수술 기술은 원래 조지아 메디컬 대학에서 박사 윌리엄 D. 힐의 연구실에 인수되었다. 저자는 또한 절개 카메라의 사용에 대한 데이빗 A. Rempe 및 란다 Prifti 감사하고 싶습니다. 이 연구는 NIH NS041744, NS051279, F31 NS064700 및 AHA 30815697D 지원했다.

이 프로토콜은 연구에 동물의 윤리적 사용 (UCAR)에 전념 로체스터위원회의 대학에 의해 승인되었습니다. 무균 기법은 프로토콜 동안에 따라야한다. 무균 장갑과 마스크의 사용이 필요합니다. 프로토콜 동안 사용되는 모든 장비, 재료, 화학 물질 및 도구는 표 1에 설명되어 있습니다. 1. 사전 수술 준비 <ol><li> 즉시 수술 후 후 처음 24 H 포스트 수술 기간 동안 매일 3-5 H, buprenorphine (0.05mg/kg) 수술 전에 2 H와 subcutaneously 쥐를 주사. </li><li> 압력솥의 모든 수술기구, 거즈 스폰지, 그리고 면화 팁 applicators을 소독. 바로 사용하기 전에 멸균 거즈에 건조 수술과 공기 중에 70 % 에탄올에 수술 도구를 보관하십시오. 70 % 에탄올로 작업 영역을 스프레이. </li><li> 마취 기화기를 사용하여 3 % isofluorane - 20 %의 산소 기체 혼합물로 마우스를 마취. 유량계와 산소 금액을 조정합니다. 발가락이나 꼬리 곤란 (그들은 응답한다)에 의해 마취의 수준을 테스트합니다. 2퍼센트 (V / V) isofluorane과 마취의 수준을 유지합니다. </li><li> 마우스의 눈에 인공 눈물을 적용하고 수술 중에 눈을 손상을 방지하기 위해주의를 사용합니다. 가로 위치의 오른쪽에있는 마우스를 놓습니다. </li><li> 왼쪽 눈과 동물 가위를 사용하여 왼쪽 귀의 기본 사이에 왼쪽에있는 영역을 쉐이브. betadine 솔루션과 면화 팁 applicators과 가볍게 지역을 바르고을 사용하여 70 % 에탄올 사이의 교류에 의해이 지역을 정화. 필요에 따라 반복합니다. </li><li> 37 체온을 유지하기 위해 직장 탐침에 연결된 손난로 ° C.에 마우스를 놓고 미네랄 오일을 사용하여 직장 프로브를 삽입합니다. </li><li> 현미경의 오른쪽에 마우스를 위치하고 테이프로 고정하십시오. 거즈 스폰지에 창문을 잘라 외과 영역을 커버. </li></ol> 2. 수술 및 MCA 내고 <ol><li> 왼쪽 눈 및 좋은 직선 가위를 사용하여 왼쪽 귀의 기지 사이에 수직 절개를합니다. 외과 영역을 넓히려고 곡선 hemostats를 사용합니다. </li><li> 스프링 가위를 사용하여 시간적 근육에 수평 절개를 만들고 약간 천천히 포셉로 당겨 두개골에서 관자놀이 근육을 분리. </li><li> 굽​​은 집게와 턱뼈를 누른 상태에서 18G 바늘을 사용하여 광대뼈 아치와 squamosal 뼈의 교차점에 작은 subtemporal craniotomy하십시오. </li><li> 뼈 rongeurs와 함께 두개골의 작은 조각을 제거하여 MCA을 폭로. 광대뼈 아치와 궤도 내용은이 과정에서 손상되지 않습니다. 조직의 과도한 건조는이 과정에서 발생하면, 면화 팁 applicators과 멸균 PBS를 적용합니다. </li><li> 작은 선박 cauterizer를 사용하여 MCA의 말초 부분을 Ligate. </li><li> 원래 위치로 다시 관자놀이 근육을 놓고 수술 5-0 나일론 봉합과 절개 사이트를 닫습니다. </li><li> 마취를 중단하고 직장 프로브를 제거합니다. subcutaneously buprenorphine (0.05mg/kg)의 두번째 복용량으로 마우스를 주사. 난방 패널 ° C 37 보관했던 케이지 마우스를 반환합니다. </li><li> 밀접하게 어떤 불편 (감소 식욕 / 물 소비, hunched 자세, 호흡 증가, 머리카락을 pilo가 - 건설)에 대한 향후 24 H에 대한 마우스를 모니터합니다. 최대 24 시간에 buprenorphine subcutaneously 매일 3-5 H 수술로 마우스를 주사. 이 기간 동안 복구 젤로 마우스를 제공합니다. </li></ol> 3. TTC의 스테 이닝과 뇌졸중 볼륨의 결정 <ol><li> 수술 후 스물넷 시간 깊이 마우스를 마취, transcardially 인산의 4% TTC (W / V)을 사용하여 10 분에 대한 4 % paraformaldehyde 솔루션 후 15 분 및 위해 호수 (PBS)를 버퍼로 마우스를 perfuse 중간 유량의 미니 연동이 펌프. 두뇌를 제거하고 밤새 4 % paraformaldehyde 용액에 넣습니다. </li><li> sectioning 블록에 머리를 놓습니다. 면도날을 사용하여 1mm 두께의 조각에 머리를 슬라이스. </li><li> 밀리미터 스케일 통치자 옆에있는 슬라이스를 놓습니다. 해부 현미경에 연결된 디지털 카메라를 사용하여 조각을 사진에 담아보세요. </li><li> 이미지 J 소프트웨어 (http://rsbweb.nih.gov/ij/)를 사용하여 contralateral 사이트의 전체 영역에서 ipsilateral 사이트의 비 infarcted 지역을 빼서하여 스트로크 영역을 계산합니다. 조각 22 스트로크 영역을 쌓아하여 스트로크 볼륨을 계산합니다. </li></ol> 4. 방향 이미지 J 소프트웨어를 사용하는 <ol><li> 파일 메뉴를 클릭하여 이미지 J 소프트웨어의 분석에 이미지 파일을 엽니다. </li><li> 프로그램의 &#39;직선&#39;탭을 클릭하십시오. 눈금자에있는 두 개의 여백 사이에 직선을 그립니다. &#39;분석&#39;메뉴를 클릭하고 &#39;설정 규모&#39;를 선택합니다. 1로 알려진 거리, mm로 길이의 단위를 설정합니다. </li><li> &#39;프리핸드 원&#39;탭을 클릭하십시오. contralateral 반구를 대략적으로 원을 그립니다. &#39;분석&#39;메뉴를 클릭하고 &#39;측정&#39;을 선택합니다. 계산 창이 그려진 동그라미의 영역을 보여주는 팝업됩니다. </li><li> ipsilateral 반구 제외 뇌졸중 (흰색) 주변 다른 원을 그립니다. 로 단계 4.3에 표시된 영역을 측정합니다. 새로운 값은 계산 창에 추가됩니다. 1, 2 가치의 차이는 아래의 수식으로 표시됩니다 경색의 영역을 나타냅니다. </li><li> 4.2-4.4 단계를 반복하여 각 슬라이스에서 스트로크 영역을 계산합니다. 각 슬라이스 (ΣA N)으로 계산 스트로크 지역의 변론을 가져가라. 이것은 각 슬라이스의 두께는 1mm있다는 사실을 알고 뇌졸중 볼륨을 나타냅니다. ΣA N X 1mm (각 슬라이스의 두께) = 뇌졸중 볼륨 </li></ol> 5. 대표 결과 : 마우스의 영구 MCA의 결합에 의해 얻은 infarcts는 대부분 대뇌 피질 있습니다. 그러나, 그것은 MCA가 lenticulostriate 지점 근위 출혈도 잡았 경우 subcortical 병변을 얻을 수 있습니다. MCA 내고 후 뇌졸중 볼륨 10mm ³에서 35mm ³ 19 23 달라질 수 있습니다. 스트로크 볼륨 23 35mm 20mm ³ ³하는 사이 Filiano 외의 MRI 이미지에서 결정하면서 Moyanova 외의 TTC의 얼룩과 계산 스트로크 볼륨. 19 22mm 10mm ³ ³로 사이에있다. 아마 내고의 정확한 위치 및 스트로크 볼륨을 측정하는 데 사용되는 다른 방법이 차이에 사용할 수있는 이유. 그림 1에서, 야생의 유형 C57/BL6 마우스에서 TTC 자국 두뇌 24 시간 게시물 영구 MCA의 결합이 표시됩니다. 뇌졸중 사이트 모양 (그림 1A, B)에 흰색입니다. 그림 1A와 B는 서로 다른 각도에서 경색 사이트를 보여줍니다. 그림 2는 (그림 2A - G) 후부에 앞쪽에에서 TTC 스테인드 스트로크 두뇌의 1mm 두께의 조각 보여줍니다. 경색 부피 24 H 수술 후 계산되었다. 경색의 볼륨 ~ 23mm ³이다. <img src="/files/ftp_upload/3039/3039fig1.jpg" alt="그림 1"/> 그림 1. 스트로크 사이트입니다. AB의 표현, TTC는 MCA의 결합 후 전체 뇌 이미지, 24 H를 더럽 혔다. 화이트 영역 경색을 나타냅니다. <img src="/files/ftp_upload/3039/3039fig2.jpg" alt="그림 2"/> 그림 2. 대표 결과입니다. (A&gt; G), TTC 스테인드 1mm 두뇌 섹션, MCAL 후 24 H. 조각은 뒷부분에 앞쪽에에서 정렬됩니다. 화이트 영역 경색을 나타냅니다.

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J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rodent+models+of+focal+cerebral+ischemia.">Rodent models of focal cerebral ischemia.</a> Current Protocols in Neuroscience. , 9.6.1-9.6.32 (2000).</li><li>Lin, T. N., He, Y. Y., Wu, G., Khan, M., Hsu, C. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+brain+edema+on+infarct+volume+in+a+focal+cerebral+ischemia+model+in+rats.">Effect of brain edema on infarct volume in a focal cerebral ischemia model in rats.</a> Stroke. 24, 117-121 (1993).</li><li>Filiano, A. J., Tucholski, J., Dolan, P. J., Colak, G., Johnson, G. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transglutaminase+2+protects+against+ischemic+stroke.">Transglutaminase 2 protects against ischemic stroke.</a> Neurobiol Dis. 39, 334-343 (2010).</li></ol>

관심 없음 충돌 선언하지 않습니다.

영구 MCA의 결합 방법은 높은 재현성 경색 볼륨과 사용 가능한 다른 방법에 비해 증가 이후 수술 생존 속도를 제공합니다. 절차의 간편하고 짧은 기간 (~ 30 분)는 더욱 실용적합니다. 방법은 널리 마우스 및 쥐 모두에서 사용됩니다. 이 기술은 stereomicroscope 아래 침략 수술을 필요로합니다. 따라서 현미경으로 운영하고 성공적인 craniotomy을 완성 경험이 필수적입니다. 실험이 수?...

<table border="1"><tbody><tr><th> NAME </th><th> 회사 및 카탈로그 번호 </th></tr><tr><td> A. 솔루션 및 화학 물질 </td><td></td></tr><tr><td> Povidone - 요오드 솔루션, 외과 스크럽 </td><td> Aplicare, 82-209 </td></tr><tr><td> 70 % 에탄올 </td><td> Koptec, V1001 </td></tr><tr><td> 2,3,5 Triphenyltetrazolium 염화 (TTC) </td><td> 시그마, T8877 </td></tr><tr><td> 4 % Paraformaldehyde </td><td> 전자 현미경 과학, 159SP </td></tr><tr><td> 7.4 : 인산이 살린, 산도를 버퍼 </td><td> 만든 실험실에서 살균. </td></tr><tr><td> 미네랄 오일 </td><td> PML Microbiologicals, R6570 </td></tr><tr><td> Isofluorane </td><td> 버틀러 회사, 029,405 </td></tr><tr><td> Buprenorphine (0.3mg/ml) </td><td> Hospira, NDC 0409-2012-32 </td></tr><tr><td> 인공 눈물 </td><td> 버틀러 회사, 007,312 </td></tr><tr><td> 복구 젤 </td><td> 지우기 H 2 O, 72-01-5022 </td></tr><tr><td> B. 외과 재료 및 장비 </td><td></td></tr><tr><td> 10-12주 오래된 C57/BL6 마우스, </td><td> 식민지는 대학 동식물 사육장에서 자랐다는 </td></tr><tr><td> 100 % 산소 탱크 </td><td> Airgas </td></tr><tr><td> 마취 기화기 및 유량계 </td><td> Surgivet, 모델 100 </td></tr><tr><td> 직장 프로브와 함께 가열 패드 </td><td> 파인 과학 도구, TR200 </td></tr><tr><td> 해부 현미경 </td><td> 자이스 혈구, Stemi 2000 </td></tr><tr><td> 난방 패널 </td><td> 페트코 서비스 (주) 0307-013 </td></tr><tr><td> 소형 선박 Cauterizer 세트 </td><td> 파인 과학 도구 18000-00 </td></tr><tr><td> 미니 연동 펌프 </td><td> 하버드 장치, MPII </td></tr><tr><td> 1ml 주사기 </td><td> BD, 329,650 </td></tr><tr><td> 18G의 바늘 </td><td> 백톤 디킨슨, 305,196 </td></tr><tr><td> 외과 5.0 나일론 봉합사 </td><td> Ethilon, 698H </td></tr><tr><td> 블록 Sectioning </td><td> 켄트 과학, 1mm </td></tr><tr><td> 단일 에지 면도날 </td><td> 전자 현미경 과학, 71962 </td></tr><tr><td> 라벨 테이프 </td><td> Scienceware, 벨 - 예술 제품 F13463 - 2075 </td></tr><tr><td> 거즈 스폰지 </td><td> 켄들 큐리티 2252, 존슨 앤드 존슨 6,415 </td></tr><tr><td> 커튼 팁 Applicators </td><td> 청교도 806 - WCL </td></tr><tr><td> C. 외과 도구 </td><td></td></tr><tr><td> 외과 스프링 가위 </td><td> 파인 과학 도구 15003-08 </td></tr><tr><td> Microdissecting 포셉는 직각 </td><td> 파인 과학 도구 45분의 5 </td></tr><tr><td> Microdissecting 포셉는 곡선 </td><td> 좋은 과학 도구 5 티 </td></tr><tr><td> 파인 직선 가위 </td><td> 파인 과학 도구 14568-12 </td></tr><tr><td> 곡선 뼈 rongeur </td><td> 파인 과학 도구 16021-14 </td></tr><tr><td> 하트먼 Hemostat가 휘어 </td><td> 파인 과학 도구 13003-10 </td></tr></tbody></table> 표 1. MCA의 결합 절차에 사용되는 재료, 도구 및 화학 물질의 목록입니다. A. 솔루션 및 화학 물질, B. 외과 재료 및 장비, C. 외과 공구를 추천하십시오.

the application of permanent middle cerebral artery ligation in the mouse

초점 대뇌 국소 빈혈 환자에서 볼 뇌졸중의 가장 일반적인 유형 중 하나입니다. 때문에 임상 중요성에 허혈성 모욕하는 동안 펼쳐 이벤트를 연구하기 위해 적절한 동물 모델을 개발하기 위해 장기간 노력하고있다. 이러한 기법은 가장 일반되는 설치류와 다양한 동물 모델을 사용하여 과도하거나 영구적인 초점 또는 글로벌 국소 빈혈 모델을 포함합니다. 또한 문학 pMCAo로 불리는 영구 MCA의 결합 방법은 설치류 1-6에 초점 국소 빈혈 모델로 광범위하게 사용됩니다. 이 방법은 원래 타무라 외 의해 쥐에 대해 설명했다. 1981 7인치 이 프로토콜에서는 craniotomy은 MCA와 근위 지역이 electrocoagulation에 의해 occluded되었습니다 액세스하는 데 사용되었다. infarcts는 폐색의 위치에 따라 대부분 피질과 때로는 striatal 지역을 포함하고 있습니다. 이 기술은 이제 잘 설립하여 많은 실험실 8-13에서 사용됩니다. 이 기술의 조기 사용 &quot;경색 코어&quot;와 &quot;penumbra&quot;14-16의 정의 및 설명을 주도하고, 그것은 종종 잠재 neuroprotective 화합물 10, 12, 13, 17을 평가하는 데 사용됩니다. 초기 연구는 쥐에서 수행 되었으나, 영구 MCA의 결합은 약간의 수정을 18-20로 생쥐에서 성공적으로 사용되었습니다. 이 모델은 재현성 infarcts 증가 후 생존 속도를 산출. 인간의 허혈성 뇌졸중의 약 80 %가 MCA 지역 21 일이되므로이 뇌졸중 연구의 관련성이 매우 모델입니다. 현재 뇌졸중 환자에 사용할 효과적인 치료법의 소수가, 따라서 잠재적인 약리 화합물을 테스트하고 생리적 결과를 평가하는 좋은 모델 필요가있다. 이 방법은 생체내 세포에서 hypoxia 응답 메커니즘을 연구하고 사용할 수 있습니다. 여기, 우리는 C57/BL6 마우스에서 MCA의 내고 수술을 제시한다. 우리는 미리 수술 준비, MCA의 결합 수술과 경색 볼륨 부량 위해 2,3,5 Triphenyltetrazolium 염화물 (TTC) 얼룩을 설명합니다.

Watch this Scientific Journal Video about The Application Of Permanent Middle Cerebral Artery Ligation in the Mouse at JoVE.com

The Application Of Permanent Middle Cerebral Artery Ligation in the Mouse

중간 대뇌 동맥 (MCA) 내고 동물 모델에 초점 대뇌 국소 빈혈을 연구하는 기술입니다. 이 방법에서는, 중간 대뇌 동맥 craniotomy에 의해 노출 소작된에 의해 출혈도 잡았. 이 방법은 높은 재현성 경색 볼륨과 사용 가능한 다른 방법에 비해 증가 이후 수술 생존 속도를 제공합니다.

마우스에 영구 중간 대뇌 동맥 내고의 응용

Focal cerebral ischemia is among the most common type of stroke seen in patients. Due to the clinical significance there has been a prolonged effort to ...

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19.5K 조회수.  University of Rochester. 중간 대뇌 동맥 (MCA) 내고 동물 모델에 초점 대뇌 국소 빈혈을 연구하는 기술입니다. 이 방법에서는, 중간 대뇌 동맥 craniotomy에 의해 노출 소작된에 의해 출혈도 잡았. 이 방법은 높은 재현성 경색 볼륨과 사용 가능한 다른 방법에 비해 증가 이후 수술 생존 속도를 제공합니다.

비디오: The Application Of Permanent Middle Cerebral Artery Ligation in the Mouse

Modeling Stroke in Mice - Middle Cerebral Artery Occlusion with the Filament Model

Stroke is among the most frequent causes of death and adult disability, especially in highly developed countries. However, treatment options to date are very limited. To meet the need for novel therapeutic approaches, experimental stroke research frequently employs rodent models of focal cerebral ischaemia. Most researchers use permanent or transient occlusion of the middle cerebral artery (MCA) in mice or rats.
Proximal occlusion of the middle cerebral artery (MCA) via the intraluminal suture technique (so called filament or suture model) is probably the most frequently used model in experimental stroke research. The intraluminal MCAO model offers the advantage of inducing reproducible transient or permanent ischaemia of the MCA territory in a relatively non-invasive manner. Intraluminal approaches interrupt the blood flow of the entire territory of this artery. Filament occlusion thus arrests flow proximal to the lenticulo-striate arteries, which supply the basal ganglia. Filament occlusion of the MCA results in reproducible lesions in the cortex and striatum and can be either permanent or transient. In contrast, models inducing distal (to the branching of the lenticulo-striate arteries) MCA occlusion typically spare the striatum and primarily involve the neocortex. In addition these models do require craniectomy. In the model demonstrated in this article, a silicon coated filament is introduced into the common carotid artery and advanced along the internal carotid artery into the Circle of Willis, where it blocks the origin of the middle cerebral artery. In patients, occlusions of the middle cerebral artery are among the most common causes of ischaemic stroke. Since varying ischemic intervals can be chosen freely in this model depending on the time point of reperfusion, ischaemic lesions with varying degrees of severity can be produced. Reperfusion by removal of the occluding filament at least partially models the restoration of blood flow after spontaneous or therapeutic (tPA) lysis of a thromboembolic clot in humans.
In this video we will present the basic technique as well as the major pitfalls and confounders which may limit the predictive value of this model.

Filamentous occlusion of the Middle cerebral artery is a common model for studying ischemic stroke in mice.

마우스의 모델링 스트로크 - 필라멘트 모델 중간 대뇌 동맥 폐색

Stroke is among the most frequent causes of death and adult disability, especially in highly developed countries. However, treatment options to date are ...

연구

의학

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 ...

Utilizing a Cranial Window to Visualize the Middle Cerebral Artery During Endothelin-1 Induced Middle Cerebral Artery Occlusion

Creation of a cranial window is a method that allows direct visualization of structures on the cortical surface of the brain1-3. This technique can be performed in many locations overlying the rat cerebrum, but is most easily carried out by creating a craniectomy over the readily accessible frontal or parietal bones. Most frequently, we have used this technique in combination with the endothelin-1 middle cerebral artery occlusion model of ischemic stroke to quantify the changes in middle cerebral artery vessel diameter that occur with injection of endothelin-1 into the brain parenchyma adjacent to the proximal MCA4, 5. In order to visualize the proximal portion of the MCA during endothelin -1 induced MCAO, we use a technique to create a cranial window through the temporal bone on the lateral aspect of the rat skull (Figure 1). Cerebral arteries can be visualized either with the dura intact or with the dura incised and retracted. Most commonly, we leave the dura intact during visualization since endothelin-1 induced MCAO involves delivery of the vasoconstricting peptide into the brain parenchyma. This bypasses the need to incise the dura directly over the visualized vessels for drug delivery. This protocol will describe how to create a cranial window to visualize cerebral arteries in a step-wise fashion, as well as how to avoid many of the potential pitfalls pertaining to this method.

This article describes a method for visualizing rat cerebral arteries through a cranial window using temporal craniectomy in order to view proximal portions of the middle cerebral artery (Figure 1). This versatile method can be combined with various techniques of drug delivery to measure cerebral artery reactivity in vivo.

엔도 테린-1 유도 중간 뇌 동맥 폐색 동안 중동 뇌동맥을 시각화하기 위해 두개골 창 활용

Creation of a cranial window is a method that allows direct visualization of structures on the cortical surface of the brain1-3. This technique can be ...

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 ...

Modeling Stroke in Mice: Permanent Coagulation of the Distal Middle Cerebral Artery

Stroke is the third most common cause of death and a main cause of acquired adult disability in developed countries. Only very limited therapeutical options are available for a small proportion of stroke patients in the acute phase. Current research is intensively searching for novel therapeutic strategies and is increasingly focusing on the sub-acute and chronic phase after stroke because more patients might be eligible for therapeutic interventions in a prolonged time window. These delayed mechanisms include important pathophysiological pathways such as post-stroke inflammation, angiogenesis, neuronal plasticity and regeneration. In order to analyze these mechanisms and to subsequently evaluate novel drug targets, experimental stroke models with clinical relevance, low mortality and high reproducibility are sought after. Moreover, mice are the smallest mammals in which a focal stroke lesion can be induced and for which a broad spectrum of transgenic models are available. Therefore, we describe here the mouse model of transcranial, permanent coagulation of the middle cerebral artery via electrocoagulation distal of the lenticulostriatal arteries, the so-called &#8220;coagulation model&#8221;. The resulting infarct in this model is located mainly in the cortex; the relative infarct volume in relation to brain size corresponds to the majority of human strokes. Moreover, the model fulfills the above-mentioned criteria of reproducibility and low mortality. In this video we demonstrate the surgical methods of stroke induction in the &#8220;coagulation model&#8221; and report histological and functional analysis tools.

Various murine models of middle cerebral artery occlusion (MCAO) are widely used in experimental brain research. Here, we demonstrate the model of transcranial permanent distal MCAO which produces consistent cortical infarction of a size corresponding to damage imposed by the majority of human ischemic strokes.

마우스의 모델링 스트로크 : 말초 중간 대뇌 동맥의 영구 응고

Stroke is the third most common cause of death and a main cause of acquired adult disability in developed countries. Only very limited therapeutical ...

Permanent Ligation of the Left Anterior Descending Coronary Artery in Mice: A Model of Post-myocardial Infarction Remodelling and Heart Failure

Heart failure is a syndrome in which the heart fails to pump blood at a rate commensurate with cellular oxygen requirements at rest or during stress. It is characterized by fluid retention, shortness of breath, and fatigue, in particular on exertion. Heart failure is a growing public health problem, the leading cause of hospitalization, and a major cause of mortality. Ischemic heart disease is the main cause of heart failure.
Ventricular remodelling refers to changes in structure, size, and shape of the left ventricle. This architectural remodelling of the left ventricle is induced by injury (e.g., myocardial infarction), by pressure overload (e.g., systemic arterial hypertension or aortic stenosis), or by volume overload. Since ventricular remodelling affects wall stress, it has a profound impact on cardiac function and on the development of heart failure. A model of permanent ligation of the left anterior descending coronary artery in mice is used to investigate ventricular remodelling and cardiac function post-myocardial infarction. This model is fundamentally different in terms of objectives and pathophysiological relevance compared to the model of transient ligation of the left anterior descending coronary artery. In this latter model of ischemia/reperfusion injury, the initial extent of the infarct may be modulated by factors that affect myocardial salvage following reperfusion. In contrast, the infarct area at 24 hr after permanent ligation of the left anterior descending coronary artery is fixed. Cardiac function in this model will be affected by 1) the process of infarct expansion, infarct healing, and scar formation; and 2) the concomitant development of left ventricular dilatation, cardiac hypertrophy, and ventricular remodelling.
Besides the model of permanent ligation of the left anterior descending coronary artery, the technique of invasive hemodynamic measurements in mice is presented in detail.

Heart failure is the leading cause of hospitalization and a major cause of mortality. A model of permanent ligation of the left anterior descending coronary artery in mice is applied to investigate ventricular remodelling and cardiac dysfunction post-myocardial infarction. The technique of invasive hemodynamic measurements in mice is presented.

마우스의 왼쪽 전방 내림차순 관상 동맥의 영구 결찰 : 심근 경색 리모델링과 심장 마비의 모델

Heart failure is a syndrome in which the heart fails to pump blood at a rate commensurate with cellular oxygen requirements at rest or during stress. It ...

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, ...

Transient Middle Cerebral Artery Occlusion Model of Neonatal Stroke in P10 Rats

A number of animal models have been used to study hypoxic-ischemic injury, traumatic injury, global hypoxia, or permanent ischemia in both the immature and mature brain. Stroke occurs commonly in the perinatal period in humans, and transient ischemia-reperfusion is the most common form of stroke in neonates. The reperfusion phase is a critical component of injury progression, which occurs over a period of days to weeks, and of the endogenous response to injury. This postnatal day 10 (p10) rat model of transient middle cerebral artery occlusion (tMCAO) creates a unilateral, non-hemorrhagic focal ischemia-reperfusion injury that can be utilized to study the mechanisms of focal injury and repair in the full-term-equivalent brain. The injury pattern that is produced by tMCAO is consistent and highly reproducible and can be confirmed with MRI or histological analyses. The severity of injury can be manipulated through changes in occlusion time and other methods that will be discussed.

Neonatal stroke is a significant cause of early brain injury requiring a translational model with consistent focal injury patterns and high reproducibility in order to enable study. This study describes the detailed surgical procedure for creating a non-hemorrhagic, unilateral focal ischemia-reperfusion injury in full-term-equivalent rodents.

P10 쥐에서 신생아 뇌졸중의 과도 중간 대뇌 동맥 폐색 모델

A number of animal models have been used to study hypoxic-ischemic injury, traumatic injury, global hypoxia, or permanent ischemia in both the immature ...

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 ...