption of the intracranial environment. We present a method to form a polished and reinforced thinned skull (PoRTS) window in the mouse skull that spans several millimeters in diameter and is stable for months. The skull is thinned to 10 to 15 &mu;m in thickness with a hand held drill to achieve optical clarity, and is then overlaid with cyanoacrylate glue and a cover glass to: 1) provide rigidity, 2) inhibit bone regrowth and 3) reduce light scattering from irregularities on the bone surface. Since the skull is not breached, any inflammation that could affect the process being studied is greatly reduced. Imaging depths of up to 250 &mu;m below the cortical surface can be achieved using two-photon laser scanning microscopy. This window is well suited to study cerebral blood flow and cellular function in both anesthetized and awake preparations. It further offers the opportunity to manipulate cell activity using optogenetics or to disrupt blood flow in targeted vessels by irradiation of circulating photosensitizers.

We present a method to form an imaging window in the mouse skull that spans millimeters and is stable for months without inflammation of the brain. This method is well suited for longitudinal studies of blood flow, cellular dynamics, and cell/vascular structure using two-photon microscopy.

マウス脳の長期イメージングのためのポリッシュと強化間引き頭蓋骨ウィンドウ

In vivo imaging of cortical function requires optical access to the brain without disruption of the intracranial environment. We present a method to form ...

Fiber-optic Implantation for Chronic Optogenetic Stimulation of Brain Tissue

Elucidating patterns of neuronal connectivity has been a challenge for both clinical and basic neuroscience. Electrophysiology has been the gold standard for analyzing patterns of synaptic connectivity, but paired electrophysiological recordings can be both cumbersome and experimentally limiting. The development of optogenetics has introduced an elegant method to stimulate neurons and circuits, both in vitro1 and in vivo2,3. By exploiting cell-type specific promoter activity to drive opsin expression in discrete neuronal populations, one can precisely stimulate genetically defined neuronal subtypes in distinct circuits4-6. Well described methods to stimulate neurons, including electrical stimulation and/or pharmacological manipulations, are often cell-type indiscriminate, invasive, and can damage surrounding tissues. These limitations could alter normal synaptic function and/or circuit behavior. In addition, due to the nature of the manipulation, the current methods are often acute and terminal. Optogenetics affords the ability to stimulate neurons in a relatively innocuous manner, and in genetically targeted neurons. The majority of studies involving in vivo optogenetics currently use a optical fiber guided through an implanted cannula6,7; however, limitations of this method include damaged brain tissue with repeated insertion of an optical fiber, and potential breakage of the fiber inside the cannula. Given the burgeoning field of optogenetics, a more reliable method of chronic stimulation is necessary to facilitate long-term studies with minimal collateral tissue damage. Here we provide our modified protocol as a video article to complement the method effectively and elegantly described in Sparta et al.8 for the fabrication of a fiber optic implant and its permanent fixation onto the cranium of anesthetized mice, as well as the assembly of the fiber optic coupler connecting the implant to a light source. The implant, connected with optical fibers to a solid-state laser, allows for an efficient method to chronically photostimulate functional neuronal circuitry with less tissue damage9 using small, detachable, tethers. Permanent fixation of the fiber optic implants provides consistent, long-term in vivo optogenetic studies of neuronal circuits in awake, behaving mice10 with minimal tissue damage.

The development of optogenetics now provides the means to precisely stimulate genetically defined neurons and circuits, both in vitro and in vivo. Here we describe the assembly and implantation of a fiber optic for chronic photostimulation of brain tissue.

脳組織の慢性Optogenetic刺激のための光ファイバ注入

Elucidating patterns of neuronal connectivity has been a challenge for both clinical and basic neuroscience. Electrophysiology has been the gold standard ...

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 ...

Direct Visualization of the Murine Dorsal Cochlear Nucleus for Optogenetic Stimulation of the Auditory Pathway

Investigation into the use of virus-mediated gene transfer to arrest or reverse hearing loss has largely been relegated to the peripheral auditory system. Few studies have examined gene transfer to the central auditory system. The dorsal cochlear nucleus (DCN) of the brainstem, which contains second order neurons of the auditory pathway, is a potential site for gene transfer. In this protocol, a technique for direct and maximal exposure of the murine DCN via a posterior fossa approach is demonstrated. This approach allows for either acute or survival surgery. Following direct visualization of the DCN, a host of experiments are possible, including injection of opsins into the cochlear nucleus and subsequent stimulation by an optical fiber coupled to a blue light laser.&#160;Other neurophysiology experiments, such as electrical stimulation and neural injector tracings are also feasible.&#160;The level of visualization and the duration of stimulation achievable make this approach applicable to a wide range of experiments.

The goal of this protocol is to outline a surgical approach to provide direct access to the dorsal cochlear nucleus in a murine model.

聴覚路の光遺伝学的刺激のためのマウス背側蝸牛神経核の直接可視化

Investigation into the use of virus-mediated gene transfer to arrest or reverse hearing loss has largely been relegated to the peripheral auditory system. ...

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 ...

Cannula Implantation into the Cisterna Magna of Rodents

Cisterna magna cannulation (CMc) is a straightforward procedure that enables direct access to the cerebrospinal fluid (CSF) without operative damage to the skull or the brain parenchyma. In anesthetized rodents, the exposure of the dura mater by blunt dissection of the neck muscles allows the insertion of a cannula into the cisterna magna (CM). The cannula, composed either by a fine beveled needle or borosilicate capillary, is attached via a polyethylene (PE) tube to a syringe. Using a syringe pump, molecules can then be injected at controlled rates directly into the CM, which is continuous with the subarachnoid space. From the subarachnoid space, we can trace CSF fluxes by convective flow into the perivascular space around penetrating arterioles, where solute exchange with the interstitial fluid (ISF) occurs. CMc can be performed for acute injections immediately following the surgery, or for chronic implantation, with later injection in anesthetized or awake, freely moving rodents. Quantitation of tracer distribution in the brain parenchyma can be performed by epifluorescence, 2-photon microscopy, and magnetic resonance imaging (MRI), depending on the physico-chemical properties of the injected molecules. Thus, CMc in conjunction with various imaging techniques offers a powerful tool for assessment of the glymphatic system and CSF dynamics and function. Furthermore, CMc can be utilized as a conduit for fast, brain-wide delivery of signaling molecules and metabolic substrates that could not otherwise cross the blood brain barrier (BBB).

Here we describe a protocol to perform cisterna magna cannulation (CMc), a minimally invasive way to deliver tracers, substrates and signaling molecules into the cerebrospinal fluid (CSF). Combined with different imaging modalities, CMc enables glymphatic system and CSF dynamics assessment, as well as brain-wide delivery of various compounds.

齧歯動物の槽注入カニューレ

Cisterna magna cannulation (CMc) is a straightforward procedure that enables direct access to the cerebrospinal fluid (CSF) without operative damage to ...

Chronic Implantation of Whole-cortical Electrocorticographic Array in the Common Marmoset

Electrocorticography (ECoG) allows the monitoring of electrical field potentials from the cerebral cortex with high spatiotemporal resolution. Recent development of thin, flexible ECoG electrodes has enabled conduction of stable recordings of large-scale cortical activity. We have developed a whole-cortical ECoG array for the common marmoset. The array continuously covers almost the entire lateral surface of cortical hemisphere, from the occipital pole to the temporal and frontal poles, and it captures whole-cortical neural activity in one shot. This protocol describes a chronic implantation procedure of the array in the epidural space of the marmoset brain. Marmosets have two advantages regarding ECoG recordings, one being the homologous organization of anatomical structures in humans and macaques, including frontal, parietal, and temporal complexes. The other advantage is that the marmoset brain is lissencephalic and contains a large number of complexes, which are more difficult to access in macaques with ECoG, that are exposed to the brain surface.These features allow direct access to most cortical areas beneath the surface of the brain. This system provides an opportunity to investigate global cortical information processing with high resolutions at a sub-millisecond order in time and millimeter order in space.

We have developed a whole-cortical electrocorticographic array for the common marmoset that continuously covers almost the entire lateral surface of cortex, from the occipital pole to the temporal and frontal poles. This protocol describes a chronic implantation procedure of the array in the epidural space of the marmoset brain.

一般的なセットの全皮質のシェットランドシープドッグ配列の慢性注入

Electrocorticography (ECoG) allows the monitoring of electrical field potentials from the cerebral cortex with high spatiotemporal resolution. Recent ...

Assessment of Long-term Depression Induction in Adult Cerebellar Slices

Synaptic plasticity provides a mechanism for learning and memory. For cerebellar motor learning, long-term depression (LTD) of synaptic transmissions from parallel fibers (PF) to Purkinje cells (PC) is considered the basis for motor learning, and deficiencies of both LTD and motor learning are observed in various gene-manipulated animals. Common motor learning sets, such as adaptation of the optokinetic reflex (OKR), the vestibular-ocular reflex (VOR), and rotarod test were used for evaluation of motor learning ability. However, results obtained from the GluA2-carboxy terminus modified knock-in mice demonstrated normal adaptation of the VOR and the OKR, despite lacking PF-LTD. In that report, induction of LTD was only attempted using one type of stimulation protocol at room temperature. Thus, conditions to induce cerebellar LTD were explored in the same knock-in mutants using various protocols at near physiological temperature. Finally, we found stimulation protocols, by which LTD could be induced in these gene-manipulated mice. In this study, a set of protocols are proposed to evaluate LTD-induction, which will more accurately allow examination of the causal relationship between LTD and motor learning. In conclusion, experimental conditions are crucial when evaluating LTD in gene-manipulated mice.

In some gene-manipulated animals, using a single protocol may fail to induce LTD in cerebellar Purkinje cells, and there may be a discrepancy between LTD and motor learning. Multiple protocols are necessary to assess LTD-induction in gene-manipulated animals. Standard protocols are shown.

成人小脳スライスにおける長期うつ病誘導の評価

Synaptic plasticity provides a mechanism for learning and memory. For cerebellar motor learning, long-term depression (LTD) of synaptic transmissions from ...