Name: a repetitive concussive head injury model in mice
Uploaded: 2016-10-12T14:00:00.000+00:00
Duration: 5 min 42 s
Description: Watch this Scientific Journal Video about A Repetitive Concussive Head Injury Model in Mice at JoVE.com

This works was supported by funding from a Florida Health grant (Brain and spinal cord injury research fund) (KKW).

すべての手順は、フロリダ大学の施設内動物管理使用委員会によって、および実験動物の管理と使用に関する健康ガイドの国立研究所に従って承認されたプロトコル番号201207692下で行いました。 1.動物ケア<ol><li> 3-4ヶ月齢の雄のC57BL / 6Jマウスを使用してください。寝具、ネスティング材料、食品、および水を自由に提供します。一定の12時間明/ 12時間暗サイクルで22°C - 20で制御された周囲温度にマウスを保管してください。 </li></ol> 2.事前衝突準備<ol><li>電磁定位インパクトデバイスにカスタムメイドのシリコーンゴムでコーティングされた金属チップを取り付けます。先端の平坦な底部は、プローブ先端（ 図1A）の表面に平行であることを確認してください。 </li><li> 2.5％イソフルランの維持麻酔に続いて4％イソフルランでマウスを麻酔。流量計を介して麻酔を確認してください。 Monito動物がペダル撤退反射の消失を示すことによって、麻酔の外科的なレベルに達するまで、麻酔レベルrを。 </li><li>加熱パッド上の腹臥位でマウスを置きます。麻酔下でマウスを維持するために漏斗状のノーズコーンを使用してください。完全にトリマーを使用して頭を剃ります。麻酔下ながら乾燥を防ぐために、マウスの目にワセリン眼軟膏を使用してください。 </li></ol> 3.インパクトパラメータ設定注：影響システムは、衝撃パラメータを設定するための制御ボックス、衝突を実行するためのアクチュエータ、及び3の移動軸を有するデジタル定位フレームを含みます。 <ol><li> 4メートル/秒の衝撃装置の速度を事前に設定し、コントロールボックスに240ミリ秒に滞留時間。 </li></ol> 4.ポジショニングインパクトセンター<ol><li> 39℃付近で体温を維持するために、動物の体の下に柔らかい加熱パッドを置きます。 PRONに定位フレームにマウスをマウント平滑末端耳バー付きの電子の位置。 </li><li> Z-ドライバを移動することで、マウスの頭部に近い衝撃先端を下げます。ターゲットは矢状縫合の上座標に途中でX軸とYドライバを移動させることにより、フラットなインパクトチップ（直径9mm）を調整します。 </li><li>衝撃先端の1辺が上下に2耳（ 図 1C）の間に引かれた仮想の水平線に平行であることを確認してください。インパクトの中心はinterfrontalとラムダ状縫合（0ミリメートル、横4.5ミリメートルを耳にする耳9ミリメートル）の間の中央矢状縫合の中間に相当します。 </li></ol> 5.インパクトの深さの設定<ol><li>正しくインパクトの深さを設定するには、絶縁されたシリコーンゴムでコーティングされたインパクトチップを交換するために、追加のプローブ・チップを使用しています。 </li><li>ヒントを切り替えた後の衝撃中心のないシフトがないことを確認するために、ヒントを切り替える前にゼロにデジタル定位コントロールパネルにXとYのチャンネルを設定します。 </li><li> PROBを移動手動でX-とY-ドライブを移動することにより、インパクトエリアの中心に電子チップ。 </li><li>マウスの尾のクリップ接触センサ。 </li><li>プローブ先端が衝突部位の表面に触れるまでインパクタ（Zドライブ）を下に移動します。 </li><li>ゼロに定位コントロールパネルのZチャンネルを設定します。 </li><li>手動でX軸とYドライバまで、X軸とYドライバ（NOTデジタル定位コントロールパネル上のゼロボタン）を調整することにより、バックインパクトエリアへの衝撃先端を移動（衝撃先端が以前に配置された）はゼロです。 </li><li>コントロールボックスに後退スイッチを動かすことによって、アクチュエータを撤回。手動で4ミリメートルによって（Zドライバ）インパクターを下に移動。 </li></ol> 6.影響<ol><li>コントロールボックスへの影響スイッチをクリックすることで、衝撃をトリガし、4ミリメートルの変形の深さを実現。 </li></ol> 7.ポストインパク<ol><li>タイマーを使用して、マウスの最初の呼吸までの衝撃からの時間を測定します。 </li><li&gt;楽器から動物を削除し、自分の体の温度を維持するために暖かいパッド上に置きます。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。 </li><li>きれいなケージに戻って動物を返す前に、リカバリを可能にします。完全に回復するまで、他の動物の会社に動物を返しません。 </li><li>毎日マウスを観察し、重量を量ります。マウスは、痛みの兆候を示した場合は、腹腔内に1でメロキシカムでそれらを注入 - 24時間 - 2ミリグラム/毎12 kgです。 </li></ol> 8.繰り返しインパク<ol><li>日4、7日にマウスを追加傷害を与え、最初の損傷（インパクトの間に72時間間隔）後の10。 </li></ol> 9.免疫組織化学（IHC） <ol><li> Transcardial灌流 <ol><li> 200メートル/ kgのペントバルビタールで腹腔内注射によりマウスを麻酔。 </li><li>評価し、つま先のピンチによって外科的平面麻酔を確保します。秒URE優しく化学ヒュームフードの内側に発泡スチロールの作業面に前足と後足をテーピングすることにより、仰臥位でマウス。 </li><li>ちょうど鎖骨の剣状突起下から胸部正中線に沿って皮膚を通して切開を行います。剣状突起の2つの追加の皮膚切開を行い、横方向に腹側胸郭のベースに沿って進みます。 </li><li>胸腔を開き、胸部の筋肉や肋骨を切断することにより、心臓を露出。 </li><li>鈍鉗子で鼓動している心臓を固定し、1作る - 左心室中2ミリメートルの切開を。 </li><li>すぐに右心房に翼状針を挿入します。ゆっくりとシリンジを押して20ミリリットルの生理食塩水の注入を開始します。 </li><li>生理食塩水から4％パラホルムアルデヒドに切り替えます。パラホルムアルデヒドの20ミリリットルで灌流を続けます。 </li><li>マウスを刎ねるとハサミで皮膚を取り除きます。骨カッターを使用して、頭蓋骨から脳を分離します。 </li></ol></li><li> C言語切片ryostat <ol><li> -80℃における最適切断温度（OCT）製剤、凍結中の埋め込み脳組織。矢状方向にクライオスタットに脳を置きます。カット脳切片厚さ5μm。 </li></ol></li><li> 染色 <ol><li>室温で1時間凍結切片を乾燥させます。 </li><li>室温で1時間、リン酸緩衝生理食塩水（PBS）中の2％ヤギ血清および0.1％トリトンX-100100μlでスライドをインキュベートします。 </li><li> 300μlのPBSでスライドを3回洗浄。 4℃で別々に一晩：（200 1）または抗フェリチン抗体：その後、抗GFAP（200 1）でスライドをインキュベートします。 </li><li> 300μlのPBSでスライドを3回洗浄。その後、ビオチン結合二次抗体とともに室温で2時間、スライドをインキュベートします。 </li><li> 300μlのPBSでスライドを3回洗浄。その後、室温で30分間アビジン - ビオチン複合体（ABC）溶液（午前1時50分）でスライドをインキュベートします。 </li><li>300μlのPBSでスライドを3回洗浄。 8分-そして5用の3,3&#39;-ジアミノベンジジン（DAB）基質溶液（50ミリリットルPBS、10μlのH 2 O 2、10ミリグラムのDABピル、使用する前に、フィルタ）でインキュベートします。陽性細胞が現れるまで顕微鏡下でスライドを観察します。 </li><li> 5分間の実行速度の遅いスライドを水道水ですすぎます。ラボ・ワイプできれいにスライドします。その後、封入剤とカバーガラスとのセクションをマウントします。 </li></ol></li></ol>

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The authors have no financial interest to disclose.

臨床症状と形態学的に類似した脳損傷を模倣するために、ポスト脳震盪の症状が期待されています。ポスト脳震盪の症状は、一般的に頭痛、めまい、めまい、疲労感、メモリや睡眠の問題、トラブル集中だけでなく、不安、および抑うつ気分が含まれます。身体症状はまだ動物モデルにおいて測定可能ではないかもしれないので、運動および認知機能および感情行動の変化は、合理的に、動?...

脳震盪は、とも呼ばれる軽度の外傷性脳損傷（MTBI）、外傷性脳損傷（TBI）の中で最も頻繁に発生であり、米国で何百万人もの人々に影響を与えます。脳震盪は、診断するのが難しいことができ、脳震盪のための具体的な治療法はありません。成長している認識とスポーツ傷害、戦闘、および他の物理的に魅力的な追求に起因する軽度の機械的外傷が累積し、慢性神経結果1,2を有していてもよいことを、いくつかの証拠があります。しかし、脳震盪とその効果に関する知識の欠如が依然として存在します。唯一の神経学的評価とイメージング評価は臨床診断のために利用可能であるため、現在の方法論は、ヒトの病態と治療の研究を制限します。動物モデルは、MTBIのさらなる診断と治療の希望を持って、効率的で厳格な、かつ制御された方法で脳震盪を研究するための手段を提供します。 研究は、伝統的なTBIを適応していますこのような制御皮質衝撃（CCI）、流体パーカッションのインパクト（FPI）のようなモデルは、重量MTBIを実行し、けがのパラメータを変更することにより、低損傷の重大度を刺激するために、損傷、およびブラスト傷害をドロップします。これらのモデルは、原因の臨床症状と形態学的に類似した脳外傷を複製する能力を使用することは有益です。しかしながら、それらはまた、独自の制限を有します。加速損傷（重量の低下）によって誘発される損傷の重症度は、多くの場合、非常に可変です。軽度のCCIの二つの結果 - くも膜下出血と焦点挫傷は - 典型的な人間の脳震盪と比較することはできません。爆風損傷が異なる露光位置とピーク圧力測定だけでなく、露出3-6中の変数二次的損傷に関して、より論争のモデルでありながら、CCIとFPIは、臨床的に関連していない開頭術を必要としています。更新震とう動物モデルをそれは、臨床セッティに前臨床研究を翻訳することができますngが研究に必要です。 軽度のTBIのモデル化における重要な問題は、最も密接に臨床設定で怪我を複製する実験的な傷害の重症度を、定義することです。最近、別の研究グループは、閉鎖性頭部外傷または震盪頭部外傷（CHI）モデル7-10を開発しました。 CHIは開頭せずにCCIの変形であるが、それはまだ頭部衝撃を生成するために、従来の電子、磁気インパクト方式を使用しています。 CHIはインパクトパラメータを調整することによって、軽度から中等度の範囲で脳震盪を誘導することができます。意識消失（LOC）は、すぐに呼吸速度または呼吸の一時的停止の減少を検出することにより、衝撃後に観察することができます。 LOCの期間は、損傷の重症度を決定するために使用されます。本稿では、詳細なステップバイステップのプロトコルおよび代表的な結果と一緒に、マウスにおける反復CHI（rCHI）モデルのわずかに改善および更新されたバージョンが含まれています。 rCHIモデルの研究戦略のA脳震盪誘発性の病理学的変化の全てを模倣することのできるいかなる個々の動物モデルが存在しない、特に以来、MTBIの効果と潜在的な治療法を決定する上で有益な再。

<table><tbody><tr><td>anesthesia machine</td><td>Eagle Eye Anesthesia, Inc</td><td>Model 150&amp;nbsp;</td><td>anesthesia</td></tr><tr><td>Electromagnetic Impactor</td><td>LeicaBiosystems</td><td>Impact One Stereotaxic Impactor</td><td>perform impaction</td></tr><tr><td>Digital Stereotaxic instrument</td><td>LeicaBiosystems</td><td>39462501</td><td>mount mouse and positioning tips</td></tr><tr><td>Sicilone rubber-coated metal tip</td><td>Precision Tool &amp;amp; Engineering, Gainesvill FL</td><td>custom-made</td><td>impact tip</td></tr><tr><td>Lithium Ion All-in-One Trimmer</td><td>WAHL Home Products</td><td>9854-600</td><td>shave mouse hair</td></tr><tr><td>paper clips</td><td>custom-made</td><td></td><td>probe tip</td></tr><tr><td>Cotton tipped applicators</td><td>MEDLINE</td><td>MDS202055</td><td>scrub head with saline</td></tr><tr><td>Tissue Tek O.C.T.</td><td>ASKURA FINETEK USA INC</td><td>4583</td><td>tissue embedding</td></tr><tr><td>anti-GFAP</td><td>Dako</td><td>CA93013</td><td>antibody for IHC</td></tr><tr><td>anti Ferritin</td><td>Sigma</td><td>F6136</td><td>antibody for IHC</td></tr><tr><td>VECTASTAIN Elite ABC&amp;nbsp; kit</td><td>Vector laboratories</td><td>PK-6100</td><td>IHC detection system</td></tr><tr><td>Permount Mounting Medium</td><td>Fisher Scientific</td><td>SP15-100</td><td></td></tr><tr><td>Aperio XT ScanScope scanner</td><td>Leica Microsystems Inc,</td><td></td><td>slides scanning</td></tr><tr><td>Leica AutoStainer XL</td><td>Leica the pathology Company</td><td>ST2010</td><td>H&amp;amp;E staining</td></tr><tr><td>DAB&amp;nbsp;</td><td>sigma</td><td>D3939</td><td>IHC detection system</td></tr></tbody></table>

a repetitive concussive head injury model in mice

Despite the concussion/ mild traumatic brain injury (mTBI) being the most frequent occurrence of traumatic brain injury, there is still a lack of knowledge on the injury and its effects. To develop a better understanding of concussions, animals are often used because they provide a controlled, rigorous, and efficient model. Studies have adapted traditional animal models to perform mTBI to stimulate mild injury severity by changing the injury parameters. These models have been used because they can produce morphologically similar brain injuries to the clinical condition and provide a spectrum of injury severities. However, they are limited in their ability to present the identical features of injuries in patients. Using a traditional impact system, a repetitive concussive injury (rCHI) model can induce mild to moderate human-like concussion. The injury degree can be determined by measuring the period of loss of consciousness (LOC) with a sign of a transient termination of breathing. The rCHI model is beneficial to use for its accuracy and simplicity in determining mTBI effects and potential treatments.

このモデル（ 図1 AC）では、あえぎ呼吸や浅い呼吸の短い期間ではありました。意識（無意識）の損失は、呼吸速度または正常な呼吸を再開する前に、呼吸の一時的停止の減少として定義されます。頭の中心への影響は短期的無意識（個別インパクト時の7.5±4.7、7.8±5.5、10.2±8.8、9.5±8.0秒、 図1D）が発生しました。マウスの脳には明らかな構?...

Watch this Scientific Journal Video about A Repetitive Concussive Head Injury Model in Mice at JoVE.com

A Repetitive Concussive Head Injury Model in Mice

Concussion presents the most common type of traumatic brain injury. Therefore, a repetitive concussive animal model, which replicates the important features of an injury in patients, may provide a means to study concussion in a rigorous, controlled, and efficient manner.

マウスにおける反復震とう頭部損傷モデル

Despite the concussion/ mild traumatic brain injury (mTBI) being the most frequent occurrence of traumatic brain injury, there is still a lack of ...

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Research

JoVE Journal

Medicine

11.8K ビュー.  University of Florida. Concussion presents the most common type of traumatic brain injury. Therefore, a repetitive concussive animal model, which replicates the important features of an injury in patients, may provide a means to study concussion in a rigorous, controlled, and efficient manner.

ビデオ: A Repetitive Concussive Head Injury Model in Mice

Lateral Fluid Percussion: Model of Traumatic Brain Injury in Mice

Traumatic brain injury (TBI) research has attained renewed momentum due to the increasing awareness of head injuries, which result in morbidity and mortality. Based on the nature of primary injury following TBI, complex and heterogeneous secondary consequences result, which are followed by regenerative processes 1,2. Primary injury can be induced by a direct contusion to the brain from skull fracture or from shearing and stretching of tissue causing displacement of brain due to movement 3,4. The resulting hematomas and lacerations cause a vascular response 3,5, and the morphological and functional damage of the white matter leads to diffuse axonal injury 6-8. Additional secondary changes commonly seen in the brain are edema and increased intracranial pressure 9. Following TBI there are microscopic alterations in biochemical and physiological pathways involving the release of excitotoxic neurotransmitters, immune mediators and oxygen radicals 10-12, which ultimately result in long-term neurological disabilities 13,14. Thus choosing appropriate animal models of TBI that present similar cellular and molecular events in human and rodent TBI is critical for studying the mechanisms underlying injury and repair.
Various experimental models of TBI have been developed to reproduce aspects of TBI observed in humans, among them three specific models are widely adapted for rodents: fluid percussion, cortical impact and weight drop/impact acceleration 1. The fluid percussion device produces an injury through a craniectomy by applying a brief fluid pressure pulse on to the intact dura. The pulse is created by a pendulum striking the piston of a reservoir of fluid. The percussion produces brief displacement and deformation of neural tissue 1,15. Conversely, cortical impact injury delivers mechanical energy to the intact dura via a rigid impactor under pneumatic pressure 16,17. The weight drop/impact model is characterized by the fall of a rod with a specific mass on the closed skull 18. Among the TBI models, LFP is the most established and commonly used model to evaluate mixed focal and diffuse brain injury 19. It is reproducible and is standardized to allow for the manipulation of injury parameters. LFP recapitulates injuries observed in humans, thus rendering it clinically relevant, and allows for exploration of novel therapeutics for clinical translation 20.
We describe the detailed protocol to perform LFP procedure in mice. The injury inflicted is mild to moderate, with brain regions such as cortex, hippocampus and corpus callosum being most vulnerable. Hippocampal and motor learning tasks are explored following LFP.

Lateral fluid percussion (LFP), an established model of traumatic brain injury in mice, is demonstrated. LFP fulfills three major criteria for animal models: validity, reliability and clinical relevance. The procedure, consisting of surgical craniotomy, fixation of hub followed by induction of injury, resulting in focal and diffuse injuries, is described.

横方向の流体のパーカッション：マウスにおける外傷性脳損傷のモデル

Traumatic brain injury (TBI) research has attained renewed momentum due to the increasing awareness of head injuries, which result in morbidity and ...

神経科学

A Mouse Model of Single and Repetitive Mild Traumatic Brain Injury

Mild Traumatic Brain Injury (mTBI) can result in the acute loss of brain function, including a period of confusion, a loss of consciousness (LOC), focal neurological deficits and even amnesia. Athletes participating in contact sports are at high risk of exposure to large number of mTBIs. In terms of the level of injury in a sporting athlete, a mTBI is defined as a mild injury that does not cause gross pathological changes, but does cause short-term neurological deficits that are spontaneously resolved. Despite previous attempts to model mTBI in mice and rats, many have reported gross adverse effects including skull fractures, intracerebral bleeding, axonal injury and neuronal cell death. Herein, we describe our highly reproducible animal model of mTBI that reproduces clinically relevant symptoms. This model uses a custom made pneumatic impactor device to deliver a closed-head trauma. This impact is made under precise velocity and deformation parameters, creating a reliable and reproducible model to examine the mechanisms that contribute to effects of single or repetitive concussive mTBI.

Athletes absorb several hundred mild traumatic brain injuries (mTBI)/concussions every year; however, the consequence of these on the brain is poorly understood. Therefore, an animal model of single and repetitive mTBI that consistently replicates clinically relevant symptoms provides the means to advance the study of mTBI and concussion.

片性および反復性軽度外傷性脳損傷のマウスモデル

Mild Traumatic Brain Injury (mTBI) can result in the acute loss of brain function, including a period of confusion, a loss of consciousness (LOC), focal ...

医学

A Novel and Translational Rat Model of Concussion Combining Force and Rotation with In Vivo Cerebral Microdialysis

Persistent cognitive and motor symptoms are known consequences of concussions/mild traumatic brain injury (mTBIs) that can be partly attributable to altered neurotransmission. Indeed, cerebral microdialysis studies in rodents have demonstrated an excessive extracellular glutamate release in the hippocampus within the first 10 min following trauma. Microdialysis offers the clear advantage of in vivo neurotransmitter continuous sampling while not having to sacrifice the animal. In addition to the aforementioned technique, a closed head injury model that exerts rapid acceleration and deceleration of the head and torso is needed, as such a factor is not available in many other animal models. The Wayne State weight-drop model mimics this essential component of human craniocerebral trauma, allowing the induction of an impact on the head of an unrestrained rodent with a falling weight. Our novel and translational rat model combines cerebral microdialysis with the Wayne State weight-drop model to study, in lightly anesthetized and unrestrained adult rats, the acute changes in extracellular neurotransmitter levels following concussion. In this protocol, the microdialysis probe was inserted inside the hippocampus as region of interest, and was left inserted in the brain at impact. There is a high density of terminals and receptors in the hippocampus, making it a relevant region to document altered neurotransmission following concussion. When applied to adult Sprague-Dawley rats, our combined model induced increases in hippocampal extracellular glutamate concentrations within the first 10 min, consistent with the previously reported post-concussion symptomology. This combined weight-drop model provides a reliable tool for researchers to study early therapeutic responses to concussions in addition to repetitive brain injury, since this protocol induces a closed-head mild trauma.

Neurotransmitter alteration is a mechanism of neural dysfunction that occurs after concussion and contributes to the sometimes-catastrophic long-term consequences. This rat model combines microdialysis, allowing in vivo neurotransmitter quantification, with a weight-drop technique exerting rapid acceleration and deceleration of the head and torso, an important factor of human craniocerebral trauma.

生体内脳微小透析における力と回転を組み合わせた脳震盪の新規および翻訳ラットモデル

Persistent cognitive and motor symptoms are known consequences of concussions/mild traumatic brain injury (mTBIs) that can be partly attributable to ...

Murine Model of Controlled Cortical Impact for the Induction of Traumatic Brain Injury

The Centers for Disease Control and Injury Prevention estimate that almost 2 million people sustain a traumatic brain injury (TBI) every year in the United States. In fact, TBI is a contributing factor to over a third of all injury-related mortality. Nonetheless, the cellular and molecular mechanisms underlying the pathophysiology of TBI are poorly understood. Thus, preclinical models of TBI capable of replicating the injury mechanisms pertinent to TBI in human patients are a critical research need. The controlled cortical impact (CCI) model of TBI utilizes a mechanical device to directly impact the exposed cortex. While no model can full recapitulate the disparate injury patterns and heterogeneous nature of TBI in human patients, CCI is capable of inducing a wide range of clinically applicable TBI. Furthermore, CCI is easily standardized allowing investigators to compare results across experiments as well as across investigative groups. The following protocol is a detailed description of applying a severe CCI with a commercially available impacting device in a murine model of TBI.

Here we describe a protocol for the induction of murine traumatic brain injury via an open-head controlled cortical impact.

外傷性脳損傷の誘導に対する制御された皮質影響のマウスモデル

The Centers for Disease Control and Injury Prevention estimate that almost 2 million people sustain a traumatic brain injury (TBI) every year in the ...

Low-intensity Blast Wave Model for Preclinical Assessment of Closed-head Mild Traumatic Brain Injury in Rodents

Traumatic brain injury (TBI) is a large-scale public health problem. Mild TBI is the most prevalent form of neurotrauma and accounts for a large number of medical visits in the United States. There are currently no FDA-approved treatments available for TBI. The increased incidence of military-related, blast-induced TBI further accentuates the urgent need for effective TBI treatments. Therefore, new preclinical TBI animal models that recapitulate aspects of human blast-related TBI will greatly advance the research efforts into the neurobiological and pathophysiological processes underlying mild to moderate TBI as well as the development of novel therapeutic strategies for TBI.
Here we present a reliable, reproducible model for the investigation of the molecular, cellular, and behavioral effects of mild to moderate blast-induced TBI. We describe a step-by-step protocol for closed-head, blast-induced mild TBI in rodents using a bench-top setup consisting of a gas-driven shock tube equipped with piezoelectric pressure sensors to ensure consistent test conditions. The benefits of the setup that we have established are its relative low-cost, ease of installation, ease of use and high-throughput capacity. Further advantages of this non-invasive TBI model include the scalability of the blast peak overpressure and the generation of controlled reproducible outcomes. The reproducibility and relevance of this TBI model has been evaluated in a number of downstream applications, including neurobiological, neuropathological, neurophysiological and behavioral analyses, supporting the use of this model for the characterization of processes underlying the etiology of mild to moderate TBI.

We present here a protocol of a blast wave model for rodents to investigate neurobiological and pathophysiological effects of mild to moderate traumatic brain injury. We established a gas-driven, bench-top setup equipped with pressure sensors allowing for reliable and reproducible generation of blast-induced mild to moderate traumatic brain injury.

げっ歯類における閉鎖性軽度外傷性脳損傷の前臨床評価のための低強度ブラスト波モデル

Traumatic brain injury (TBI) is a large-scale public health problem. Mild TBI is the most prevalent form of neurotrauma and accounts for a large number of ...

Electromagnetic Controlled Closed-Head Model of Mild Traumatic Brain Injury in Mice

Highly reproducible animal models of traumatic brain injury (TBI), with well-defined pathologies, are needed for testing therapeutic interventions and understanding the mechanisms of how a TBI alters brain function. The availability of multiple animal models of TBI is necessary to model the different aspects and severities of TBI seen in people. This manuscript describes the use of a midline closed head injury (CHI) to develop a mouse model of mild TBI. The model is considered mild because it does not produce structural brain lesions based on neuroimaging or gross neuronal loss. However, a single impact creates enough pathology that cognitive impairment is measurable at least 1 month after injury. A step-by-step protocol to induce a CHI in mice using a stereotaxically guided electromagnetic impactor is defined in the paper. The benefits of the mild midline CHI model include the reproducibility of the injury-induced changes with low mortality. The model has been temporally characterized up to 1 year after the injury for neuroimaging, neurochemical, neuropathological, and behavioral changes. The model is complementary to open skull models of controlled cortical impact using the same impactor device. Thus, labs can model both mild diffuse TBI and focal moderate-to-severe TBI with the same impactor.

The protocol describes mild traumatic brain injury in a mouse model. In particular, a step-by-step protocol to induce a mild midline closed head injury and the characterization of the animal model is fully explained.

マウスにおける軽度外傷性脳損傷の電磁制御クローズドヘッドモデル

Highly reproducible animal models of traumatic brain injury (TBI), with well-defined pathologies, are needed for testing therapeutic interventions and ...

クローズドヘッドケガ法によるマウスのrmTBI導入

Modified Mouse Model of Repetitive Mild Traumatic Brain Injury Incorporating Thinned-Skull Window and Fluid Percussion

Mild traumatic brain injury is a clinically highly heterogeneous neurological disorder. Highly reproducible traumatic brain injury (TBI) animal models with well-defined pathologies are urgently needed for studying the mechanisms of neuropathology after mild TBI and testing therapeutics. Replicating the entire sequelae of TBI in animal models has proven to be a challenge. Therefore, the availability of multiple animal models of TBI is necessary to account for the diverse aspects and severities seen in TBI patients. CHI is one of the most common methods for fabricating rodent models of rmTBI. However, this method is susceptible to many factors, including the impact method used, the thickness and shape of the skull bone, animal apnea, and the type of head support and immobilization utilized. The aim of this protocol is to demonstrate a combination of the thinned-skull window and fluid percussion injury (FPI) methods to produce a precise mouse model of CHI-associated rmTBI. The primary objective of this protocol is to minimize factors that could impact the accuracy and consistency of CHI and FPI modeling, including skull bone thickness, shape, and head support. By utilizing a thinned-skull window method, potential inflammation due to craniotomy and FPI is minimized, resulting in an improved mouse model that replicates the clinical features observed in patients with mild TBI. Results from behavior and histological analysis using hematoxylin and eosin (HE) staining suggest that rmTBI can lead to a cumulative injury that produces changes in both behavior and gross morphology of the brain. Overall, the modified CHI-associated rmTBI presents a useful tool for researchers to explore the underlying mechanisms that contribute to focal and diffuse pathophysiological changes in rmTBI.

著者スポットライト:軽度外傷性脳損傷の繰り返しにおけるメカニズムとリハビリテーションを調査するためのげっ歯類モデルの最適化

This protocol presents a modified mouse model of repetitive mild traumatic brain injury (rmTBI) induced via a closed-head injury (CHI) method. The approach features a thinned-skull window and fluid percussion to reduce the inflammation commonly caused by meninges exposure, along with improved reproducibility and accuracy in modeling rmTBI in rodents.

間伐頭蓋窓と流体パーカッションを組み込んだ反復性軽度外傷性脳損傷の改良マウスモデル

Mild traumatic brain injury is a clinically highly heterogeneous neurological disorder. Highly reproducible traumatic brain injury (TBI) animal models ...

<meta charset="utf8"></head><body>CHILDモデル — マウスにおける若年性脳震盪と長期転帰の再現

A Pediatric Concussion Model in Mice: Closed Head Injury with Long-Term Disorders (CHILD)

Concussions and mild traumatic brain injuries (mTBI) during childhood represent a significant health endangerment, with many patients later in life exhibiting debilitating physiological, neurological, and psychosocial outcomes. The cellular and molecular pathophysiological mechanisms are relatively unknown, and this significant gap effectively precludes investigations into specific therapeutic strategies to mitigate chronic sequelae. No single animal concussion model recapitulates all the features reported in human subjects, and current models are designed to answer specific research questions. We set out to develop a juvenile concussion that recapitulates clinical early, and long-term symptomology encountered, termed Closed Head Injury with Long-term Disorder (CHILD). In this model, a concussive force via an electromagnetic impactor is delivered to the head of a lightly anesthetized unrestrained post-natal day 17 (P17) mouse, allowing free rotation of the head. Mice are placed on a tightly stretched tin foil across a stereotactic device, and the impactor is carefully aligned with the targeted cortical region. The electromagnetic impactor facilitates the selection of impact depth, dwell, and duration to determine injury severity. A strength of this model is that it allows head rotation, an important clinical feature. After impact, mice are immediately monitored for righting time and time to explore their environment, followed by their return to their dam. Increasing the severity of concussion results in intracranial bleeds in a subset of mice. Mice can be routinely monitored for behavior and neuroimaging over their lifespan, as desired. Various injury severities mimic the heterogeneous nature of juvenile concussions. The current standard CHILD model exhibits no skull fracture, no observable conventional neuroimaging change (similar to clinical), but leads to progressive and persistent neuronal death, altered diffusion MRI, modified neuronal activity and plasticity, increased gliosis, and progressive behavioral perturbations with age. In summary, this CHILD model mimics the early and long-term features observed in many clinical concussion patients.

<meta charset="utf8"></head><body>マウスの小児脳震盪モデル:長期障害を伴う閉鎖性頭部外傷(CHILD)

Pediatric concussions can result in long-term sequelae with physiological and psychological perturbations. This pediatric concussion mouse model results in long-term neuroinflammation, white matter alterations, and neuronal modifications with behavioral dysfunction. The model can be adapted to differential impact locations or repeated concussions to assess lifetime changes.

マウスの小児脳震盪モデル:長期障害を伴う閉鎖性頭部外傷(CHILD) 

Concussions and mild traumatic brain injuries (mTBI) during childhood represent a significant health endangerment, with many patients later in life ...

マウスにおける反復震とう頭部損傷モデル

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横方向の流体のパーカッション：マウスにおける外傷性脳損傷のモデル

片性および反復性軽度外傷性脳損傷のマウスモデル

生体内脳微小透析における力と回転を組み合わせた脳震盪の新規および翻訳ラットモデル

外傷性脳損傷の誘導に対する制御された皮質影響のマウスモデル

げっ歯類における閉鎖性軽度外傷性脳損傷の前臨床評価のための低強度ブラスト波モデル

マウスにおける軽度外傷性脳損傷の電磁制御クローズドヘッドモデル

間伐頭蓋窓と流体パーカッションを組み込んだ反復性軽度外傷性脳損傷の改良マウスモデル

間伐頭蓋窓と流体パーカッションを組み込んだ反復性軽度外傷性脳損傷の改良マウスモデル

間伐頭蓋窓と流体パーカッションを組み込んだ反復性軽度外傷性脳損傷の改良マウスモデル

マウスの小児脳震盪モデル:長期障害を伴う閉鎖性頭部外傷(CHILD)