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.

Bu protokol Emory Üniversitesi Kurumsal Hayvan Bakım ve Kullanım Komitesi (IACUC) tarafından onaylanmış ve Laboratuvar Hayvanları Bakım ve Kullanım Sağlık Talimatı&#39;nın Ulusal Enstitüleri izler. 1. Kur <ol><li> Ameliyat öncesi en az 15 dakika süreyle 37 ° C&#39;de ısı pompası ile bağlantılı ısınma pad cerrahi yatak hazırlayın. Cerrahi yatakta 3 ml şırınga varil kullanarak boyun rulo yerleştirin. Tıbbi havada% 2 izofluran ile anestezi gazı hazırlayın. </li><li> Otoklavlanmış forseps, makas, iğne mikro sahipleri, hemostat, pamuklu ve sütür hazırlayın. Doku yapıştırıcısı ve göz merhemi hazırlayın. </li><li> Isıtma lambası ve rektal prob ile hipoksi sistemi ve sıcaklık kontrol ayarlayın. % 92.5 N 2 göre dengeli 7.5% O 2% 2 izofluran ile hipoksi gazı hazırlayın. </li><li> Bir saat ameliyattan önce, fareler yavaş salınım Meloksikam (4.0 mg / kg) deri altından enjeksiyonu ile analgesized edilir. </li></ol> &lt;p class = &quot;jove_title&quot;&gt; 2. Geçici Serebral Hipoksi-iskemi (Şekil 1B) <ol><li> 10-13 haftalık erkek C57BL anestezisi / hayvan ayak sıkmak yanıt vermeyen ve daha sonra bir elektronik tıraş makinesi kullanarak doğru boyun saç kaldırmak kadar% 3 izofluran ile anestezi indüksiyon odasında 22 ila 30 g ağırlığındaki 6 fareleri. </li><li> 2 litre / dakika akış hızında, tıbbi hava içinde% 2 izofluran ile bağlantılı cerrahi yatak fareler yerleştirin. Güvenli ön ayakları tıbbi bant kullanarak yanlarda boyun rulo boyunca uzanmış. </li><li> Alkol ve daha sonra pamuklu tarafından takip betadin kesi cerrahi siteyi temizleyin. </li><li> Diseksiyon mikroskop altında düz bir forseps ve yaklaşık 0.2 cm orta hat cilt yanal mikro makas kullanarak 0.5 cm sağ servikal kesi yapmak. </li><li> Sağ karotis arter (RCCA) maruz fasya ve doku dışında çekin ince dişli forseps bir çift kullanın. Dikkatle ince yumuşak forseps bir çift kullanarak vagus sinirden RCCA ayrı. </li><li> Canlı RCCA üzerinde (serbestleştirilebilen) iki precut 5-0 ipek sütür düğüm, sonra 4-0 Naylon monofilament sütür (Şekil 1C) kullanarak cilt dikmek. </li><li> Kuruluğunu önlemek için her iki göze göz merhemi sürün. </li><li> Hızla hipoksi sistemine fareler aktarmak ve 30 dakika boyunca / 0.5-1 L akış hızında% 7.5% 2 izofluran ile yüz maskesi olarak min O 2 burun ve ağız koydu. <ol><li> Hipoksi sırasında, 37.5 ± 0.5 ° C&#39;de rektal sıcaklığı kontrol etmek için ısıtma lambalı sıcaklık kontrolörleri kullanın. / Dk 80-120 nefesler solunum hızını izleyin. hipoksi sırasında 37 ° C&#39;nin üzerinde vücut ısısı bakım tutarlı beyin enfarktüsü oluşturmak için önemlidir. Düşük solunum hızı genellikle 20 dakika hipoksi sonra olur. Yüz maskesi çıkarın ve solunum hızı Bu 1-2 dakika sürer ve 30 dakika hipoksi süresi içine sayılmaz 40&#39;a altına düşerse, normal hava girişini sağlar. </li></ol></li><li> Hipoksi sonra, aktarmakcerrahi yatağa fareler ve RCCA iki sütür serbest bırakın. Kafesine fareler dönün doku yapıştırıcı kullanarak yara kapatın ve. İki canlı knot hem beklenmedik hipoksi sonra yayımlanan ise hayvanların dışla. </li><li> Hipoksi ve anestezi kurtarmak için 5-10 dakika boyunca fareler izleyin. Kafeste ıslanan yiyecek koyun ve hayvan bakım tesisi iade. Not: Thi 24 saat sonra şiddetli çizerek davranışı hafif gösteren hayvanlar beyin kural ihlali ile ilişkilidir. Nöbet semptomları olan çoğu hayvanlar THI sonra 24 saat timepoint gelmeden ölmektedir. </li></ol> 3. Lazer Benek Kontrast Görüntüleme Not: Bu thi modelin önemli bir işlem değildir, ancak, iki boyutlu bir lazer benek kontrast görüntüleme sistemi 16 içinde ya da geçici hipoksi-iskemi sonrası beyin kan akışının (CBF) değişiklikleri karakterize etmek için kullanılabilir. St hemen sonra, THI altında rekor CBF değişiklikler belgelemek içinep 2.6. Alternatif olarak, bu işlemler adım 2.10 Aşağıdaki yapılabilir, THI hakaret sonra CBF kurtarma karşılaştırmak. <ol><li> Yüzüstü pozisyonda bir anestezi fare yerleştirin ve maruz ancak açılmamış kafatası ile saçlı deride 1 cm uzunluğunda orta hat kesi yapmak. </li><li> Üreticinin protokolüne göre bir kan akışı görüntüleyici altında hem serebral hemisferde CBF Monitör ve CCAO cerrahisi (adım 2.6) hemen sonra serebral kan akımını kaydetmeye başlayın. 50 dakika devam edin. </li><li> 16 renk paleti keyfi birimleri ile CBF görüntüyü göstermek ve gerçek zamanlı olarak analiz üreticinin talimatlarına (Şekil 2) Aşağıdaki MoorFLPI yazılımını kullanarak seçilen bölgeler. </li><li> CBF görüntüyü kaydettikten sonra, doku yapıştırıcısı ile kafa derisi kapatın ve kafesine hayvan dönün. </li></ol> 4. tPA İdaresi <ol><li> (220-300 ve kg yeniden birleştirici tPA / çözücü ya da 10 mg kuyruk damarından hayvanların enjekte956. 1 mg / m tPA) l, 0.5, 1 ya da tCCAo artı hipoksi sonra 4 saat sonra (Şekil 4). </li></ol> Birkaç Farklı Seçenekleri ile 5. Brain Damage Algılama Beyin örnekleri toplamak için Thi sonra 1, 4 veya 24 saatte fareler euthanize: Not. <ol><li> Açıklanan önceki olarak Thi hakaret sonra 24 saat in vivo 2,3,5-Triphenyltetrazolium klorür enfarktüs hacminin ölçümü yapın (TTC) yöntemi. 17 <ol><li> İntraperitoneal 1.4 M mannitol çözeltisi (~ 0.1 ml / g vücut ağırlığı) transcardial perfüzyon 30 dakika önce hayvanlara enjekte edilir. PBS ile perfüze Transcardial farenin% 2 TTC 10 ml eklenmiştir. </li><li> 10 dakika sonra, cerrahi aletler ile hayvanların beyin çıkarın ve bir vibratome 1 mm kalınlığında içine gece boyunca tespit ve bölüm için% 4 paraformaldehid içine koyun. </li><li> Dijital mikroskop ve kuvantı dört kesitli beyin slaytlar bir dizi çekinImageJ yazılımı kullanarak hasarsız, kontralateral yarımkürenin alanına enfarkte alana (sağ tarafta beyaz alan) oranı olarak enfarktüs hacmini ify. </li></ol></li><li> Alternatif olarak, THI hakaret sonra 1 saat sonra immünofloresans ile tromboz oluşumunu gerçekleştirin. <ol><li> Ekim bileşik ve bölüm bir Kriyostat kullanılarak 12 mikron kalınlığında beyinleri sabit beyin dondurun. </li><li> Bir floresan mikroskop üzerinde floresan gözlemlemek için: keçi anti-tavşan Alexa Floro 488 boya (200 1) ile: (1: 100), aşağıdaki tavşan anti-fibrinojen antikor ile beyin slayt inkübe edin. </li></ol></li><li> Alternatif olarak, thi akıntıdan sonra 4 saat sonra, 100 ul% 2 Evans mavi boya kuyruk damarı enjeksiyonu ile damar tıkanıklığı gerçekleştirin. <ol><li> Evans mavisi enjeksiyondan sonra% 4 paraformaldehid içine beyinleri kaldırmak için başını kesip hızla fareler Euthanize ve. Not: Hem fotoğrafların ön mavi renk ile Evans mavisi dolaşım için 5-10 dakika sürer ve bacaklarda gerisinde kalınmıştır. </li><li> Secti100 mikron kalınlığında sabit beyinleri üzerinde kayan bir mikrotom kullanılarak ve bir floresan mikroskop üzerinde 680 nm emisyon filtre kullanarak floresan gözlemleyin. </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>

İnme bir yaşlanma nüfusa sahip herhangi bir toplum için önemi giderek artan önemli bir sağlık sorunudur. Küresel olarak, inme, tüm ölümlerin 18% 11,1 eşdeğer 2010 yılında tahmini 5,9 milyon ölümcül olaylar ile ikinci ölüm önde gelen nedenidir. İnme da sakatlığa ayarlanmış yaşam yıllık üçüncü önde gelen nedenidir (DALY) 1990, 19 beşinci pozisyona yükselen, 2010 yılında küresel kaybetti. Bu epidemiyolojik veriler akut (iskemik) inme daha etkili tedavilerin ihti...

Tromboliz ve rekanalizasyonun klinik pratikte 1 akut iskemik inme en etkili tedavidir. Oysa, klinik öncesi araştırma nöro çoğunluğu damar tıkanıklığı çıkarılması üzerine serebral kan akımının hızlı iyileşme üreten ve tPA lizisle hiçbir fayda az gösteren bir geçici mekanik tıkanıklık modeli (lümen dikiş orta serebral arter tıkanıklığı) uygulandı. Inme modelini şüpheli seçimi hastanın 2,3 için nöro-koruyucu tedavisi çeviri zorluk, en azından kısmen, katkıda bulunduğu öne sürülmüştür. Bu nedenle, klinik öncesi araştırma tPA duyarlı tromboembolik inme modelleri istihdam için artan bir çağrı var, ama bu tür modelleri de teknik sorunlar (Tartışma) 4-7 var. Burada tek taraflı geçici hipoksik-iskemik (ti) hakaret ve intravenöz tPA tedavisine 8 yanıtları dayalı yeni bir trombotik inme modeli açıklar. Thi inme modeli 1960 9 yetişkin sıçan deneyleri için icat edilmiştir Levine prosedürü (bir odasında geçici hipoksi maruz kalma ve ardından tek taraflı karotis arter kalıcı ligasyonu) dayalı olarak geliştirilmiştir. Orijinal Levine prosedürü karanlığa gömüldüğü, çünkü sadece değişken beyin hasarı üretti, fakat 1981 10 neonatal hipoksik-iskemik ensefalopati (HİE) bir model olarak Robert Vannucci ve arkadaşları tarafından yeniden tanıtıldı aynı hakaret kemirgen yavrular tutarlı nevropatolojisi neden oldu. Son yıllarda, bazı Araştırmacılar hipoksik odasına 11 sıcaklığı ayarlayarak yetişkin farelere Levine-Vannucci modeli adapte yeniden. Orijinal Levine prosedüründe tutarsız beyin lezyonları hipoksik odasında yetişkin kemirgenler vücut sıcaklıklarını dalgalanan doğabilecek düşünülebilir. Bu hipotezi test etmek için, hipoksik gaz verilerek Levine prosedürünü modifiyeBir yüz maskesi vasıtasıyla, ameliyat masasına 12, 37 ° C&#39;de, kemirgen temel sıcaklık muhafaza edilirken. Beklendiği gibi, katı vücut ısısı kontrolü büyük ölçüde HI-kaynaklı beyin patolojisi tekrarlanabilirliği artmıştır. HI hakaret de pıhtılaşmayı, Otofaji ve gri-beyaz-madde hasarı 13 tetikler. Diğer araştırmacılar da inme sonrası inflamatuar yanıtlar 14 araştırmak için HI modeli kullandık. HI inme modelinin benzersiz bir özelliği de yakından kan akımı, endotel hasarı (örn nedeniyle HI-kaynaklı oksidatif strese) ve hiperkoagübilite (HI-indüklenen platelet aktivasyonu) staz dahil trombüs oluşumu Virchow&#39;un üçlü, (aşağıda olduğunu Şekil 1A) 15. Bu nedenle, HI modeli gerçek dünya iskemik inme ile ilgili bazı patofizyolojik mekanizmaları yakalayabilir. Bu düşünceyle, biz daha un tersine çevrilebilir ligasyonu ile HI modeli rafineilateral karotis arter (dolayısıyla geçici HI hakaret oluşturmak için), ve ya Edaravone olmadan tPA trombolitik onun yanıtları test edilmiştir. Edaravone zaten başlangıç ​​9 24 saat içinde iskemik inme tedavisinde Japonya&#39;da onaylanmış serbest radikal temizleyicisidir. Bizim deneyler kadar kısa 30 dakika kadar geçici HI trombotik enfarktüs tetikler gösterdi ve bu kombine tPA-Edaravone tedavi sinerjik faydalar 8 bahşeder. Burada detaylı cerrahi girişimler ve akut iskemik inme reperfüzyon tedavileri optimize etmek için kullanılabilecek Thi modelinin metodolojik hususlar, açıklar.

<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>
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		<tr>
			<td>Isoflurane</td>
			<td>Piramal Healthcare</td>
			<td>NDC 66794-013-25</td>
			<td>anesthesia</td>
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		<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

İnme araştırması klinik uygulamaya nöroprotektif tedavilerin tercüme birçok aksiliklere dayandı. Buna karşılık, gerçek dünya tedavisi (tPA trombolitik) nadiren preklinik inme araştırma hakim mekanik tıkanıklığı tabanlı deneysel modellerde faydaları üretir. Tezgah ve Yatağımın arasındaki bu bölünme klinik öncesi inme araştırma tPA-duyarlı modeller istihdam ihtiyacını göstermektedir. Bu amaçla, basit ve tPA-reaktif trombotik inme modeli icat burada tarif edilmektedir. 37.5 ± 0.5 ° C &#39;de, hayvan, rektal sıcaklık muhafaza edilirken bu model, 30 dakika boyunca, yetişkin farelerde bir yüz maskesi vasıtasıyla tek taraflı karotis ve% 7.5 oksijen teslimat geçici oklüzyonu meydana gelir. Tek taraflı karotis arter veya hipoksi geri dönüşümlü ligasyonu her biri sadece geçici serebral kan akımı bastırılmış olsa da, her iki hakaret kombinasyonu reperfüzyon açıkları, fibrin ve trombosit birikimi ve büyük infar kalıcı nedenorta serebral arter tarafından sağlanan bölgede ct. Önemli olarak, kuyruk ven 0.5, 1, ya da 4 saat sonra-Thi (10 mg / kg) yeniden birleştirici tPA enjeksiyonunu bir ölüm oranı ve enfarkt boyutunun zamana bağımlı bir azalma sağladı. Bu yeni model, inme basit ve deney sonuçlarını karşılaştırmak için laboratuvarlar arasında standardize edilebilir. Bundan başka, bu kraniektomi ya da önceden-oluşturulmuş emboli yol açmadan tromboz indükler. Bu eşsiz yararları göz önüne alındığında, THI modeli klinik öncesi inme araştırma repertuvarına bir kullanışlı bir ektir.

İki boyutlu lazer benek kontrast görüntüleme (LSCI) 16 30 dk geçici tek taraflı karotis tıkanıklığı (tCCAO), hipoksi 30 dakika maruz kalma (% 7.5 oksijen) ve 30 dakika tek taraflı serebral kan akımı (CBF) ve değişiklikleri karşılaştırmak için kullanıldı hipoksi (ti) altında ligasyonu karotis. Bu deney tCCAO normoxia altında karotis bağlandı yarımkürede CBF bastırılmış ortaya için ~ hızla karotis tıkanıklığı (Şekil 2A R) yayımlandıktan sonra 85 üzeri...

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.

Geçici Serebral Hipoksi-iskemi Dayalı Bir Trombotik İnme Modeli

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

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14.7K Görüntüleme.  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. 

Video: 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.

Fare Modelleme İnme - Filament Model Orta Serebral Arter Tıkanıklığı

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

Tıp

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.

Fare lazer Doppler flowmetre Rehberlik İskemik İnme intraluminal Orta Serebral Arter Tıkanıklığı (MCAO) Model

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.

Odak Serebral İskemi Erken İskemik Tolerans tetikleyebilecek bir Yeterli Önkoşullama Uyarıcı olarak bilateral Ortak Karotis Arter Tıkanıklığı

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.

Rat Lazer Doppler Flowmetri Rehberlik ile İskemik İnme için Endotelin-1 Bağlı Orta Serebral Arter Tıkanıklığı Modeli

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.

Intraluminar Orta Serebral Arter Tıkanıklığı ve Sıçan Strok Model Serebral Perfüzyon İzleme için Optimize Sistemi

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.

Subaraknoid Kanama bir fare modeli

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.

Bir olarak Beyin Microslices arasında excitotoxic uyarılması<em&gt; In vitro</em&gt; İnme Model

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.

Farelerde Modelleme İnme: Distal Orta Serebral Arter Daimi Pıhtılaşma

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.

Sıçanlar Homolog Kan pıhtısı ile İskemik İnme emboli Orta Serebral Arter Tıkanıklığı (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.

Fare Serebral Hipoksi-iskemi sırasında Eşzamanlı PET / MR Görüntüleme

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