The present study was supported by grants from Kidney Research UK (ST4/2011), the Cunningham Trust (CT11/14) and the Mrs EA Hogg's Charitable Trust.

注：動物実験は動物（科学的処置）法1986によって課せられたガイドラインおよび規制に準拠して実施された手続きは、滅菌（オートクレーブ処理）手術器具や消耗品を使用して行われた。最適な年齢た状態で、15週 - 腎臓IRIのマウスモデルがここに提示されている間は、8週齢の雄、それが再現可能通常は7歳までの男女どちらかのマウス系統のさまざまな実行することができますBALB / cマウスで実施した8週間。代表的な結果の項に示されたデータは、BALB / cマウスおよびFVBマウスの両方から得た。温めた生理食塩水の適用は、湿った腸および手術領域を保持するために使用されるが、流体の過剰なアプリケーションが重要である血清または血漿クレアチニン濃度の低下アーチファクトを引き起こし得るように注意深く監視され、最小に保たれるべきである実験読み出し。 1。動物の準備および開腹 &lt;OL&gt; <li>腹腔内に塩酸ケタミン（70 mg / kg）を、およびメデトミジン塩酸塩（1 mg / kg）をして、マウスを麻酔し、反射神経などつま先のピンチの損失によって、麻酔の深さを確認してください。得られた麻酔薬面の持続時間は4時間である。これは、全体の外科的処置を行うのに十分であり、まだ補足麻酔を必要としない。 </li><li> 、切開領域を取り巻く髪を削除するエリアは抜け毛がないことを確認し、希薄クロルヘキシジン液を用いて腹部の皮膚を消毒。 </li><li>仰臥位で加熱された手術用パッドの上にマウスを置き、低粘着性粘着テープを使用してパッドに上下の手足を固定します。上肢が肺の圧縮を防ぐために、通常の位置に維持されていることを確認します。手順では熱傷のためのマウスを監視します。可能であればそれは非電気熱源を使用することをお勧めします。 </li><li>鎮痛剤ブプレノルフィン塩酸塩（0.06 mg / kgの）を管理皮下および角膜乾燥を防ぐために、目に目の潤滑剤を塗布。鎮痛薬は、手術後の回復を助けるために手術前に投与される。 </li><li>組織分離ハサミやメスの刃を使用して、正中開腹切開を行い、ぶっきらぼう腹膜から肌を分離する。これは、皮膚と腹膜の創傷閉鎖の際に、別々に縫合することができます。無血管白線の切開を腹腔へのアクセスを与えることになる。 </li><li>手術領域の明確なビューを作成するには、リトラクターを挿入し、マウスを羽織る。 </li></ol> 2。尿管本部、右腎摘出<ol><li>静かに右の腎臓と尿管を露出させ、生理食塩水で湿らせたオートクレーブした滅菌綿棒を使用して腹腔の左側に向かって腸を押してください。乾燥を防ぐために湿らせたドレープと腸をカバーしています。 </li><li>角度のついたピンセットで右尿管を持ち上げます。 6/0 SILを使用して二度、右尿管を結紮K-編組縫合。長期的な実験のために、すべての腹部手術のための吸収性縫合糸を使用しています。 </li><li>縫合糸の間に尿管を分割。膀胱尿管接合部が膀胱から腹膜に漏れる尿を防ぐ必要がありながら、尿管は、長期実験中、手術後の漏洩に対する保護手段として連結する。 </li><li>右腎摘出術を行う際に簡単にアクセスを許可するように、ゆっくりと湿らせた綿棒を使用して、右の肝臓を押して右腎動脈と静脈を露出させ湿らせたガーゼを使用して所定の位置に保持する。 </li><li>ぶっきらぼうな限り右腎動脈と静脈、右腎の内側面に沿って結合組織と脂肪を解剖。 </li><li>慎重に血管の下に角度のついた鉗子をスライドさせることで腎動脈と静脈の下にチャネルを作成します。ヒントを閉じて静かにチャネルの形成を容易にするために開いたヒントを削除して下に鉗子を導く。</li><li>鉗子の先端がちょうど腎動脈と静脈上記の結合組織を通して見えるまで、ステップ2.6を繰り返します。一回目に見える優しく優しく結合組織を破壊するために角度のついた鉗子の別のセットの先端に対する鉗子の先端をこする。 </li><li>腎動脈と静脈を明確にアクセスできると、彼らは今閉塞することができます。慎重にヒントを閉じた状態で、腎動脈と静脈の下に角度のついた鉗子をスライドさせます。一度場所で鉗子の先端を開き、先端間の止血クリップアプリケータを導き、しっかりと腎動脈と腎臓に近い静脈に止血クリップを適用します。あるいは腎動脈と静脈は、6/0シルク編組縫合糸を用いて結紮することができる。 </li><li>今残っている接着性の結合組織を除去することができ、腎臓の近くに閉塞した腎動脈と静脈を分割します。 </li><li>肝臓を保持し、綿棒で腸を置き換えるために使用される任意のガーゼを取り除きます。 </li></ol><ol><li>そっと左腎臓と尿管を露出し、湿らせたドレープでカバーするために綿棒と腹腔の右側に向かって腸を押してください。必要であれば、膵臓はより容易なアクセスを可能にするために湿らせたガーゼで偏向させることができる。 </li><li>優しく結合組織が前方および後方左腎動脈と静脈をした後、尿管本部、右腎摘出ステップ2.4​​と同様の方法で船の下にチャンネルを作成破る鈍い切​​開を使用しています。 </li><li>腎動脈と静脈にマイクロserrafineクリップを適用することで、左腎臓の虚血を誘発する。成功した虚血は、視覚的に腎臓が徐々に均一な暗色化することによって確認することができる。左腎動脈および静脈に加えて、血管が時折腎臓を供給することができる。存在する場合、これらの追加の血管にも成功するために、マイクロserrafineクリップを使用して閉塞する必要があります全体腎臓の虚血を誘発する。 </li><li>腸を交換して、慎重に腸の虚血の妥協の結果につながる可能性が急激ねじれがないことを確認してください。一時的に、単一の縫合糸で腹膜を閉じます。 </li><li>虚血誘導の後にはすぐに虚血に必要な期間を37℃で体温を維持する制御装置に接続され、直腸サーミスタ恒温ブランケット上にマウスを置きます。腎障害や腎不全の所望の程度を誘導するために必要な虚血の適切な長さを定義するには、滴定を各菌株と術者のために実行することをお勧めします。 </li><li>まもなく虚血期間の終了前に腹膜を再度開き、慎重にクランプへのアクセスを可能にし、腎臓を表示するには、腸を配置。開創器の挿入は、ビデオに示すように、必要ではなく、単に表象的な目的のために実施した。 </li><li> CLを削除虚血期間の後アンペアは締結しています。すぐに除去した後に、腎臓は急速に成功した再灌流を示す健全な濃いピンクに濃い栗色から色が変わります。 </li><li>再び腸前6/0編組絹縫合糸を使用したブランケットステッチで腹膜を閉じるにねじれていないことを確認してください。その後、金属の皮膚クリップを用いて皮膚を閉じます。 </li><li>術後感染のリスクを最小限に抑えるために、このような外科領域へのヨウ素/アルコール溶液などの消毒を適用します。 </li></ol> 4。術後の回復とケア<ol><li>アチパメゾール塩酸塩（2 mg / kg）の皮下投与と部分的に逆麻酔し、1mlの皮下注射で流体を投与手術後の脱水を防ぐために生理食塩水を加温した。 </li><li>彼らは意識を回復するまで、慎重にマウスを監視、警告が表示され、自分自身を正しくすることができます。 </li><li> 29℃、LOCに保ったマウスは、加熱ボックスに回復することができ24時間、静かな環境でated。マウスは、麻酔に温度調節する能力の障害を持っているし、それは彼らが効果的な復旧を可能にするために高温で収納されていることが非常に重要です。湿らせた食品はまた、流体と栄養摂取を促進するために提供することができる。 </li><li>長期的な回収実験は、継続的な鎮痛薬を提供し、手術後7日間、皮膚のクリップを削除するため。 </li></ol> 5。機能的および構造的腎損傷と再生の評価<ol><li>血液は、実験終了時に安楽死の時点で実験中または心臓穿刺により尾静脈から採取してもよい。先に説明したように7生化学遠心分析に基づく方法クレアチナーゼ、および血中尿素窒素（BUN）を用いて、血清クレアチニンによって腎機能を測定する。正常な健康なマウスの腎機能がこのような腎不全のモデルを確立する前に決定する必要があることdependi著しく変化することができるヤッフェ反応およびクレアチニンを測定するためのクレアチナーゼ基づく方法例えば使用した分析方法のNGは異なる結果を与える。 </li><li>ヘマトキシリン​​およびエオシン（H＆E）または過ヨウ素酸シッフ（PAS）で染色したパラフィン包埋腎臓組織切片上の組織病理で腎の構造を調べます。各マウスについて外部髄質（OSOM）の外側のストライプ内200Xの倍率で10枚の画像 - 5の間にキャプチャします。この領域は、このモデルで負傷したように、それが低酸素症に非常に脆弱であるため、OSOMにおける損傷のレベルを評価する。 </li><li>急性尿細管壊死（ATN）または再生を測定するために、以下に詳述2スコアリングシステムのいずれかを使用します。これらのシステムで使用される分類の代表的な形態は、 図1に示されている。 </li><li> ATNを獲得する単純なバイナリシステムを使用してください。細胞の完全性と形態マークを基に、生（無傷の細胞形態学）または壊死（妥協細胞統合のいずれかとして細管を数えるgrity、異常な細胞形態や細胞の喪失）。 <ol><li>細管の合計数（壊死性細管の％）の割合として、壊死細管の数を表現しています。 </li></ol></li><li>細胞の完全性、形態および核数に基づいて、別のシステムを使用して再生を分類します。マークとカウント細管など、健康的な壊死性、負傷したか、次の手順に記載されている基準に従って回収することを含む。 <ol><li> （ステップ5.4で詳述）ATNスコアリングシステムと同様に危険にさらさ細胞の完全性を示す尿細管ながら健康な無傷の正常細胞とマーク細管は、異常な細胞形態や明白な細胞の損失は、壊死したとして採点されるべきである。 </li><li>負傷したように、彼らはいくつかの核を含む薄くなった細胞質を持っている場合細管を分類する。これとは対照的に、回復し、より正常な細胞形態を有するより多くの核を含む細管を指定します。 </li><li>総細管数の割合として各細管分類を表現しています。 </li></ol></li></ol>

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The authors have no competing or conflicting interests to disclose.

腎臓IRIが使用可能な特定の治療とAKIの重要な原因である。腎臓IRIの実験的研究は、マクロファージ、樹状細胞、リンパ球、制御性T細胞だけでなく、他の細胞および急性損傷と位相5,8癒しの両方の誘導におけるメディエーターの役割を示す以前の研究で非常に有益でした- 16。また、実験的腎IRIは、様々な治療薬4,17-19の効果を評価するために使用されている。 <p class...

虚血再灌流障害（IRI）は、腎臓、心臓や脳を含む複数臓器傷害の一般的なモードです。腎臓IRIが患者の急性腎障害（AKI）につながる可能性や特定の治療法はありません。 IRIの結果、AKIは自然免疫と獲得免疫応答の1の両方を含む複雑な病因を持っています。腎臓IRIの実験モデルは、AKIの病因と同様に、その後の日にわたって続いて起こるその後の腎再生に関与する重要な細胞やメディエーターを解剖する可能性を提供しています。さらに、疾患プロセスの際に新規な治療剤の効果を評価することができる。 腎臓IRIの実験モデルの全体的な目標は、ここで説明し、急性機能的および構造的腎障害を誘発することがある。一部の研究者は、両側IRI 2の誘導を伴うモデルを利用している。両側腎IRIモデルは、使い一方的RENですが、らIRIモデルは、手術時に実施されて右腎摘出術の利点を有する。右腎摘出組織は、そのいずれか又は誘導する遺伝子またはタンパク質の発現を抑制する前処理工程を含む研究における貴重な対照組織として機能する。例えば、我々はヘムアルギン酸（HA）24時間の腎臓IRI 3前の注射の前処理効果を評価するためにこのモデルを使用している。 IRIの前にHAによって細胞保護酵素ヘムオキシゲナーゼ-1（HO-1）の成功した誘導は、右腎摘出コントロール組織4で確認された。 HAは、HO-1依存性機構を介して部分的には、老齢マウスでは腎臓IRIを減少させた。同様に、我々は、腎臓IRI 5におけるマクロファージの役割を調べるためにマクロファージ枯渇研究においてモデルを使用している。右腎摘出組織の免疫組織化学的分析は、アブレーションの方法の有効性を確認するために使用することができる。右腎摘出組織は、そのための確認の両方に使用し、私のレベルを定量化することができますnductionまたは個々の実験動物における目的の分子の阻害。このモデルは、前の腎臓IRIの誘導遺伝子またはタンパク質などの発現を調節する薬物または他の化合物を使用している研究者にとって興味深いであろう。 他の研究は、腎臓へのアクセスにフランク切開を使用している。ここで説明するモデルは、右腎摘出術の両方を実行し、左腎臓の虚血再灌流を誘導するために、単一の正中腹部の手術を使用しています。この外科的アプローチは、腎茎クランプに続く腎茎や色の変更など、術野の優れた可視化を提供します。モデル4-6と当方の経験では、マウスはすぐにほぼ100％の生存率は手術から回復ことを示しています。 最後に、7日間にわたってモデルの動力学的分析は、このモデルは腎機能および管状の整合性の両方の回復を呈することを示す、と重要な管状の細胞増殖。

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renal ischaemia reperfusion injury: a mouse model of injury and regeneration

腎臓虚血再灌流障害（IRI）は、患者の急性腎臓損傷（AKI）の一般的な原因であり、腎臓の血流の閉塞は、腎移植の際に避けられない。正確かつ再現性腎IRIを再現実験モデルは、AKIの病態生理及び新規治療薬の開発の分析において重要である。ここで紹介するには、再現可能な、AKI、その結果、腎IRIのマウスモデルである。これは、左腎臓の虚血を誘発する左腎椎弓のコントロール組織とクランプを提供して右腎摘出術の両方を許可する1切開で手術のための正中開腹アプローチによって達成される。虚血期間中のクランプ位置及び体温を注意深く監視することによって、このモデルは、再現性の機能的および構造的損傷を達成する。マウスは、血清または血漿クレアチニン値の上昇だけでなくて、腎機能の喪失を実証手術後24時間を犠牲に構造明らか急性尿細管壊死とtural腎臓損傷。腎機能は改善し、急性組織損傷は、このモデルは、腎臓再生を研究するために使用することができるように、腎臓IRI次の7日間の過程の間に解決されます。腎臓IRIのこのモデルは、分子·細胞AKIの病態だけでなく、その後の腎再生の分析を研究するために利用されている。

細管障害および回復は、腎臓IRI後の組織切片のH＆E又はPAS染色により評価することができる。 OSOM内にある細管は、細胞の形態、整合性と核の数（ 図1）によると、健康的な怪我、壊死または回復に分類される。このモデルでは機能的および構造的損傷は虚血の持続期間に依存している。虚血の持続時間が（ 図2）の 2分ずつ増加されるように、血漿クレアチニンおよ?...

Watch this Scientific Journal Video about Renal Ischaemia Reperfusion Injury: A Mouse Model of Injury and Regeneration at JoVE.com

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

renal-ischaemia-reperfusion-injury-mouse-model-injury

Research

JoVE Journal

Medicine

49.3K ビュー.  University of Edinburgh. 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.

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

Normothermic Cardiac Arrest and Cardiopulmonary Resuscitation: A Mouse Model of Ischemia-Reperfusion Injury

Acute Kidney Injury (AKI) is a common, highly lethal, complication of critical illness which has a high mortality1-4 and which is most 
frequently caused by whole-body hypoperfusion.5,6 Successful reproduction of whole-body hypoperfusion in rodent models has been fraught with 
difficulty.7-9,9,10 Models which employ focal ischemia have repeatedly demonstrated results which do not translate to the clinical setting, and larger 
animal models which allow for whole body hypoperfusion lack access to the full toolset of genetic manipulation possible in the mouse.11,12 However, in recent 
years a mouse model of cardiac arrest and cardiopulmonary resuscitation has emerged which can be adapted to model AKI.13 This model reliably reproduces 
physiologic, functional, anatomic, and histologic outcomes seen in clinical AKI, is rapidly repeatable, and offers all of the significant advantages of a murine surgical 
model, including access to genetic manipulative techniques, low cost relative to large animals, and ease of use. Our group has developed extensive experience 
with use of this model to assess a number of organ-specific outcomes in AKI.14,15

A powerful model for perioperative and critical care related acute kidney injury is presented. Using whole body hypoperfusion induced by cardiac arrest it is possible to nearly replicate the histologic and functional changes of clinical AKI.

正常体温心停止と心肺蘇生法：虚血再灌流障害のマウスモデル

Acute Kidney Injury (AKI) is a common, highly lethal, complication of critical illness which has a high mortality1-4 and which is most 
frequently caused ...

医学

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 ...

A Neonatal Mouse Spinal Cord Compression Injury Model

Spinal cord injury (SCI) typically causes devastating neurological deficits, particularly through damage to fibers descending from the brain to the spinal cord. A major current area of research is focused on the mechanisms of adaptive plasticity that underlie spontaneous or induced functional recovery following SCI. Spontaneous functional recovery is reported to be greater early in life, raising interesting questions about how adaptive plasticity changes as the spinal cord develops. To facilitate investigation of this dynamic, we have developed a SCI model in the neonatal mouse. The model has relevance for pediatric SCI, which is too little studied. Because neural plasticity in the adult involves some of the same mechanisms as neural plasticity in early life1, this model may potentially have some relevance also for adult SCI. Here we describe the entire procedure for generating a reproducible spinal cord compression (SCC) injury in the neonatal mouse as early as postnatal (P) day 1. SCC is achieved by performing a laminectomy at a given spinal level (here described at thoracic levels 9-11) and then using a modified Yasargil aneurysm mini-clip to rapidly compress and decompress the spinal cord. As previously described, the injured neonatal mice can be tested for behavioral deficits or sacrificed for ex vivo physiological analysis of synaptic connectivity using electrophysiological and high-throughput optical recording techniques1. Earlier and ongoing studies using behavioral and physiological assessment have demonstrated a dramatic, acute impairment of hindlimb motility followed by a complete functional recovery within 2 weeks, and the first evidence of changes in functional circuitry at the level of identified descending synaptic connections1.

This article describes a method for generating a reproducible spinal cord compression injury (SCI) in the neonatal mouse. The model provides an advantageous platform for studying mechanisms of adaptive plasticity that underlie spontaneous functional recovery.

新生児マウス脊髄圧縮損傷モデル

Spinal cord injury (SCI) typically causes devastating neurological deficits, particularly through damage to fibers descending from the brain to the spinal ...

Murine Model of Intestinal Ischemia-reperfusion Injury

Intestinal ischemia is a life-threatening condition associated with a broad range of clinical conditions including atherosclerosis, thrombosis, hypotension, necrotizing enterocolitis, bowel transplantation, trauma and chronic inflammation. Intestinal ischemia-reperfusion (IR) injury is a consequence of acute mesenteric ischemia, caused by inadequate blood flow through the mesenteric vessels, resulting in intestinal damage. Reperfusion following ischemia can further exacerbate damage of the intestine. The mechanisms of IR injury are complex and poorly understood. Therefore, experimental small animal models are critical for understanding the pathophysiology of IR injury and the development of novel therapies.
Here we describe a mouse model of acute intestinal IR injury that provides reproducible injury of the small intestine without mortality. This is achieved by inducing ischemia in the region of the distal ileum by temporally occluding the peripheral and terminal collateral branches of the superior mesenteric artery for 60 min using microvascular clips. Reperfusion for 1 hr, or 2 hr after injury results in reproducible injury of the intestine examined by histological analysis. Proper position of the microvascular clips is critical for the procedure. Therefore the video clip provides a detailed visual step-by-step description of this technique. This model of intestinal IR injury can be utilized to study the cellular and molecular mechanisms of injury and regeneration.

Here we describe the detailed procedure of intestinal ischemia-reperfusion in mice which results in reproducible injury without mortality to encourage the standardization of this technique across the field. This model of intestinal ischemia-reperfusion injury can be utilized to study the cellular and molecular mechanisms of injury and regeneration.

腸管虚血 - 再灌流傷害のマウスモデル

Intestinal ischemia is a life-threatening condition associated with a broad range of clinical conditions including atherosclerosis, thrombosis, ...

A Mouse Model of Retinal Ischemia-Reperfusion Injury Through Elevation of Intraocular Pressure

Retinal ischemia-reperfusion (I/R) is a pathophysiological process contributing to cellular damage in multiple ocular conditions, including glaucoma, diabetic retinopathy, and retinal vascular occlusions. Rodent models of I/R injury are providing significant insights into mechanisms and treatment strategies for human I/R injury, especially with regard to neurodegenerative damage in the retinal neurovascular unit. Presented here is a protocol for inducing retinal I/R injury in mice through elevation of intraocular pressure (IOP). In this protocol, the ocular anterior chamber is cannulated with a needle, through which flows the drip of an elevated saline reservoir. Using this drip to raise IOP above systolic arterial blood pressure, a practitioner temporarily halts inner retinal blood flow (ischemia). When circulation is reinstated (reperfusion) by removal of the cannula, severe cellular damage ensues, resulting ultimately in retinal neurodegeneration. Recent studies demonstrate inflammation, vascular permeability, and capillary degeneration as additional elements of this model. Compared to alternative retinal I/R methodologies, such as retinal arterial ligation, retinal I/R injury by elevated IOP offers advantages in its&#160;anatomical specificity, experimental tractability, and technical accessibility, presenting itself as a valuable tool for examining neuronal pathogenesis and therapy in the retinal neurovascular unit.

This article describes a procedure for inducing retinal ischemia-reperfusion injury by elevated intraocular pressure in mice. Retinal ischemia-reperfusion injury by elevated intraocular pressure serves to model human pathologies characterized by compromised oxygen and nutrient delivery in the retina, enabling researchers to examine potential cellular mechanisms and treatments for human diseases of the retinal neurovascular unit.

眼圧の上昇を通じて、網膜虚血再灌流障害のマウスモデル

Retinal ischemia-reperfusion (I/R) is a pathophysiological process contributing to cellular damage in multiple ocular conditions, including glaucoma, ...

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 ...

Evaluation of Coronary Flow Reserve After Myocardial Ischemia Reperfusion in Rats

Coronary artery disease is the leading cause of death worldwide. After an acute myocardial infarction, early and successful myocardial intervention via recanalization of the coronary artery is the most effective strategy for reducing the size of ischemic myocardium. The coronary microvasculature cannot be visualized and imaged&#160;in vivo, but there are several invasive and noninvasive techniques that can be used to assess parameters which depend directly on coronary microvascular function. The endothelial function after ischemia reperfusion can be assessed also at the level of the coronary circulation via the coronary flow reserve (CFR).&#160;In this study, peak velocity of left anterior descending (LAD) coronary arteries was measured in rats in vivo via Transthoracic Doppler Echocardiography during resting and stress challenge (induced by Dobutamine). A normal heart can increase its coronary blood flow up to four times above the resting values during stress induction. Following ischemia reperfusion, we found a significantly diminished CFR, which can be used as a marker of coronary microvascular dysfunction. CFR has opened a window on the importance of microvascular dysfunction and has been shown to predict cardiovascular risk independent of whether the severe obstructive disease is present.

Coronary flow reserve (CFR), is defined as the ratio of maximal coronary blood flow to the resting coronary blood flow. We present a protocol for evaluating CFR in rats via ultrasound, which offers the opportunity to predict cardiovascular risk factors in the absence of obstructive coronary disease.

ラットにおける心筋虚血再灌流後の冠動脈流量の評価

Coronary artery disease is the leading cause of death worldwide. After an acute myocardial infarction, early and successful myocardial intervention via ...

A Pre-clinical Rat Model for the Study of Ischemia-reperfusion Injury in Reconstructive Microsurgery

Ischemia-reperfusion injury is the main cause of flap failure in reconstructive microsurgery. The rat is the preferred preclinical animal model in many areas of biomedical research due to its cost-effectiveness and its translation to humans. This protocol describes a method to create a preclinical free skin flap model in rats with ischemia-reperfusion injury. The described 3 cm x 6 cm rat free skin flap model is easily obtained after the placement of several vascular ligatures and the section of the vascular pedicle. Then, 8 h after the ischemic insult and completion of the microsurgical anastomosis, the free skin flap develops the tissue damage. These ischemia-reperfusion injury-related damages can be studied in this model, making it a suitable model for evaluating therapeutic agents to address this pathophysiological process. Furthermore, two main monitoring techniques are described in the protocol for the assessment of this animal model: transit-time ultrasound technology and laser speckle contrast analysis.

Here, we describe a pre-clinical animal model for studying the pathophysiology of ischemia-reperfusion injury in reconstructive microsurgery. This free skin flap model based on the superficial caudal epigastric vessels in the rat may also allow for the evaluation of different therapies and compounds to counteract ischemia-reperfusion injury-related damage.

再建微小外科における虚血再灌流損傷の研究のための前臨床ラットモデル

Ischemia-reperfusion injury is the main cause of flap failure in reconstructive microsurgery. The rat is the preferred preclinical animal model in many ...

Delayed Intramyocardial Delivery of Stem Cells after Ischemia Reperfusion Injury in a Murine Model

There is significant interest in the use of stem cells (SCs) for the recovery of cardiac function in individuals with myocardial injuries. Most commonly, cardiac stem cell therapy is studied by delivering SCs concurrently with the induction of myocardial injury. However, this approach presents two significant limitations: the early hostile pro-inflammatory ischemic environment may affect the survival of transplanted SCs, and it does not represent the subacute infarction scenario where SCs will likely be used. Here we describe a two-part series of surgical procedures for the induction of ischemia-reperfusion injury and delivery of mesenchymal stem cells (MSCs). This method of stem cell administration may allow for the longer viability and retention around damaged tissue by circumventing the initial immune response. A model of ischemia reperfusion injury was induced in mice accompanied by the delivery of mesenchymal stem cells (3.0 x 105), stably expressing the reporter gene firefly luciferase under the constitutively expressed CMV promoter, intramyocardially 7 days later. The animals were imaged via ultrasound and bioluminescent imaging for confirmation of injury and injection of cells, respectively. Importantly, there was no added complication rate when performing this two-procedure approach for SC delivery. This method of stem cell administration, collectively with the utilization of state-of-the-art reporter genes, may allow for the in vivo study of viability and retention of transplanted SCs in a situation of chronic ischemia commonly seen clinically, while also circumventing the initial pro-inflammatory response. In summary, we established a protocol for the delayed delivery of stem cells into the myocardium, which can be used as a potential new approach in promoting regeneration of the damaged tissue.

Stems cells are continuously investigated as potential treatments for individuals with myocardial damage, however, their decreased viability and retention within injured tissue can impact their long-term efficacy. In this manuscript we describe an alternative method for stem cell delivery in a murine model of ischemia reperfusion injury.

マウスモデルにおける虚血再灌流損傷後の幹細胞の筋内内出産の遅延

There is significant interest in the use of stem cells (SCs) for the recovery of cardiac function in individuals with myocardial injuries. Most commonly, ...

A Rat Lung Transplantation Model of Warm Ischemia/Reperfusion Injury: Optimizations to Improve Outcomes

From our experience with rat lung transplantation, we have found several areas for improvement. Information in the existing literature regarding methods for choosing appropriate cuff sizes for the pulmonary vein (PV), pulmonary artery (PA), or bronchus (Br) are varied, thus making the determination of proper cuff size during rat lung transplantation an exercise of trial and error. By standardizing the cuffing technique to use the smallest effective cuff appropriate for the size of the vessel or bronchus, one can make the transplantation procedure safer, faster, and more successful. Since diameters of the PV, PA, and Br are related to the body weight of the rat, we present a strategy to choosing an appropriate size using a weight-based guide. Since lung volume is also related to body weight, we recommend that this relationship should also be considered when choosing the proper volume of air for donor lung inflation during warm ischemia as well as for the proper volume of PBS to be instilled during bronchoalveolar lavage (BAL) fluid collection. We also describe methods for 4th intercostal space dissection, wound closure, and sample collection from both the native and transplanted lobes.

Here, we present optimizations to a rat lung transplantation model that serve to improve outcomes. We provide a size guide for cuffs based on body weight, a measurement strategy to ascertain the 4th intercostal space, and methods of wound closure and BAL (bronchoalveolar lavage) fluid and tissue collection.

温虚血/再灌流損傷のラット肺移植モデル:転帰を改善するための最適化

From our experience with rat lung transplantation, we have found several areas for improvement. Information in the existing literature regarding methods ...