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>

<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 border="0" cellpadding="0" cellspacing="0" width="831">
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			<td style="width:188px;">Eagle Eye Anesthesia, Inc</td>
			<td style="width:137px;">Model 150&#160;</td>
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			<td style="width:137px;">Impact One Stereotaxic Impactor</td>
			<td>perform impaction</td>
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			<td height="21" style="height:21px;width:216px;">Digital Stereotaxic instrument</td>
			<td style="width:188px;">LeicaBiosystems</td>
			<td style="width:137px;">39462501</td>
			<td>mount mouse and positioning tips</td>
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			<td height="42" style="height:42px;width:216px;">Sicilone rubber-coated metal tip</td>
			<td style="width:188px;">Precision Tool &#38; Engineering, Gainesvill FL</td>
			<td style="width:137px;">custom-made</td>
			<td>impact tip</td>
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			<td height="21" style="height:21px;width:216px;">Lithium Ion All-in-One Trimmer</td>
			<td style="width:188px;">WAHL Home Products</td>
			<td style="width:137px;">9854-600</td>
			<td>shave mouse hair</td>
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			<td height="23" style="height:23px;width:216px;">paper clips</td>
			<td style="width:188px;">custom-made</td>
			<td style="width:137px;"></td>
			<td>probe tip</td>
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			<td style="width:188px;">MEDLINE</td>
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			<td>scrub head with saline</td>
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			<td height="21" style="height:21px;">Tissue Tek O.C.T.</td>
			<td style="width:188px;">ASKURA FINETEK USA INC</td>
			<td style="width:137px;">4583</td>
			<td>tissue embedding</td>
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			<td height="21" style="height:21px;width:216px;">anti-GFAP</td>
			<td style="width:188px;">Dako</td>
			<td style="width:137px;">CA93013</td>
			<td>antibody for IHC</td>
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			<td style="width:188px;">Sigma</td>
			<td style="width:137px;">F6136</td>
			<td>antibody for IHC</td>
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			<td style="width:137px;">PK-6100</td>
			<td>IHC detection system</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Permount Mounting Medium</td>
			<td style="width:188px;">Fisher Scientific</td>
			<td style="width:137px;">SP15-100</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Aperio XT ScanScope scanner</td>
			<td style="width:188px;">Leica Microsystems Inc,</td>
			<td style="width:137px;"></td>
			<td>slides scanning</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Leica AutoStainer XL</td>
			<td style="width:188px;">Leica the pathology Company</td>
			<td style="width:137px;">ST2010</td>
			<td>H&#38;E staining</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">DAB&#160;</td>
			<td style="width:188px;">sigma</td>
			<td style="width:137px;">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 ...

a-repetitive-concussive-head-injury-model-in-mice

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

The Spared Nerve Injury (SNI) Model of Induced Mechanical Allodynia in Mice

Peripheral neuropathic pain is a severe chronic pain condition which may result from trauma to sensory nerves in the peripheral nervous system. The spared nerve injury (SNI) model induces symptoms of neuropathic pain such as mechanical allodynia i.e. pain due to tactile stimuli that do not normally provoke a painful response [1]. 
The SNI mouse model involves ligation of two of the three branches of the sciatic nerve (the tibial nerve and the common peroneal nerve), while the sural nerve is left intact [2]. The lesion results in marked hypersensitivity in the lateral area of the paw, which is innervated by the spared sural nerve. The non-operated side of the mouse can be used as a control. The advantages of the SNI model are the robustness of the response and that it doesn&#x2019;t require expert microsurgical skills.
The threshold for mechanical pain response is determined by testing with von Frey filaments of increasing bending force, which are repetitively pressed against the lateral area of the paw [3], [4]. A positive pain reaction is defined as sudden paw withdrawal, flinching and/or paw licking induced by the filament. A positive response in three out of five repetitive stimuli is defined as the pain threshold. 
As demonstrated in the video protocol, C57BL/6 mice experience profound allodynia as early as the day following surgery and maintain this for several weeks.

The Spared Nerve Injury SNI Model of Induced Mechanical Allodynia in Mice

The Spared Nerve Injury animal model is described here as a mouse model of peripheral neuropathic pain following partial denervation of the sciatic nerve by lesioning the tibial and common peroneal nerve branches, leaving the remaining sural nerve intact. Behavioral modification resulting from mechanical allodynia is quantified by von Frey filaments.

マウスで誘導機械的異痛症の倹約神経損傷（SNI）モデル

Peripheral neuropathic pain is a severe chronic pain condition which may result from trauma to sensory nerves in the peripheral nervous system. The spared ...

医学

Controlled Cervical Laceration Injury in Mice

Use of genetically modified mice enhances our understanding of molecular mechanisms underlying several neurological disorders such as a spinal cord injury (SCI). Freehand manual control used to produce a laceration model of SCI creates inconsistent injuries often associated with a crush or contusion component and, therefore, a novel technique was developed. Our model of cervical laceration SCI has resolved inherent difficulties with the freehand method by incorporating 1) cervical vertebral stabilization by vertebral facet fixation, 2) enhanced spinal cord exposure, and 3) creation of a reproducible laceration of the spinal cord using an oscillating blade with an accuracy of &plusmn;0.01 mm in depth without associated contusion. Compared to the standard methods of creating a SCI laceration such as freehand use of a scalpel or scissors, our method has produced a consistent lesion. This method is useful for studies on axonal regeneration of corticospinal, rubrospinal, and dorsal ascending tracts.

A novel technique to create a reproducible in vivo model of cervical spinal cord laceration injury in the mouse is described. This technique is based on spine stabilization by fixation of the cervical facets and laceration of the spinal cord using an oscillating blade with an accuracy of &plusmn;0.01 mm.

マウスで制御子宮裂傷の傷害

Use of genetically modified mice enhances our  understanding of molecular mechanisms underlying several neurological disorders  such as a spinal cord ...

A Contusion Model of Severe Spinal Cord Injury in Rats

The translational potential of novel treatments should be investigated in severe spinal cord injury (SCI) contusion models. A detailed methodology is described to obtain a consistent model of severe SCI. Use of a stereotactic frame and computer controlled impactor allows for creation of reproducible injury. Hypothermia and urinary tract infection pose significant challenges in the post-operative period. Careful monitoring of animals with daily weight recording and bladder expression allows for early detection of post-operative complications. The functional results of this contusion model are equivalent to transection models. The contusion model can be utilized to evaluate the efficacy of both neuroprotective and neuroregenerative approaches.

A contusion model of severe spinal cord injury is described. Detailed pre-operative, operative and post-operative steps are described to obtain a consistent model.

ラットにおける重度の脊髄損傷挫傷モデル

The translational potential of novel treatments should be investigated in severe spinal cord injury (SCI) contusion models. A detailed methodology is ...

Ischemia-reperfusion Model of Acute Kidney Injury and Post Injury Fibrosis in Mice

Ischemia-reperfusion induced acute kidney injury (IR-AKI) is widely used as a model of AKI in mice, but results are often quite variable with high, often unreported mortality rates that may confound analyses. Bilateral renal pedicle clamping is commonly used to induce IR-AKI, but differences between effective clamp pressures and/or renal responses to ischemia between kidneys often lead to more variable results. In addition, shorter clamp times are known to induce more variable tubular injury, and while mice undergoing bilateral injury with longer clamp times develop more consistent tubular injury, they often die within the first 3 days after injury due to severe renal insufficiency. To improve post-injury survival and obtain more consistent and predictable results, we have developed two models of unilateral ischemia-reperfusion injury followed by contralateral nephrectomy. Both surgeries are performed using a dorsal approach, reducing surgical stress resulting from ventral laparotomy, commonly used for mouse IR-AKI surgeries. For induction of moderate injury BALB/c mice undergo unilateral clamping of the renal pedicle for 26 min and also undergo simultaneous contralateral nephrectomy. Using this approach, 50-60% of mice develop moderate AKI 24 hr after injury but 90-100% of mice survive. To induce more severe AKI, BALB/c mice undergo renal pedicle clamping for 30 min followed by contralateral nephrectomy 8 days after injury. This allows functional assessment of renal recovery after injury with 90-100% survival. Early post-injury tubular damage as well as post injury fibrosis are highly consistent using this model.

We describe models of moderate and severe ischemia-reperfusion-induced kidney injury in which the mice undergo unilateral renal pedicle clamping followed by simultaneous or delayed contralateral nephrectomy, respectively. These models consistently give rise to renal dysfunction and post-injury fibrosis, but injury severity and survival are dependent on mouse background, age and surgical equipment.

マウスにおける急性腎障害とポスト傷害線維の虚血再灌流モデル

Ischemia-reperfusion induced acute kidney injury (IR-AKI) is widely used as a model of AKI in mice, but results are often quite variable with high, often ...

Renal Ischaemia Reperfusion Injury: A Mouse Model of Injury and Regeneration

Renal ischaemia reperfusion injury (IRI) is a common cause of acute kidney injury (AKI) in patients and occlusion of renal blood flow is unavoidable during renal transplantation. Experimental models that accurately and reproducibly recapitulate renal IRI are crucial in dissecting the pathophysiology of AKI and the development of novel therapeutic agents. Presented here is a mouse model of renal IRI that results in reproducible AKI. This is achieved by a midline laparotomy approach for the surgery with one incision allowing both a right nephrectomy that provides control tissue and clamping of the left renal pedicle to induce ischaemia of the left kidney. By careful monitoring of the clamp position and body temperature during the period of ischaemia this model achieves reproducible functional and structural injury. Mice sacrificed 24 hr&#160;following surgery demonstrate loss of renal function with elevation of the serum or plasma creatinine level as well as structural kidney damage with acute tubular necrosis evident. Renal function improves and the acute tissue injury resolves during the course of 7 days following renal IRI such that this model may be used to study renal regeneration. This model of renal IRI has been utilized to study the molecular and cellular pathophysiology of AKI as well as analysis of the subsequent renal regeneration.

The mouse model of renal ischaemia reperfusion injury described here comprises of a right nephrectomy that provides control tissue and clamping of the left renal pedicle to induce ischaemia that results in acute kidney injury. This model uses a midline laparotomy approach with all steps performed via one incision.

腎臓虚血再灌流障害：障害のマウスモデルおよび再生

Renal ischaemia reperfusion injury (IRI) is a common cause of acute kidney injury (AKI) in patients and occlusion of renal blood flow is unavoidable ...

A Simple Critical-sized Femoral Defect Model in Mice

While bone has a remarkable capacity for regeneration, serious bone trauma often results in damage that does not properly heal. In fact, one tenth of all limb bone fractures fail to heal completely due to the extent of the trauma, disease, or age of the patient. Our ability to improve bone regenerative strategies is critically dependent on the ability to mimic serious bone trauma in test animals, but the generation and stabilization of large bone lesions is technically challenging. In most cases, serious long bone trauma is mimicked experimentally by establishing a defect that will not naturally heal. This is achieved by complete removal of a bone segment that is larger than 1.5 times the diameter of the bone cross-section. The bone is then stabilized with a metal implant to maintain proper orientation of the fracture edges and allow for mobility. 
Due to their small size and the fragility of their long bones, establishment of such lesions in mice are beyond the capabilities of most research groups. As such, long bone defect models are confined to rats and larger animals. Nevertheless, mice afford significant research advantages in that they can be genetically modified and bred as immune-compromised strains that do not reject human cells and tissue.
Herein, we demonstrate a technique that facilitates the generation of a segmental defect in mouse femora using standard laboratory and veterinary equipment. With practice, fabrication of the fixation device and surgical implantation is feasible for the majority of trained veterinarians and animal research personnel. Using example data, we also provide methodologies for the quantitative analysis of bone healing for the model.

Animal models are frequently employed to mimic serious bone injury in biomedical research. Due to their small size, establishment of stabilized bone lesions in mice are beyond the capabilities of most research groups. Herein, we describe a simple method for establishing and analyzing experimental femoral defects in mice.

マウスの簡単な重要なサイズの大腿欠陥モデル

While bone has a remarkable capacity for regeneration, serious bone trauma often results in damage that does not properly heal. In fact, one tenth of all ...

Facial Nerve Axotomy in Mice: A Model to Study Motoneuron Response to Injury

The goal of this surgical protocol is to expose the facial nerve, which innervates the facial musculature, at its exit from the stylomastoid foramen and either cut or crush it to induce peripheral nerve injury. Advantages of this surgery are its simplicity, high reproducibility, and the lack of effect on vital functions or mobility from the subsequent facial paralysis, thus resulting in a relatively mild surgical outcome compared to other nerve injury models. A major advantage of using a cranial nerve injury model is that the motoneurons reside in a relatively homogenous population in the facial motor nucleus in the pons, simplifying the study of the motoneuron cell bodies. Because of the symmetrical nature of facial nerve innervation and the lack of crosstalk between the facial motor nuclei, the operation can be performed unilaterally with the unaxotomized side serving as a paired internal control. A variety of analyses can be performed postoperatively to assess the physiologic response, details of which are beyond the scope of this article. For example, recovery of muscle function can serve as a behavioral marker for reinnervation, or the motoneurons can be quantified to measure cell survival. Additionally, the motoneurons can be accurately captured using laser microdissection for molecular analysis. Because the facial nerve axotomy is minimally invasive and well tolerated, it can be utilized on a wide variety of genetically modified mice. Also, this surgery model can be used to analyze the effectiveness of peripheral nerve injury treatments. Facial nerve injury provides a means for investigating not only motoneurons, but also the responses of the central and peripheral glial microenvironment, immune system, and target musculature. The facial nerve injury model is a widely accepted peripheral nerve injury model that serves as a powerful tool for studying nerve injury and regeneration.

We present a surgical protocol detailing how to perform a cut or crush axotomy on the facial nerve in the mouse. The facial nerve axotomy can be employed to study the physiological response to nerve injury and test therapeutic techniques.

マウスにおける顔面神経軸索切断：傷害に運動ニューロン応答を研究するためのモデル

The goal of this surgical protocol is to expose the facial nerve, which innervates the facial musculature, at its exit from the stylomastoid foramen and ...

Quantification of Neurovascular Protection Following Repetitive Hypoxic Preconditioning and Transient Middle Cerebral Artery Occlusion in Mice

Experimental animal models of stroke are invaluable tools for understanding stroke pathology and developing more effective treatment strategies. A 2 week protocol for repetitive hypoxic preconditioning (RHP) induces long-term protection against central nervous system (CNS) injury in a mouse model of focal ischemic stroke. RHP consists of 9 stochastic exposures to hypoxia that vary in both duration (2 or 4 hr) and intensity (8% and 11% O2). RHP reduces infarct volumes, blood-brain barrier (BBB) disruption, and the post-stroke inflammatory response for weeks following the last exposure to hypoxia, suggesting a long-term induction of an endogenous CNS-protective phenotype. The methodology for the dual quantification of infarct volume and BBB disruption is effective in assessing neurovascular protection in mice with RHP or other putative neuroprotectants. Adult male Swiss Webster mice were preconditioned by RHP or duration-equivalent exposures to 21% O2 (i.e. room air). A 60 min transient middle cerebral artery occlusion (tMCAo) was induced 2 weeks following the last hypoxic exposure. Both the occlusion and reperfusion were confirmed by transcranial laser Doppler flowmetry. Twenty-two hr after reperfusion, Evans Blue (EB) was intravenously administered through a tail vein injection. 2 hr later, animals were sacrificed by isoflurane overdose and brain sections were stained with 2,3,5- triphenyltetrazolium chloride (TTC). Infarcts volumes were then quantified. Next, EB was extracted from the tissue over 48 hr to determine BBB disruption after tMCAo. In summary, RHP is a simple protocol that can be replicated, with minimal cost, to induce long-term endogenous neurovascular protection from stroke injury in mice, with the translational potential for other CNS-based and systemic pro-inflammatory disease states.

This protocol describes repetitive hypoxic preconditioning, or brief exposures to systemic hypoxia that reduce infarct volumes and blood-brain barrier disruption following transient middle cerebral artery occlusion in mice. It also details dual quantification of infarct volume and blood-brain barrier disruption after stroke to assess the efficacy of neurovascular protection.

マウスにおける反復低酸素プレコンディショニングと過渡中大脳動脈閉塞後に神経血管保護の定量

Experimental animal models of stroke are invaluable tools for understanding stroke pathology and developing more effective treatment strategies. A 2 week ...

Photothrombosis-induced Focal Ischemia as a Model of Spinal Cord Injury in Mice

Spinal cord injury (SCI) is a devastating clinical condition causing permanent changes in sensorimotor and autonomic functions of the spinal cord (SC) below the site of injury. The secondary ischemia that develops following the initial mechanical insult is a serious complication of the SCI and severely impairs the function and viability of surviving neuronal and non-neuronal cells in the SC. In addition, ischemia is also responsible for the growth of lesion during chronic phase of injury and interferes with the cellular repair and healing processes. Thus there is a need to develop a spinal cord ischemia model for studying the mechanisms of ischemia-induced pathology. Focal ischemia induced by photothrombosis (PT) is a minimally invasive and very well established procedure used to investigate the pathology of ischemia-induced cell death in the brain. Here, we describe the use of PT to induce an ischemic lesion in the spinal cord of mice. Following retro-orbital sinus injection of Rose Bengal, the posterior spinal vein and other capillaries on the dorsal surface of SC were irradiated with a green light resulting in the formation of a thrombus and thus ischemia in the affected region. Results from histology and immunochemistry studies show that PT-induced ischemia caused spinal cord infarction, loss of neurons and reactive gliosis. Using this technique a highly reproducible and relatively easy model of SCI in mice can be achieved that would serve the purpose of scientific investigations into the mechanisms of ischemia induced cell death as well as the efficacy of neuroprotective drugs. This model will also allow exploration of the pathological changes that occur following SCI in live mice like axonal degeneration and regeneration, neuronal and astrocytic Ca2+ signaling using two-photon microscopy.

Photothrombosis is a minimally invasive and highly reproducible procedure to induce focal ischemia in the spinal cord and serves as a model of spinal cord injury in mice.

マウスの脊髄損傷のモデルとしての光血栓誘発性局所虚血

Spinal cord injury (SCI) is a devastating clinical condition causing permanent changes in sensorimotor and autonomic functions of the spinal cord (SC) ...

Induction of Accelerated Atherosclerosis in Mice: The "Wire-Injury" Model

Atherosclerosis is a proliferative fibro-inflammatory disease developing in the arterial wall, inducing a deficient blood flow or a lack of blood flow. Moreover, by rupture of the defective vascular wall, atherosclerosis induces occlusive thrombus formation, which represents the main cause of myocardial infarction or stroke and the most frequent cause of death. Despite the advances in the cardiovascular field, many questions remain unanswered, and additional basic research is essential to improve our understanding of the molecular mechanisms during atherosclerosis and its effects. Due to limited clinical studies, there is a need for representative animal models recreating atherosclerotic conditions such as neointima formation after stent implantation, balloon angioplasty, or endarterectomy. Since the mouse presents many advantages and is the most frequently used model for studying molecular processes, the current study proposes an invasive procedure of endothelial denudation, also known as the wire-injury model, which is representative of the human condition of neointima formation in arteries after revascularization procedures.

This study describes an invasive procedure for the induction of accelerated atherosclerosis in mice. In comparison to other methods using electric- or cryo-induced injury, mechanical-induced injury mimics the human condition of restenosis after revascularization therapies and is ideal for the study of the molecular mechanisms involved.

マウスにおける加速アテローム性動脈硬化症の誘導:ワイヤー傷害モデル

Atherosclerosis is a proliferative fibro-inflammatory disease developing in the arterial wall, inducing a deficient blood flow or a lack of blood flow. ...