This work was supported by National Institutes of Health grants HL087990 (G.L.).

윤리 선언문 : (330~380g 무게) 남성 성인 흰쥐이 프로토콜에 사용되었다. 이 프로토콜은 LSU 건강 과학 센터 - 슈 리브 포트에서 기관 동물 관리 및 사용위원회 (IACUC)의 승인 (제 8 판, 국립 과학 아카데미 2011 &#39;실험실 동물의 관리 및 사용을위한 설명서&#39;을 준수하였습니다 ). 수정 된 PE-50 튜브의 1 준비 <ol><li> 점차적으로 튜브를 부드럽게, 손에 의해 가스 불 위의 30cm 긴 PE-50 튜브를 잡습니다. 부드럽게 튜브를 스트레칭. </li><li> 디지털 캘리퍼스 (그림 1)를 사용하여 연신 튜브에 지점을 선택하고, 그것을 표시하고 (외경 : 0.30-0.34 mm) 1cm 길이 끝이 25cm 긴 세그먼트로 수정 된 PE-50 튜브를 잘라. </li></ol> 동종 혈액 응고 2 준비 <ol><li> 70 % N 2 O 30 % O의 이소 플루 란 (유도 5 %, 유지 보수를 위해 2 ~ 3 %)로 쥐를 마취 &lt;채혈하기 전에 얼굴에 마스크에 의해 서브&gt; 2. 발가락 핀치에 의해 마취를 확인합니다. </li><li> 발행 방법 4 기증자 쥐의 대퇴 동맥 삽관을 수행하고 20cm 긴 PE-50 튜브에 직접 대퇴 동맥의 혈액을 전송합니다. 혈전이 혈액을 실온에서 2 시간 동안 튜브를 놓은 다음 4 ℃에서 22 시간 동안 튜브를 유지합니다. 참고 : 삽관 단계 3.1에 ​​설명 된대로 무균 기술을 사용하여 수행됩니다. 기증자 쥐 더욱 샘플링 회수한다. </li><li> 50mm 세그먼트로 PE-50 튜브를 잘라 생리 식염수가 들어있는 멸균 페트리 접시에 관에서 혈전을 세척하십시오. </li><li> 60mm 긴 PE-10 튜브에 50mm에게 긴 혈전을 전송하고 23 G 바늘을 생리 식염수를 포함하는 1 ML의 주사기에 연결된 20cm 긴 PE-50 관에 PE-10 튜브의 각 끝을 연결합니다 (그림 2) . </li><li> 혈액 세포를 제거 할 수있는, 5 분 동안 다른 하나의 주사기로부터의 연속 교호 이동에 의해 응고 시프트응고와 섬유소 코어를 방해하지 않고, 혈관 외 응고 된 혈전의 허약을 최소화 할 수 있습니다. </li><li> 40mm 긴 세그먼트로 혈전을 자르고 무균 조건 하에서 단계 1.2에 설명 된대로 수정 된 PE-50 카테터로 혈전 세그먼트를 전송합니다. 참고 : 모든 PE 튜브가 산화 에틸렌 살균된다. </li></ol> 3 색전 중간 대뇌 동맥 폐색 (그림 3A, B) <ol><li> (최소 121 ° C, 15 분 15 PSI를) 고압 증기 멸균하여 모든 수술 도구를 소독. 70 % 에탄올을 사용하여 수술 테이블과 연관된 수술 장비를 살균. </li><li> 2.1 단계에 설명 된대로 이소 플루 란과 함께 쥐를 마취. 모두 뒤 발에 발가락 핀치를 수행하여 마취의 깊이를 테스트하고 (발을 인출) 모든 관찰 운동은 동물이 수술을 할 충분히 마취가 아님을 나타냅니다. 마취 동안 건조를 방지하기 위해 두 눈에 수의사 연고의 작은 금액을 적용합니다. 복부 목과 머리 재에 털을 면도전기 면도기와 gions은 피부 영역을 노출시킨다. </li><li> 가열 패드에 누운 자세에서 쥐를 놓습니다. 직장 프로브, 컨트롤을 삽입하고 homeothermic 담요 제어 장치를 사용하여 36.5-37.5 ° C 사이의 체온을 유지한다. </li><li> 70 % 에탄올이어서 10 % 포비돈 - 요오드와 면도 피부와 주변 모피 소독. </li><li> 해부 현미경 목에 2 cm 길이의 중간 선 절개를합니다. 수술 부위를 노출하고 오른쪽 경동맥 (CCA), 외부 경동맥 (ECA), 내 경동맥 (ICA) 및 pterygopalatine 동맥 (PPA) 주변 신경 및 근막 (그림 3A)에서 무료 해부 견인기를 사용합니다. 참고 : 길잡이 동물을 준비하는 데 도움이없는 경우 수술 전에 멸균 장갑을 변경합니다. </li><li> (미주 신경에 손상을주지 않고) 주변 신경에서 CCA 무료 해부 및 동맥에서 멸균 5-0 실크 봉합사를 배치합니다. 슬립 매듭 (# 1)을 묶어 작은 HEMOS을 사용하여 봉합을 파악문신과는 몸으로 가르쳐 당깁니다. </li><li> ECA와 그 두 가지, 후두 동맥 (OA) 및 갑상선 동맥 (STA)을 해부하다. 수의학 전기 수술 장치를 사용하여 두 가지를 응고. ECA에서 멸균 6-0 실크 봉합사의 두 조각을 놓습니다. </li><li> 동맥, 머리를 향해 한 조각 (선단) 몸으로 다른 아래에 위치이 실크를 분리합니다. (2 #) 머리에 가장 가까운 측면에 작은 지혈제와 실크를 잡고 머리를 향해 가르쳐 당겨 꽉 합자 타이. 나중에 사용하기 위해 다른 실크 느슨한 매듭 (# 3)를 준비합니다. </li><li> (미주 신경에 손상을주지 않고) 주변 신경에서 ICA와 PPA를 해부하다. 6-0 봉합사 (# 4)와 PPA를 묶어. ICA (# 5) 약 6-0 봉합과 슬립 매듭을합니다. 다르게는, 미세 혈관 클립을 사용하여 ICA 클립. </li><li> Vannas 스타일의 봄 가위로 꽉 (# 2) 느슨한 (# 3) 합자 사이의 ECA에 작은 구멍 (를 통해 2)를 잘라. 수정 된 PE-50 욕조를 삽입CCA의 분기점에 절개를 사전에 혈전이 포함 된 전자. (3 #) 충분한 루멘 주변에 거주하는 튜브의 이동성을 보존 확보하기 위해 느슨한 합자를 조입니다. </li><li> 그루터기를 해제하고 ECA와 ICA의 분기점 아래의 그루터기를 배치하는 작은 구멍의 사이트에서 ECA를 잘라; 이 개질 관 쉽게 ICA로 밀어 허용 할 것이다. ICA를 열고 카테터의 끝이 MCA의 근원에 도달 할 때까지 부드럽게 (~ 분기점에서 17mm를) ICA에 ECA의 루멘에서 튜브를 진행합니다. 참고 : 전에 ECA, 70 % 에탄올로 튜브의 멸균 외부 표면에 튜브의 끝 부분을 삽입 한 다음 멸균 생리 식염수로 씻어. </li><li> 100 μL 해밀턴 주사기 (그림 3B)를 사용하여 10 초 이상 생리 식염수 10 ㎕를 함께 수정 된 PE-50 카테터를 통해 혈전을 주입한다. </li><li> 5 분 후에 ECA에서 카테터를 철회. ECA를 묶어 CCA를 다시 엽니 다. 목에 절개를 봉합. </li></ol> (4) 모니터링 지역 대뇌 혈류 (rCBF) <ol><li> 이전 MCA 폐색으로, 두개골의 뼈를 노출 두피에 1.2 cm 긴 중간 선 절개를합니다. 치과 스크레이퍼 및 멸균면 스왑과 두개골 뼈의 조직을 제거합니다. 참고 : 수술 전에 노출 된 두피 지역 및 주변 모피는 70 % 에탄올 다음 10 % 포비돈 요오드로 소독한다. </li><li> 2mm 후방 및 0.7 mm 구형 스테인레스 버어 (도 4)를 이용하여 5mm 브레 그마 측방에 위치한 직경 1.5 mm 버 홀을 뚫는다. 주 : 그대로 두라을 유지합니다. </li><li> 프로브에게 경질 표면 위의 0.5 mm를 놓습니다. 0 (베이스 라인)에서 rCBF, 5, 15, 30, 60, 90을 모니터링하고 색전술 후 및 120 분에서 계속 5, 15, 30, 45, 및 tPA의 정맥 내 투여 후 60 분. rCBF의 마지막 측정 후, 두피 절개는 봉합에 의해 폐쇄된다. 참고 : 목 절개를하기 전에 우리가 MCA의 occlus 전 기준으로 rCBF을 측정이온. 포스트 MCA 폐색 우리는 목 절개 봉합 폐쇄 후 rCBF을 측정한다. 따라서, 목 절개 무균 유지된다. </li></ol> (5) 수술 후 케어 <ol><li> 탈수를 방지하고 즉시 수술과 통증 완화에 필요한 모든 6 ~ 12 시간 후 Buprenex (0.05 ㎎ / ㎏, SC)를 주입하는 식염수를 피하의 2.5 ml에 주입한다. </li><li> 이소 플루 란 마취를 중지합니다. 37 ° C 수의학 회수 챔버에 쥐를 놓고 (동물을 따뜻하게 유지) 관찰을 유지합니다. 쥐가 마취에서 회복하는 것은 일반적으로 10 분입니다. 그런 다음, 멸균 케이지에 동물을 넣어 장에 페트리 접시에 약간 젖은 음식을 놓고 동물 살균 방에 케이지를 반환합니다. </li><li> 뇌졸중 후 24 시간이 나트륨 펜토 바르 비탈 (100 ㎎ / ㎏ 체중, IP)의 과다로 래트를 안락사. </li></ol> (6) 신경 적자 점수 <ol><li> 전 색전술 후 2 시간에서 Bederson 점수를 수행합니다. 등급의를 사용하여0-3의 케일이 아니라 이전에 5 설명 : 꼬리 쥐를 올리는 공을 바닥에 두 앞발을 확장하고 다른 적자를 나타내지 않는 동물 (정상 동작); 한, 꼬리 쥐를 제기 반대측 앞다리 플렉스; 도 2는 선회하지 않고 횡 푸시 (및 앞다리 굴곡)에 대한 내성을 감소; 3, 돌고와 등급이 동일한 동작. 참고 : 색전술 후 2 시간에서 Bederson 점수 = 0 (적자)을 보여주는 쥐가 더 많은 연구에서 제외됩니다. </li></ol>

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We do not have any potential conflicting interests to disclose.

본 연구에서 우리는 MCA의 원점이 풍부한 피브린 응고에 의해 폐색 된 쥐에서 색 전성 뇌졸중 MCAO 모델을 수행하기위한 표준 방법을 설명했다. 이 모델의 주요 장점은 : 피브린 풍부한 혈병과 MCA의 줄기의 폐색은 인간에서 혈전 색 전성 뇌졸중 유사 색 전성 뇌졸중 모델 혈전 용해 요법의 전임상 연구를 수행하기에 적합하고,이 모델을 개발할 수 있음 MCA에 의해 공급 영토 내에서 재현하고 예측 경?...

A correction was made to <a href="/video/51956">Embolic Middle Cerebral Artery Occlusion (MCAO) for Ischemic Stroke with Homologous Blood Clots in Rats</a>. The institution information was updated.
The institution "Louisiana State University Health Science Center" was changed to "Louisiana State University Health Science Center, Shreveport".

A correction was made to <a href="/video/51956">Embolic Middle Cerebral Artery Occlusion (MCAO) for Ischemic Stroke with Homologous Blood Clots in Rats</a>. The institution information was updated.

Erratum: Embolic Middle Cerebral Artery Occlusion (MCAO) for Ischemic Stroke with Homologous Blood Clots in Rats

뇌졸중은 미국에서 사망의 세 번째 주요 원인이지만, 급성 뇌졸중의 치료 옵션이 제한 남아있다. 현재, 혈전을 용해하는 재조합 조직 플라스 미노 겐 활성제 (TPA)의 정맥 주입은 급성 허혈성 뇌졸중에 대한 가장 효과적인 치료법이다. 그러나, tPA의 사용은 좁은 치료 창이과 뇌출혈의 위험 증가에 의해 제한됩니다. 따라서 혈전 용해 연구에 적합한 행정의 모델을 긴급하게 필요합니다. 중대 뇌동맥 (MCA)는 가장 자주 인간 스트로크에 가려 동맥이다. 이 동맥에 초점을 맞춘, 허혈성 뇌졸중의 여러 동물 모델이 설정되어있는 것. 현재, MCA를 폐색에 의해 쥐 초점 허혈 모델의 두 가지 주요 유형이 개발되어 있습니다 : 봉합 MCAO 모델과 전성 MCAO 모델. 관내 봉합 MCAO 기법을 통해 모델 널리기구 구동 스트로크 연구에 사용되어 왔지만, 이러한 봉합 모델 몰라인간의 스트로크의 80 %까지로 모방 인간의 스트로크는, 혈전이나 색전에 의해 발생합니다. 그러나, 혈액 응고를 사용하여 색 전성 뇌졸중 모델은 밀접하게 인간의 스트로크를 모방하고 혈전 용해 연구에 적합한 것으로 간주됩니다. 이 전성 모델은 1992 년에 처음 Overgaard 등. 1 쥐에서 개발 및 추가 장 등을 특징으로했다. 1,997 두와 디 나폴리 등에 의해. 2,006 3. 전성 MCAO 증가 주목을 받았다 있지만, 직면 한 기술적 문제가 많은 실험실에 의해. 이 문서에서는, 우리는 MCA 영토 내에서 예측 경색을 개발할 수 있습니다 성인 쥐에서 동종 혈액 응고와 색전 MCAO을 생산하기위한 표준 방법을 보여줍니다. 이 문서에 제시된 기술은 연구자 뇌졸중 연구를위한이 모델을 설정하기위한 기술적 인 문제를 극복하는 데 도움이됩니다.

<table><tbody><tr><td>rh-tPA</td><td>Chemical</td><td>Genentech</td><td></td></tr><tr><td>2,3,5-triphenyltetrazolium chloride</td><td>Chemical</td><td>Sigma</td><td>T8877</td></tr><tr><td>Anesthesia vaporizer</td><td>Equipment</td><td>Soma Technology</td><td>Drager Vapor 19.1&amp;nbsp;&amp;nbsp;</td></tr><tr><td>Rechargeable high speed micro drill</td><td>Equipment</td><td>Fine Science Tools</td><td>18000-17</td></tr><tr><td>Curved scissors</td><td>Equipment</td><td>Fine Science Tools</td><td>14117-14</td></tr><tr><td>Dumont forceps (Medical #7)</td><td>Equipment</td><td>Fine Science Tools</td><td>11270-20</td></tr><tr><td>Dumont forceps (Medical #5)</td><td>Equipment</td><td>Fine Science Tools</td><td>11251-35</td></tr><tr><td>Vannas-style spring scissors</td><td>Equipment</td><td>Fine Science Tools</td><td>15000-03</td></tr><tr><td>Veterinary recovery chamber&amp;nbsp;</td><td>Equipment&amp;nbsp;</td><td>Peco Services&amp;nbsp;</td><td>V1200&amp;nbsp;&amp;nbsp;</td></tr><tr><td>Genie plus syringe pump</td><td>Equipment</td><td>Kent Scientific Corporation</td><td></td></tr><tr><td>Rat brain matrix</td><td>Equipment</td><td>Kent Scientific Corporation</td><td>RBMA-310C</td></tr><tr><td>Digital caliper&amp;nbsp;</td><td>Equipment</td><td>World Precision Instruments</td><td>501601</td></tr><tr><td>Dissecting microscope</td><td>Equipment</td><td>World Precision Instruments</td><td>PZMTIII-BS-LWD</td></tr><tr><td>Hamilton syringe</td><td>Equipment</td><td>Hamilton</td><td>model 710</td></tr><tr><td>Homeothermic blanket control unit</td><td>Equipment</td><td>Harvard Apparatus</td><td></td></tr><tr><td>Electric clipper</td><td>Equipment</td><td>Braintree Scientific</td><td>CLP-9931</td></tr><tr><td>Veterinary electrosurgical unit</td><td>Equipment</td><td>MACAN Manufacturing Company</td><td>MV-9</td></tr><tr><td>Blood flowmeter</td><td>Equipment</td><td>Adinstruments</td><td></td></tr><tr><td>PowerLab 4/30</td><td>Equipment</td><td>Adinstruments</td><td></td></tr><tr><td>LabChart 7.2</td><td>software</td><td>Adinstruments</td><td></td></tr><tr><td>1 ml syringe</td><td>Consumable</td><td>Becton, Dickinson and Company</td><td>309659</td></tr><tr><td>23 G needle</td><td>Consumable</td><td>Becton, Dickinson and Company</td><td>305143</td></tr><tr><td>30 G needle</td><td>Consumable</td><td>Becton, Dickinson and Company</td><td>305106</td></tr><tr><td>PE-50 tubing</td><td>Consumable</td><td>Becton, Dickinson and Company</td><td>427517</td></tr><tr><td>PE-10 tubing</td><td>Consumable</td><td>Becton, Dickinson and Company</td><td>427400</td></tr><tr><td>6-0 Silk suture</td><td>Consumable</td><td>Harvard apparatus</td><td>723287</td></tr><tr><td>5-0 Silk suture</td><td>Consumable</td><td>Harvard Apparatus</td><td>517607</td></tr></tbody></table>

embolic middle cerebral artery occlusion (mcao) for ischemic stroke with homologous blood clots in rats

재조합 조직 플라스 미노 겐 활성제 (TPA)의 사용과 임상, 혈전 용해 치료는 급성 허혈성 뇌졸중의 가장 효과적인 치료 남아있다. 그러나, tPA의 사용은 좁은 치료 윈도우 의해 출혈성 변형의 위험 증가에 의해 제한된다. 허혈성 뇌졸중의 치료를위한 새로운 혈전 용해제 및 전략을 연구하기 적합한 뇌졸중 모델을 개발하기위한 시급한 필요성이있다. 관내 봉합 MCAO 및 색전 MCAO : 현재, 허혈성 뇌졸중 모델의 두 가지 주요 유형은 쥐와 생쥐에서 개발되었다. 관내 봉합 MCAO 기법을 통해 모델 널리기구 구동 스트로크 연구에 사용되어 왔지만, 이러한 봉합 모델 임상 상황을 모방하지 혈전 및 연구에 적합하지 않는다. 이러한 모델 중 전성 MCAO 모델은 밀접하게 인간의 허혈성 뇌졸중을 모방하고 혈전 용해 치료의 임상 연구에 적합하다. 이 전성 모델은 첫째 OV에 의해 쥐에서 개발 된1992 년. 등 1 ergaard 또한 장 등이 특징. 1,997 2. 전성 MCAO가 관심을 증가 얻고있다하더라도, 많은 실험실에서 직면 기술적 인 문제가 있습니다. 혈전 용해 연구에 대한 증가하는 요구를 충족하기 위해, 우리는 MCA 영토 내에서 예측 경색 볼륨을 개발할 수 쥐에 전성 MCAO의 높은 재현성 모델을 제시한다. 요컨대, 변성 PE-50 튜브 부드럽게 MCA의 원점에 도달 카테터의 팁까지 내 경동맥 (ICA)의 내강에 외부 경동맥 (ECA)에서 전진된다. 카테터를 통해 하나의 동종 혈액 응고는 MCA의 기원에 배치됩니다. MCA 폐색의 성공을 식별하는 지역 뇌 혈류량을 모니터링하고, 신경 학적 결손 및 경색 부피를 측정 하였다. 이 논문에서 제시 한 기술은 뇌졸중 연구를위한 모델을 구축하기위한 기술적 인 문제를 극복하기 위해 수사관을 도움이 될 것이다. &lt;/ P&gt;

레이저 도플러 flowmetry (LDF)는 뇌허혈 6,7의 유도시 rCBF를 모니터링하는 데 사용 하였다. 우리 연구실을 포함한 많은 실험실이 성공 MCA 폐색 동물을 식별 할 수 rCBF를 사용하고 있지만,베이스 라인의 임계 값은 측정 사이트에 관련된 실험실 사이에서 변화. LDF의 프로브는 2mm의 후방에 배치되고, 이전에 설명한 바와 같이 6 5mm는 브레 그마 측방. rCBF 0에서 모니터 된 5, 15, 30, 60, 90, 및 120...

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Embolic Middle Cerebral Artery Occlusion MCAO for Ischemic Stroke with Homologous Blood Clots in Rats

In this paper, we demonstrate a standard method for producing an embolic middle cerebral artery occlusion with homologous blood clots (fibrin-rich) in adult rat. This model closely mimics human ischemic stroke and is suitable for preclinical study of thrombolytic therapy for ischemic stroke.

흰쥐의 동종 혈액 혈전과 허혈성 뇌졸중에 대한 전성 중간 대뇌 동맥 폐색 (MCAO)

Clinically, thrombolytic therapy with use of recombinant tissue plasminogen activator (tPA) remains the most effective treatment for acute ischemic ...

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Embolic Middle Cerebral Artery Occlusion (MCAO) for Ischemic Stroke with Homologous Blood Clots in Rats

26.9K 조회수.  Louisiana State University Health Science Center, Shreveport. In this paper, we demonstrate a standard method for producing an embolic middle cerebral artery occlusion with homologous blood clots (fibrin-rich) in adult rat. This model closely mimics human ischemic stroke and is suitable for preclinical study of thrombolytic therapy for ischemic stroke. 

비디오: Embolic Middle Cerebral Artery Occlusion MCAO for Ischemic Stroke with Homologous Blood Clots in Rats

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.

쥐 중간 대뇌 동맥의 폐색의 Endovascular 봉합사에 의해 초점 대뇌 국소 빈혈 모델

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 ...

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신경과학

Mouse Model of Middle Cerebral Artery Occlusion

Stroke is the most common fatal neurological disease in the United States 1. The majority of strokes (88%) result from blockage of blood vessels in the brain (ischemic stroke) 2. Since most ischemic strokes (~80%) occur in the territory of middle cerebral artery (MCA) 3, many animal stroke models that have been developed have focused on this artery. The intraluminal monofilament model of middle cerebral artery occlusion (MCAO) involves the insertion of a surgical filament into the external carotid artery and threading it forward into the internal carotid artery (ICA) until the tip occludes the origin of the MCA, resulting in a cessation of blood flow and subsequent brain infarction in the MCA territory 4. The technique can be used to model permanent or transient occlusion 5. If the suture is removed after a certain interval (30 min, 1 h, or 2 h), reperfusion is achieved (transient MCAO); if the filament is left in place (24 h) the procedure is suitable as a model of permanent MCAO. This technique does not require craniectomy, a neurosurgical procedure to remove a portion of skull, which may affect intracranial pressure and temperature 6. It has become the most frequently used method to mimic permanent and transient focal cerebral ischemia in rats and mice 7,8. To evaluate the extent of cerebral infarction, we stain brain slices with 2,3,5-triphenyltetrazolium chloride (TTC) to identify ischemic brain tissue 9. In this video, we demonstrate the MCAO method and the determination of infarct size by TTC staining.

We demonstrate in the video a method for producing a middle cerebral artery occlusion in adult mice using an intraluminal monofilament. We also show how to evaluate the extent of cerebral infarction by 2,3,5-triphenyltetrazolium chloride (TTC) staining.

중간 대뇌 동맥 폐색의 마우스 모델

Stroke is the most common fatal neurological disease in the United States 1. The majority of strokes (88%) result from blockage of blood vessels in the ...

의학

Endothelin-1 Induced Middle Cerebral Artery Occlusion Model for Ischemic Stroke with Laser Doppler Flowmetry Guidance in Rat

Stroke is the number one cause of disability and third leading cause of death in the world, costing an estimated $70 billion in the United States in 20091, 2. Several models of cerebral ischemia have been developed to mimic the human condition of stroke. It has been suggested that up to 80% of all strokes result from ischemic damage in the middle cerebral artery (MCA) area3. In the early 1990s, endothelin-1 (ET-1) 4 was used to induce ischemia by applying it directly adjacent to the surface of the MCA after craniotomy. Later, this model was modified 5 by using a stereotaxic injection of ET-1 adjacent to the MCA to produce focal cerebral ischemia. The main advantages of this model include the ability to perform the procedure quickly, the ability to control artery constriction by altering the dose of ET-1 delivered, no need to manipulate the extracranial vessels supplying blood to the brain as well as gradual reperfusion rates that more closely mimics the reperfusion in humans5-7. On the other hand, the ET-1 model has disadvantages that include the need for a craniotomy, as well as higher variability in stroke volume8. This variability can be reduced with the use of laser Doppler flowmetry (LDF) to verify cerebral ischemia during ET-1 infusion. Factors that affect stroke variability include precision of infusion and the batch of the ET-1 used6. Another important consideration is that although reperfusion is a common occurrence in human stroke, the duration of occlusion for ET-1 induced MCAO may not closely mimic that of human stroke where many patients have partial reperfusion over a period of hours to days following occlusion9, 10. This protocol will describe in detail the ET-1 induced MCAO model for ischemic stroke in rats. It will also draw attention to special considerations and potential drawbacks throughout the procedure.

Several animal models of cerebral ischemia have been developed to simulate the human condition of stroke. This protocol describes the endothelin-1 (ET-1) induced middle cerebral artery occlusion (MCAO) model for ischemic stroke in rats. In addition, important considerations, advantages, and shortcomings of this model are discussed.

쥐의 레이저 도플러 Flowmetry 안내와 허혈성 뇌졸중에 대한 엔도 테린-1 유도 중간 뇌 동맥 폐색 모델

Stroke is the number one cause of disability and third leading cause of death in the world, costing an estimated $70 billion in the United States in ...

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 혈관 폐색 / 저혈압 : 세계적인 두뇌 국소 빈혈의 쥐 모델

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

Optimized System for Cerebral Perfusion Monitoring in the Rat Stroke Model of Intraluminal Middle Cerebral Artery Occlusion

The translational potential of pre-clinical stroke research depends on the accuracy of experimental modeling. Cerebral perfusion monitoring in animal models of acute ischemic stroke allows to confirm successful arterial occlusion and exclude subarachnoid hemorrhage. Cerebral perfusion monitoring can also be used to study intracranial collateral circulation, which is emerging as a powerful determinant of stroke outcome and a possible therapeutic target. Despite a recognized role of Laser Doppler perfusion monitoring as part of the current guidelines for experimental cerebral ischemia, a number of technical difficulties exist that limit its widespread use. One of the major issues is obtaining a secure and prolonged attachment of a deep-penetration Laser Doppler probe to the animal skull. In this video, we show our optimized system for cerebral perfusion monitoring during transient middle cerebral artery occlusion by intraluminal filament in the rat. We developed in-house a simple method to obtain a custom made holder for twin-fibre (deep-penetration) Laser Doppler probes, which allow multi-site monitoring if needed. A continuous and prolonged monitoring of cerebral perfusion could easily be obtained over the intact skull.

Cerebral perfusion monitoring has been demonstrated to improve accuracy in ischemic stroke models. Technical difficulties often limit the use of this essential tool for cerebrovascular research. In this video, an optimized system is shown to obtain a single or multi-site hemodynamic monitoring during intraluminal middle cerebral artery occlusion in rats.

Intraluminal 중간 대뇌 동맥 폐색의 쥐 뇌졸중 모델에서 뇌 관류 모니터링에 최적화 된 시스템

The translational potential of pre-clinical stroke research depends on the accuracy of experimental modeling. Cerebral perfusion monitoring in animal ...

Permanent Cerebral Vessel Occlusion via Double Ligature and Transection

Stroke is a leading cause of death, disability, and socioeconomic loss worldwide. The majority of all strokes result from an interruption in blood flow (ischemia) 1. Middle cerebral artery (MCA) delivers a great majority of blood to the lateral surface of the cortex 2, is the most common site of human stroke 3, and ischemia within its territory can result in extensive dysfunction or death 1,4,5. Survivors of ischemic stroke often suffer loss or disruption of motor capabilities, sensory deficits, and infarct. In an effort to capture these key characteristics of stroke, and thereby develop effective treatment, a great deal of emphasis is placed upon animal models of ischemia in MCA.
Here we present a method of permanently occluding a cortical surface blood vessel. We will present this method using an example of a relevant vessel occlusion that models the most common type, location, and outcome of human stroke, permanent middle cerebral artery occlusion (pMCAO). In this model, we surgically expose MCA in the adult rat and subsequently occlude via double ligature and transection of the vessel. This pMCAO blocks the proximal cortical branch of MCA, causing ischemia in all of MCA cortical territory, a large portion of the cortex. This method of occlusion can also be used to occlude more distal portions of cortical vessels in order to achieve more focal ischemia targeting a smaller region of cortex. The primary disadvantages of pMCAO are that the surgical procedure is somewhat invasive as a small craniotomy is required to access MCA, though this results in minimal tissue damage. The primary advantages of this model, however, are: the site of occlusion is well defined, the degree of blood flow reduction is consistent, functional and neurological impairment occurs rapidly, infarct size is consistent, and the high rate of survival allows for long-term chronic assessment.

Permanent Cerebral Vessel Occlusion via Double Ligature and Transection

We describe a highly reproducible method for the permanent occlusion of a rodent major cerebral blood vessel. This technique can be accomplished with very little peripheral damage, minimal blood loss, a high rate of long-term survival, and consistent infarct volume commensurate with the human clinical population.

영구적 인 뇌 혈관 폐색<em&gt;를 통해</em&gt; 더블 합자 및 절개

Stroke is a leading cause of death, disability, and socioeconomic loss worldwide. The majority of all strokes result from an interruption in blood flow ...

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.

세 쥐의 경동맥 폐색으로 영구 말초 중간 대뇌을 수행하면 두피 허혈 지속적인 장애를 연구하는

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

Middle Cerebral Artery Occlusion Allowing Reperfusion via Common Carotid Artery Repair in Mice

The ischemic stroke is a major cause of adult long-term disability and death worldwide. The current treatments available are limited, with only tissue plasminogen activator (tPA) as an approved drug treatment to target ischemic strokes. Current research in the field of ischemic stroke focuses on better understanding the pathophysiology of stroke, to develop and investigate novel pharmaceutical targets. Reliable experimental stroke models are crucial for the progression of potential treatments. The middle cerebral artery occlusion (MCAO) model is clinically relevant and the most frequently used surgical model of ischemic stroke in rodents. However, the outcomes of this model, such as lesion volume, are associated with high levels of variability, particularly in mice. The alternative MCAO model described here allows the reperfusion of the common carotid artery (CCA) and the increased perfusion of the middle cerebral artery (MCA) territory, using a tissue pad with fibrinogen-based sealant to repair the vessel, and the improved welfare of the mice by avoiding external carotid artery (ECA) ligation. This reduces the reliance on the Circle of Willis, which is known to be highly anatomically variable in mice. Representative data show that using this alternative surgical approach decreases the variability in lesion volumes between the traditional MCAO approach and the alternative approach described here.

Intraluminal filament occlusion of the middle cerebral artery is the most frequently used in vivo model of experimental stroke in rodents. An alternative surgical approach to allow common carotid artery repair is performed here, which allows the reperfusion of the common carotid artery and a full reperfusion to the middle cerebral artery territory.

쥐에 있는 일반적인 경 동맥 수리 통해 Reperfusion를 허용 하는 중간 대뇌 동맥 폐색

The ischemic stroke is a major cause of adult long-term disability and death worldwide. The current treatments available are limited, with only tissue ...

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>쥐 MCAO 뇌졸중 모델의 세포 사멸을 연구하기 위한 면역형광 염색

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>허혈성 뇌졸중의 쥐 모델에서 면역형광에 의한 세포 사멸 신호 평가

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.

허혈성 뇌졸중의 쥐 모델에서 면역형광에 의한 세포 사멸 신호 평가

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 ...