Yazarlar NHMRC ve NHF teknik Joy Yao Dr. Karen Alt destek yanı sıra, fon kabul etmek istiyorum.

Hayvanlarla ilgili tüm deneyler Alfred Tıbbi Araştırma ve Eğitim Semt Hayvan Etik Komitesi (E / 1534/2015 / B) tarafından onaylanmıştır. Tüm cerrahi manipülasyonlar tam anestezi altında gerçekleştirildi ve hayvanlar herhangi bir aşamada ağrı deneyimi yoktu. Tarif edilen tüm deneyler olmayan kazanımı vardır. 1. Hazırlık <ol><li> Filtre kağıdı (1 mm x 2 mm), ince şeritler kesilir. </li><li> Taze% 4 demir klorür 2 çözüm hazırlanması (a / h) ve% 6 (w / v) deiyonize su içinde seyreltilmiştir. 0.22 um ile filtrelenmiştir, PBS içinde rodamin 6G solüsyon 0.3% hazırlayın. </li><li> Şırınga sargının beyaz plastikten küçük bir parça (5 mm x 1 cm) kesin. </li></ol> Intravital Mikroskopi tarafından Gözlenen 2. Mezenter Arteriole trombüs oluşumu <ol><li> 10-12 haftalık erkek C57BL / 6 vahşi tip fare tartılır ve intraperitoneal enjeksiyon olsa ketamin karışımı (100 mg / kg) ve ksilazin (10 mg / kg) ile buna göre anestezi. Ayak, kuyruk ve / veya cilt tutam, kornea ve palpebral tutam gelen reaktivite ve bıyıkları hareketin olmaması tepki ile anestezi derinliği izleyin. Gerekirse, hayvanın anesteziyi muhafaza etmek üzere ketamin ikinci bir doz (50 mg / kg) enjekte edilir. Anestezi altında iken kuruluğunu önlemek için gözleri veteriner merhem sürün. 37 ° C&#39;ye ayarlanmış bir ısıtma pedi üzerine yerleştirilmiş küçük bir petri fare yerleştirin. NOT: IP enjeksiyon hayvanlar için ağrıya neden olabilir, ancak bu rahatsızlık süresi asgari (en az 3 sn) olacaktır. Enjeksiyonlar ile ilişkili ağrı ve rahatsızlık deneyimli ve yetkin personeli ve uygun iğne boyutu (25 G) kullanımı ile minimize edilecektir. Tüm prosedürler izlenerek, sonra servikal dislokasyon yapılmıştır ketamin ve ksilazin bir doz aşımı ile tüm hayvanlar ötenazi. </li><li> Deride yaklaşık 3 cm&#39;lik bir abdominal orta hat kesi gerçekleştirin ve dikkatli Periton kesti. </li><li> Pet tarafında fare yerleştirinri çanak, yavaşça bağırsak exteriorize ve dikkatle Petri kabı yüzeyine uygun bir kap getirmek için 2 pamuk tomurcukları ile mezenter yayıldı. Hassas bir silecek ile düzgün kurulayın. Not: mezenterik damar hareketini sınırlamak için, papaverin bağırsak peristalsis inhibe etmek için de kullanılabilir. </li><li> Damarlarını dilate ve lökosit ve trombosit sayısında etiketlemek için bir 29 G şırınga ile fare kuyruğu damar içine Rodamin 6G (% 0.3) 30 ul enjekte ılık suda fare kuyruğu ıslatın. </li><li> Ters bir mikroskop altında Petri kabı yerleştirin ve parlak saha kanalı kullanılarak seçilen arteriole odaklanın. </li><li> % 6 (ağırlık / hacim), demir (III) klorid ile filtre kağıdı bir bant emmek ve iki forseps ile arteriol filtre kağıdı uygulanır; İlki filtre kağıdı yavaşça ilgi alanı üzerine basın ikinci bir tutun. Filtre kağıdı çökelmesi sonraki ilk 10 saniye içinde trombus oluşumu dikkate alınmalıdır. NOT: Bu oldukça comm olduğunuçevrili mikrovaskuleterini ve filtre kağıdı birikimi ve yavaşça basınç hassasiyeti yaralama bu sorunu sınırlamak için önemlidir. </li><li> Filtre kağıdı ile: floresan mikroskop ile trombüs oluşumunu (tepe uyarma 557 nm, pik emisyon 576 nm TRITC kanalı) dikkate alınmalıdır. Trombüs içine Rodamin 6G ve agregasyonu kadar almış dolaşan lökosit ve trombosit sayısında uyun dolayısıyla tespit etmek kolaydır. </li><li> Demir (III) klorür maruz kalma 1 dakika sonra filtre kağıdı çıkarın ve trombüs oluşumunu izlemeye devam edin. PBS ile kap yıkayın. </li><li> Gözlemleyin ve Rodamin 6G ile trombositlerin ve lökositlerin etiketleme ile vurgulanan trombüs dinamik oluşumunu kaydedin. Görüntü yakalamak ve trombüs boyutunu ölçmek. Bu tarifnamede sunulan resimler TRITC floresan kanal, objektif 4X da, ters intravital mikroskobu ile elde edildi. </li><li> Ftüm işlemleri ollowing, servikal dislokasyon tarafından takip ketamin ve ksilizin aşırı dozda kullanarak hayvan euthanize. </li></ol> 3. Karotis Arter trombüs oluşumu Kan Akış Hızı Ölçümü tarafından Değerlendirilen <ol><li> 10-12 haftalık erkek C57BL / 6 fare tartılır ve intraperitoneal enjeksiyon da ketamin (100 mg / kg) ve ksilazin (10 mg / kg) karışımı ile anestezi buna göre. Anestezi altında iken kuruluğunu önlemek için gözleri veteriner merhem sürün. </li><li> 37 ° C ayarlanmış bir ısıtma yastığı, bacaklarda yapışkan bant kullanarak bir işletim mikroskop altında fare sabitleyin. </li><li> Tek silecek dışında küçük bir yastık yapmak ve hafifçe başını yükseltmek için fare başının altında bantlayın. (Üst dişleri kullanın) burun aşağı çekmek için maşa ile bir iplik döngü kullanın. Bu kolay erişim için karotis arter bölgesini gösterecektir. </li><li> Aşağı sternum, doğrudan çene altında küçük bir deri 5 mm derinliğinde kesi gerçekleştirin. </li><li> Fa teşrihscia çatallanma üzerinde sağ veya sol ortak karotid arter ya da bir fragmanını izole eder. </li><li> Dikkatle-in arasındaki arter ve onları ayırmak için sinir cımbız tanıtmak. Yakın artere çalışan sinir rahatsız ve o olabilir gemiye zarar olarak karotis arter çok dokunmamak etmeyin. Arterin en az 5 mm&#39;lik bir bölümü izole edin. </li><li> Herhangi bir sıvı FeCl3 müdahale önlemek için düzgün silecekler arterin alanını kurutun. </li><li> Çevre dokularda emmek değil FeCl 3 çok yaygın karotid izole bir parçası altında küçük beyaz plastik parçası koyun. Bu amaç için, arter altında plastik kağıt sonra yavaş yavaş ilk bir parçası getirmek kaymasına ikinci forseps kullanır. </li><li> % 4 (a / h) ya da% 6 (ağırlık / hacim), ferrik klorid ile filtre kağıdı parçası daldırın ve arter çevresindeki tüm yerleştirin. </li><li> 3 dk maruz kaldıktan sonra, filtre kağıdı çıkarmak PBS ile durulayın ve kurulayın w alanıi silecekler. </li><li> Yaralı alanda geminin etrafında Doppler probu yerleştirin ve akış değişiklikleri kaydetmeye başlayın. Yetişkin farelerin sağlıklı genel karotis, akış genellikle yaklaşık 1 ml / dk&#39;dır. Dikkat! Herhangi bir temas kaçınılmalıdır böylece demir klorür ile prob temas probu zarar verecektir. Burada sunulan veriler, bir nano Doppler akış sondası 0.5 PBS ile donatılmış bir Transonic System Inc. Akış ölçer, TS420 perivasküler modülü ile elde edilmiştir. NOT: demir klorid konsantrasyonu, farklı bir oklüzyon zamanlarda elde edilen trombus oluşumunun kinetiği değiştirebilir. Böylece,% 6&#39;lık bir maruziyet (a / h), ferrik klorür, ferrik klorür (w / v)% 4 maruz daha hızlı bir oklüzyon verir. </li><li> Tüm prosedürler izlenerek, servikal dislokasyon tarafından takip ketamin ve ksilizin aşırı dozda kullanan tüm hayvanlar euthanize ve dikkatle Doppler probu temizleyin. </li></ol>

<ol>
	<li>Leadley, R. J., Chi, L., Rebello, S. S., Gagnon, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Contribution+of+in+vivo+models+of+thrombosis+to+the+discovery+and+development+of+novel+antithrombotic+agents.">Contribution of in vivo models of thrombosis to the discovery and development of novel antithrombotic agents.</a> J Pharmacol Toxicol Methods. 43 (2), 101-116 (2000).</li><li>Johnson, G. J., Griggs, T. R., Badimon, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+utility+of+animal+models+in+the+preclinical+study+of+interventions+to+prevent+human+coronary+artery+restenosis:+analysis+and+recommendations.+On+behalf+of+the+Subcommittee+on+Animal,+Cellular+and+Molecular+Models+of+Thrombosis+and+Haemostasis+of+the+Scientific+and+Standardization+Committee+of+the+International+Society+on+Thrombosis+and+Haemostasis.">The utility of animal models in the preclinical study of interventions to prevent human coronary artery restenosis: analysis and recommendations. On behalf of the Subcommittee on Animal, Cellular and Molecular Models of Thrombosis and Haemostasis of the Scientific and Standardization Committee of the International Society on Thrombosis and Haemostasis.</a> Thromb Haemost. 81 (5), 835-843 (1999).</li><li>Day, S. M., Reeve, J. L., Myers, D. D., Fay, W. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Murine+thrombosis+models.">Murine thrombosis models.</a> Thromb Haemost. 92 (3), 486-494 (2004).</li><li>Sachs, U. J. H., Nieswandt, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+thrombus+formation+in+murine+models.">In vivo thrombus formation in murine models.</a> Circ Res. 100 (7), 979-991 (2007).</li><li>Furie, B., Furie, B. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanisms+of+thrombus+formation.">Mechanisms of thrombus formation.</a> N Engl J Med. 359 (9), 938-949 (2008).</li><li>Pierangeli, S. S., Liu, X. W., Barker, J. H., Anderson, G., Harris, E. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induction+of+thrombosis+in+a+mouse+model+by+IgG,+IgM+and+IgA+immunoglobulins+from+patients+with+the+antiphospholipid+syndrome.">Induction of thrombosis in a mouse model by IgG, IgM and IgA immunoglobulins from patients with the antiphospholipid syndrome.</a> Thromb Haemost. 74 (5), 1361-1367 (1995).</li><li>Kikuchi, S., Umemura, K., Kondo, K., Saniabadi, A. R., Nakashima, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Photochemically+induced+endothelial+injury+in+the+mouse+as+a+screening+model+for+inhibitors+of+vascular+intimal+thickening.">Photochemically induced endothelial injury in the mouse as a screening model for inhibitors of vascular intimal thickening.</a> Arterioscler Thromb Vasc Biol. 18 (7), 1069-1078 (1998).</li><li>Rosen, E. D., Raymond, S., 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=Laser-induced+noninvasive+vascular+injury+models+in+mice+generate+platelet-+and+coagulation-dependent+thrombi.">Laser-induced noninvasive vascular injury models in mice generate platelet- and coagulation-dependent thrombi.</a> Am J Pathol. 158, 1613-1622 (2001).</li><li>Kurz, K. D., Main, B. W., Sandusky, G. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rat+model+of+arterial+thrombosis+induced+by+ferric+chloride.">Rat model of arterial thrombosis induced by ferric chloride.</a> Thromb Res. 60 (4), 269-280 (1990).</li><li>Eckly, A., Hechler, B., 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=Mechanisms+underlying+FeCl3-induced+arterial+thrombosis.">Mechanisms underlying FeCl3-induced arterial thrombosis.</a> J Thromb Haemost. 9 (4), 779-789 (2011).</li><li>Darbousset, 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=Involvement+of+neutrophils+in+thrombus+formation+in+living+mice.">Involvement of neutrophils in thrombus formation in living mice.</a> Pathol Biol (Paris). 62 (1), 1-9 (2014).</li><li>Denis, C., Methia, N., 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+mouse+model+of+severe+von+Willebrand+disease:+defects+in+hemostasis+and+thrombosis).">A mouse model of severe von Willebrand disease: defects in hemostasis and thrombosis).</a> Proc Natl Acad Sci U S A. 95 (16), 9524-9529 (1998).</li><li>Wang, X., Hagemeyer, C. E., 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=Novel+single-chain+antibody-targeted+microbubbles+for+molecular+ultrasound+imaging+of+thrombosis:+validation+of+a+unique+noninvasive+method+for+rapid+and+sensitive+detection+of+thrombi+and+monitoring+of+success+or+failure+of+thrombolysis+in+mice.">Novel single-chain antibody-targeted microbubbles for molecular ultrasound imaging of thrombosis: validation of a unique noninvasive method for rapid and sensitive detection of thrombi and monitoring of success or failure of thrombolysis in mice.</a> Circulation. 125 (25), 3117-3126 (2012).</li><li>Wang, X., Smith, P. L., Hsu, M. -. Y., Ogletree, M. L., Schumacher, W. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Murine+model+of+ferric+chloride-induced+vena+cava+thrombosis:+evidence+for+effect+of+potato+carboxypeptidase+inhibitor.">Murine model of ferric chloride-induced vena cava thrombosis: evidence for effect of potato carboxypeptidase inhibitor.</a> J Thromb Haemost. 4 (2), 403-410 (2006).</li><li>Karatas, H., Erdener, S. E., 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=Thrombotic+distal+middle+cerebral+artery+occlusion+produced+by+topical+FeCl(3)+application:+a+novel+model+suitable+for+intravital+microscopy+and+thrombolysis+studies.">Thrombotic distal middle cerebral artery occlusion produced by topical FeCl(3) application: a novel model suitable for intravital microscopy and thrombolysis studies.</a> J Cereb Blood Flow Metab. 31 (6), 1452-1460 (2011).</li><li>Jirouskov&#225;, M., Chereshnev, I., V&#228;&#228;n&#228;nen, H., Degen, J. L., Coller, B. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antibody+blockade+or+mutation+of+the+fibrinogen+gamma-chain+C-terminus+is+more+effective+in+inhibiting+murine+arterial+thrombus+formation+than+complete+absence+of+fibrinogen.">Antibody blockade or mutation of the fibrinogen gamma-chain C-terminus is more effective in inhibiting murine arterial thrombus formation than complete absence of fibrinogen.</a> Blood. 103 (6), 1995-2002 (2004).</li><li>Dubois, C., Panicot-Dubois, L., Merrill-Skoloff, G., Furie, B., Furie, B. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glycoprotein+VI-dependent+and+-independent+pathways+of+thrombus+formation+in+vivo.">Glycoprotein VI-dependent and -independent pathways of thrombus formation in vivo.</a> Blood. 107 (10), 3902-3906 (2006).</li><li>Navarrete, A. -. M., Casari, 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=A+murine+model+to+characterize+the+antithrombotic+effect+of+molecules+targeting+human+von+Willebrand+factor.">A murine model to characterize the antithrombotic effect of molecules targeting human von Willebrand factor.</a> Blood. 120 (13), 2723-2732 (2012).</li><li>Rayes, J., Hollestelle, M. 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=Mutation+and+ADAMTS13-dependent+modulation+of+disease+severity+in+a+mouse+model+for+von+Willebrand+disease+type+2B.">Mutation and ADAMTS13-dependent modulation of disease severity in a mouse model for von Willebrand disease type 2B.</a> Blood. 115 (23), 4870-4877 (2010).</li><li>Lamrani, L., Lacout, 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=Hemostatic+disorders+in+a+JAK2V617F-driven+mouse+model+of+myeloproliferative+neoplasm.">Hemostatic disorders in a JAK2V617F-driven mouse model of myeloproliferative neoplasm.</a> Blood. 124 (7), 1136-1145 (2014).</li><li>Zhang, Y., Ye, 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=Increased+platelet+activation+and+thrombosis+in+transgenic+mice+expressing+constitutively+active+P2Y12.">Increased platelet activation and thrombosis in transgenic mice expressing constitutively active P2Y12.</a> J Thromb Haemost. 10 (10), 2149-2157 (2012).</li><li>Sch&#228;fer, K., Konstantinides, S., 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=Different+mechanisms+of+increased+luminal+stenosis+after+arterial+injury+in+mice+deficient+for+urokinase-+or+tissue-type+plasminogen+activator.">Different mechanisms of increased luminal stenosis after arterial injury in mice deficient for urokinase- or tissue-type plasminogen activator.</a> Circulation. 106 (14), 1847-1852 (2002).</li><li>Wang, X., Palasubramaniam, 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=Towards+effective+and+safe+thrombolysis+and+thromboprophylaxis:+preclinical+testing+of+a+novel+antibody-targeted+recombinant+plasminogen+activator+directed+against+activated+platelets.">Towards effective and safe thrombolysis and thromboprophylaxis: preclinical testing of a novel antibody-targeted recombinant plasminogen activator directed against activated platelets.</a> Circ Res. 114 (7), 1083-1093 (2014).</li><li>Decrem, 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=Ir-CPI,+a+coagulation+contact+phase+inhibitor+from+the+tick+Ixodes+ricinus,+inhibits+thrombus+formation+without+impairing+hemostasis.">Ir-CPI, a coagulation contact phase inhibitor from the tick Ixodes ricinus, inhibits thrombus formation without impairing hemostasis.</a> J Exp Med. 206 (11), 2381-2395 (2009).</li><li>Ma, D., 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=Desmolaris,+a+novel+factor+XIa+anticoagulant+from+the+salivary+gland+of+the+vampire+bat+(Desmodus+rotundus)+inhibits+inflammation+and+thrombosis+in+vivo.">Desmolaris, a novel factor XIa anticoagulant from the salivary gland of the vampire bat (Desmodus rotundus) inhibits inflammation and thrombosis in vivo.</a> Blood. 122 (25), 4094-4106 (2013).</li><li>Lei, X., 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=Anfibatide,+a+novel+GPIb+complex+antagonist,+inhibits+platelet+adhesion+and+thrombus+formation+in+vitro+and+in+vivo+in+murine+models+of+thrombosis.">Anfibatide, a novel GPIb complex antagonist, inhibits platelet adhesion and thrombus formation in vitro and in vivo in murine models of thrombosis.</a> Thromb Haemost. 111 (2), 279-289 (2014).</li><li>Waisberg, M., 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=Plasmodium+falciparum+infection+induces+expression+of+a+mosquito+salivary+protein+(Agaphelin)+that+targets+neutrophil+function+and+inhibits+thrombosis+without+impairing+hemostasis.">Plasmodium falciparum infection induces expression of a mosquito salivary protein (Agaphelin) that targets neutrophil function and inhibits thrombosis without impairing hemostasis.</a> PLoS Pathog. 10 (9), e1004338 (2014).</li><li>Owens, A. P., Lu, Y., Whinna, H. C., Gachet, C., Fay, W. P., Mackman, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Towards+a+standardization+of+the+murine+ferric+chloride-induced+carotid+arterial+thrombosis+model.">Towards a standardization of the murine ferric chloride-induced carotid arterial thrombosis model.</a> J Thromb Haemost. 9 (9), 1862-1863 (2011).</li><li>Wang, X., Xu, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+optimized+murine+model+of+ferric+chloride-induced+arterial+thrombosis+for+thrombosis+research.">An optimized murine model of ferric chloride-induced arterial thrombosis for thrombosis research.</a> Thromb Res. 115 (1-2), 95-100 (2005).</li><li>Tseng, M. T., Dozier, A., Haribabu, B., Graham, U. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transendothelial+migration+of+ferric+ion+in+FeCl3+injured+murine+common+carotid+artery.">Transendothelial migration of ferric ion in FeCl3 injured murine common carotid artery.</a> Thromb Res. 118 (2), 275-280 (2006).</li><li>Bonnard, T., 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=Leukocyte+mimetic+polysaccharide+microparticles+tracked+in+vivo+on+activated+endothelium+and+in+abdominal+aortic+aneurysm.">Leukocyte mimetic polysaccharide microparticles tracked in vivo on activated endothelium and in abdominal aortic aneurysm.</a> Acta Biomater. 10 (8), 3535-3545 (2014).</li><li>Boulaftali, Y., Lamrani, L., 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=The+mouse+dorsal+skinfold+chamber+as+a+model+for+the+study+of+thrombolysis+by+intravital+microscopy.">The mouse dorsal skinfold chamber as a model for the study of thrombolysis by intravital microscopy.</a> Thromb Haemost. 107 (5), 962-971 (2012).</li><li>Konstantinides, S., Sch&#228;fer, K., Thinnes, T., Loskutoff, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Plasminogen+activator+inhibitor-1+and+its+cofactor+vitronectin+stabilize+arterial+thrombi+after+vascular+injury+in+mice.">Plasminogen activator inhibitor-1 and its cofactor vitronectin stabilize arterial thrombi after vascular injury in mice.</a> Circulation. 103 (4), 576-583 (2001).</li><li>Li, W., McIntyre, T. M., Silverstein, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ferric+chloride-induced+murine+carotid+arterial+injury:+A+model+of+redox+pathology.">Ferric chloride-induced murine carotid arterial injury: A model of redox pathology.</a> Redox Biol. 1 (1), 50-55 (2013).</li></ol>

The authors have nothing to disclose.

ferrik klorür trombozu modeli mükemmel bir araştırma aracıdır. Bu çalışmada gösterildiği gibi, uygulanması son derece kolay ve intravital mikroskobiyle veya doppler akış ölçer ile kombinasyon halinde kullanıldığında, bu trombus oluşumu için iyi bir gerçek zamanlı izlenmesini sağlar. Zaman pozlama ve FeCl3 konsantrasyonu ayarlanması, aynı zamanda tıkayıcı-olmayan ya da tıkayıcı ya trombüs üretmek için imkan sunuyor. Bununla birlikte, bu yöntem, ...

tromboza ve anti-trombotik ilaçların etkinliğinin değerlendirilmesinde ilgili mekanizmaların çalışma ile deneysel hayvan modelleri kurulmuştur gerektirir. Onlar kemirgenler 1 den insanlara büyük gemiler daha benzer sağlamak gibi büyük hayvan modellerinde kullanılmak üzere ilk edildi. Ancak, yüksek maliyet, gerekli büyük tesisler ve onları manipüle zorluk genetik kullanımlarına önemli dezavantajları vardır ve kemirgenler üzerinde ön testler kesin sonuç 2 verdik kez büyük hayvanlar artık geç preklinik çalışmalarda sınırlıdır. In vivo test edilmesi için gerekli olan anti-trombotik ilaçlarının miktarı en aza indirir transgenik ve boşaltma suşlar ve bunların küçük boyutu geniş durumu ile, fareler esas olarak tromboz araştırma için kullanılmıştır. Bu nedenle, trombotik bozuklukların ve çeşitli modelleri, farelerde 3 geliştirilmiştir. Birçok kurulan tromboz modellerinde intim bozabilirkan pıhtılaşması 4 oluşumunu uyaran kan akışına alt endotel hücre dışı matrisin maruz bırakılarak, ardından damar duvarının bir tabaka. trombüsler ve / pıhtılaşma kaskadını 5 aktive doku faktörünün maruz veya trombosit aktivasyonunu tetikleyen kollajen maruz neden olabilir. Çeşitli teknikler sonra başlangıç ​​damar yaralanması elde etmek için kullanılır. Pierangeli ve ark. Femoral ven 6 bir mikrocerrahi aracıyla mekanik bozulma modeli geliştirmiştir. Kikushi ve ark., Yeşil ışık (540 nm), 7 ile çıkar damar duvarı belirli bir uyarım ardından endoteliyal hücrelerin lipid çift tabakası içinde biriken bir verilmedi reaktif bileşik (Rose Bengal) uygulanmasından oluşur, bir modeli tarif. yaralanması aynı zamanda, kısa bir yüksek yoğunluklu darbe lazer aydınlatma 8 tarafından indüklenebilir. Başka bir teknik öncelikle sıçanların karotis arter üzerine kurulmuşdemir klorür, topikal uygulama (FeCl3), 9 oluşur. Bu durumda, FeCl3 ile yaratılan serbest köklerin alınan damar denüdasyon sonuçları olan endotel hücreleri 10 lipid peroksidasyonu ve tahribata yol açar. yaralanma lökositlerin işe gibi trombosit yapışması ve yığılmasını tetikleme birçok yapışma moleküllerinin ifadesini indükler. Bu lökosit, özellikle nötrofiller, tromboz 11 giden kan pıhtılaşma kaskadının aktivasyonuna çok önemli bir rol oynadığı gösterilmiştir. Bu yöntem de koagulasyon akışını üretmek için uygundur; Araştırmacılar, insanlarda tromboz çoğunlukla hastalıklı örneğin meydana oysa bu fare modelinde, tromboz genellikle sağlıklı damarlar indüklenir, akılda tutmak gerekir. aterosklerotik gemiler. Bu modeli uygulamak için çok basit ve aynı zamanda farelerde etkili olduğu gibi, bu en çok kullanılan tromboz moduin vivo çalışmalar küçük hayvan için l. Buna ek olarak, bu teknik, gemilerin çeşitli trombüs oluşumunu uyarmak için imkanı sunar. Hedef gemiler arterler veya büyük çaplı (karotis, femoral, vena kava) ya da küçük çaplı (mezenter, cremaster) 12-14 damarları olabilir. Daha yakın bir zamanda, aynı zamanda, inme 15 bir model geliştirmek için proksimal orta serebral arter kullanıldı. tromboz oluşumu ile doğrudan trombosit ve lökosit flüoresan etiketleme sonra intravital mikroskopi ile gözlemlenmiştir veya sıcaklık sensörü veya Doppler prob 12,16,17 kan akışı azalma ölçülerek izlenebilir. Örneğin kapama süresi, trombus oluşumu zaman veya trombüs büyüklüğü gibi çeşitli parametreler araştırılabilir. gemiler arasındaki fizyolojik farklılıklar elde trombüsün önemli farklılıklar sonucu araştırıldı. Bu nedenle, araştırmacılar genellikle measu isteyen parametrelere göre hedef geminin seçinyeniden ve / veya araştırmak istediğiniz ayarı hastalığı. Tipik olarak, karotis arter model vena kava üzerinde çalışmalar derin ven trombozu araştırma için daha uygun iken, miyokard infarktüsü veya inme ile ilişkili aterotromboz araştırma için daha uygun olduğunu. Farklı damarların erişilebilirlik de trombüs gelişimini ölçmek için kullanılan bir yöntem belirler. Örneğin, mezenterik damarlar intravital mikroskopik gözlem ve trombüs oluşumu dinamikleri çalışması için bu model de uygun hale erişmek kolaydır. Karotis arter daha az erişilebilir ama sağlayan kan akımı ölçümleri daha büyük ve tıkayıcı trombozu incelemek için mükemmel bir model oluşturabilir. ferrik klorür trombozu modeli bu patolojinin anlayışında muazzam bir ilerleme sağlamıştır. Bu tromboz oluşumunun 18,19 von Willebrand faktör rolü üzerinde duruluyor birçok çalışmada kullanılmıştır. Genetik modi ile birliktenağı teknikleri, bu trombotik bozukluklar dahil birçok spesifik genin belirlenmesini sağladı. Lamrani ve diğ. Örneğin JAK2 V617F geninin bir knock-istikrarsız pıhtı 20 hızlanan oluşumu ile ilişkili olduğunu göstermiştir. Zhang ve ark. P2Y12 trombosit reseptörü fizyolojik ima incelendi ve trombositler özel olarak bu reseptörü aşırı ifade eden transgenik fareler, sadece, FeCl3 21 yaralı mezenterik arter içinde daha hızlı ve kararlı bir trombüs oluşumunu gösterildi olduğunu göstermiştir. doku-tipi plazminojen aktivatörü (tPA) ve fibrin bozunma süreci ürokinaz tipi plazminojen aktivatörü (uPA) önemli bir rol de bu yöntemle 22 olarak araştırılmıştır. Bundan başka, bu model, aynı zamanda, in vivo olarak pek çok yeni ilaçların fibrinolitik kapasiteleri test basit ve doğru bir şekilde sağlar. Örneğin, Wang ve diğ. Th Bu model kullanmışaktive trombositler 23 karşı hedeflenen yeni rekombinant plazminojen aktivatörü e preklinik doğrulama. Bu yöntem aynı zamanda kene, vampir yarasalar ve sivrisinek tükürük ya da hedefin 24-27 spesifik tanımlama ile yılan zehirinden izole terapötik proteinlerin doğrulama sağladı. Bu örnekler, ferrik klorür modelinin çok yönlülüğünü göstermektedir. Bu yazıda iki yöntem üzerinde durulacak ve iki farklı damar tipine demir klorür kaynaklı tromboz çalışma; mezenterik damar ve karotid arter.

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			<td>Iron (III) chloride 40 % (w/v)</td>
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			<td>Inverted microscope&#160;</td>
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			<td>Digital black-and-white camera&#160;</td>
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			<td>0409-2051-05</td>
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			<td>75313&#160;</td>
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			<td>Petri dish</td>
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			<td>Insulin syringe (29 G)</td>
			<td>BD Ultra-Fine</td>
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			<td>Cotton tipped applicators</td>
			<td>BSN medical</td>
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			<td>Dulbecco&#39;s phosphate buffer saline (PBS)</td>
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ferric chloride-induced thrombosis mouse model on carotid artery and mesentery vessel

Severe thrombosis and its ischemic consequences such as myocardial infarction, pulmonary embolism and stroke are major worldwide health issues. The ferric chloride injury is now a well-established technique to rapidly and accurately induce the formation of thrombi in exposed veins or artery of small and large diameter. This model has played a key role in the study of the pathophysiology of thrombosis, in the discovery and validation of novel antithrombotic drugs and in the understanding of the mechanism of action of these new agents. Here, the implementation of this technique on a mesenteric vessel and carotid artery in mice is presented. The method describes how to label circulating leukocytes and platelets with a fluorescent dye and to observe, by intravital microscopy on the exposed mesentery, their accumulation at the injured vessel wall which leads to the formation of a thrombus. On the carotid artery, the occlusion caused by the clot formation is measured by monitoring the blood flow with a Doppler probe.

Rodamin 6G birikimini ortaya koyacaktır mezenter floresan intravital mikroskopi gözlem FeCl3 yaralandı damar duvarı boyunca trombosit ve lökositlerin etiketli. kısmi trombüs aşamalı oluşumu 200 um mezenter kabına (Şekil 1) takip edilir. Bir trombüs yavaş görünür ve açıkça FeCl 3 maruz kalma ilk dakikadan sonra tanımlanabilir (Şekil 1, t = 60 sn). 40 saniye FeCl3 ile ıslatılmış filtre kağıdı çıkarılmasından sonra, tromboz ...

Watch this Scientific Journal Video about Ferric Chloride-induced Thrombosis Mouse Model on Carotid Artery and Mesentery Vessel at JoVE.com

Ferric Chloride-induced Thrombosis Mouse Model on Carotid Artery and Mesentery Vessel

The FeCl3 induced thrombosis model in mice is described herein. A method to monitor thrombus growth by intravital microscopy observation on a mesenteric vessel and by blood flow measurement in the carotid artery is presented.

Ferric Klorür kaynaklı Karotis Arter ve Mezenter Gemi Tromboz Fare Modeli

Severe thrombosis and its ischemic consequences such as myocardial infarction, pulmonary embolism and stroke are major worldwide health issues. The ferric ...

ferric-chloride-induced-thrombosis-mouse-model-on-carotid-artery

Research

JoVE Journal

Medicine

17.3K Görüntüleme.  Baker IDI Heart and Diabetes Institute. The FeCl3 induced thrombosis model in mice is described herein. A method to monitor thrombus growth by intravital microscopy observation on a mesenteric vessel and by blood flow measurement in the carotid artery is presented.

Video: Ferric Chloride-induced Thrombosis Mouse Model on Carotid Artery and Mesentery Vessel

A Model of Disturbed Flow-Induced Atherosclerosis in Mouse Carotid Artery by Partial Ligation and a Simple Method of RNA Isolation from Carotid Endothelium

Despite the well-known close association, direct evidence linking disturbed flow to atherogenesis has been lacking. We have recently used a modified version of carotid partial ligation methods [1,2] to show that it acutely induces low and oscillatory flow conditions, two key characteristics of disturbed flow, in the mouse common carotid artery. Using this model, we have provided direct evidence that disturbed flow indeed leads to rapid and robust atherosclerosis development in Apolipoprotein E knockout mouse [3]. We also developed a method of endothelial RNA preparation with high purity from the mouse carotid intima [3]. Using this mouse model and method, we found that partial ligation causes endothelial dysfunction in a week, followed by robust and rapid atheroma formation in two weeks in a hyperlipidemic mouse model along with features of complex lesion formation such as intraplaque neovascularization by four weeks. This rapid in vivo model and the endothelial RNA preparation method could be used to determine molecular mechanisms underlying flow-dependent regulation of vascular biology and diseases. Also, it could be used to test various therapeutic interventions targeting endothelial dysfunction and atherosclerosis in considerably reduced study duration.

This describes a partial carotid ligation surgery, which causes disturbed flow conditions and subsequent atherosclerosis development (in two weeks) with intraplaque neo-vascularization (in four weeks) in the mouse common carotid artery. We also describe a novel method of RNA isolation from the carotid intima, providing high purity endothelial RNA.

Kısmi ligasyon Fare Karotis Arter Disturbed Akış Kaynaklı Ateroskleroz ve Karotis Endotel RNA İzolasyonu Basit Bir Yöntem Bir Model

Despite the well-known close association, direct evidence linking disturbed flow to atherogenesis has been lacking. We have recently used a modified ...

Tıp

Mouse Complete Stasis Model of Inferior Vena Cava Thrombosis

Venous thromboembolism (VTE) includes both deep vein thrombosis (DVT) and pulmonary embolism (PE). In the United States (U.S.), the high morbidity and mortality rates make VTE a serious health concern 1-2. After heart disease and stroke, VTE is the third most common vascular disease 3. In the U.S. alone, there is an estimated 900,000 people affected each year, with 300,000 deaths occurring annually 3. A reliable in vivo animal model to study the mechanisms of this disease is necessary.
The advantages of using the mouse complete stasis model of inferior vena cava thrombosis are several. The mouse model allows for the administration of very small volumes of limited availability test agents, reducing costs dramatically. Most promising is the potential for mice with gene knockouts that allow specific inflammatory and coagulation factor functions to be delineated. Current molecular assays allow for the quantitation of vein wall, thrombus, whole blood, and plasma for assays. However, a major concern involving this model is the operative size constraints and the friability of the vessels. Also, due to the small IVC sample weight (mean 0.005 grams) it is necessary to increase animal numbers for accurate statistical analysis for tissue, thrombus, and blood assays such as real-time polymerase chain reaction (RT-PCR), western blot, enzyme-linked immunosorbent (ELISA), zymography, vein wall and thrombus cellular analysis, and whole blood and plasma assays 4-8.
The major disadvantage with the stasis model is that the lack of blood flow inhibits the maximal effect of administered systemic therapeutic agents on the thrombus and vein wall.

The mouse complete stasis model of inferior vena cava thrombosis yields quantifiable amounts of vein wall tissue and thrombus. It has proven useful for evaluating interactions between the vein wall and the occlusive thrombus and in assessing the progression from acute to chronic inflammation.

İnferior Vena Kava Trombozu Fare Komple Durağanlık Model

Venous thromboembolism (VTE) includes both deep vein thrombosis (DVT) and pulmonary embolism (PE).  In the United States (U.S.), the high morbidity and ...

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.

Orta Serebral Arter Tıkanıklığı Fare Modeli

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 ...

A Murine Model of Stent Implantation in the Carotid Artery for the Study of Restenosis

Despite the considerable progress made in the stent development in the last decades, cardiovascular diseases remain the main cause of death in western countries. Beside the benefits offered by the development of different drug-eluting stents, the coronary revascularization bears also the life-threatening risks of in-stent thrombosis and restenosis. Research on new therapeutic strategies is impaired by the lack of appropriate methods to study stent implantation and restenosis processes. Here, we describe a rapid and accessible procedure of stent implantation in mouse carotid artery, which offers the possibility to study in a convenient way the molecular mechanisms of vessel remodeling and the effects of different drug coatings.

A model of stent implantation in mouse carotid artery is described. Compared to other similar methods, this procedure is very rapid, simple and accessible, offering the possibility to study in a convenient way the vascular wall reaction to different drug-eluting stents and the molecular mechanisms of restenosis.

Restenoz Çalışmaları Karotis Arter Stent İmplantasyonu bir murin Modeli

Despite  the considerable progress made in the stent development in the last decades,  cardiovascular diseases remain the main cause of death in western ...

Adult Mouse Venous Hypertension Model: Common Carotid Artery to External Jugular Vein Anastomosis.

The understanding of the pathophysiology of brain arteriovenous malformations and arteriovenous fistulas has improved thanks to animal models. A rat model creating an artificial fistula between the common carotid artery (CCA) and the external jugular vein (EJV) has been widely described and proved technically feasible. This construct provokes a consistent cerebral venous hypertension (CVH), and therefore has helped studying the contribution of venous hypertension to formation, clinical symptoms, and prognosis of brain AVMs and dural AVFs. Equivalent mice models have been only scarcely described and have shown trouble with stenosis of the fistula. An established murine model would allow the study of not only pathophysiology but also potential genetic therapies for these cerebrovascular diseases.
We present a model of arteriovenous fistula that produces a durable intracranial venous hypertension in the mouse. Microsurgical anastomosis of the murine CCA and EJV can be difficult due to diminutive anatomy and frequently result in a non-patent fistula. In this step-by-step protocol we address all the important challenges encountered during this procedure. Avoiding excessive retraction of the vein during the exposure, using 11-0 sutures instead of 10-0, and making a carefully planned end-to-side anastomosis are some of the critical steps. Although this method requires advanced microsurgical skills and a longer learning curve that the equivalent in the rat, it can be consistently developed.
This novel model has been designed to integrate transgenic mouse techniques with a previously well-established experimental system that has proved useful to study brain AVMs and dural AVFs. By opening the possibility of using transgenic mice, a broader spectrum of valid models can be achieved and genetic treatments can also be tested. The experimental construct could also be further adapted to the study of other cerebrovascular diseases related with venous hypertension such as migraine, transient global amnesia, transient monocular blindness, etc.

We describe a method for creating a reliable model of cerebral venous hypertension in the adult mouse. This model has been widely described and tested in the rat. This new counterpart in the mice opens the possibility of using genetic modified animals and thereby broadens the applications of the model.

Yetişkin Fare Venöz Hipertansiyon Modeli: Dış Boyun Ven Anastomoza Ortak Karotis Arter.

The understanding of the pathophysiology of brain arteriovenous malformations and arteriovenous fistulas has improved thanks to animal models. A rat model ...

Vascular Balloon Injury and Intraluminal Administration in Rat Carotid Artery

The carotid artery balloon injury model in rats has been well established for over two decades. It remains an important method to study the molecular and cellular mechanisms involved in vascular smooth muscle dedifferentiation, neointima formation and vascular remodeling. Male Sprague-Dawley rats are the most frequently employed animals for this model. Female rats are not preferred as female hormones are protective against vascular diseases and thus introduce a variation into this procedure. The left carotid is typically injured with the right carotid serving as a negative control. Left carotid injury is caused by the inflated balloon that denudes the endothelium and distends the vessel wall. Following injury, potential therapeutic strategies such as the use of pharmacological compounds and either gene or shRNA transfer can be evaluated. Typically for gene or shRNA transfer, the injured section of the vessel lumen is locally transduced for 30 min with viral particles encoding either a protein or shRNA for delivery and expression in the injured vessel wall. Neointimal thickening representing proliferative vascular smooth muscle cells usually peaks at 2 weeks after injury. Vessels are mostly harvested at this time point for cellular and molecular analysis of cell signaling pathways as well as gene and protein expression. Vessels can also be harvested at earlier time points to determine the onset of expression and/or activation of a specific protein or pathway, depending on the experimental aims intended. Vessels can be characterized and evaluated using histological staining, immunohistochemistry, protein/mRNA assays, and activity assays. The intact right carotid artery from the same animal is an ideal internal control. Injury-induced changes in molecular and cellular parameters can be evaluated by comparing the injured artery to the internal right control artery. Likewise, therapeutic modalities can be evaluated by comparing the injured and treated artery to the control injured only artery.

This protocol uses a balloon catheter to cause an intraluminal injury on the rat carotid artery and henceforth elicit neointimal hyperplasia. This is a well-established model for studying the mechanisms of vascular remodeling in response to injury. It is also widely used to determine the validity of potential therapeutic approaches.

Sıçan Karotis Arter Vasküler Balon Yaralanma ve intraluminal İdaresi

The carotid artery balloon injury model in rats has been well established for over two decades. It remains an important method to study the molecular and ...

A Rat Carotid Balloon Injury Model to Test Anti-vascular Remodeling Therapeutics

The rat carotid balloon injury is a well-established surgical model that has been used to study arterial remodeling and vascular cell proliferation. It is also a valuable model system to test, and to evaluate therapeutics and drugs that negate maladaptive remodeling in the vessel. The injury, or barotrauma, in the vessel lumen caused by an inflated balloon via an inserted catheter induces subsequent neointimal growth, often leading to hyperplasia or thickening of the vessel wall that narrows, or obstructs the lumen. The method described here is sufficiently sensitive, and the results can be obtained in relatively short time (2 weeks after the surgery). The efficacy of the drug or therapeutic against the induced-remodeling can be evaluated either by the post-mortem pathological and histomorphological analysis, or by ultrasound sonography in live animals. In addition, this model system has also been used to determine the therapeutic window or the time course of the administered drug. These studies can leadto the development of a better administrative strategy and a better therapeutic outcome. The procedure described here provides a tool for translational studies that bring drug and therapeutic candidates from bench research to clinical applications.

The rat carotid balloon injury model described below allows researchers to evaluate drugs or therapeutics that negate injury-induced arterial hyperplasia. Detailed pre-surgical preparation, surgical procedure, and post-surgical cares of the animal are described.

Rat Karotis Balon Yaralanma Modeli Anti-vasküler Tadilat Therapeutics Test

The rat carotid balloon injury is a well-established surgical model that has been used to study arterial remodeling and vascular cell proliferation. It is ...

Ferric Chloride-induced Murine Thrombosis Models

Arterial thrombosis (blood clot) is a common complication of many systemic diseases associated with chronic inflammation, including atherosclerosis, diabetes, obesity, cancer and chronic autoimmune rheumatologic disorders. Thrombi are the cause of most heart attacks, strokes and extremity loss, making thrombosis an extremely important public health problem. Since these thrombi stem from inappropriate platelet activation and subsequent coagulation, targeting these systems therapeutically has important clinical significance for developing safer treatments. Due to the complexities of the hemostatic system, in vitro experiments cannot replicate the blood-to-vessel wall interactions; therefore, in vivo studies are critical to understand pathological mechanisms of thrombus formation. To this end, various thrombosis models have been developed in mice. Among them, ferric chloride (FeCl3) induced vascular injury is a widely used model of occlusive thrombosis that reports platelet activation and aggregation in the context of an aseptic closed vascular system. This model is based on redox-induced endothelial cell injury, which is simple and sensitive to both anticoagulant and anti-platelets drugs. The time required for the development of a thrombus that occludes blood flow gives a quantitative measure of vascular injury, platelet activation and aggregation that is relevant to thrombotic diseases. We have significantly refined this FeCl3-induced vascular thrombosis model, which makes the data highly reproducible with minimal variation. Here we describe the model and present representative data from several experimental set-ups that demonstrate the utility of this model in thrombosis research.

We report a refined procedure of the ferric chloride (FeCl3)-induced thrombosis models on carotid and mesenteric artery as well as vein, characterized efficiently using intravital microscopy to monitor time to occlusive thrombi formation.

Ferrik klorit indüklenen fare tromboz modellerinde

Arterial thrombosis (blood clot) is a common complication of many systemic diseases associated with chronic inflammation, including atherosclerosis, ...

Phase Contrast Magnetic Resonance Imaging in the Rat Common Carotid Artery

Phase contrast magnetic resonance imaging (PC-MRI) is a noninvasive approach that can quantify flow-related parameters such as blood flow. Previous studies have shown that abnormal blood flow may be associated with systemic vascular risk. Thus, PC-MRI can facilitate the translation of data obtained from animal models of cardiovascular diseases to pertinent clinical investigations. In this report, we describe the procedure for measuring blood flow in the common carotid artery (CCA) of rats using cine-gated PC-MRI and discuss relevant analysis methods. This procedure can be performed in a live, anesthetized animal and does not require euthanasia after the procedure. The proposed scanning parameters yield repeatable measurements for blood flow, indicating excellent reproducibility of the results. The PC-MRI procedure described in this article can be used for pharmacological testing, pathophysiological assessment, and cerebral hemodynamics evaluation.

The overall goal of this procedure is to measure the blood flow in the rat common carotid artery by using noninvasive phase contrast magnetic resonance imaging.

Faz kontrast manyetik rezonans görüntüleme sıçan ortak karotis arter

Phase contrast magnetic resonance imaging (PC-MRI) is a noninvasive approach that can quantify flow-related parameters such as blood flow. Previous ...

Ferric Chloride-induced Canine Carotid Artery Thrombosis: A Large Animal Model of Vascular Injury

Occlusive arterial thrombosis leading to cerebral ischemic stroke and myocardial infarction contributes to ~13 million deaths every year globally. Here, we have translated a vascular injury model from a small animal into a large animal (canine), with slight modifications that can be used for pre-clinical screening of prophylactic and thrombolytic agents. In addition to the surgical methods, the modified protocol describes the step-by-step methods to assess carotid artery canalization by angiography, detailed instructions to process both the brain and carotid artery for histological analysis to verify carotid canalization and cerebral hemorrhage, and specific parameters to complete an assessment of downstream thromboembolic events by utilizing magnetic resonance imaging (MRI). In addition, specific procedural changes from the previously well-established small animal model necessary to translate into a large animal (canine) vascular injury are discussed.

Here, we present the modifications necessary to a well characterized and commonly used small animal ferric chloride-induced (FeCl3) carotid artery injury model for use in a large animal vascular injury model. The resulting model can be utilized for pre-clinical trial assessment of both prophylactic and thrombolytic pharmacological and mechanical interventions.

Ferrik klorid kaynaklı köpek karotid arter trombozu: Bir büyük hayvan modeli vasküler yaralanma

Occlusive arterial thrombosis leading to cerebral ischemic stroke and myocardial infarction contributes to ~13 million deaths every year globally. Here, ...