Name: quantification of neurovascular protection following repetitive hypoxic preconditioning and transient middle cerebral artery occlusion in mice
Uploaded: 2015-05-04T12:00:00.000+00:00
Duration: 9 min 48 s
Description: Watch this Scientific Journal Video about Quantification of Neurovascular Protection Following Repetitive Hypoxic Preconditioning and Transient Middle Cerebral Artery Occlusion in Mice at JoVE.com

Special thanks to the Gidday lab for their work in developing the RHP protocol, as well as the Neuro-Models Facility (UTSW) for their assistance in the tMCAo surgeries. This work was supported by grants from the American Heart Association (AMS), The Haggerty Center for Brain Injury and Repair (UTSW; AMS), and The Spastic Paralysis Research Foundation of the Illinois-Eastern Iowa District of Kiwanis International (JMG).

The authors have nothing to disclose.

全身低酸素症マウスでの（ すなわち、2時間11％O 2の）への単回暴露は「一過性」 -短い持続である低酸素プレコンディショニングの課題へのエピジェネティックな応答を意味tMCAo、29から脳を保護し、ベースラインの表現型は、内に復元され、日。低酸素プレコンディショニング刺激の繰り返しプレゼンテーションは劇的に神経保護表現型の持続時間を延ばす。6</sup...

成人障害と死因の第4位の主要な原因としては、脳卒中は、米国の成人人口が直面する最も衰弱性疾患状態の一つである。脳卒中の1動物モデルは、虚血性傷害を軽減する新しい方法の実験的調査を可能にし、脳卒中後の回復を改善します。そのようなトランスレーショナルリサーチのための新規な一道がプリコンディショニングされます。前処理は、その後、より重度の傷害からの損傷を低減するために、非損傷刺激の意図的な使用である。2低酸素プレコンディショニングは、 インビボおよびインビトロ試験の両方において脳卒中に対する保護を提供する脳内の多面的な変化をもたらすことが示されています。低比重低酸素しかし、低酸素に一回の露光のみ成体マウスにおける虚血に対する耐性未満の72時間の誘導、短期的な神経保護を提供しています3。でも14時間毎日のエクスポージャーの4週間後の4、Linら。 FOウントその神経保護は、一週間持続した。5繰り返し低酸素プレコンディショニング（RHP）頻度、期間、および低酸素曝露の強度の確率的変動によって特徴付けられます。単一プレコンディショニング課題とは対照的に、RHPはマウスで8週間まで持続脳保護表現型を誘導する。6 RHP、最終低酸素暴露後週間梗塞体積、血液脳関門（BBB）破壊、血管炎症、および白血球漏出を減少。 RHPは、具体的には、虚血性半球におけるB細胞集団を維持しながら、T細胞、単球、およびマクロファージ集団を減少させることにより、虚血性脳の炎症を減少させた。 図7は、実際には、RHPは、脳卒中を含む、任意のCNS ​​傷害の前に、マウスにおける免疫表現型を誘導しました。 RHP処理された健康なマウスから単離したRHP処理されたB細胞は、抗原提示および抗体産生の両方のダウンレギュレーションと、ユニークな抗炎症性の表現型を示しました。ザ炎症誘発性適応免疫機構の全体的な減少だけでなく、中枢神経系に特異的な炎症性疾患のための内因性の免疫抑制を誘導するためにRHPに優れた方法論になりますが、また前炎症性病変を含む全身損傷または疾患モデル。 RHPは一過性の中大脳動脈閉塞（tMCAo）以下の梗塞体積およびBBB破壊の両方を低減します。このような一般的に使用されるtMCAoとして脳卒中の動物モデルは、劇的に脳卒中の病態生理の理解だけでなく、より効果的な神経治療薬の設計を改善します。まず、小泉らによって開発された1986年に、8 tMCAo手順は、広く使用されているげっ歯類において脳卒中を誘導する方法と、再灌流後の炎症を調査するための好ましい方法の一つです。 tMCAoための方法が進化するにつれて、シリコーンコートフィラメントのより最近の使用は、他のモデル9,10 &lt;と比較して、くも膜下出血の危険性を減らします/ SUP&gt;残念ながらtMCAoは、多くの場合、梗塞量に大きなばらつきを生成してもして、信頼性を向上させる。11-13これらの研究のほとんどは、2,3,5-トリフェニルクロリド（TTC）で染色することによって冠状脳切片における梗塞領域の輪郭を描く、考え梗塞定量化のためのゴールドスタンダードは、鮮やかな、複製可能な結果を​​生成するための簡単​​で安価な方法であるためです。 TTCは、ミトコンドリア内に存在する脱水素酵素の基質として機能します。脳切片をTTC溶液にさらされると、ホルマザン、非水溶性の還元生成物は、生存可能なミトコンドリアの深い赤色の沈殿ところ、TTCを選択的に生細胞内に取り込まれます。そのため、虚血組織におけるミトコンドリア機能障害のため、この組織が ​​損傷を受け、健康な組織の分化を可能にする、白のまま。14 RHPはまた、虚血性半球におけるBBB破壊を減少させる。6したがって、同じB内のBBBの整合性の二重の定量化をTTCベース梗塞ボリュームと雨は、内因性の保護の完全な有効性に関する有用な情報、および未処理と処理された動物におけるBBB破壊および梗塞の間の潜在的な因果関係を提供する15の測定します。脳卒中の二次破砕BBBを介して末梢血の流入は、最終的には虚血性脳卒中の患者における感染と死亡率を増加させ、虚血性半球で白血球集団、炎症性サイトカイン、酸化ストレス、血管原性浮腫、出血性転換を増加させます。16,17動物モデルにおいてBBB破壊を測定する一般的な方法は、脳へのエバンスブルー（EB）色素漏出の定量化によるものである。15,18-21 EBを選択的にアルブミン、球状タンパク質を血清に結合する（MW = 65kDaの）それは無傷の動物でBBBを通過していません。22虚血性脳卒中後、EBは、脳に浸透し、光学濃度withiの測定を可能にする、620 nmで蛍光を発しますnは灌流が実質を負傷。22光学密度はEBは経心臓灌流によって死後皮質血管系から洗い流されたBBBの透過性に正比例します。 EBを投与した動物におけるTTC染色脳の即時処理により、梗塞容積及びBBB破壊の両方を効果的に定量することができます。これは、神経損傷およびBBBの破壊は、脳卒中後の脳の同時処理ではないことに留意すべきである、23,24ので屠殺時の選択は重要な考慮事項です。 詳細RHP方法、そのモデルヒト患者において、中大脳動脈閉塞、神経および血管脳卒中損傷エンドポイントを決定するための二重の組織学的方法を、一時的な動脈閉塞を誘導するためのtMCAo方法を以下のプロトコル。 TTCは、全体の梗塞体積の定量化を可能にする、細胞死および累積的組織損傷を測定します梅、EBはBBB損傷の半球定量化を提供します。

<table><tbody><tr><td>Flowmeters, regulators</td><td>VetEquip, Inc</td><td>Specialty order</td><td>Four flowmeters are attached to 6.0 mm flexible PVC tubing which connects to the inlet port on each induction chamber with a plastic female connector. These flowmeters are bolted to a 6.5&quot; x 1&quot; x 1&quot; metal bar. This metal bar is bolted to a MI-246-P pressure gauge with a DISS outlet. This pressure gauge and flowmeter equipment can be attached to each new gas cylinder with a wrench.</td></tr><tr><td>21% O&lt;sub&gt;2&lt;/sub&gt; tank</td><td>AirGas</td><td>OX USP200</td><td></td></tr><tr><td>11% O&lt;sub&gt;2 &lt;/sub&gt;tank</td><td>AirGas</td><td>Specialty order</td><td></td></tr><tr><td>8% O&lt;sub&gt;2&lt;/sub&gt; tank&amp;nbsp;</td><td>AirGas</td><td>Specialty order</td><td></td></tr><tr><td>15 L induction chambers</td><td>VetEquip</td><td>941454</td><td></td></tr><tr><td>Moor Laber Dopper Flow&amp;nbsp;</td><td>Moor Instruments&amp;nbsp;</td><td>moorVMS-LDF1-HP</td><td>0.8 mm diameter probe&amp;nbsp;</td></tr><tr><td>High Intensity Illuminator&amp;nbsp;</td><td>Nikon</td><td>NI-150</td><td></td></tr><tr><td>Zoom Stereo Microscope&amp;nbsp;</td><td>NIkon</td><td>SMZ800</td><td>Other surgical microscopes may be used.&amp;nbsp;</td></tr><tr><td>Kent Scientific Right Temperature CODA</td><td>Kent Scientific Corporation</td><td>Discontinued</td><td>Recommended replacement is PhysioSuite with RightTemp Temperature Monitoring and Homeothermic Control (Kent Scientific, #PS-RT).</td></tr><tr><td>Hovabator Incubator</td><td>Stromberg&#039;s</td><td>2362-E</td><td>Our model is the 2362N. 2362E is a later model and includes an electronic thermostat.&amp;nbsp;</td></tr><tr><td>V010 Anesthesia system&amp;nbsp;</td><td>VetEquip</td><td>901807</td><td>Includes: ten foot high-pressure oxygen hose, frame, flowmeter, oxygen flush assembly, vaporizer, breathing circuit, chamber, nosecones, waste gas evacuation tubing and two VapoGuard filters</td></tr><tr><td>250 ml isoflurane&amp;nbsp;</td><td>Butler Schein</td><td>NDC-11695</td><td></td></tr><tr><td>D-6 Vet Trim Animal Cordless Trimmer&amp;nbsp;</td><td>Andis&amp;nbsp;</td><td>#23905</td><td>Replacement blades are available from Andis (#23995)</td></tr><tr><td>Betadine&amp;nbsp;</td><td>Fisher Scientific</td><td>19-898-867&amp;nbsp;</td><td></td></tr><tr><td>Q-tips</td><td>Multiple sellers&amp;nbsp;</td><td>Catalog number not available&amp;nbsp;</td><td></td></tr><tr><td>Gauze Pads</td><td>Fisher Scientific</td><td>67622</td><td></td></tr><tr><td>Surgical drape</td><td>Fisher Scientific</td><td>GM300&amp;nbsp;</td><td></td></tr><tr><td>Silk Sutures&amp;nbsp;</td><td>Look/Div Surgical Specialties</td><td>SP115</td><td></td></tr><tr><td>Nylon Sutures</td><td>Look/Div Surgical Specialties</td><td>SP185</td><td></td></tr><tr><td>Durmont #5 forceps (2)&amp;nbsp;</td><td>Fine Science Tools&amp;nbsp;</td><td>11251-35</td><td>Angled 45&amp;deg;</td></tr><tr><td>Surgical Scissors</td><td>Fine Science Tools&amp;nbsp;</td><td>14028-10</td><td></td></tr><tr><td>3 mm Vannas</td><td>Kent Scientific Corporation</td><td>INS600177</td><td>Straight blade</td></tr><tr><td>Hartman Hemostats&amp;nbsp;</td><td>Fine Scientific Tools</td><td>13002-10</td><td></td></tr><tr><td>Occluding filaments</td><td>Washington University</td><td>Specialty order</td><td>Filaments are silicone coated at Washington Univeristy and provided to UTSW facilities for a fee.&amp;nbsp;</td></tr><tr><td>Evans Blue</td><td>Sigma Aldritch</td><td>E2129-10G</td><td></td></tr><tr><td>Filter Paper&amp;nbsp;</td><td>Sigma Aldritch</td><td>WHA1001150</td><td>150 mm, circles, Grade 1&amp;nbsp;</td></tr><tr><td>Weigh Boats</td><td>Fisher Scientific</td><td>02-202-101</td><td>2.5&quot; diameter</td></tr><tr><td>0.9% Sodium Chloride Injection USP&amp;nbsp;</td><td>Baxter Pharmaceutics&amp;nbsp;</td><td>2B1321</td><td></td></tr><tr><td>0.3 cc insulin syringe with 29 gauge needle</td><td>Becton Dickinson Labware</td><td>309301</td><td></td></tr><tr><td>Flat bottom restrainer&amp;nbsp;</td><td>Braintree Scientific&amp;nbsp;</td><td>FB&amp;nbsp;M</td><td>2.0&quot; diameter</td></tr><tr><td>TTC</td><td>Sigma</td><td>T8877</td><td></td></tr><tr><td>10x PBS, pH 7.4</td><td>Fisher Scientific</td><td>BP399-20</td><td></td></tr><tr><td>Water Bath</td><td>Multiple sellers&amp;nbsp;</td><td>Catalog number not available&amp;nbsp;</td><td>Scintillation tubes with TTC may be manually held under running warm water as an alternative to the water bath.</td></tr><tr><td>Styrofoam board</td><td>Multiple sellers&amp;nbsp;</td><td>Catalog number not available&amp;nbsp;</td><td></td></tr><tr><td>Large Syringe Kit</td><td>PumpSystems Inc</td><td>P-SYRKIT-LG</td><td></td></tr><tr><td>Perfusion Pump</td><td>PumpSystems Inc</td><td>NE-300&amp;nbsp;</td><td></td></tr><tr><td>60 cc syringe</td><td>Fisher Scientific</td><td>NC9203256</td><td></td></tr><tr><td>27 gauge winged infusion set</td><td>Kawasumi Laboratories, Inc</td><td>D3K1-25G 1</td><td></td></tr><tr><td>20 ml scintillation vial</td><td>Fisher Scientific</td><td>50-367-126</td><td></td></tr><tr><td>Stainless steel spatula</td><td>Fisher Scientific</td><td>14-373-25A</td><td></td></tr><tr><td>Alto acrylic 1.0 mm mouse brain, coronal</td><td>CellPoint Scientific&amp;nbsp;</td><td>Catalog number not available&amp;nbsp;</td><td></td></tr><tr><td>0.21 mm stainless steel blades, 25 pk</td><td>CellPoint Scientific&amp;nbsp;</td><td>Catalog number not available&amp;nbsp;</td><td>Reusable cryostat blades are an inexpensive alternative.</td></tr><tr><td>4% paraformaldehyde</td><td>Santa Cruz Biotechnology&amp;nbsp;</td><td>SC-281692</td><td></td></tr><tr><td>Superfrost microscope slides&amp;nbsp;</td><td>Fisher Scientific</td><td>12-550-15</td><td></td></tr><tr><td>HP Scanjet G4050</td><td>Multiple sellers&amp;nbsp;</td><td>Catalog number not available&amp;nbsp;</td><td>Other commercial scanners are suitable for this step in the protocol.</td></tr><tr><td>ImageJ&amp;nbsp;</td><td>National Institute of Health</td><td>Catalog number not available&amp;nbsp;</td><td></td></tr><tr><td>Analytical Balance</td><td>Mettler Toledo&amp;nbsp;</td><td>XSE 205U</td><td></td></tr><tr><td>Precision Compact Oven&amp;nbsp;&amp;nbsp;</td><td>Thermo Scientific&amp;nbsp;</td><td>PR305225M</td><td></td></tr><tr><td>1.7 ml microcentrifuge tubes (Eppendorfs)</td><td>Denville Scientific&amp;nbsp;</td><td>C2170</td><td></td></tr><tr><td>Formamide</td><td>Fisher Scientific</td><td>BP228-100</td><td></td></tr><tr><td>96-well plates</td><td>Fisher Scientific</td><td>07-200-9</td><td></td></tr><tr><td>Epoch Microplate Spectrophotometer&amp;nbsp;</td><td>BioTek&amp;nbsp;</td><td>Catalog number not available&amp;nbsp;</td><td></td></tr></tbody></table>

quantification of neurovascular protection following repetitive hypoxic preconditioning and transient middle cerebral artery occlusion in mice

脳卒中の実験動物モデルは、ストロークの病態を理解し、より効果的な治療戦略を開発するための貴重なツールです。反復的な低酸素プレコンディショニング（RHP）2週間のプロトコルは、焦点虚血性脳卒中のマウスモデルにおいて、中枢神経系（CNS）損傷に対する長期保護を誘導します。 RHPは期間（2または4時間）と強度（8％と11％のO 2）の両方に変化する低酸素症に9確率的エクスポージャーで構成されています。 RHPは、内因性CNS-保護表現型の長期的な誘導を示唆し、梗塞体積、血液脳関門（BBB）の破壊、および低酸素への最後の暴露後の数週間のための脳卒中後の炎症反応を減少させます。梗塞体積及びBBB破壊の二重定量化のための方法は、RHPまたは他の推定上の神経保護剤を有するマウスにおける神経血管保護を評価するのに有効です。成人男性のスイスウェブスターマウスを21％OにRHPまたは期間と同等の曝露によって予め調整しました。 <suB&gt; 2（ すなわち、室内空気）。 60分一過中大脳動脈閉塞（tMCAo）は、最後の低酸素曝露後の2週間誘導しました。閉塞および再灌流の両方が経頭蓋レーザドップラー流量測定によって確認しました。再灌流後の二十二時間、エバンスブルー（EB）を尾静脈注射を介して投与しました。 2時間後、動物をイソフルランの過剰投与、脳切片を2,3,5-塩化トリフェニルテトラゾリウム（TTC）で染色したことにより屠殺しました。梗塞容積を定量しました。次に、EBをtMCAo後にBBB破壊を決定するために、48時間かけて組織から抽出しました。要約すると、RHPは他のCNSベースおよびプロ炎症性全身疾患状態のための翻訳可能性のある、マウスにおける脳卒中損傷から長期内因性神経血管保護を誘導するために、最小限のコストで、複製することができる単純なプロトコルです。

本研究では、10週齢がRHPに無作為化の開始時であった（N = 10）25の雄Swiss Websterマウスまたは21％のO 2（N = 15）のグループが含まれていました。二週間最終RHP曝露後、外科的処置は盲検基により行い、日の間相殺します。 tMCAoに続いて、1マウスは、術後の回復中に死亡し、再灌流CBF基準を満たしていなかったので、1マウスは研究から除外しました。両方除外マウスは、21％O 2群?...

Watch this Scientific Journal Video about Quantification of Neurovascular Protection Following Repetitive Hypoxic Preconditioning and Transient Middle Cerebral Artery Occlusion in Mice at JoVE.com

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

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 ...

quantification-neurovascular-protection-following-repetitive-hypoxic

Research

JoVE Journal

Medicine

13.7K ビュー.  University of Texas Southwestern Medical Center. 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.

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

Bilateral Common Carotid Artery Occlusion as an Adequate Preconditioning Stimulus to Induce Early Ischemic Tolerance to Focal Cerebral Ischemia

There is accumulating evidence, that ischemic preconditioning - a non-damaging ischemic challenge to the brain - confers a transient protection to a subsequent damaging ischemic insult. We have established bilateral common carotid artery occlusion as a preconditioning stimulus to induce early ischemic tolerance to transient focal cerebral ischemia in C57Bl6/J mice. In this video, we will demonstrate the methodology used for this study.

There is accumulating evidence, that ischemic preconditioning (PC) &#x2013; a non-damaging ischemic challenge to the brain - confers a transient protection to a subsequent damaging ischemic insult. We established bilateral common carotid artery occlusion (BCCAO) as a preconditioning stimulus to induce early ischemic tolerance (IT) to transient focal cerebral ischemia (induced by middle cerebral artery occlusion, MCAO) in C57Bl6/J mice.

局所脳虚血への早期虚血耐性を誘導するために適切な前処理刺激として二国間の総頚動脈閉塞

There is accumulating evidence, that ischemic preconditioning - a non-damaging ischemic challenge to the brain - confers a transient protection to a ...

医学

2-Vessel Occlusion/Hypotension: A Rat Model of Global Brain Ischemia

Cardiac arrest followed by resuscitation often results in dramatic brain damage caused by ischemia and subsequent reperfusion of the brain. Global brain ischemia produces damage to specific brain regions shown to be highly sensitive to ischemia 1. Hippocampal neurons have higher sensitivity to ischemic insults compared to other cell populations, and specifically, the CA1 region of the hippocampus is particularly vulnerable to ischemia/reperfusion 2.
The design of therapeutic interventions, or study of mechanisms involved in cerebral damage, requires a model that produces damage similar to the clinical condition and in a reproducible manner. Bilateral carotid vessel occlusion with hypotension (2VOH) is a model that produces reversible forebrain ischemia, emulating the cerebral events that can occur during cardiac arrest and resuscitation. We describe a model modified from Smith et al. (1984) 2, as first presented in its current form in Sanderson, et al. (2008) 3, which produces reproducible injury to selectively vulnerable brain regions 3-6. The reliability of this model is dictated by precise control of systemic blood pressure during applied hypotension, the duration of ischemia, close temperature control, a specific anesthesia regimen, and diligent post-operative care. An 8-minute ischemic insult produces cell death of CA1 hippocampal neurons that progresses over the course of 6 to 24 hr of reperfusion, while less vulnerable brain regions are spared. This progressive cell death is easily quantified after 7-14 days of reperfusion, as a near complete loss of CA1 neurons is evident at this time.
In addition to this brain injury model, we present a method for CA1 damage quantification using a simple, yet thorough, methodology. Importantly, quantification can be accomplished using a simple camera-mounted microscope, and a free ImageJ (NIH) software plugin, obviating the need for cost-prohibitive stereology software programs and a motorized microscopic stage for damage assessment.

Bilateral carotid occlusion coupled with systemic hypotension produces global brain ischemia in the rat, resulting in damage to the hippocampus with reproducible severity. Animal subjects are impaired with predictable patterns of brain damage, they recover expediently, and mortality rates are comparatively low.

2血管閉塞/低血圧：グローバル脳虚血のラットモデル

Cardiac arrest followed by resuscitation often results in dramatic brain damage caused by ischemia and subsequent reperfusion of the brain. Global brain ...

Lateral Chronic Cranial Window Preparation Enables In Vivo Observation Following Distal Middle Cerebral Artery Occlusion in Mice

Focal cerebral ischemia (i.e., ischemic stroke) may cause major brain injury, leading to a severe loss of neuronal function and consequently to a host of motor and cognitive disabilities. Its high prevalence poses a serious health burden, as stroke is among the principal causes of long-term disability and death worldwide1. Recovery of neuronal function is, in most cases, only partial. So far, treatment options are very limited, in particular due to the narrow time window for thrombolysis2,3. Determining methods to accelerate recovery from stroke remains a prime medical goal; however, this has been hampered by insufficient mechanistic insights into the recovery process. Experimental stroke researchers frequently employ rodent models of focal cerebral ischemia. Beyond the acute phase, stroke research is increasingly focused on the sub-acute and chronic phase following cerebral ischemia. Most stroke researchers apply permanent or transient occlusion of the MCA in mice or rats. In patients, occlusions of the MCA are among the most frequent causes of ischemic stroke4. Besides proximal occlusion of the MCA using the filament model, surgical occlusion of the distal MCA is probably the most frequently used model in experimental stroke research5. Occlusion of a distal (to the branching of the lenticulo-striate arteries) MCA branch typically spares the striatum and primarily affects the neocortex. Vessel occlusion can be permanent or transient. High reproducibility of lesion volume and very low mortality rates with respect to the long-term outcome are the main advantages of this model. Here, we demonstrate how to perform a chronic cranial window (CW) preparation lateral to the sagittal sinus, and afterwards how to surgically induce a distal stroke underneath the window using a craniotomy approach. This approach can be applied for sequential imaging of acute and chronic changes following ischemia via epi-illuminating, confocal laser scanning, and two-photon intravital microscopy.

Lateral Chronic Cranial Window Preparation Enables In Vivo Observation Following Distal Middle Cerebral Artery Occlusion in Mice

Surgical occlusion of a distal middle cerebral artery branch (MCAo) is a frequently used model in experimental stroke research. This manuscript describes the basic technique of permanent MCAo, combined with the insertion of a lateral cranial window, which offers the opportunity for longitudinal intravital microscopy in mice.

横慢性頭蓋窓の準備を有効<em&gt;インビボ</em&gt;マウスの遠位中大脳動脈閉塞後に観測

Focal cerebral ischemia (i.e., ischemic stroke) may cause major brain injury, leading to a severe loss of neuronal function and consequently to a host of ...

神経科学

Surgical Approach for Middle Cerebral Artery Occlusion and Reperfusion Induced Stroke in Mice

Stroke is a leading cause of death worldwide and continues to be one of the major causes of long-term adult disabilities. About 87% of strokes are ischemic in origin and occur in the territory of the middle cerebral artery (MCA). Currently the only Food and Drug Administration (FDA) approved drug for the treatment of this devastating disease is tissue plasminogen activator (tPA). However, tPA has a small therapeutic window for administration (3 - 6 hr), and is only effective in 4% of the patients who actually receive it. Current research focuses on understanding the pathophysiology of stroke in order to find potential therapeutic targets. Thus, reliable models are crucial, and the MCA occlusion (MCAo) model (also termed the intraluminal filament or suture model) is deemed to be the most clinically relevant surgical model of ischemic stroke, and is fairly non-invasive and easily reproducible. Typically the MCAo model is used with rodents, especially with mice due to all the genetic variations available for this species. Here we describe (and present in the video) how to successfully perform the MCAo model (with reperfusion) in mice to generate reliable and reproducible data.

In order to understand the pathophysiology of stroke, it is important to use reliable models. This paper will describe one of the most frequently used stroke models in mice, termed the middle cerebral artery occlusion (MCAo) model (also termed the intraluminal filament or suture model) with reperfusion.

マウスの中大脳動脈閉塞再灌流誘発性の脳卒中のための外科的アプローチ

Stroke is a leading cause of death worldwide and continues to be one of the major causes of long-term adult disabilities. About 87% of strokes are ...

Induction of Ischemic Stroke and Ischemia-reperfusion in Mice Using the Middle Artery Occlusion Technique and Visualization of Infarct Area

Cerebrovascular disease is highly prevalent in the global population and encompasses several types of conditions, including stroke. To study the impact of stroke on tissue injury and to evaluate the effectiveness of therapeutic interventions, several experimental models in a variety of species were developed. They include complete global cerebral ischemia, incomplete global ischemia, focal cerebral ischemia, and multifocal cerebral ischemia. The model described in this protocol is based on the middle cerebral artery occlusion (MCAO) and is related to the focal ischemia category. This technique produces consistent focal ischemia in a strictly defined region of the hemisphere and is less invasive than other methods. The procedure described is performed on mice, given the availability of several genetic variants and the high number of tests standardized for mice to aid in the behavioral and neurodeficit evaluation.

We describe a mouse model of stroke induced by the occlusion of the middle cerebral artery using a silicone coated suture. The protocol can be applied to induce permanent occlusion or a temporary ischemia, followed by reperfusion.

梗塞エリアの中央動脈閉塞技術と可視化を使用したマウスにおける虚血性脳卒中および虚血 - 再灌流の誘導

Cerebrovascular disease is highly prevalent in the global population and encompasses several types of conditions, including stroke. To study the impact of ...

Experimental Models to Study the Neuroprotection of Acidic Postconditioning Against Cerebral Ischemia

Stroke is one of the leading causes of mortality and disability worldwide, with limited therapeutic approaches. As an endogenous strategy for neuroprotection, postconditioning treatments have proven to be promising therapies against cerebral ischemia. However, complicated procedures and potential safety issues limit their clinical application. To overcome these disadvantages, we have developed acidic postconditioning (APC) as a therapy for experimental focal cerebral ischemia. APC refers to the mild acidosis treatment by inhaling CO2 during reperfusion following ischemia. Here we present two models to execute APC in vitro and in vivo, respectively. The oxygen-glucose deprivation (OGD) treatment of mice and the corticostriatal occlusion and middle cerebral artery occlusion (MCAO) of mice were employed to mimic cerebral ischemia. APC can be simply achieved by transferring brain slices to acidic buffer bubbled with 20% CO2, or by mice inhaling 20% CO2. APC showed significant protective effects against cerebral ischemia, as reflected by tissue viability and brain infarct volume.

Acidic postconditioning protects against cerebral ischemia. Here we present two models to execute APC. They are achieved respectively by transferring corticostriatal slices to acidic buffer after oxygen-glucose deprivation in vitro and by inhaling 20% CO2 after middle cerebral artery occlusion in vivo.

脳虚血に対する酸性ポストコンディショニングの神経保護を研究するための実験モデル

Stroke is one of the leading causes of mortality and disability worldwide, with limited therapeutic approaches. As an endogenous strategy for ...

Assessing Neuroprotective Effects of Glycyrrhizae Radix et Rhizoma Extract Using a Transient Middle Cerebral Artery Occlusion Mouse Model

Ischemia followed by reperfusion of cerebral blood flow after a stroke leads to the death of nerve cells and loss of brain tissue. The most commonly used animal model for studying stroke is the middle cerebral artery occlusion (MCAO) model. Previous research studies have reported different infarct sizes even when the same experimental animal species was used under similar MCAO conditions. Therefore, we developed an improved experimental method to address this discrepancy. Mice were subjected to MCAO using a filament as the occlusion material to mimic human stroke conditions and filament thickness was optimized to establish more reproducible infarction volume. Mice treated with a methanol extract of Glycyrrhizae Radix et Rhizome (GRex) following stroke induction showed a significantly decreased total infarction volume and increased number of surviving cells relative to the untreated control group. This modified experimental protocol successfully and reproducibly demonstrated the beneficial effect of GRex on ischemic stroke.

In this study, we modify an existing experimental method to obtain more reproducible results, by establishing a middle cerebral artery occlusion (MCAO) mouse model. Oral administration of Glycyrrhizae Radix et Rhizome (GR) methanol extract (GRex), following stroke induction, significantly decreased total infarction volume relative to the untreated control group.

芩の神経保護効果を評価する et 大黄抽出一過性中大脳動脈閉塞モデルマウスを使用して

Ischemia followed by reperfusion of cerebral blood flow after a stroke leads to the death of nerve cells and loss of brain tissue. The most commonly used ...

Two-vessel Occlusion Mouse Model of Cerebral Ischemia-reperfusion

In this study, a middle cerebral artery (MCA) occlusion mouse model is employed to study cerebral ischemia-reperfusion. A reproducible and reliable mouse model is useful for investigating the pathophysiology of cerebral ischemia-reperfusion and determining potential therapeutic strategies for patients with stroke. Variations in the anatomy of the circle of Willis of C57BL/6 mice affects their infarct volume after cerebral-ischemia-induced injury. Studies have indicated that distal MCA occlusion (MCAO) can overcome this problem and result in a stable infarct size. In this study, we establish a two-vessel occlusion mouse model of cerebral ischemia-reperfusion through the interruption of the blood flow to the right MCA. We distally ligate the right MCA and right common carotid artery (CCA) and restore blood flow after a certain period of ischemia. This ischemia-reperfusion injury induces an infarct of stable size and a behavioral deficit. Peripheral immune cells infiltrate the ischemic brain within the 24 h infiltration period. Additionally, the neuronal loss in the cortical area is less for a longer reperfusion duration. Therefore, this two-vessel occlusion model is suitable for investigating the immune response and neuronal recovery during the reperfusion period after cerebral ischemia.

A mouse model of cerebral ischemia-reperfusion is established to investigate the pathophysiology of stroke. We distally ligate the right middle cerebral artery and right common carotid artery and restore blood flow after 10 or 40 min of ischemia.

2 血管閉塞マウス脳虚血・再灌流モデル

In this study, a middle cerebral artery (MCA) occlusion mouse model is employed to study cerebral ischemia-reperfusion. A reproducible and reliable mouse ...

A Murine Model of Ischemic Retinal Injury Induced by Transient Bilateral Common Carotid Artery Occlusion

Diverse vascular diseases such as diabetic retinopathy, occlusion of retinal veins or arteries and ocular ischemic syndrome can lead to retinal ischemia. To investigate pathological mechanisms of retinal ischemia, relevant experimental models need to be developed. Anatomically, a main retinal blood supplying vessel is the ophthalmic artery (OpA) and OpA originates from the internal carotid artery of the common carotid artery (CCA). Thus, disruption of CCA could effectively cause retinal ischemia. Here, we established a mouse model of retinal ischemia by transient bilateral common carotid artery occlusion (tBCCAO) to tie the right CCA with 6-0 silk sutures and to occlude the left CCA transiently for 2 seconds via a clamp, and showed that tBCCAO could induce acute retinal ischemia leading to retinal dysfunction. The current method reduces reliance on surgical instruments by only using surgical needles and a clamp, shortens occlusion time to minimize unexpected animal death, which is often seen in mouse models of middle cerebral artery occlusion, and maintains reproducibility of common retinal ischemic findings. The model can be utilized to investigate the pathophysiology of ischemic retinopathies in mice and further can be used for in vivo drug screening.

Here, we describe a mouse model of retinal ischemia by transient bilateral common carotid artery occlusion using simple sutures and a clamp. This model can be useful for understanding the pathological mechanisms of retinal ischemia caused by cardiovascular abnormalities.

一過性両側性頸動脈閉塞による虚血性筋炎損傷のマウスモデル

Diverse vascular diseases such as diabetic retinopathy, occlusion of retinal veins or arteries and ocular ischemic syndrome can lead to retinal ischemia. ...

中大脳動脈の閉塞による一過性脳虚血のモデル

Transient Middle Cerebral Artery Occlusion Model of Stroke

Stroke stands as a major cause of death or chronic disability globally. Nevertheless, existing optimal treatments are limited to reperfusion therapies during the acute phase of ischemic stroke. To gain insights into stroke physiopathology and develop innovative therapeutic approaches, in vivo rodent models of stroke play a fundamental role. The availability of genetically modified animals has particularly propelled the use of mice as experimental stroke models.
In stroke patients, occlusion of the middle cerebral artery (MCA) is a common occurrence. Consequently, the most prevalent experimental model involves intraluminal occlusion of the MCA, a minimally invasive technique that doesn't require craniectomy. This procedure involves inserting a monofilament through the external carotid artery (ECA) and advancing it through the internal carotid artery (ICA) until it reaches the branching point of the MCA. After a 45 min arterial occlusion, the monofilament is removed to allow reperfusion. Throughout the process, cerebral blood flow is monitored to confirm the reduction during occlusion and subsequent recovery upon reperfusion. Neurological and tissue outcomes are evaluated using behavioral tests and magnetic resonance imaging (MRI) studies.

著者スポットライト:中大脳動脈閉塞モデルによる虚血性脳卒中損傷の評価 

This protocol describes the model of transient focal cerebral ischemia in mice through intraluminal occlusion of the middle cerebral artery. Additionally, examples of outcome assessment are shown using magnetic resonance imaging and behavioral tests.

脳卒中の一過性中大脳動脈閉塞モデル

Stroke stands as a major cause of death or chronic disability globally. Nevertheless, existing optimal treatments are limited to reperfusion therapies ...