Name: reproducible motor deficit following aortic occlusion in a rat model of spinal cord ischemia
Uploaded: 2017-07-22T13:00:00.000+00:00
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Description: Watch this Scientific Journal Video about Reproducible Motor Deficit Following Aortic Occlusion in a Rat Model Of Spinal Cord Ischemia at JoVE.com

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このプロトコールは、ソウル大学分立病院動物実験センターの承認を受けた。動物のケアおよび実験は、実験動物のケアおよび使用のための米国国立衛生研究所のガイドラインに従って行われた。 1.外科的準備<ol><li>手術の前に、開存性を確実にするために滅菌生理食塩水でカテーテルを洗い流す。 </li><li>手術用テーブルに暖房用毛布を置き、無菌ドレープでテーブルを覆う。 </li><li>雄Sprague-Dawleyラット（270-330g）を、100％酸素中3.0％〜4.0％のイソフルランを含むアクリルボックスに入れる。 </li><li>ラットの目に潤滑剤を塗布する。 </li><li>ラットを仰臥位の手術台に置き、吸入イソフルラン（1.0％〜2.5容積％）を連続的に投与したフェイシャルマスクを用いて麻酔を維持する。 </li><li>直腸プローブを配置し、体温を37.0〜38.0℃の間で監視および維持する。 </li&gt; </ol> 2.大腿動脈カテーテル法<ol><li>右の鼠径部をベタディンと70％エタノールを使って静かにこすります。 </li><li>右鼠径部のはさみを使って、2cmの水平な皮膚切開を行います。 </li><li>開創器を使用して、手術野を露出させる。 </li><li>周囲の静脈および神経から大腿動脈を解剖する。湾曲した鉗子と鈍い鉗子を使用して、動脈の1cmの部分を分離する。 </li><li> 4.0黒い絹の縫合糸を使用して、暴露を最大にするために、動脈の近位端および遠位端の両方に緩い結び目を置く。 </li><li>大腿動脈をマイクロはさみで切開する。 </li><li> 2-Frを挿入します。マイクロ鉗子を用いて大腿動脈内にバルーンチップカテーテルを挿入した。近位結紮でカテーテルヘッドから約1cmのところでカテーテルを血管に固定し、遠位結紮糸を結ぶ。 </li></ol> 3.頚動脈カテーテル法<ol><li> betadを使用して右前頸部を擦るineと70％エタノールを入れ、皮膚切開を行います。 </li><li>開創器を使用して、手術野を露出させる。動脈の1cmの部分を分離し、暴露を最大にするために、動脈の近位端および遠位端の両方に絹の縫合糸を用いて緩く結ぶ。 </li><li> 24 Gの静脈内カテーテルを用いて頸動脈を穿刺し、カテーテルの近位1cmを心臓に向けて前進させる。 </li><li>近位結紮でカテーテルを固定し、遠位結紮糸を結ぶ。 </li><li>三方活栓を使用して、カテーテルの遠位端を生理食塩水で満たされたマイクロチューブおよび外部リザーバーに接続する。 </li></ol> 4.尾部動脈カテーテル法<ol><li>ベタディンと70％エタノールを使用して尾の腹側を擦り、2cmの皮膚切開を行います。 </li><li>周囲の構造から尾動脈を解剖し、湾曲鉗子と鈍端鉗子を用いて動脈の1cm切片を分離する。 </li><li> 4.0黒の絹の縫合糸を使用して、緩く置く暴露を最大限にするために動脈の近位端および遠位端の両方を結ぶ。 </li><li> 24 Gの静脈内カテーテルを用いて尾動脈を突き刺し、カテーテルを動脈内に進める。 </li><li>近位の結紮具でカテーテルを血管（カテーテルヘッドから約1cm）に固定し、遠位結紮糸を結ぶ。 </li><li>カテーテルの遠位部分を動脈圧監視装置に接続する。 </li></ol> 5.脊髄虚血の誘導<ol><li>カテーテル挿入が完了したら、2 Fr.を進める。カテーテルの先端が左鎖骨下動脈に達するように、バルーンチップカテーテルを下降する胸部大動脈に挿入する。 </li><li>挿入部位から10 cmの深さまで挿入します。 </li><li> 100ユニット/ mLの濃度の150単位のヘパリンを頸動脈カテーテルに注入する。 </li><li> 0.05mLの生理食塩水でカテーテルバルーンを膨らませる。 </li><li>同時に、血液を外部血液コンテナにcか​​ら排出させる近位動脈圧を80mmHgに調節するための頚動脈動脈。 </li><li>動脈圧モニタリングシステムを3方向ストップコックの残りの管腔に接続し、動脈圧を監視して排液量を制御する。 </li><li>突然の減少と遠位動脈圧の連続的な損失によって、大動脈閉塞の成功を確認する。 </li><li> 9、10、または11分の大動脈閉塞の後、Fogartyカテーテルバルーンを収縮させ、排出された血液を再注入する。 </li></ol> 6.手術後のケアおよび神経学的評価<ol><li>尾動脈を通る動脈圧を監視しながら、大腿動脈および頸動脈からカテーテルを取り出し、絹の縫合糸で創傷を閉じる。 </li><li>動脈圧が正常範囲に回復したことを確認した後、尾動脈からカテーテルを取り出し、創傷を閉じる。 </li><li>麻酔から回復した後、ラットをケージに戻す。 </li><li>評価する手術後24時間で運動不足指数を用いて肢体運動機能を評価した（ 表1 ）。運動不足指数は、歩行スコアと配置/歩行反射スコアの合計として定義される。 6のモーター赤字指数は最大赤字を示します。 </li></ol> 7.組織病理学的評価<ol><li>神経学的評価後、ラットをマスク送達イソフルランで麻酔し、麻酔下でヘパリン処理された生理食塩水100mLで心筋灌流によりそれらを屠殺し、脊髄を抽出する。 8 </li><li>パラフィンに腰椎3-5のレベルからコードセクションを埋め込みます。 </li><li>ヘマトキシリンおよびエオシン（H＆E）で横断切片を染色する。 </li><li>顕微鏡下での運動ニューロン損傷を観察する。 9 </li></ol> 8.統計分析<ol><li> Social Sciencesバージョン20の統計パッケージを使用して統計分析を実行します。モータdefKruskal-Wallis検定、続いてMann-Whitney U検定を用いて、3つの群の奏功指数を比較した。 0.05未満のP値は統計的に有意であると考えられた。 </li></ol>

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著者には競合する金銭的利益はない。この研究は、韓国政府の国立研究財団からの助成金2012R1A1A3014010によって支援された。

今回の研究では、大動脈閉塞時間に依存して後肢に様々な程度の運動障害を誘発するTaira and Marsalaの方法5に基づく脊髄虚血のラットモデルを実証した。 大動脈閉塞の長さは、運動不足の程度に影響し得る。大動脈閉塞時間が長い場合、運動障害はより深刻になる。したがって、研究者は、このモデルで大動脈閉塞時間を制御することによって、ある程?...

対麻痺は、胸腹部大動脈瘤手術の致命的な合併症である。これは、大動脈のクロスクランプおよびアンクランプ中に起こる脊髄虚血 - 再灌流損傷に起因する。怪我の影響を受けて全身低体温症と脳脊髄ドレナージ脊髄を保護するために導入されている、2、3、4しかし、多くの患者が残っているなど、1つのいくつかの戦略。 いくつかの動物の脊髄虚血モデルがその病因を調査し、損傷に対する防御戦略を考案するために導入されている。今回の研究では、平良とマルサラの方法に基づいた脊髄虚血のラットモデルを概説する。サイズに多少の違いはあるものの5ラットにおける脊髄循環システムは、ヒトでの脊髄の血管および担保システムに非常に類似しており、ロケーション。 図6、 図7はこのように、ラットは、病因、合併症、および脊髄虚血の治療を調査する実験モデルのために利用する解剖学的に適切な動物です。さらに、この脊髄虚血モデルは、胸部大動脈の血管内バルーン閉塞を利用することにより最小限の介入で確実な大動脈閉塞を生じる。 本研究では、この脊髄虚血ラットモデルが、大動脈閉塞時間に応じて重症度が異なる後肢における再現性運動障害を誘発することを実証した。

<table><tbody><tr><td>Fogarty Arterial Embolectomy catheter</td><td>Edward Life Sciences</td><td>120602F</td><td>a balloon-tipped catheter inserted into the femoral artery</td></tr><tr><td>BD Insyte-N Autoguard Shielded IV catheter&amp;nbsp;</td><td>BD&amp;nbsp;</td><td>381411</td><td>24-gauge intravenous catheter</td></tr><tr><td>50 mL syringe</td><td>KOREA VACCINE&amp;nbsp;</td><td>KOVAX-SYRINGE 50mL</td><td>Facial mask</td></tr><tr><td>1 mL syringe</td><td>KOREA VACCINE</td><td>KOVAX-SYRINGE 1ml</td><td></td></tr><tr><td>Recal probe</td><td>HARVARD APPARATUS</td><td>50-7221F</td><td>Rectal probe for temperature monitoring</td></tr><tr><td>Micro dissecting spring scissor</td><td>Jeung do bio &amp;amp; Plant co.LTD.</td><td>JD-S-10</td><td>Micro-scissor</td></tr><tr><td>SCISSOR (SHARP-SHARP)</td><td>Jeung do bio &amp;amp; Plant co.LTD.</td><td>S-51-12-S</td><td>Scissors</td></tr><tr><td>Retractor</td><td>Jeung do bio &amp;amp; Plant co.LTD.</td><td>JD-S-74A</td><td>Retractor</td></tr><tr><td>Micro forcep&amp;nbsp;</td><td>Jeung do bio &amp;amp; Plant co.LTD.</td><td>JD-S-29</td><td>Micro-forceps</td></tr><tr><td>MOSQUITO FORCEP (Curved)</td><td>Jeung do bio &amp;amp; Plant co.LTD.</td><td>S-44-CPK</td><td>Curved forceps</td></tr><tr><td>DRESSING FORCEP&amp;nbsp;</td><td>Jeung do bio &amp;amp; Plant co.LTD.</td><td>S-37-16S</td><td>Blunted forceps</td></tr><tr><td>4/0 black silk&amp;nbsp;</td><td>Woori Medical</td><td>S431</td><td>4.0 black silk suture</td></tr><tr><td>3-WAY STOCK</td><td>Seonwon Medcal</td><td>D-98-01</td><td>3-way stopcock</td></tr><tr><td>Patient monitor</td><td>PHILIPS</td><td>MP20</td><td>The arterial pressure monitoring device.&amp;nbsp;</td></tr><tr><td>Heating blanket</td><td>Self production</td><td></td><td>Heating blanket</td></tr><tr><td>Microtube and external reservoir</td><td>Self production</td><td></td><td>Microtube and external reservoir</td></tr><tr><td>Heparin</td><td>JW Pharmaceutical</td><td></td><td>Heparin</td></tr><tr><td>0.9% NS 1000ml</td><td>JW Pharmaceutical</td><td></td><td>Normal saline</td></tr><tr><td>Isoflurane</td><td>Hana Med</td><td></td><td>Isoflurane</td></tr></tbody></table>

reproducible motor deficit following aortic occlusion in a rat model of spinal cord ischemia

<html>脊髄虚血は、胸腔腹部大動脈瘤手術後の致命的な合併症である。研究者は、脊髄虚血の実験モデルを用いてこの合併症を予防および治療するための戦略を調査することができる。ここに記載されたモデルは、ラットの脊髄虚血モデルにおける胸部大動脈閉塞後の閉塞の長さに関連する様々な程度の対麻痺を示す。 2-Fr。バルーン先端カテーテルを、麻酔した雄Sprague-Dawleyラットの左鎖骨下動脈にカテーテル先端を配置するまで、大腿動脈を通して下行胸部大動脈に前進させた。カテーテルバルーンを膨張させることによって脊髄虚血を誘発した。一定の閉塞（9,10または11分）後、バルーンは収縮した。手術後24時間で運動異常を用いて神経学的評価を行い、病理組織学的検査のために脊髄を採取した。 10分の大動脈閉塞を受けたラットは、中程度ではあるが可逆的な運動障害を示した。大動脈閉塞の11分間に曝されたラットは、完全で持続的であった大動脈閉塞の期間がより短いラットにおいて、脊髄切片における運動ニューロンはより保存された。 研究者は、この脊髄虚血モデルを用いて胸部大動脈閉塞後に再現性のある後肢運動障害を達成することができる。

脊髄虚血の期間中、大動脈閉塞を9分（n = 3）、10分（n = 3）、または11分（n = 3）行った。ラットにおける運動障害指数を表2に示す。大動脈閉塞を9分間受けたラットは、後肢に軽度の可逆性運動障害を示した。 10分間の大動脈閉塞を受けたラットは、中等度の運動障害を示したが、完全な麻痺はなかった。 11分の閉塞時間を経たラットは、完全で持続的な麻痺を示した。 ...

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Reproducible Motor Deficit Following Aortic Occlusion in a Rat Model Of Spinal Cord Ischemia

この研究は、ラットにおいて最小侵襲性で容易に再現可能な脊髄虚血モデルを作製する技術を実証している。大動脈閉塞時間を制御することにより、様々な程度の後肢運動欠損を生じさせることができる。

脊髄虚血ラットモデルにおける大動脈閉塞後の再現性のある運動障害

Spinal cord ischemia is a fatal complication following thoracoabdominal aortic aneurysm surgery. Researchers can investigate the strategies for preventing ...

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JoVE Journal

Medicine

7.1K ビュー.  SMG-SNU Boramae Medical Center, Seoul National University College of Medicine, Seoul. この研究は、ラットにおいて最小侵襲性で容易に再現可能な脊髄虚血モデルを作製する技術を実証している。大動脈閉塞時間を制御することにより、様々な程度の後肢運動欠損を生じさせることができる。 

ビデオ: Reproducible Motor Deficit Following Aortic Occlusion in a Rat Model Of Spinal Cord Ischemia

Focal Cerebral Ischemia Model by Endovascular Suture Occlusion of the Middle Cerebral Artery in the Rat

Stroke is the leading cause of disability and the third leading cause of death in adults worldwide1. In human stroke, there exists a highly variable clinical state; in the development of animal models of focal ischemia, however, achieving reproducibility of experimentally induced infarct volume is essential. The rat is a widely used animal model for stroke due to its relatively low animal husbandry costs and to the similarity of its cranial circulation to that of humans2,3. In humans, the middle cerebral artery (MCA) is most commonly affected in stroke syndromes and multiple methods of MCA occlusion (MCAO) have been described to mimic this clinical syndrome in animal models. Because recanalization commonly occurs following an acute stroke in the human, reperfusion after a period of occlusion has been included in many of these models. In this video, we demonstrate the transient endovascular suture MCAO model in the spontaneously hypertensive rat (SHR). A filament with a silicon tip coating is placed intraluminally at the MCA origin for 60 minutes, followed by reperfusion. Note that the optimal occlusion period may vary in other rat strains, such as Wistar or Sprague-Dawley. Several behavioral indicators of stroke in the rat are shown. Focal ischemia is confirmed using T2-weighted magnetic resonance images and by staining brain sections with 2,3,5-triphenyltetrazolium chloride (TTC) 24 hours after MCAO.

Surgical induction of ischemic brain damage in the rat is a widely used model for stroke research. Here we demonstrate the induction of focal cerebral ischemia by occlusion of the middle cerebral artery. Visualization of the resulting infarct by histological staining and magnetic resonance imaging is also shown.

ラット中大脳動脈の血管縫合閉塞による局所脳虚血モデル

Stroke is the leading cause of disability and the third leading cause of death in adults worldwide1. In human stroke, there exists a highly variable ...

神経科学

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 ...

医学

Reproducable Paraplegia by Thoracic Aortic Occlusion in a Murine Model of Spinal Cord Ischemia-reperfusion

Background 
Lower extremity paralysis continues to complicate aortic interventions. The lack of understanding of the underlying pathology has hindered advancements to decrease the occurrence this injury.&#160;The current model demonstrates reproducible lower&#160;extremity paralysis following thoracic aortic occlusion.
Methods 
Adult male C57BL6 mice were anesthetized with isoflurane. Through a cervicosternal incision the aorta was exposed. The descending thoracic aorta and left subclavian arteries were identified without entrance into pleural space. Skeletonization of these arteries was followed by immediate closure (Sham) or occlusion for 4 min (moderate ischemia) or 8 min (prolonged ischemia). The sternotomy and skin were closed and the mouse was transferred to warming bed for recovery.&#160; Following recovery, functional analysis was obtained at 12 hr intervals until 48 hr.
Results 
Mice that underwent sham surgery showed no observable hind&#160;limb deficit. Mice subjected to moderate ischemia for 4 min had minimal functional deficit at 12 hr followed by progression to complete paralysis at 48 hr. Mice subjected to prolonged ischemia had an immediate paralysis with no observable hind-limb movement at any point in the postoperative period. There was no observed intraoperative or post&#160;operative mortality.
Conclusion 
Reproducible lower extremity paralysis whether immediate or delayed can be achieved in a murine model. Additionally, by using a median sternotomy and careful dissection, high survival rates, and reproducibility can be achieved.

The lack of mechanistic understanding of spinal cord ischemia-reperfusion injury has hindered further adjuncts to prevent paraplegia following high risk aortic operations. Thus, the development of animal models is imperative. This manuscript demonstrates reproducible lower extremity paralysis following thoracic aortic occlusion in a murine model.

脊髄虚血再灌流のマウスモデルにおいて胸部大動脈閉塞によって再現可能対麻痺

Background
Lower extremity paralysis continues to complicate aortic interventions. The lack of understanding of the underlying pathology has hindered ...

Performing Permanent Distal Middle Cerebral with Common Carotid Artery Occlusion in Aged Rats to Study Cortical Ischemia with Sustained Disability

Stroke typically occurs in elderly people with a range of comorbidities including carotid (or other arterial) atherosclerosis, high blood pressure, obesity and diabetes. Accordingly, when evaluating therapies for stroke in animals, it is important to select a model with excellent face validity. Ischemic stroke accounts for 80% of all strokes, and the majority of these occur in the territory of the middle cerebral artery (MCA), often inducing infarcts that affect the sensorimotor cortex, causing persistent plegia or paresis on the contralateral side of the body. We demonstrate in this video a method for producing ischemic stroke in elderly rats, which causes sustained sensorimotor disability and substantial cortical infarcts. Specifically, we induce permanent distal middle cerebral artery occlusion (MCAO) in elderly female rats by using diathermy forceps to occlude a short segment of this artery. The carotid artery on the ipsilateral side to the lesion was then permanently occluded and the contralateral carotid artery was transiently occluded for 60 min. We measure the infarct size using structural T2-weighted magnetic resonance imaging (MRI) at 24 hr and 8 weeks after stroke. In this study, the mean infarct volume was 4.5% &#177; 2.0% (standard deviation) of the ipsilateral hemisphere at 24 hr (corrected for brain swelling using Gerriet&#8217;s equation, n = 5). This model is feasible and clinically relevant as it permits the induction of sustained sensorimotor deficits, which is important for the elucidation of pathophysiological mechanisms and novel treatments.

Here we present a protocol to produce permanent distal middle cerebral artery occlusion in elderly female rats with simultaneous occlusion of the carotid arteries to generate large cortical infarcts and sustained deficits. We show confirmation of the lesion size using structural MRI at 24 hr and 8 weeks after stroke.

持続的な障害と皮質虚血を研究するために高齢ラットにおける一般的な頸動脈閉塞と永久遠位中大脳の実行

Stroke typically occurs in elderly people with a range of comorbidities including carotid (or other arterial) atherosclerosis, high blood pressure, ...

Modeling Stroke in Mice: Transient Middle Cerebral Artery Occlusion via the External Carotid Artery

Stroke is the third most common cause of mortality and the leading cause of acquired adult disability in developed countries. To date, therapeutic options are limited to a small proportion of stroke patients within the first hours after stroke. Novel therapeutic strategies are being extensively investigated, especially to prolong the therapeutic time window. These current investigations include the study of important pathophysiological pathways after stroke, such as post-stroke inflammation, angiogenesis, neuronal plasticity, and regeneration. Over the last decade, there has been increasing concern about the poor reproducibility of experimental results and scientific findings among independent research groups. To overcome the so-called &#8220;replication crisis&#8221;, detailed standardized models for all procedures are urgently needed. As an effort within the &#8220;ImmunoStroke&#8221; research consortium (https://immunostroke.de/), a standardized mouse model of transient middle cerebral artery occlusion (MCAo) is proposed. This model allows the complete restoration of the blood flow upon removal of the filament, simulating the therapeutic or spontaneous clot lysis that occurs in a large proportion of human strokes. The surgical procedure of this &#8220;filament&#8221; stroke model and tools for its functional analysis are demonstrated in the accompanying video.

Different models of middle cerebral artery occlusion (MCAo) are used in experimental stroke research. Here, an experimental stroke model of transient MCAo via the external carotid artery (ECA) is described, which aims to mimic human stroke, in which the cerebrovascular thrombus is removed due to spontaneous clot lysis or therapy.

マウスにおける脳卒中のモデリング:外頸動脈を介した過渡中脳動脈閉塞

Stroke is the third most common cause of mortality and the leading cause of acquired adult disability in developed countries. To date, therapeutic options ...

Real-Time Assessment of Spinal Cord Microperfusion in a Porcine Model of Ischemia/Reperfusion

Spinal cord injury is a devastating complication of aortic repair. Despite developments for the prevention and treatment of spinal cord injury, its incidence is still considerably high and therefore, influences patient outcome. Microcirculation plays a key role in tissue perfusion and oxygen supply and is often dissociated from macrohemodynamics. Thus, direct evaluation of spinal cord microcirculation is essential for the development of microcirculation-targeted therapies and the evaluation of existing approaches in regard to spinal cord microcirculation. However, most of the methods do not provide real-time assessment of spinal cord microcirculation. The aim of this study is to describe a standardized protocol for real-time spinal cord microcirculatory evaluation using laser-Doppler needle probes directly inserted in the spinal cord. We used a porcine model of ischemia/reperfusion to induce deterioration of the spinal cord microcirculation. In addition, a fluorescent microsphere injection technique was used. Initially, animals were anesthetized and mechanically ventilated. Thereafter, laser-Doppler needle probe insertion was performed, followed by the placement of cerebrospinal fluid drainage. A median sternotomy was performed for exposure of the descending aorta to perform aortic cross-clamping. Ischemia/reperfusion was induced by supra-celiac aortic cross-clamping for a total of 48 min, followed by reperfusion and hemodynamic stabilization. Laser-Doppler Flux was performed in parallel with macrohemodynamic evaluation. In addition, automated cerebrospinal fluid drainage was used to maintain a stable cerebrospinal pressure. After completion of the protocol, animals were sacrificed, and the spinal cord was harvested for histopathological and microsphere analysis. The protocol reveals the feasibility of spinal cord microperfusion measurements using laser-Doppler probes and shows a marked decrease during ischemia as well as recovery after reperfusion. Results showed comparable behavior to fluorescent microsphere evaluation. In conclusion, this new protocol might provide a useful large animal model for future studies using real-time spinal cord microperfusion assessment in ischemia/reperfusion conditions.

Spinal cord microcirculation plays a pivotal role in spinal cord injury. Most methods do not allow real-time assessment of spinal cord microcirculation, which is essential for the development of microcirculation-targeted therapies. Here, we propose a protocol using Laser-Doppler-Flow Needle probes in a large animal model of ischemia/reperfusion.

イシュ血症/再灌流の豚モデルにおける脊髄微小灌流のリアルタイム評価

Spinal cord injury is a devastating complication of aortic repair. Despite developments for the prevention and treatment of spinal cord injury, its ...

Real-Time Monitoring and Modulation of Blood Pressure in a Rabbit Model of Ischemic Stroke

Control of blood pressure, in terms of both absolute values and its variability, affects outcomes in ischemic stroke patients. However, it remains challenging to identify the mechanisms that lead to poor outcomes or evaluate measures by which these effects can be mitigated because of the prohibitive limitations inherent to human data. In such cases, animal models can be utilized to conduct rigorous and reproducible evaluations of diseases. Here we report refinement of a previously described model of ischemic stroke in rabbits that is augmented with continuous blood pressure recording to assess the impacts of modulation on blood pressure. Under general anesthesia, femoral arteries are exposed through surgical cutdowns to place arterial sheaths bilaterally. Under fluoroscopic visualization and roadmap guidance, a microcatheter is advanced into an artery of the posterior circulation of the brain. An angiogram is performed by injecting the contralateral vertebral artery to confirm occlusion of the target artery. With the occlusive catheter remaining in position for a fixed duration, blood pressure is continuously recorded to allow for tight titration of blood pressure manipulations, whether through mechanical or pharmacological means. At the completion of the occlusion interval, the microcatheter is removed, and the animal is maintained under general anesthesia for a prescribed length of reperfusion. For acute studies, the animal is then euthanized and decapitated. The brain is harvested and processed to measure the infarct volume under light microscopy and further assessed with various histopathological stains or spatial transcriptomic analysis. This protocol provides a reproducible model that can be utilized for more thorough preclinical studies on the effects of blood pressure parameters during ischemic stroke. It also facilitates effective preclinical evaluation of novel neuroprotective interventions that might improve care for ischemic stroke patients.

Continuous arterial blood pressure recording allows the investigation of impacts of various hemodynamic parameters. This report demonstrates the application of continuous arterial blood pressure monitoring in a large animal model of ischemic stroke for determination of stroke pathophysiology, impact of different hemodynamic factors, and the assessment of novel treatment approaches.

虚血性脳卒中のウサギモデルにおける血圧のリアルタイムモニタリングと調節

Control of blood pressure, in terms of both absolute values and its variability, affects outcomes in ischemic stroke patients. However, it remains ...

Middle Cerebral Artery Occlusion in Rats with Preserved Vascular Anatomy

A Rat Model of Middle Cerebral Artery Occlusion/Reperfusion Without Damaging the Anatomical Structure of Cerebral Vessels

Ischemic stroke stands as the primary cause of long-term disability and mortality among adults worldwide. Animal models of ischemic stroke have significantly contributed to our understanding of its pathological mechanisms and the development of potential treatments. Presently, there are two common methods involving filament (endovascular suture) techniques to induce animal models of cerebral ischemia. However, these methods have inherent limitations, such as reduced blood perfusion to the brain, damage to the external carotid artery system, impaired food and/or water intake, and sensory dysfunction of the face. This article introduces a new method for inducing a rat ischemic stroke model without compromising the cerebral vascular anatomy. In this study, the common carotid artery (CCA) of Sprague-Dawley rats was exposed, and an incision was made. A filament was then inserted through the incision into the internal carotid artery to occlude the middle cerebral artery. After 1.5 h of induced ischemia, the occluding filament was fully removed from both the internal carotid artery and the CCA. The incision in the CCA was subsequently sutured using 11-0 microsurgical sutures under a microscope (magnification 4x). Through the utilization of microsurgical techniques to repair the CCA, this study successfully developed a unique method to induce an ischemic stroke model in rats while preserving the anatomical integrity of cerebral blood vessels.

The current protocol describes a rat model of middle cerebral artery occlusion/reperfusion that preserves the anatomical structure of cerebral vessels without causing damage.

Ischemic stroke stands as the primary cause of long-term disability and mortality among adults worldwide. Animal models of ischemic stroke have ...

<meta charset="utf8"></head><body>ラットMCAO脳卒中モデルにおける細胞死を研究するための免疫蛍光染色

Evaluating Cell Death Signaling by Immunofluorescence in a Rat Model of Ischemic Stroke

Stroke is a leading cause of death and disability worldwide. Most cases of stroke are ischemic and result from the occlusion of the middle cerebral artery (MCA). Current pharmacological approaches for the treatment of ischemic stroke are limited; therefore, novel therapies providing effective neuroprotection against ischemic injury following stroke are urgently needed. In the brain tissue following ischemic stroke, the area of the ischemic penumbra is salvageable but at risk of progressing to irreversible damage. The penumbral area surrounding the infarcted core is a conceptual target for neuroprotection. Since the blood flow is partially maintained in the penumbral area, neuronal and non-neuronal cells in this area transiently survive after stroke, and these cells that are still viable may be rescued by appropriate medical interventions. Understanding the pathophysiology of the penumbra is important in the development of neuroprotective therapies because the cell death pathways activated in the ischemic penumbra may indicate therapeutic targets, such as RUNX1 and cathepsins. These protein targets functioning as mediators of programmed cell death can be further exploited in translational research. With moderate size and similarity to the human brain, the in vivo rat model of MCA occlusion (MCAO) mimics the human ischemic stroke and offers an applicable tool to investigate the penumbral pathology, examine the cell death signaling, and evaluate the effects of potential targets in the context of MCAO. Here, we describe how to induce MCAO in rats and how to perform immunofluorescence staining for the detection of cell death signaling in the rat brain following MCAO.

<meta charset="utf8"></head><body>虚血性脳卒中のラットモデルにおける免疫蛍光法による細胞死シグナル伝達の評価

Rat in vivo models are indispensable tools to investigate the pathological mechanisms and therapeutic targets for ischemic stroke. This work will describe performing immunofluorescence staining in infarcted brain slices following middle cerebral artery occlusion (MCAO) in rats.

虚血性脳卒中のラットモデルにおける免疫蛍光法による細胞死シグナル伝達の評価

Stroke is a leading cause of death and disability worldwide. Most cases of stroke are ischemic and result from the occlusion of the middle cerebral artery ...

脊髄虚血ラットモデルにおける大動脈閉塞後の再現性のある運動障害

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ラット中大脳動脈の血管縫合閉塞による局所脳虚血モデル

2血管閉塞/低血圧：グローバル脳虚血のラットモデル

脊髄虚血再灌流のマウスモデルにおいて胸部大動脈閉塞によって再現可能対麻痺

持続的な障害と皮質虚血を研究するために高齢ラットにおける一般的な頸動脈閉塞と永久遠位中大脳の実行

マウスにおける脳卒中のモデリング:外頸動脈を介した過渡中脳動脈閉塞

イシュ血症/再灌流の豚モデルにおける脊髄微小灌流のリアルタイム評価

虚血性脳卒中のウサギモデルにおける血圧のリアルタイムモニタリングと調節

A Rat Model of Middle Cerebral Artery Occlusion/Reperfusion Without Damaging the Anatomical Structure of Cerebral Vessels

虚血性脳卒中のラットモデルにおける免疫蛍光法による細胞死シグナル伝達の評価

虚血性脳卒中のラットモデルにおける免疫蛍光法による細胞死シグナル伝達の評価