This research was supported by Basic Science Research Program through the NRF funded by the Ministry of Education, Science and Technology (2010-0022362) and (2012R1A2A2A02046812).

倫理の声明： この実験は、慶北大学校施設内動物管理使用委員会によって承認され、実験動物の管理と使用（国立衛生研究所、1996）のための案内に従って行きました。実験後、動物は、二酸化炭素の安楽死の下で屠殺しました。 ラット用頭部固定バーの1デザイン注：この手順は、固定バー設計のための一般的なガイドラインです。寸法および材料についての変更は、動物やインプラントに応じて可能です。 CADソフトウェアを使用して、ヘッドの固定バーを設計する必要は推奨されないが。 <ol><li>固定バーを設計するためのCADソフトウェアを使用してください。 <ol><li>長方形（123×35×6ミリメートル）（ 図1A）の固定バーを設計します。固定バーの中央に、長方形のノッチ（ポート16×20ミリメートル）を行います。斜面やくぼみを追加whicHは、ポートの境界にガイドレール（ 図1A-A、 図1Bは -arrows）として機能します。 注：ポートの面積は開頭を通じて脳領域のほとんどに定位マニピュレータを使用して垂直方向のアクセスを可能にするために、脳の上にラットの頭蓋骨の推定矩形をカバーしていることを確認してください。 </li><li>固定バーの両側に2つの細長いアンカー穴とプレート（50×20×6 mm）を取り付けます。これらの穴は直径6mm（ 図1A-D）のネジを渡したことを確認してください。ロックフック（ 図1A-B）のためのポートの前に直径3mmの穴を作成します。 </li><li>箱が床（135×305ミリメートル）と側壁（305×76ミリメートル）を有することを確認します。 図1A（挿入図）に示すように、2つの直径6mmの穴をドリル。固定バー（ 図1A-E）に支持壁を追加します。 </li></ol></li><li>設計バーとボックスを製造し、としてシアノアクリレートを使用して、それらのそれぞれを組み立てます図1A（挿入図）に示します。直径6mmのネジを使用してボックスにバーを組み立てます。人間工学的な頭部固定（ 図1A、挿入図 ）は、バーのネジ位置を調整することによって、勾配を適用します。 注：マウスの​​場合、固定バーの寸法は、（厚さ2mmのアクリル板の二層からなる）60×20×4mmです。ポートは8 mmの幅（上部）および図1（c）に示すように 9×10 mmの（下側）の正方形です。アンカープレートは、ラットのと同じサイズです。ボックスプレートとサイドの壁はそれぞれ100×72ミリメートルと100×50ミリメートル、です。 </li></ol>移植手術中に2頭マウンター建設注意：歯科アクリル樹脂を扱うときは注意してください。液体は皮膚の炎症を引き起こす可能性があります。マスクを着用し、手袋を着用してください。換気の良い場所で作業してください。 <ol><li>手術前の準備<ol><li>しっかりと固定バー使用して、APのポートの境界をラップバーに適用されたアクリル樹脂の付着を防止するaraffin膜（ 図1B及び図2Aを参照）。コー​​トアクリル樹脂の薄層でパラフィンフィルムで覆われたポートの境界。 </li><li>滅菌手術器具を使用してください。完全に定位フレームとその周辺用いて70％のエチルアルコールを消毒します。 </li></ol></li><li>定位移植手術を行います<ol><li> 10 mlのケタミン（50 mg / mlで）、キシラジン塩酸1.5ミリリットル（23.32 mg / mlで）、および生理食塩水2.5ミリリットルのカクテル混合物（2ミリリットル/ kg）を腹腔内注射によって動物を麻酔。動物が完全に麻酔し、尾とつま先ピンチに応答しないことを確認してください。必要な場合に手術中に1ミリリットル/ kgでのカクテル混合物を注入します。 </li><li>定位固定装置に動物を修正。手術中の乾燥を防ぐために、目に眼軟膏を適用します。 70％エタノールで頭皮を剃るし、清掃してください。 2％のリドカインの皮下注射を与え、SCAを切開頭蓋骨の正中線に沿ってのlp。完全に骨膜を削り取ります。生理食塩水で数回すすいでください。 </li><li>頭蓋骨のリム付近にバリ穴加工を行います。硬膜の違反を防止するために、頭蓋骨の表面からインプラントのネジ（1ミリメートルの深さまで）4-5ターン。ネジ移植後頭蓋表面上CSFおよび血液のことを確認漏れを行いません。生理食塩水ですすぎ、空気パフを使用して、頭蓋骨の表面を乾燥、特にネジの周り。 </li><li> 「歯科用接着剤（スーパーボンドC＆B）」の薄層でコート頭蓋骨の表面。追加の電極注入が予定されている場合、完全にブレグマ含む頭蓋骨の表面を覆わないでください。開頭術座標をマークするためにブレグマを使用してください。インプラント電極および/またはマイクロドライブとその周りにアクリル樹脂を塗布することにより頭蓋骨​​に固定します。 </li><li>歯科用セメントの上に付加的なアクリル系樹脂を適用します。バーでコーティングされた樹脂のANとの間に十分なフィルアウトスペースを残すためにアクリル樹脂の適切な量を使用しますねじおよび/ ​​または電極（ 図2A）を固定する際の取り付け樹脂を頭文字D。 </li></ol></li><li> 5mm以上頭蓋骨表面上置きヘッド固定バーと、その中心（ 図2A -left）を合わせます。 </li><li>頭蓋骨と初期の実装系樹脂のベースとの間の隙間にアクリル樹脂を適用します。 （ 図2A -center）。樹脂が硬くなるまで待ちます。 </li><li>耳バーを取り外し、固定バーポートから成形マウンタを引き出します。 （ 図2A右回転）。必要に応じて成形マウンタのエッジレールの周りに追加のアクリル樹脂を慎重に適用されます。 </li><li>滅菌生理食塩水で頭皮の傷の表面を洗浄します。手術直後抗生物質（エンロフロキサシン、5ミリグラム/ kgの皮下）と鎮痛剤（酒石酸ブトルファノール、2ミリグラム/ kgの皮下）注入します。ハウス個別に1ケージ内の動物。 </li></ol> 3.頭部固定に動物をHabituating 注：この手順のデ著者の経験的に成功した評価に基づい慣れスケジュールをスクライブ。ヘッド固定用慣れ手順の概要については、シュワルツの10または郭の11作品を参照してください。 <ol><li>動物への十分な回復期間（週またはそれ以上）を与えます。 1日目に、頻繁に分割短いセッションで動物を処理します。 </li><li>次の日（2日目）には、固定環境（ボックス）に動物を公開します。固定バーのポートに自分の頭マウンタを挿入しないでください。環境に慣れるために動物のままにしておきます。しっかり1〜2秒間、手を使ってヘッドマウンタを取得し、すぐにそれを解放。ハンドグラブ数回短期固定を繰り返します。 </li><li> 3日目、頭部固定に動物を慣らします。 <ol><li>ホームケージから動物を持参し、固定バーのポートに向かって頭を向けるボックスでそれらを決済します。ヘッドマウンタをつかみ、固定バーのポートに挿入します。 </li><li> （動物のマウンタをバインドゴムバンドを使用して、固定バーのロッキングフックの頭）。 5分間放置。 注：きしむ音、動物のストレスを増加苦労した動物をリードする、固定後の動物のトランクまたは尾に触れないでください。動物は1分以上に苦労し続けた場合、それを解放し、後で試してください。頭部固定にさらされるたびに、このステップから、動物は尿と糞排泄を示すが、これは徐々に慣れが進むにつれて減少します。 </li><li>ロックゴムバンドを取得し、ポートからマウンタを取り出します。バックホームケージに動物を入れて、報酬としてホームケージに穀物の一部またはヒマワリ種子を提供しています。次の試験は30分に開始されます。日以内3.4.4 5以上に3.4.1を繰り返します。 </li></ol></li><li> 7日目4日、プロトコル3.4で説明したように固定を頭ために動物を公開します。徐々に（10分、20分、30分、および馴化が進むにつれて50分）ヘッド固定の持続時間を増加させます。 注：動物が行うとなしtはそれがストレスの高レベルの徴候である可能性があり、食品の報酬を養います。定着時間を増加させないでください。動物は繰り返し苦労挙動を示す場合は、より長い期間のためにそれらを慣らします。 </li><li>毎日の動物の体重をチェックしてください。体重が減少している失敗慣れを考慮してください。成功慣れた後、頭部固定と行動のタスクに進みます。 注：成功慣れは、その後の行動訓練（ 図3EおよびF）を容易にします。 </li></ol>

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The authors have nothing to disclose.

This protocol demonstrates a simple and convenient head fixation method using materials at hand. For physically successful head fixation, the first critical steps during surgery is to rinse and dry the skull surface clearly before applying the dental adhesive resin cement. Second, this method does not uses screws implanting under temporal muscles and only depends on the screws on the skull, their perpendicular implantation against the surface of skull rim is crucial10. This reduces the possibility of accidenta...

頭部固定や頭の拘束は、剛性クランプ用の頭蓋骨上のフレームまたはプレートを取り付けることによって、動物の頭を固定化する技術です。技術は1-3目を覚まし動物および睡眠-覚醒サイクル 4の間juxtacellular標識のための神経記録の有無にかかわらず、正確な感覚刺激のための行動センシングを採用しています。また、 インビボでの脳イメージング5,6のために使用されます。今頭部固定技術は、神経科学と行動研究で一般的に使用されるツールです。 基本的には、従来の頭部固定は頭蓋骨6-8の上にメタルヘッド・ポストの外科用アタッチメントが必要です。添付ファイルは、その後、ヘッドの固定化の結果、静止プラットフォームにクランプされます。このような神経および/または光記録、脳回路の介入として覚醒した動物における神経生理学的研究は、脳イメージングは​​、歯科用アクリルを使用して移植手術を伴っています。そのため、それは、実験が必要であるときに、これらの実験室で頭部固定を実現するための簡単​​な作業です。フレームは軽量かつホームケージ内の動物の自然な行動を邪魔しない程度に小さくなるように設計する必要があります。静止プラットフォームにクランプする際にも機械的剛性を提供します。さらに、実験の目的に応じて、生体適合性を有しています。したがって、一般的な実験装置にかかる頭柱を設計し、製作することは技術的に困難です。 本研究では、新規な頭部固定方法は、金属ヘッドのポストを使用することなく、便利なクランプを介して機械的剛性を提供し、記載されています。この方法を用いることにより、ヘッドの固定は、移植手術は、通常、脳の研究のためにげっ歯類で行われる一般的な実験室の条件下で行うことができます。メタルヘッド・ポストはimplan中に行われたカスタムメイドの頭部固定バーと成形プロセスによって部分的に成形されたアクリル樹脂セメントに置き換えられますテーション手術。固定バーは、ニューラル記録プローブ、刺激電極として他のインプラント、との高い空間の互換性を提供します。また、それはネジ止めを除去することにより、固定プラットフォーム上の動物の頭をクランプする工程を簡素化します。私たちは前の行動の研究では、この方法を使用し、我々は、この方法は、実装が容易で実用的な実験9で便利であると結論しました。

<table border="0" cellpadding="0" cellspacing="0" width="901">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">Dental acrylic resin</td>
			<td style="width:199px;">Lang Dental Manufacturing</td>
			<td style="width:129px;">Ortho-Jet</td>
			<td style="width:343px;">to make molded mounter</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">Paraffin film</td>
			<td style="width:199px;">Bemis</td>
			<td style="width:129px;">Parafilm</td>
			<td>to prevent adhesion of acrylic resin to the bar</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">Dental adhesive</td>
			<td style="width:199px;">Sun Medical</td>
			<td style="width:129px;">Super-Bond C&#38;B</td>
			<td>to bond between acrylic resin and skull</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">Plexglass or other acrylic plate</td>
			<td style="width:199px;"></td>
			<td style="width:129px;"></td>
			<td>to make fixation bar and box platform</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:231px;">Cyanoacrylate</td>
			<td style="width:199px;"></td>
			<td style="width:129px;"></td>
			<td>to bond acrylic plates</td>
		</tr>
	</tbody>
</table>

acrylic resin molding based head fixation technique in rodents

Head fixation is a technique of immobilizing animal's head by attaching a head-post on the skull for rigid clamping. Traditional head fixation requires surgical attachment of metallic frames on the skull. The attached frames are then clamped to a stationary platform resulting in immobilization of the head. However, metallic frames for head fixation have been technically difficult to design and implement in general laboratory environment. In this study, we provide a novel head fixation method. Using a custom-made head fixation bar, head mounter is constructed during implantation surgery. After the application of acrylic resin for affixing implants such as electrodes and cannula on the skull, additional resins applied on top of that to build a mold matching to the port of the fixation bar. The molded head mounter serves as a guide rails, investigators conveniently fixate the animal&#8217;s head by inserting the head mounter into the port of the fixation bar. This method could be easily applicable if implantation surgery using dental acrylics is necessary and might be useful for laboratories that cannot easily fabricate CNC machined metal head-posts.

このヘッドの固定方法は、ラットとマウスの両方に適用可能です。合計8匹のラットは、内側前脳束に電極を刺激する二国間の注入を受けました。それらのうち4については、カスタマイズされたタングステンワイヤ電極（マイクロドライブ）を追加的にM1前肢領域（ 図2C）に移植しました。マウスについて（N = 2）は、4つのステンレス鋼ワイヤ電極を、前頭/後頭部皮質領域に移植...

Watch this Scientific Journal Video about Acrylic Resin Molding Based Head Fixation Technique in Rodents at JoVE.com

Acrylic Resin Molding Based Head Fixation Technique in Rodents

Traditional head fixation techniques have used metallic frames attached on the skull as an interface for head fixation apparatus. This protocol demonstrates a frameless, acrylic resin molding based head fixation technique for behavioral tasks in rodents.

げっ歯類でのアクリル樹脂成形ベースヘッド固定法

Head fixation is a technique of immobilizing animal's head by attaching a head-post on the skull for rigid clamping. Traditional head fixation requires ...

acrylic-resin-molding-based-head-fixation-technique-in-rodents

Research

JoVE Journal

Neuroscience

11.7K ビュー.  Kyungpook National University. Traditional head fixation techniques have used metallic frames attached on the skull as an interface for head fixation apparatus. This protocol demonstrates a frameless, acrylic resin molding based head fixation technique for behavioral tasks in rodents.

ビデオ: Acrylic Resin Molding Based Head Fixation Technique in Rodents

Preparing Undercut Model of Posttraumatic Epileptogenesis in Rodents

Partially isolated cortex ("undercut") is an animal model of posttraumatic epileptogenesis. The surgical procedure involves cutting through the sensorimotor cortex and the underneath white matter (undercut) so that a specific region of the cerebral cortex is largely isolated from the neighboring cortex and subcortical regions1-3. After a latency of two or more weeks following the surgery, epileptiform discharges can be recorded in brain slices from rodents1; and electrical or behavior seizures can be observed in vivo from other species such as cat and monkey4-6. This well established animal model is efficient to generate and mimics several important characteristics of traumatic brain injury. However, it is technically challenging attempting to make precise cortical lesions in the small rodent brain with a free hand. Based on the procedure initially established in Dr. David Prince's lab at the Stanford University1, here we present an improved technique to perform a surgery for the preparation of this model in mice and rats. We demonstrate how to make a simple surgical device and use it to gain a better control of cutting depth and angle to generate more precise and consistent results. The device is easy to make, and the procedure is quick to learn. The generation of this animal model provides an efficient system for study on the mechanisms of posttraumatic epileptogenesis.

Partially isolated cortex (&#x201C;undercut&#x201D;) is an efficient animal model of posttraumatic epileptogenesis. Here we demonstrate how to make a novel surgical device and use it to make more precise and consistent lesions to generate this model.

げっ歯類で心的外傷後てんかん原のアンダーカットモデルを準備

Partially isolated cortex ("undercut") is an animal model of posttraumatic epileptogenesis. The surgical procedure involves cutting through the ...

神経科学

Whole Animal Perfusion Fixation for Rodents

The goal of fixation is to rapidly and uniformly preserve tissue in a life-like state. While placing tissue directly in fixative works well for small pieces of tissue, larger specimens like the intact brain pose a problem for immersion fixation because the fixative does not reach all regions of the tissue at the same rate 5,7. Often, changes in response to hypoxia begin before the tissue can be preserved 12. The advantage of directly perfusing fixative through the circulatory system is that the chemical can quickly reach every corner of the organism using the natural vascular network. In order to utilize the circulatory system most effectively, care must be taken to match physiological pressures 3. It is important to note that physiological pressures are dependent on the species used. Techniques for perfusion fixation vary depending on the tissue to be fixed and how the tissue will be processed following fixation. In this video, we describe a low-cost, rapid, controlled and uniform fixation procedure using 4% paraformaldehyde perfused via the vascular system: through the heart of the rat to obtain the best possible preservation of the brain for immunohistochemistry. The main advantage of this technique (vs. gravity-fed systems) is that the circulatory system is utilized most effectively.

Here we describe a low-cost, rapid, controlled and uniform fixation procedure using 4% paraformaldehyde perfused via the vascular system: through the heart of the rat to obtain the best possible preservation of the brain.

齧歯類のために全体の動物灌流固定

The goal of fixation is to rapidly and uniformly preserve tissue in a life-like state. While placing tissue directly in fixative works well for small ...

Large-scale Recording of Neurons by Movable Silicon Probes in Behaving Rodents

A major challenge in neuroscience is linking behavior to the collective activity of neural assemblies. Understanding of input-output relationships of neurons and circuits requires methods with the spatial selectivity and temporal resolution appropriate for mechanistic analysis of neural ensembles in the behaving animal, i.e. recording of representatively large samples of isolated single neurons. Ensemble monitoring of neuronal activity has progressed remarkably in the past decade in both small and large-brained animals, including human subjects1-11. Multiple-site recording with silicon-based devices are particularly effective because of their scalability, small volume and geometric design.
Here, we describe methods for recording multiple single neurons and local field potential in behaving rodents, using commercially available micro-machined silicon probes with custom-made accessory components. There are two basic options for interfacing silicon probes to preamplifiers: printed circuit boards and flexible cables. Probe supplying companies (<a href="http://www.neuronexustech.com/" >http://www.neuronexustech.com/</a>; <a href="http://www.sbmicrosystems.com/">http://www.sbmicrosystems.com/</a>; <a href="http://www.acreo.se/">http://www.acreo.se/</a>) usually provide the bonding service and deliver probes bonded to printed circuit boards or flexible cables. Here, we describe the implantation of a 4-shank, 32-site probe attached to flexible polyimide cable, and mounted on a movable microdrive. Each step of the probe preparation, microdrive construction and surgery is illustrated so that the end user can easily replicate the process.

We describe methods for large-scale recording of multiple single units and local field potential in behaving rodents with silicon probes. Drive fabrication, probe attachment to the drive and probe implantation processes are illustrated in sufficient details for easy replication.

動作げっ歯類の可動シリコンプローブによるニューロンの大規模なレコーディング

A major challenge in neuroscience is linking behavior to the collective activity of neural assemblies. Understanding of input-output relationships of ...

Recording and Analysis of Circadian Rhythms in Running-wheel Activity in Rodents

When rodents have free access to a running wheel in their home cage, voluntary use of this wheel will depend on the time of day1-5. Nocturnal rodents, including rats, hamsters, and mice, are active during the night and relatively inactive during the day. Many other behavioral and physiological measures also exhibit daily rhythms, but in rodents, running-wheel activity serves as a particularly reliable and convenient measure of the output of the master circadian clock, the suprachiasmatic nucleus (SCN) of the hypothalamus. In general, through a process called entrainment, the daily pattern of running-wheel activity will naturally align with the environmental light-dark cycle (LD cycle; e.g. 12 hr-light:12 hr-dark). However circadian rhythms are endogenously generated patterns in behavior that exhibit a ~24 hr period, and persist in constant darkness. Thus, in the absence of an LD cycle, the recording and analysis of running-wheel activity can be used to determine the subjective time-of-day. Because these rhythms are directed by the circadian clock the subjective time-of-day is referred to as the circadian time (CT). In contrast, when an LD cycle is present, the time-of-day that is determined by the environmental LD cycle is called the zeitgeber time (ZT).
Although circadian rhythms in running-wheel activity are typically linked to the SCN clock6-8, circadian oscillators in many other regions of the brain and body9-14 could also be involved in the regulation of daily activity rhythms. For instance, daily rhythms in food-anticipatory activity do not require the SCN15,16 and instead, are correlated with changes in the activity of extra-SCN oscillators17-20. Thus, running-wheel activity recordings can provide important behavioral information not only about the output of the master SCN clock, but also on the activity of extra-SCN oscillators. Below we describe the equipment and methods used to record, analyze and display circadian locomotor activity rhythms in laboratory rodents.

Circadian rhythms in voluntary wheel-running activity in mammals are tightly coupled to the molecular oscillations of a master clock in the brain. As such, these daily rhythms in behavior can be used to study the influence of genetic, pharmacological, and environmental factors on the functioning of this circadian clock.

げっ歯類におけるランニングホイール活動の概日リズムの記録および分析

When rodents have free access to a running wheel in their home cage, voluntary use of this wheel will depend on the time of day1-5. Nocturnal rodents, ...

Transplantation of Olfactory Ensheathing Cells to Evaluate Functional Recovery after Peripheral Nerve Injury

Olfactory ensheathing cells (OECs) are neural crest cells which allow growth and regrowth of the primary olfactory neurons. Indeed, the primary olfactory system is characterized by its ability to give rise to new neurons even in adult animals. This particular ability is partly due to the presence of OECs which create a favorable microenvironment for neurogenesis. This property of OECs has been used for cellular transplantation such as in spinal cord injury models. Although the peripheral nervous system has a greater capacity to regenerate after nerve injury than the central nervous system, complete sections induce misrouting during axonal regrowth in particular after facial of laryngeal nerve transection. Specifically, full sectioning of the recurrent laryngeal nerve (RLN) induces aberrant axonal regrowth resulting in synkinesis of the vocal cords. In this specific model, we showed that OECs transplantation efficiently increases axonal regrowth.
OECs are constituted of several subpopulations present in both the olfactory mucosa (OM-OECs) and the olfactory bulbs (OB-OECs). We present here a model of cellular transplantation based on the use of these different subpopulations of OECs in a RLN injury model. Using this paradigm, primary cultures of OB-OECs and OM-OECs were transplanted in Matrigel after section and anastomosis of the RLN. Two months after surgery, we evaluated transplanted animals by complementary analyses based on videolaryngoscopy, electromyography (EMG), and histological studies. First, videolaryngoscopy allowed us to evaluate laryngeal functions, in particular muscular cocontractions phenomena. Then, EMG analyses demonstrated richness and synchronization of muscular activities. Finally, histological studies based on toluidine blue staining allowed the quantification of the number and profile of myelinated fibers.
All together, we describe here how to isolate, culture, identify and transplant OECs from OM and OB after RLN section-anastomosis and how to evaluate and analyze the efficiency of these transplanted cells on axonal regrowth and laryngeal functions.

Olfactory ensheathing cells (OECs) are neural crest cells which allow growth of the primary olfactory neurons. This specific property can be used for cellular transplantation. We present here a model of cellular transplantation based on the use of OECs in a laryngeal nerve injury model.

末梢神経損傷後の機能回復を評価する嗅神経鞘細胞の移植

Olfactory ensheathing cells (OECs) are neural crest cells which allow growth and regrowth of the primary olfactory neurons. Indeed, the primary olfactory ...

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 ...

Transcranial Electrical Brain Stimulation in Alert Rodents

Transcranial electrical brain stimulation can modulate cortical excitability and plasticity in humans and rodents. The most common form of stimulation in humans is transcranial direct current stimulation (tDCS). Less frequently, transcranial alternating current stimulation (tACS) or transcranial random noise stimulation (tRNS), a specific form of tACS using an electrical current applied randomly within a pre-defined frequency range, is used. The increase of noninvasive electrical brain stimulation research in humans, both for experimental and clinical purposes, has yielded an increased need for basic, mechanistic, safety studies in animals. This article describes a model for transcranial electrical brain stimulation (tES) through the intact skull targeting the motor system in alert rodents. The protocol provides step-by-step instructions for the surgical set-up of a permanent epicranial electrode socket combined with an implanted counter electrode on the chest. By placing a stimulation electrode into the epicranial socket, different electrical stimulation types, comparable to tDCS, tACS, and tRNS in humans, can be delivered. Moreover, the practical steps for tES in alert rodents are introduced. The applied current density, stimulation duration, and stimulation type may be chosen depending on the experimental needs. The caveats, advantages, and disadvantages of this set-up are discussed, as well as safety and tolerability aspects.

This protocol describes a surgical set-up for a permanent epicranial electrode socket and an implanted chest electrode in rodents. By placing a second electrode into the socket, different types of transcranial electrical brain stimulation can be delivered to the motor system in alert animals through the intact skull.

警告齧歯動物で経頭蓋脳の電気的刺激

Transcranial electrical brain stimulation can modulate cortical excitability and plasticity in humans and rodents. The most common form of stimulation in ...

Intracranial Pharmacotherapy and Pain Assays in Rodents

Pain is a salient sensory experience with affective and cognitive dimensions. However, central mechanisms for pain remain poorly understood, hindering the development of effective therapeutics. Intracranial pharmacology presents an important tool for understanding the molecular and cellular mechanisms of pain in the brain, as well as for novel treatments. Here we present a protocol that integrates intracranial pharmacology with pain behavior testing. Specifically, we show how to infuse analgesic drugs into a select brain region, which may be responsible for pain modulation. Furthermore, to determine the effect of the candidate drug in the central nerve system, pain assays are performed after intracranial treatment. Our results demonstrate that intracranial administration of analgesic drugs in a targeted region can provide relief of pain in rodents. Thus, our protocol successfully demonstrates that intracranial pharmacology, combined with pain behavior testing, can be a powerful tool for the study of pain mechanisms in the brain.

Here we present a protocol to perform intracranial pharmacological experiments followed by pain behavior assays in rodents. This protocol allows researchers to deliver molecular and cellular targets in the brain, for pharmacologic agents in the treatment of pain.

頭蓋内薬物療法と痛みの試金の齧歯動物

Pain is a salient sensory experience with affective and cognitive dimensions. However, central mechanisms for pain remain poorly understood, hindering the ...

The Power of Interstimulus Interval for the Assessment of Temporal Processing in Rodents

Temporal processing deficits have been implicated as a potential elemental dimension of higher-level cognitive processes, commonly observed in neurocognitive disorders. Despite the popularization of prepulse inhibition (PPI) in recent years, many current protocols promote using a percent of control measure, thereby precluding the assessment of temporal processing. The present study used cross-modal PPI and gap prepulse inhibition (gap-PPI) to demonstrate the benefits of employing a range of interstimulus intervals (ISIs) to delineate effects of sensory modality, psychostimulant exposure, and age. Assessment of sensory modality, psychostimulant exposure, and age reveals the utility of an approach varying the interstimulus interval (ISI) to establish the shape of the ISI function, including increases (sharper curve inflections) or decreases (flattening of the response amplitude curve) in startle amplitude. Additionally, shifts in peak response inhibition, suggestive of a differential sensitivity to the manipulation of ISI, are often revealed. Thus, the systematic manipulation of ISI affords a critical opportunity to evaluate temporal processing, which may reveal the underlying neural mechanisms involved in neurocognitive disorders.

Temporal processing, a preattentive process, may underlie deficits in higher-level cognitive processes, including attention, commonly observed in neurocognitive disorders. Using prepulse inhibition as an exemplar paradigm, we present a protocol for manipulating interstimulus interval (ISI) to establish the shape of the ISI function to provide an assessment of temporal processing.

刺激間隔の齧歯動物の一時的な処理の評価のための力

Temporal processing deficits have been implicated as a potential elemental dimension of higher-level cognitive processes, commonly observed in ...

A Revised Surgical Approach to Induce Endolymphatic Hydrops in the Guinea Pig

Endolymphatic hydrops is an enlargement of scala media that is most often associated with Meniere&#39;s disease, though the pathophysiologic mechanism(s) remain unclear. In order to adequately study the attributes of endolymphatic hydrops, such as the origins of low-frequency hearing loss, a reliable model is needed. The guinea pig is a&#160;good model because it hears in the low-frequency regions that are putatively affected by endolymphatic hydrops. Previous research has demonstrated that endolymphatic hydrops can be induced surgically via intradural or extradural approaches that involve drilling on the endolymphatic duct and sac. However, whether it was possible to create an endolymphatic hydrops model using an extradural approach that avoided dangerous drilling on the endolymphatic duct and sac was unknown. The objective of this study was to demonstrate a revised extradural approach to induce experimental endolymphatic hydrops at 30 days post-operatively by obliterating the endolymphatic sac and injuring the endolymphatic duct with a fine pick. The sample size consisted of seven guinea pigs. Functional measurements of hearing were made and temporal bones were subsequently harvested for histologic analysis. The approach had a success rate of 86% in achieving endolymphatic hydrops. The risk of cerebrospinal fluid leak was minimal. No perioperative deaths or injuries to the posterior semicircular canal occurred in the sample. The presented method demonstrates a safe and reliable way to induce endolymphatic hydrops at a relatively quick time point of 30 days. The clinical implications are that the presented method provides a reliable model to further explore the origins of low-frequency hearing loss that can be associated endolymphatic hydrops.

This article demonstrates an extradural approach to obliterate the guinea pig endolymphatic sac and injure the endolymphatic duct with a fine pick in order to induce experimental endolymphatic hydrops.

モルモットの内リンパ性水力を誘導する改訂された外科的アプローチ

Endolymphatic hydrops is an enlargement of scala media that is most often associated with Meniere&#39;s disease, though the pathophysiologic mechanism(s) ...