This study was supported by the NIH grant NS074559 (to C. K.). We thank all collaborators who contributed to our research articles that the present methodology report is based upon.

이 프로토콜은에 모리 대학의 기관 동물 관리 및 사용위원회 (IACUC)의 승인을 실험실 동물의 관리 및 사용을위한 건강 지침의 국립 연구소를 다음입니다. 1. 설치 <ol><li> 수술 전 적어도 15 분 동안 37 ℃에서 열 펌프와 연결 온난화 패드에 수술 침대를 준비합니다. 수술 침대에 3 ML의 주사기의 배럴을 사용하여 목 롤을 놓습니다. 의료 공기에 2 % 이소 플루 란 마취 가스를 준비합니다. </li><li> 멸균 된 집게, 가위, 마이크로 니들 홀더, 지혈제, 면봉과 봉합을 준비합니다. 조직 접착제와 눈 연고를 준비합니다. </li><li> 가열 램프와 직장 프로브 저산소증 시스템과 온도 컨트롤러를 설정합니다. 92.5 %의 N 2로 균형 7.5 %의 O 2의 2 % 이소 플루 란과 저산소증 가스를 준비합니다. </li><li> 한 시간 수술 전, 마우스는 서방 멜 록시 캄 (4.0 ㎎ / ㎏)의 피하 주사에 의해 analgesized된다. </li></ol> &lt;P 클래스 = &quot;jove_title&quot;&gt; 2. 과도 대뇌 저산소증 - 허혈 (그림 1B) <ol><li> 10~13주 세 남성 C57BL을 마취 / 동물이 발 짜기에 응답하지 않는 한 다음 전자 면도기를 사용하여 오른쪽 목에 머리를 제거 할 때까지 3 % 이소 플루 란으로 마취 유도 챔버 (22) 30g에 무게 6 마우스. </li><li> 2 L / 분의 유량으로 공기 의료 2 % 이소 플루 란으로 연결 수술 침대에 마우스를 놓는다. 안전 앞발 의료 테이프를 사용 측면에서 목 롤을 따라 밖으로 뻗어있다. </li><li> 알코올 다음 면봉 다음 betadine와 절개의 수술 부위를 청소합니다. </li><li> 해부 현미경, 직선 집게 약 0.2 센티미터 중간 선 피부에서 측 방향의 마이크로 가위를 사용하여 0.5 cm 오른쪽 자궁 절개를합니다. </li><li> 오른쪽 경동맥 (RCCA)를 노출 근막 조직을 따로 뽑아 미세 톱니 모양의 집게 한 쌍을 사용합니다. 조심스럽게 좋은 부드러운 포셉 한 쌍을 사용하여 미주 신경에서 RCCA를 구분합니다. </li><li> 라이브 RCCA에 (이형성) 개의 미리 절단 5-0 실크 봉합사 매듭 후 4-0 나일론 모노 필라멘트 봉합사 (도 1C)를 사용하여 피부를 봉합. </li><li> 건조를 방지하기 위해 두 눈에 눈 연고를 적용합니다. </li><li> 신속 저산소증 시스템에 마우스를 전송하고, 30 분간 / 0.5 L의 유속으로 7.5 %에서 2 % 이소 플루 란으로 페이스 마스크 분 O 2를 코와 입에 넣어. <ol><li> 저산소증 동안에, 37.5 ± 0.5 ° C로 직장 온도를 제어하기 위해 가열 램프와 온도 조절기를 사용한다. / 분 80-120 호흡에서 호흡을 모니터링합니다. 저산소증 동안 37 ° C 위의 체온 유지 일관된 뇌경색을 만드는 것이 중요하다. 저 호흡은 일반적으로 20 분 저산소증 후 발생합니다. 얼굴 마스크를 제거하고 호흡이 1-2 분 소요 30 분 저산소증 기간에 포함되지 않는다 (40) 이하로 떨어지면 정상 공기 공급을 할 수 있습니다. </li></ol></li><li> 저산소증 후, 전송수술 침대에 마우스와는 RCCA에서 두 개의 봉합을 놓습니다. 케이지에 마우스를 반환 한 후 조직 접착제를 사용하여 상처를 닫고. 이 라이브 노트의 모두가 예기치 않게 저산소증 이후에 출시되는 동물을 제외합니다. </li><li> 저산소증과 마취에서 회복하기 위해 5 ~ 10 분 동안 마우스를 모니터링합니다. 케이지에 젖은 음식을 놓고 동물 보호 시설에 반환. 참고 : 생 후 24 시간에서 심각한 돌고 행동에 가벼운 보여주는 동물이 뇌 위반와 관련된다. 발작 증상이있는 대부분의 동물은 생 후 24 시간 평가시기 전에 죽는다. </li></ol> 3. 레이저 스페 클 대비 이미징 주 : 이것은 THI 모델의 필수적인 절차는 아니지만, 이차원 레이저 스페 클 대비 이미징 시스템 (16) 동안 또는 일시적 저산소 허혈 뇌 혈류 (CBF)의 변화를 특징 짓는 데에 이용 될 수있다. 세인트 직후, 티에서 기록을 CBF의 변경을 문서에EP 2.6. 대안 적으로,이 절차는 다음 단계 2.10 행할 수 THI 비방 CBF 후 회복과 비교한다. <ol><li> 엎드린 자세에서 마취 마우스를 놓고 노출하지만 개봉 두개골과 두피에 1cm 길이의 중간 선 절개를 수행합니다. </li><li> 제조사의 프로토콜에 따라 혈류 메이져 하에서 모두 대뇌 반구의 CBF를 모니터링하고 CCAO 수술 (단계 260) 후 즉시 뇌 혈류를 기록하기 시작한다. 50 분 동안 계속합니다. </li><li> 16 색 팔레트의 임의 단위와 CBF 화상을 확인하고 실시간으로 분석하여 제조자의 지시 (도 2) 다음 MoorFLPI 소프트웨어를 사용하여 선택된 영역. </li><li> CBF 이미지를 촬영 한 후, 조직 접착제와 두피를 닫고 케이지에 동물을 반환합니다. </li></ol> 4. 단자 위치 보장 관리 <ol><li> (220-300 &amp; KG 재조합 tPA의 / 용매 또는 10 mg의와 꼬리 정맥에서 동물을 주입# 956; 1 밀리그램 / M TPA)의 L, 0.5, 1, 또는 tCCAo 플러스 저산소증 후 4 시간에 (그림 4). </li></ol> 여러 다른 옵션 5. 뇌 손상 탐지 뇌 샘플을 수집하기 위해 생 후 1, 4, 24 시간에서 쥐를 안락사 :합니다. <ol><li> 기술 이전과 생의 모욕 후 24 시간에서 생체 2,3,5- 트리 페닐 테트라 졸륨 클로라이드에 의해 경색 볼륨의 정량화를 수행 (TTC) 방법. (17) <ol><li> 복강 내 1.4 M 만니톨 용액 (~ 0.1 ㎖ / g 체중) transcardial 관류 전에 30 분으로 동물을 주입. PBS와 Transcardial 관류 마우스는 2 % TTC 10 ㎖로 하였다. </li><li> 10 분 후 수술 도구와 동물의 뇌를 제거하고 vibratome와 1mm 두께로 밤새 고정 및 섹션 4 % 파라 포름 알데히드로 배치합니다. </li><li> 디지털 현미경과 퀀트 포 단면 한 뇌 슬라이드 일련의 스냅ImageJ에 소프트웨어를 사용하여, 손상, 반대쪽 반구의 면적에 대한 경색 영역 (오른쪽 흰색 영역)의 비율로 경색 량을 쓸어. </li></ol></li><li> 또한, 생 모욕 후 1 시간에서 면역에 의해 혈전 형성을 수행합니다. <ol><li> 10월 화합물 및 섹션 저온 유지 장치를 사용하여 12 μm의 두께 뇌에 고정 뇌 동결. </li><li> 형광 현미경에 형광을 관찰 : 염소 항 - 토끼 알렉사 Fluro 488 염료 (200 1)에 의해 : (100 1) 다음 토끼 항 피브리노겐 항체와 뇌 슬라이드를 품어. </li></ol></li><li> 또한, 생의 모욕 후 4 시간에 100 μL 2 % 에반스 블루 염료의 꼬리 정맥에 주사하여 혈관 폐색을 수행합니다. <ol><li> 에반스 블루 주사 후 4 % 파라 포름 알데히드로 두뇌를 제거하기 위해 머리를 잘라 신속하게 쥐를 안락사합니다. 참고 : 두 ...의 앞에 푸른 색으로 에반스 블루 순환을 위해 5 ~ 10 분 소요 사지 뒷다리. </li><li> 섹션에 ì100 ㎛ 두께로 고정 뇌에 슬라이딩 마이크로톰을 사용하여 형광 현미경에 680 nm의 방출 필터를 사용하여 형광을 관찰한다. </li></ol></li></ol>

<ol>
	<li>Broderick, J. P., Hacke, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Treatment+of+acute+ischemic+stroke:+Part+I:+recanalization+strategies.">Treatment of acute ischemic stroke: Part I: recanalization strategies.</a> Circulation. 106 (12), 1563-1569 (2002).</li><li>Hossmann, K. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pathophysiological+basis+of+translational+stroke+research.">Pathophysiological basis of translational stroke research.</a> Folia Neuropathol. 47 (3), 213-227 (2009).</li><li>Hossmann, K. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+two+pathophysiologies+of+focal+brain+ischemia:+implications+for+translational+stroke+research.">The two pathophysiologies of focal brain ischemia: implications for translational stroke research.</a> J. Cereb. Blood Flow Metab. 32 (7), 1310-1316 (2012).</li><li>Macrae, I. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preclinical+stroke+research--advantages+and+disadvantages+of+the+most+common+rodent+models+of+focal+ischaemia.">Preclinical stroke research--advantages and disadvantages of the most common rodent models of focal ischaemia.</a> Br. J. Pharmacol. 164 (4), 1062-1078 (2011).</li><li>Niessen, F., Hilger, T., Hoehn, M., Hossmann, K. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differences+in+clot+preparation+determine+outcome+of+recombinant+tissue+plasminogen+activator+treatment+in+experimental+thromboembolic+stroke.">Differences in clot preparation determine outcome of recombinant tissue plasminogen activator treatment in experimental thromboembolic stroke.</a> Stroke. 34 (8), 2019-2024 (2003).</li><li>Orset, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+model+of+in+situ+thromboembolic+stroke+and+reperfusion.">Mouse model of in situ thromboembolic stroke and reperfusion.</a> Stroke. 38 (10), 2771-2778 (2007).</li><li>Watson, B. D., Dietrich, W. D., Prado, R., Ginsberg, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Argon+laser-induced+arterial+photothrombosis.+Characterization+and+possible+application+to+therapy+of+arteriovenous+malformations.">Argon laser-induced arterial photothrombosis. Characterization and possible application to therapy of arteriovenous malformations.</a> J. Neurosurgery. 66 (5), 748-754 (1987).</li><li>Sun, Y. Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synergy+of+combined+tPA-edaravone+therapy+in+experimental+thrombotic+stroke.">Synergy of combined tPA-edaravone therapy in experimental thrombotic stroke.</a> PLoS One. 9, e98807 (2014).</li><li>Levine, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anoxic-ischemic+encephalopathy+in+rats.">Anoxic-ischemic encephalopathy in rats.</a> Am. J. Pathol. 36, 1-17 (1960).</li><li>Rice, J. E. 3. r. d., Vannucci, R. C., Brierley, J. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+influence+of+immaturity+on+hypoxic-ischemic+brain+damage+in+the+rat.">The influence of immaturity on hypoxic-ischemic brain damage in the rat.</a> Annals Neurol. 9 (2), 131-141 (1981).</li><li>Vannucci, S. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+stroke+in+the+female+diabetic,+db/db,+mouse.">Experimental stroke in the female diabetic, db/db, mouse.</a> J. Cereb. Blood Flow Metab. 21 (2), 52-60 (2001).</li><li>Adhami, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cerebral+ischemia-hypoxia+induces+intravascular+coagulation+and+autophagy.">Cerebral ischemia-hypoxia induces intravascular coagulation and autophagy.</a> Am. J. Pathol. 169 (2), 566-583 (2006).</li><li>Shereen, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ex+vivo+diffusion+tensor+imaging+and+neuropathological+correlation+in+a+murine+model+of+hypoxia-ischemia-induced+thrombotic+stroke.">Ex vivo diffusion tensor imaging and neuropathological correlation in a murine model of hypoxia-ischemia-induced thrombotic stroke.</a> J. Cereb. Blood Flow Metab. 31 (4), 1155-1169 (2011).</li><li>Michaud, J. P., Pimentel-Coelho, P. M., Tremblay, Y., Rivest, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+impact+of+Ly6C+low+monocytes+after+cerebral+hypoxia-ischemia+in+adult+mice.">The impact of Ly6C low monocytes after cerebral hypoxia-ischemia in adult mice.</a> J. Cereb. Blood Flow Metab. 34 (7), e1-e9 (2014).</li><li>Zoppo, G. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Virchow's+triad:+the+vascular+basis+of+cerebral+injury.">Virchow's triad: the vascular basis of cerebral injury.</a> Rev. Neurol. Dis. 5, 12-21 (2008).</li><li>Dunn, A. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laser+speckle+contrast+imaging+of+cerebral+blood+flow.">Laser speckle contrast imaging of cerebral blood flow.</a> Annals Biomed. Eng. 40 (2), 367-377 (2012).</li><li>Sun, Y. Y., Yang, D., Kuan, 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=Mannitol-facilitated+perfusion+staining+with+2,3,5-triphenyltetrazolium+chloride+(TTC)+for+detection+of+experimental+cerebral+infarction+and+biochemical+analysis.">Mannitol-facilitated perfusion staining with 2,3,5-triphenyltetrazolium chloride (TTC) for detection of experimental cerebral infarction and biochemical analysis.</a> J. Neurosci. Methods. 203 (1), 122-129 (2012).</li><li>Lozano, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Global+and+regional+mortality+from+235+causes+of+death+for+20+age+groups+in+1990+and+2010:+a+systematic+analysis+for+the+Global+Burden+of+Disease+Study.">Global and regional mortality from 235 causes of death for 20 age groups in 1990 and 2010: a systematic analysis for the Global Burden of Disease Study.</a> Lancet. 380 (9859), 2095-2128 (2010).</li><li>Murray, C. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Disability-adjusted+life+years+(DALYs)+for+291+diseases+and+injuries+in+21+regions,+1990-2010:+a+systematic+analysis+for+the+Global+Burden+of+Disease+Study.">Disability-adjusted life years (DALYs) for 291 diseases and injuries in 21 regions, 1990-2010: a systematic analysis for the Global Burden of Disease Study.</a> Lancet. 380 (9859), 2197-2223 (2012).</li><li>Dirnagl, U., Macleod, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stroke+research+at+a+road+block:+the+streets+from+adversity+should+be+paved+with+meta-analysis+and+good+laboratory+practice.">Stroke research at a road block: the streets from adversity should be paved with meta-analysis and good laboratory practice.</a> Br. J. Pharm. 157 (7), 1154-1156 (2009).</li><li>Dirnagl, U., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+concerted+appeal+for+international+cooperation+in+preclinical+stroke+research.">A concerted appeal for international cooperation in preclinical stroke research.</a> Stroke. 44 (6), 1754-1760 (2013).</li><li>Khatri, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Revascularization+end+points+in+stroke+interventional+trials:+recanalization+versus+reperfusion+in+IMS-I.">Revascularization end points in stroke interventional trials: recanalization versus reperfusion in IMS-I.</a> Stroke. 36 (11), 2400-2403 (2005).</li><li>Rosenberg, R. D., Aird, W. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vascular-bed--specific+hemostasis+and+hypercoagulable+states.">Vascular-bed--specific hemostasis and hypercoagulable states.</a> New Eng. J. Med. 340 (20), 1555-1564 (1999).</li><li>Majid, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differences+in+vulnerability+to+permanent+focal+cerebral+ischemia+among+3+common+mouse+strains.">Differences in vulnerability to permanent focal cerebral ischemia among 3 common mouse strains.</a> Stroke. 31 (11), 2707-2714 (2000).</li></ol>

스트로크는 노화 인구 어떤 사회 중요성을 성장의 주요 건강 문제입니다. 전 세계적으로 뇌졸중이 사망 원인 (18)의 11.1 %에 해당하는 2010 년 추정 5,900,000 치명적인 이벤트와 사망의 두 번째 주요 원인이다. 스트로크는 장애 조정 생명 년의 세 번째 주요 원인이다 (DALYs)은 1990 (19)에 다섯 번째 위치에서 상승, 2010 년에 전 세계적으로 패했다.이 역학 데이터가 급성 (허혈성) 뇌졸중의?...

혈전 용해 및 개통은 임상 1 급성 허혈성 뇌졸중의 가장 효과적인 치료입니다. 그러나, 임상 신경 연구의 대부분은 혈관 폐색 제거하여 뇌 혈류의 신속한 복구가 발생하고, 단자 위치 보장 혈전에 의해 아무 효과는 거의 도시 과도 정비공 폐색 모델 (관내 봉합 중간 대뇌 동맥 폐쇄)에서 수행 하였다. 이는 뇌졸중 모델의 선택 모호한 환자에게 신경 치료 2,3- 변환하는 어려움에 적어도 부분적으로 기여한다고 제안되었다. 따라서, 전임상 연구에서 TPA에 반응 혈전 색 전성 뇌졸중 모델을 사용하는 증가 통화가 있지만, 이러한 모델은 기술적 인 문제 (토론 참조) 4-7 있습니다. 여기에서 우리는 일방적 과도 저산소 허혈성 (THI) 모욕과 정맥의 TPA 치료 8의 응답에 따라 새로운 혈전 성 뇌졸중 모델을 설명합니다. 생 스트로크 모델은 1960 9 성인 쥐 실험을 위해 발명 된 레빈 절차 (챔버 일시적인 저산소증에 노출 다음 일방적 경동맥의 영구 결찰)를 기반으로 개발되었다. 원래 레빈 절차는 무명에 머 금고 있기 때문에 그것은 단지 변수 뇌 손상을 생산하지만, 1981 10 신생아 저산소 성 허혈성 뇌증 (HIE)의 모델로 로버트 Vannucci와 그의 동료에 의해 ​​다시 도입 될 때 같은 모욕 쥐 새끼에서 일관된 신경 병리학을 일으키는 원인이되었다. 최근에는 일부 연구자들은 저산소 챔버 (11) 내의 온도를 조정함으로써 성인 마우스에 레빈-Vannucci 모델을 적응 재. 원래 레빈 절차의 일관성 뇌 병변 저산소 실에서 성인 쥐의 체온 변동으로 인해 발생할 수 있음을 그럴듯. 이 가설을 테스트하기 위해, 우리는 저산소 가스를 투여하여 레빈 절차를 수정안면 마스크를 통해 수술 테이블 (12)에서 37 ° C에서 설치류 코어 온도를 유지하면서. 예상대로, 엄격한 신체 온도 제어가 크게 HI-유도 뇌 병리학의 재현성 증가했다. HI 모욕은 응고, 자식 작용, 및 그레이 - 흰색 - 문제 부상 (13)을 트리거합니다. 다른 연구자들은 또한 뇌졸중 후 염증 반응 (14)를 조사하기 위해 HI 모델을 사용하고 있습니다. HI 뇌졸중 모델의 독특한 특징은 밀접 혈류, 내피 손상 (예 인해 HI 유도 된 산화 적 스트레스에 대한) 및 과다 응고 (HI 유도 된 혈소판 활성화)의 정체를 포함한 혈전 형성의 Virchow의 화음을 (이하 있다는 도 1A) 15. 이와 같이, HI 모델은 실제 허혈성 뇌졸중에 중요한 일부 병리 생리 학적 메카니즘을 포착 할 수있다. 마음에 이런 생각과 함께, 우리는 또한 유엔의 가역적 결찰과 HI 모델을 정제ilateral 경동맥는 (따라서 과도 HI 모욕을 만드는)와 함께 또는 Edaravone없이 단자 위치 보장 혈전 용해에의 반응을 시험했다. Edaravone은 이미 시작 9의 24 시간 이내에 허혈성 뇌졸중을 치료하기 위해 일본에서 승인 된 자유 라디칼 스 캐빈입니다. 우리의 실험은 짧은 30 분 과도 HI를 혈전 경색을 유발 것을 보여 주었다, 그 결합 된 TPA에 Edaravone 치료는 시너지 효과 8을 부여. 여기에서는 상세한 외과 적 절차, 급성 허혈성 뇌졸중의 재관류 치료를 최적화하는 데 사용될 수있는 모델의 생 방법 론적 고려 사항을 설명한다.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>adult male mice</td>
			<td>Charles River</td>
			<td>C57BL/6&#160;</td>
			<td>10~13 weeks old (22~30 g)</td>
		</tr>
		<tr>
			<td>Mobile Laboratory Animal Anesthesia System</td>
			<td>VetEquip</td>
			<td>901807</td>
			<td>anesthesia</td>
		</tr>
		<tr>
			<td>Medical air (Compressed) air tank</td>
			<td>Airgas</td>
			<td>UN1002</td>
			<td>anesthesia</td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>Piramal Healthcare</td>
			<td>NDC 66794-013-25</td>
			<td>anesthesia</td>
		</tr>
		<tr>
			<td>Multi-Station Lab Animal AnesthesiaSystem</td>
			<td>Surgivet</td>
			<td>V703501</td>
			<td>hypoxia system</td>
		</tr>
		<tr>
			<td>7.5% O2 balanced by 92.5% N2 tank</td>
			<td>Airgas</td>
			<td>UN1956</td>
			<td>hypoxia system</td>
		</tr>
		<tr>
			<td>Temperature Controller with heating lamp&#160;</td>
			<td>Cole Parmer&#160;</td>
			<td>EW-89000-10</td>
			<td>temperature controllers</td>
		</tr>
		<tr>
			<td>Rectal probe</td>
			<td>Cole Parmer&#160;</td>
			<td>NCI-00141PG</td>
			<td>temperature controllers</td>
		</tr>
		<tr>
			<td>Dissecting microscope&#160;</td>
			<td>Olympus&#160;</td>
			<td>SZ40</td>
			<td>surgical setup</td>
		</tr>
		<tr>
			<td>Heat pump with warming pad</td>
			<td>Gaymar&#160;</td>
			<td>TP700</td>
			<td>surgical setup</td>
		</tr>
		<tr>
			<td>Fine curved forceps (serrated)</td>
			<td>FST</td>
			<td>11370-31</td>
			<td>surgical instrument</td>
		</tr>
		<tr>
			<td>Fine curved forceps (smooth)</td>
			<td>FST</td>
			<td>11373-12</td>
			<td>surgical instrument</td>
		</tr>
		<tr>
			<td>micro scissors</td>
			<td>FST</td>
			<td>15000-03</td>
			<td>surgical instrument</td>
		</tr>
		<tr>
			<td>micro needle holders</td>
			<td>FST</td>
			<td>12060-01</td>
			<td>surgical instrument</td>
		</tr>
		<tr>
			<td>Halsted-Mosquito hemostats</td>
			<td>FST</td>
			<td>13008-12</td>
			<td>surgical instrument</td>
		</tr>
		<tr>
			<td>5-0 silk suture&#160;</td>
			<td>Harvard Apparatus</td>
			<td>624143</td>
			<td>surgical supplies</td>
		</tr>
		<tr>
			<td>4-0 Nylon monofilament suture</td>
			<td>LOOK</td>
			<td>766B</td>
			<td>surgical supplies</td>
		</tr>
		<tr>
			<td>Tissue glue</td>
			<td>Abbott Laboratories</td>
			<td>NC9855218</td>
			<td>surgical supplies</td>
		</tr>
		<tr>
			<td>Puralube Vet ointment</td>
			<td>Fisher</td>
			<td>NC0138063&#160;</td>
			<td>eye dryness prevention&#160;</td>
		</tr>
		<tr>
			<td>MoorFLPI-2 blood flow imager</td>
			<td>Moor</td>
			<td>780-nm laser source</td>
			<td>Laser Speckle Contrast Imaging</td>
		</tr>
		<tr>
			<td>Mannitol</td>
			<td>Sigma</td>
			<td>M4125</td>
			<td>in-vivo TTC</td>
		</tr>
		<tr>
			<td>2,3,5-triphenyltetrazolium chloride (TTC)&#160;</td>
			<td>Sigma</td>
			<td>T8877</td>
			<td>in-vivo TTC</td>
		</tr>
		<tr>
			<td>Vibratome</td>
			<td>Stoelting</td>
			<td>51425</td>
			<td>brain section for in-vivo TTC&#160;</td>
		</tr>
		<tr>
			<td>Digital microscope</td>
			<td>Dino-Lite</td>
			<td>AM2111</td>
			<td>whole-braina imaging</td>
		</tr>
		<tr>
			<td>O.C.T compound</td>
			<td>Sakura Finetek</td>
			<td>4583</td>
			<td></td>
		</tr>
		<tr>
			<td>goat anti-rabbit Alexa Fluro 488</td>
			<td>Invitrogen</td>
			<td>A11008</td>
			<td>Immunohistochemistry</td>
		</tr>
		<tr>
			<td>Cryostat</td>
			<td>Vibratome</td>
			<td>ultrapro 5000</td>
			<td>brain section for IHC</td>
		</tr>
		<tr>
			<td>Evans blue</td>
			<td>Sigma</td>
			<td>E2129</td>
			<td>Detecting vascular perfusion</td>
		</tr>
		<tr>
			<td>Microtome</td>
			<td>Electron Microscopy Sciences</td>
			<td>5000</td>
			<td>brain section for histology</td>
		</tr>
		<tr>
			<td>Avertin (2, 2, 2-Tribromoethanol)</td>
			<td>Sigma</td>
			<td>T48402</td>
			<td>euthanasia</td>
		</tr>
		<tr>
			<td>Fluorescent microscope</td>
			<td>Olympus</td>
			<td>DP73</td>
			<td></td>
		</tr>
	</tbody>
</table>

a thrombotic stroke model based on transient cerebral hypoxia-ischemia

스트로크 연구는 임상 적으로 신경 치료를 번역에 많은 좌절을 견디고있다. 반대로, 실제 치료 (TPA의 혈전)이 거의 스트로크 전임상 연구를 지배 기계적 폐색 기반 실험 모델의 이익을 생성하지 않는다. 벤치와 침대 옆 간의 분할 전임상 행정 연구 TPA에 반응하는 모델을 채용 할 필요성을 시사한다. 이를 위해, 간단하고 TPA에 반응성 혈전 성 뇌졸중 모델 발명은 여기에 설명된다. 37.5 ± 0.5 ° C에서 동물의 직장 온도를 유지하면서,이 모델은, 30 분 동안 마우스에서 성인 안면 마스크를 통해 일방적 경동맥 및 7.5 %의 산소의 전달의 일시적 폐색 이루어져있다. 일방적 경동맥 또는 저산소증 가역 결찰 각 만 일시적 뇌 혈류를 억제하지만, 양쪽의 조합 욕설 재관류 결핍, 혈소판 피브린 침착, 및 대형 infar 지속 야기중간 대뇌 동맥 제공 영토에서 CT. 중요한 것은, 꼬리 정맥 0.5, 1, 또는 4 시간 후의 생 (10 밀리그램 / kg)에서 재조합의 TPA 주입 사망률 경색 크기의 시간에 따른 감소를 제공 하였다. 이 새로운 뇌졸중 모델은 단순 및 실험 결과를 비교하는 실험실에 걸쳐 표준화 될 수있다. 또한, 두개 절제술 또는 예비 형성된 색전을 도입하지 않고, 혈전증을 유도한다. 이러한 독특한 장점을 감안할 때, 티 모델은 전임상 뇌졸중 연구의 레퍼토리에 유용한 추가이다.

이차원 레이저 스페 클 대비 이미징 (LSCI) 16을 30 분 과도 일방적 경동맥 폐색 (tCCAO), 저산소증에 30 분간 노출 (7.5 % 산소), 및 30 분 일방적으로 뇌 혈류 (CBF)의 변화를 비교하기 위해 사용 하였다 저산소증 (THI)에서 결찰을 경동맥. 이 실험은 tCCAO가 normoxia에서 경동맥 결찰 반구에서 CBF를 억제하는 것으로 나타났다에 ~ 빨리 경동맥 폐색 (그림 2A에서 R)의 출시 이후 85 % 이상으로 ?...

Watch this Scientific Journal Video about A Thrombotic Stroke Model Based On Transient Cerebral Hypoxia-ischemia at JoVE.com

A Thrombotic Stroke Model Based On Transient Cerebral Hypoxia-ischemia

Thromboembolic stroke models are vital tools for optimizing the recanalization therapy. Here we report a murine thrombotic stroke model based on transient cerebral hypoxic-ischemic (tHI) insult, which triggers thrombosis and infarction, and responds favorably to tissue plasminogen activator (tPA)-mediated fibrinolysis in a therapeutic window similar to those in stroke patients.

과도 대뇌 저산소증 - 허혈을 기반으로 혈전 성 뇌졸중 모델

Stroke research has endured many setbacks in translating neuroprotective therapies into clinical practice. In contrast, the real-world therapy (tPA ...

a-thrombotic-stroke-model-based-on-transient-cerebral-hypoxia-ischemia

Research

JoVE Journal

Medicine

14.7K 조회수.  The Emory University School of Medicine. Thromboembolic stroke models are vital tools for optimizing the recanalization therapy. Here we report a murine thrombotic stroke model based on transient cerebral hypoxic-ischemic (tHI) insult, which triggers thrombosis and infarction, and responds favorably to tissue plasminogen activator (tPA)-mediated fibrinolysis in a therapeutic window similar to those in stroke patients. 

비디오: A Thrombotic Stroke Model Based On Transient Cerebral Hypoxia-ischemia

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

Intraluminal Middle Cerebral Artery Occlusion (MCAO) Model for Ischemic Stroke with Laser Doppler Flowmetry Guidance in Mice

Stroke is the third leading cause of death and the leading cause of disability in the world, with an estimated cost of near $70 billion 
in the United States in 20091,2. The intraluminal middle cerebral artery occlusion (MCAO) model was developed by Koizumi4 in 1986 to simulate this 
impactful human pathology in the rat. A modification of the MCAO method was later presented by Longa3. Both techniques have been widely used to identify 
molecular mechanisms of brain injury resulting from ischemic stroke and potential therapeutic modalities5. This relatively noninvasive method in rats has been 
extended to use in mice to take advantage of transgenic and knockout strains6,7. To model focal cerebral ischemia, an intraluminal suture is advanced via 
the internal carotid artery to occlude the base of the MCA. Retracting the suture after a specified period of time mimics spontaneous reperfusion, but the 
suture can also be permanently retained. This video will be demonstrating the two major approaches for performing intraluminal MCAO procedure in mice in a 
stepwise fashion, as well as providing insights for potential drawbacks and pitfalls. The ischemic brain tissue will subsequently be stained by 
2,3,5-triphenyltetrazolium chloride (TTC) to evaluate the extent of cerebral infarction8.

Intraluminal Middle Cerebral Artery Occlusion MCAO Model for Ischemic Stroke with Laser Doppler Flowmetry Guidance in Mice

The intraluminal middle cerebral artery occlusion (MCAO) model is the most frequent used model among experimental ischemic stroke models. Here we will demonstrate the entire model in detail with the guide of Laser Doppler flowmetry, and its representative results.

마우스의 레이저 도플러 Flowmetry 안내와 허혈성 뇌졸중에 대한 중간 대뇌 동맥 Intraluminal 폐색 (MCAO) 모델

Stroke is the third leading cause of death and the leading cause of disability in the world, with an estimated cost of near $70 billion 
in the United ...

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

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

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

초점 대뇌 국소 빈혈에 조기 허혈성 공차를 유도하기 위해 적절한 전처리 자극으로 양측 경동맥 동맥 폐색

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

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 ...

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 ...

A Murine Model of Subarachnoid Hemorrhage

In this video publication a standardized mouse model of subarachnoid hemorrhage (SAH) is presented. Bleeding is induced by endovascular Circle of Willis perforation (CWp) and proven by intracranial pressure (ICP) monitoring. Thereby a homogenous blood distribution in subarachnoid spaces surrounding the arterial circulation and cerebellar fissures is achieved. Animal physiology is maintained by intubation, mechanical ventilation, and continuous on-line monitoring of various physiological and cardiovascular parameters: body temperature, systemic blood pressure, heart rate, and hemoglobin saturation. Thereby the cerebral perfusion pressure can be tightly monitored resulting in a less variable volume of extravasated blood. This allows a better standardization of endovascular filament perforation in mice and makes the whole model highly reproducible. Thus it is readily available for pharmacological and pathophysiological studies in wild type and genetically altered mice.

A standardized mouse model of subarachnoid hemorrhage by intraluminal Circle of Willis perforation is described. Vessel perforation and subarachnoid bleeding are monitored by intracranial pressure monitoring. In addition various vital parameters are recorded and controlled to maintain physiologic conditions.

지주막 하 출혈의 생쥐 모델

In this video publication a standardized mouse model of subarachnoid  hemorrhage (SAH) is presented. Bleeding is induced by endovascular Circle of  Willis ...

Excitotoxic Stimulation of Brain Microslices as an In vitro Model of Stroke

Examining molecular mechanisms involved in neuropathological conditions, such as ischemic stroke, can be difficult when using whole animal systems. As such, primary or &#39;neuronal-like&#39; cell culture systems are commonly utilized. While&#160;these systems are relatively easy to work with, and are useful model systems in which various functional outcomes (such as cell death) can be readily quantified, the examined outcomes and pathways in cultured immature neurons (such as excitotoxicity-mediated cell death pathways) are not necessarily the same as those observed in mature brain, or in intact tissue. Therefore, there is the need to develop models in which cellular mechanisms in mature neural tissue can be examined. We have developed an in vitro technique that can be used to investigate a variety of molecular pathways in intact nervous tissue. The technique described herein utilizes rat cortical tissue, but this technique can be adapted to use tissue from a variety of species (such as mouse, rabbit, guinea pig, and chicken) or brain regions (for example, hippocampus, striatum, etc.). Additionally, a variety of stimulations/treatments can be used (for example, excitotoxic, administration of inhibitors, etc.). In conclusion, the brain slice model described herein can be used to examine a variety of molecular mechanisms involved in excitotoxicity-mediated brain injury.

Excitotoxic Stimulation of Brain Microslices as an In vitro Model of Stroke

We have developed a brain slice model which can be used to examine molecular mechanisms involved in excitotoxicity-mediated brain injury.&#160;This technique generates viable mature brain tissue and reduces animal numbers required for experimentation, whilst keeping the neuronal circuitry, cellular interactions, and postsynaptic compartments partly intact.

같은 뇌 Microslices의 흥분 독성 자극<em&gt; 체외</em&gt; 스트로크의 모델

Examining molecular mechanisms involved in neuropathological conditions, such as ischemic stroke, can be difficult when using whole animal systems. As ...

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 ...

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

Clinically, thrombolytic therapy with use of recombinant tissue plasminogen activator (tPA) remains the most effective treatment for acute ischemic stroke. However, the use of tPA is limited by its narrow therapeutic window and by increased risk of hemorrhagic transformation. There is an urgent need to develop suitable stroke models to study new thrombolytic agents and strategies for treatment of ischemic stroke. At present, two major types of ischemic stroke models have been developed in rats and mice: intraluminal suture MCAO and embolic MCAO. Although MCAO models via the intraluminal suture technique have been widely used in mechanism-driven stroke research, these suture models do not mimic the clinical situation and are not suitable for thrombolytic studies. Among these models, the embolic MCAO model closely mimics human ischemic stroke and is suitable for preclinical investigation of thrombolytic therapy. This embolic model was first developed in rats by Overgaard et al.1 in 1992 and further characterized by Zhang et al. in 19972. Although embolic MCAO has gained increasing attention, there are technical problems faced by many laboratories. To meet increasing needs for thrombolytic research, we present a highly reproducible model of embolic MCAO in the rat, which can develop a predictable infarct volume within the MCA territory. In brief, a modified PE-50 tube is gently advanced from the external carotid artery (ECA) into the lumen of the internal carotid artery (ICA) until the tip of the catheter reaches the origin of the MCA. Through the catheter, a single homologous blood clot is placed at the origin of the MCA. To identify the success of MCA occlusion, regional cerebral blood flow was monitored, neurological deficits and infarct volumes were measured. The techniques presented in this paper should help investigators to overcome technical problems for establishing this model for stroke research.

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 ...

Simultaneous PET/MRI Imaging During Mouse Cerebral Hypoxia-ischemia

Dynamic changes in tissue water diffusion and glucose metabolism occur during and after hypoxia in cerebral hypoxia-ischemia reflecting a bioenergetics disturbance in affected cells. Diffusion weighted magnetic resonance imaging (MRI) identifies regions that are damaged, potentially irreversibly, by hypoxia-ischemia. Alterations in glucose utilization in the affected tissue may be detectable by positron emission tomography (PET) imaging of 2-deoxy-2-(18F)fluoro-&#7429;-glucose ([18F]FDG) uptake. Due to the rapid and variable nature of injury in this animal model, acquisition of both modes of data must be performed simultaneously in order to meaningfully correlate PET and MRI data. In addition, inter-animal variability in the hypoxic-ischemic injury due to vascular differences limits the ability to analyze multi-modal data and observe changes to a group-wise approach if data is not acquired simultaneously in individual subjects. The method presented here allows one to acquire both diffusion-weighted MRI and [18F]FDG uptake data in the same animal before, during, and after the hypoxic challenge in order to interrogate immediate physiological changes.

The method presented here uses simultaneous positron emission tomography and magnetic resonance imaging. In the cerebral hypoxia-ischemia model, dynamic changes in diffusion and glucose metabolism occur during and after injury. The evolving and irreproducible damage in this model necessitates simultaneous acquisition if meaningful multi-modal imaging data are to be acquired.

마우스 대뇌 저산소증 - 허혈 동안 동시 PET / MRI 영상

Dynamic changes in tissue water diffusion and glucose metabolism occur during and after hypoxia in cerebral hypoxia-ischemia reflecting a bioenergetics ...