Name: surgical approach for middle cerebral artery occlusion and reperfusion induced stroke in mice
Uploaded: 2016-10-20T13:00:00.000+00:00
Duration: 10 min 30 s
Description: Watch this Scientific Journal Video about Surgical Approach for Middle Cerebral Artery Occlusion and Reperfusion Induced Stroke in Mice at JoVE.com

This work was funded by the National Institute of Health, the National Heart Lung and Blood Institute (NIH and NHLBI; HL125572-01A1) and the LSUHSC-S start up fund to F.N.E. Gavins.

video bildirilen Bu protokol ve deneyler LSUHSC-S Kurumsal Hayvan Bakım ve Kullanım Kurulu tarafından onaylanmış ve NIH kurallarına uygundur bulundu. Not: Erkek C57BL / 25 ağırlığında 6 fareleri - 29 g Bu çalışmada kullanılmıştır. farenin tek havalandırılmış kafeslerde, 12 saat aydınlık / karanlık döngüsü altında, suya serbest erişim ile standart bir yiyecek topağı diyet ile beslendi. Prosedür steril teknikler (örneğin, steril eldiven, steril aletleri) kullanılarak steril koşullar altında yapılacaktır. 1. Ön cerrahi Hazırlıklar <ol><li> Ketamin (150 mg / kg) ve ksilazin (10 mg / kg) bir kombinasyonu kullanılarak anestezi neden intraperitonal (ip) enjekte edilmiştir. anestezi sağlanır kez 5 dakika ve her 10 dakikada - başlangıçta ayak tutam her 3 ile anestezi derinliği izleyin. hayvan anestezi altında iken, kuruluğunu önlemek için steril göz merhem yönetmek. ketamin başlangıç ​​dozu / xylazine genellikle ab sürer30-40 dakika dışarı. ayak tutam hayvanın tepki ile teyit edildiği, gerektiğinde ilave bir doz olarak uygulanabilir. </li><li> sıcaklık ayarlı ısı mat bir yatar pozisyonda fareler yerleştirin ve bir rektal sonda kullanılarak doğrulanmış olan 36.5 ± 0.1 ° C, vücut ısısını korumak. </li><li> boyun Tıraş ve% 70 etil alkol ile cilt dezenfekte edin. </li></ol> MCA 2. Oklüzyon (Şekil 1) <ol><li> İris düz makas kullanarak bir orta hat boyun kesi yapmak ve damarları açığa çıkarmak için yumuşak dokuları (retraktörler kullanarak) geri çekin. </li><li> vagus siniri zarar vermeden Dumont forseps kullanılarak çevreleyen dokudan CCA ve ECA teşrih. </li><li> 6-0 ipek kullanılarak CCA etrafında gevşek bir düğüm atarak geçici bir dikiş yapın. </li><li> ECA etrafında kalıcı bir dikiş ve sıkıca damarları (CCA çatallanma distal) bağlanarak kendisine uzanan küçük gemilere olun. </li><li> Bir dikiş aro yapınCCA çatallanma ECA proksimal und. </li><li> internal karotis arter (ICA) ve pterygopalatine arter (KİK) etrafında bir mikrovasküler klip yerleştirin. </li><li> yaylı makas diseksiyon mikro kullanarak ECA küçük bir kesi yapmak ve 180 mikron silikon uçlu monofilament yerleştirin. filamanın kolay manipülasyon için mümkün olduğunca kalıcı sütür kesme yakın olun. </li><li> takılı filament ile ECA etrafında geçici sütür sıkın ve mikrovasküler klibi çıkarın. </li><li> Kalıcı, distal sütür ve yaylı makas diseksiyon mikro kullanarak filamanın giriş noktası arasındaki ECA kesti. </li><li> MCA içinde - (CCA çatallanma ötesinde 10 mikron. Yaklaşık 9) direnç hissedene kadar ICA aracılığıyla filamanın Kılavuzu. NOT: filamanın çoğunluğu hala görünür iken çok fazla direnç hissedilir durumunda, Filament PPA girmiş olabilir. Bu durumda, yavaşça irin, çatallanma geri filamanın çekmeileri ICA kullanarak Dumont forseps içine h ve ICA görüntülenebilir kadar filamanın ilerlemek. </li></ol> 3. reperfüzyon <ol><li> 30 dakikalık oklüzyon döneminden sonra, yavaşça geri çekerek Dumont forseps kullanarak filamanın kaldırmak ve ECA açık ucundan etrafında dikiş sabitleyin. </li><li> dikkatle CCA ile sürdürülür Dumont forseps ve kan akışını kullanarak bitişik harfleri gevşeterek CCA etrafında geçici dikiş çıkarın. </li><li> Sürekli cerrahi sütür ile kesi kapatın. cilt kapatılması ya sürekli veya kesintili dikiş ile gerçekleştirilebilir. Cilt zımba da kabul edilebilir bir yöntemdir. </li><li> hacim ikmal olarak ve analjezik karprofen ile tuzlu subkutan 1 ml ile fareler enjekte (5 mg / Kg, sc) cerrahi prosedür, ek ağrı kesici geri hunched dahil gerekli işaret İşaretler, ungroomed ceket ağrı ve rahatsızlık giderilmesi için, azalmış aktivite anormalpostür ve iştah azalması. </li><li> sıcaklık kontrolörü kullanarak düzenlenmiş bir ısı lambası kullanarak ısıtılmış 30 ° C kafeste anestezi kurtarma boyunca fareler gözlemlemek ve yeme teşvik etmek için kafesin katta bir petri püresi chow yerleştirin. fareler reperfüzyon döneminde kafes başına bir yer olacaktır. </li></ol> 4. Sham Cerrahisi <ol><li> monofilament ekleme olmadan aynı prosedüre tabi fareler. </li></ol> 5. Post-operatif nörolojik değerlere göre (Tablo 1, Şekil 2) <ol><li> Nörolojik Genel değerlendiren 18 maddelik skorlama sistemi kullanılarak uygun reperfüzyon döneminden sonra fareler değerlendirmek; Motor; Duyusal; Propriyosepsiyon. Daha yüksek nörolojik puan nörolojik fonksiyonu azalmış karşılık gelir. NOT: yürümek tepkisiz ve yapamaz değerlendirilecektir Fareler ötenazi olacaktır. insani ötanazi için diğer kriterler% 20&#39;den fazla bir ağırlık kaybı içerir, solunum diCerrahi alanı çevresinde stres ve enfeksiyon. Bir CO2 odası servikal dislokasyon ya torakotomi olacak ölüm onaylamak için ötanazi ve fiziksel yöntem için kullanılır. </li></ol> Ölçüm Beyin Enfarktüs Hacminin 6. (Şekil 3) <ol><li> anestezi sağlandıktan sonra 5 dakika ve her 10 dakika - ketamin (150 mg / kg) ve ksilazin (10 mg / kg), ilk ayak tutam her 3 ile anestezi ip monitör derinliği enjekte bir kombinasyonu kullanılarak anesteziye neden. </li><li> Bir yatar pozisyonda fareler yerleştirin ve iris düz makas kullanarak periton ardından boyun karın cildi kesti. </li><li> Dumont forseps kullanarak sternum yukarı kaldırın ve kalp maruz kaburga kesti. </li><li> kalbi açığa sol ve hemostat iki çift kullanarak göğüs sağ tarafını açın. </li><li> Sol ventrikül içine bir 5 ml şırıngaya bağlanmış bir 26 G iğne takın ve sağ atriyum kesilmiş. Oda sıcaklığı normal serum fizyolojik veya Phospa serpmekte tamponlu tuz (PBS) Sıvı berrak hale gelinceye kadar (çoğunlukla 3-5 dakika). </li><li> iris düz makas ve Dumont forseps kullanarak dikkatli bir şekilde beyin uzak kafatası çıkarın. </li><li> koku ampuller ve beyin matriks içinde uyacak şekilde bir jilet kullanarak beyincik kesti. Hatta segmentler halinde beyin dilim için bu matrisi kullanın. serin doku tutmak için kullanımdan önce matrisi Chill. matriks içine beyin yerleştirin ve buz üzerinde ayarlayın. </li><li> İki Jilet kullanarak 2 mm koronal kesimleri içine beyin dilimleyin. üstten 2 mm başlayan bir jilet kullanarak ilk kesim olun ve yerine bu jilet bırakın. İlk arkasında kesilmiş başka bir 2 mm yapın. Ekli doku ile ilk jilet çıkarın. Tüm doku dilimlenmiş kadar bu işlemi tekrarlayın. NOT: 4 olmalıdır - Bittiğinde 5 segmentleri. </li><li> Daha sonra sığ bir su banyosu içine yerleştirilir% 2 2,3,5-triphenyltetrazalium klorür (TTC) içeren 24 oyuklu plaka segmentleri yerleştirin20 dakika T 37 ° C. Tek bir kuyuya her kesimi yerleştirme onlar kesildi sırayla onları tutmaya yardımcı olur. TTC çözüm tamamen doku segmentlerini kapsayan emin olun. 10 dakika tüm dilimleri üzerinde açtıktan sonra. </li><li> Yeni 24-kuyulu plakanın kuyularına% 10 formalin küçük bir miktar ve kesilmiş sırayla Bu plaka segmentleri aktarın. </li><li> (NIH 1.57 Görüntü Yazılımı) 6 bilgisayara segmentleri tarayın ve ImageJ analiz yazılımı kullanılarak tüm beyin dilim 6 bir yüzdesi olarak infarkt analiz. <ol><li> Bir alan ölçümü oluşturmak için enfarkte alanı özetlemektedir. Sonraki alanı belirlemek için tüm taraf yarımküre ölçün. kontralateral yarımkürenin alanına göre infarkt alanını bölün ve enfarktüs hacmini belirlemek için 100 ile çarpın. </li></ol></li></ol>

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The authors have nothing to disclose.

20 yıl önce anlayışı bu yana, bir filamanın sokulmasını kapsayan insan inme için MCAO modeli çalışmaları çok sayıda kullanılmıştır. Bunun nedeni o inme (yani, iskemik inme) en sık görülen klinik ne taklit gerçeği esas olarak. striatum serebral korteks daha iskemi karşı daha duyarlıdır ve bu nedenle, iskemik sürenin uzunluğu striatum ve dorsolateral korteks hem de etkilenmiş, ya da sadece striatum olup olmayacağı çevirmek olacaktır. Hem enfarktüs ve reperfüzyon süreleri buna...

İnme bir kişi hastalıktan her 4 dakikada bir ölen ile, dünya çapında ölüm beşinci önde gelen nedenidir. 800.000 üzerinde Amerikalılar sadece hasta için yıkıcı olmayan bir felç, her yıl, acı, ama aynı zamanda aileleri için. İnme yetişkin sakatlık ana nedenidir ve yıllık harcama mevcut olan çok az tedavi seçenekleri olmasına rağmen 36500000000 $ 1 düzeyinde olduğu tahmin edilmektedir. Doku plazminojen aktivatörü (tPA) Sadece Gıda ve İlaç Dairesi (FDA) iskemik inme için ilaç lisanslı. Inme 3-6 saat içinde hastaya uygulandığı ve bu gibi durumlarda bu hastaların 2% 4 yarar Ancak, bu geçerlidir. Nedenle, inme, tekrar üretilebilir, klinik olarak hayvan modelleri, bu hastalık için potansiyel bir terapötik strateji ve tedavilerin geliştirilmesine yardımcı olmak için kullanılır zorunludur. In vitro dikkat etmek önemlidir </em&gt; modelleri, serebral disfonksiyon bazı yönlerini modelleme yararlı iken, bir inme sonrası beyin ve çevrede meydana gelen karmaşık fizyolojik etkileşimleri yansıtan yeteneğine sahip değildir. Sonuç olarak, in vivo modeller gereklidir. inme en yaygın türü, toplam inmelerin% 87 için muhasebe, menşe iskemik olduğunu. Diğer vuruş intraserebral kanama (% 9) ve subaraknoid kanama (% 4), ve orta serebral arter (MCA) bir emboli en sık neden olur. Bu kesintiye hale beyin giren laminer kan akımının neden MCA kökünde belirgin eğri, atfedilebilir. MCA yanal sulkus boyunca internal karotis arter (ICA) ve yolları, doğar nerede dalları ve projeler bazal ganglionlar ve frontal, parietal ve primer motor ve duyusal korteks olmak üzere temporal lob, yanal yüzeyler. Willis Çember olma posterior serebral arterlerin tarafından oluşturulanserebral arter ve posterior iletişim arterlere bağlı. MCAO intraluminal filament veya dikiş modeli en yaygın inme araştırmalarında kullanılan biridir. Ancak, bu modele farklı varyasyonlar bir çift vardır ve bu microfilament dış karotid arter içine takılı olup olmadığını dayalı 3 (ECA, Longa yöntemi olarak adlandırılır), ya da (ICA takılı olup olmadığını Koizumi olarak adlandırılır yöntemi) 4. Filament CCA içinde kesi kanamayı önlemek için kaldırılırsa Longa yönteminde kalıcı 5 bağlanmalıdır ECA ise Koizumi yöntemde, ameliyat tarafındaki karotis arter (CCA) kalıcı olarak bağlanmalıdır . Bu çok daha üstün ve iskemik inme bir daha klinik olarak cerrahi model hissediyorum burada Longa yöntemi kullanılacaktır. Bundan başka, özellikle Longa yöntemi ile bir silikon uçlu monofilin kullanımı, çok üretirgenellikle eksik tıkanıklığı ve / veya subaraknoid kanama 6 üretmek alev körelmiş monoipliklerin, aksine tekrarlanabilir MCAO. Intraluminal filament yöntemi kalıcı veya geçici tıkanma 4,6 modeli olarak kullanılabilir. Geçici modeli gerçekleştirmek için, filament iskemi bir süre sonra çıkartılır (örneğin, 30 dakika, 60 dakika ya da 2 saat) ve reperfüzyon meydana bırakılır. Bu model, bir ölçüde, insanlarda bir tromboembolik pıhtı eritmek için kendiliğinden ya da terapötik müdahale (örneğin, tPA uygulama) sonrasında kan akımının restorasyon simüle eder. Sürekli modeller için, filament, sadece bir zaman süresi (örneğin, 24 saat) yerinde kalır, bu nedenle bir reperfüzyon oluşur. lümen içi filament yöntemin diğer bir avantajı, bir kranyotomi kafatası sağlam kalmasına olanak sağlayan ve intrakranyal basınç ve sıcaklık değişiklikleri önlemek gerçekleştirilebilir gerekmez gerçektir. <p cBu videoda lass = &quot;jove_content&quot;&gt; biz MCAO ve reperfüzyon ikna etmek Longa intraluminal filament yöntemi gerçekleştirmek için nasıl gösterilmektedir. Biz de 18 sayılık nörolojik skoru gerçekleştirmek ve 2,3,5-triphenyltetrazalium klorür (TTC) boyama kullanarak infarkt hacmini belirlemek için nasıl gösterir.

<table><tbody><tr><td>Male C57BL/6 mice</td><td>Jackson Laboratory, Bar Harbor, ME</td><td>#000664</td><td></td></tr><tr><td>Ketamine Hydrochloride</td><td>Morris &amp;amp; Dickson, Shreveport, LA</td><td>67457-108-10</td><td></td></tr><tr><td>Xylazine</td><td>Akorn, Inc, Lake Forest, IL</td><td>NADA# 139-236</td><td></td></tr><tr><td>DC temperature control system</td><td>FHC, Bowdoin, ME</td><td>40-90-8D</td><td></td></tr><tr><td>Mini rectal thermistor probe</td><td>FHC, Bowdoin, ME</td><td>40-80-5D-02</td><td></td></tr><tr><td>Heating pad</td><td>FHC, Bowdoin, ME</td><td>40-90-2-06</td><td></td></tr><tr><td>Clippers</td><td>Amazon, Bellevue, WA</td><td>#64800</td><td></td></tr><tr><td>70% ethanol</td><td>Worldwide Medical Products, Bristol, PA</td><td>#51011023</td><td></td></tr><tr><td>Dissecting microscope</td><td>Olympus, Center Valley, PA</td><td>SZ40</td><td></td></tr><tr><td>Iris scissors (straight)</td><td>Fine Science Tools, Foster City, CA</td><td>11251-20</td><td></td></tr><tr><td>Dumont forceps (45&amp;deg; bent tip)</td><td>Fine Science Tools, Foster City, CA</td><td>11297-00</td><td></td></tr><tr><td>Micro vessel clip</td><td>Fine Science Tools, Foster City, CA</td><td>18055-05</td><td></td></tr><tr><td>Micro dissecting spring scissors (straight)</td><td>Fine Science Tools, Foster City, CA</td><td>14088-10</td><td></td></tr><tr><td>Retractors (blunt)</td><td>Fine Science Tools, Foster City, CA</td><td>18200-11 (Helen used 17022-13)</td><td></td></tr><tr><td>Cotton tipped applicators</td><td>Fisher Scientific, Waltham, MA</td><td>23-400-100</td><td></td></tr><tr><td>Gauze sponges</td><td>Covidien, Mansfield, MA</td><td>#9023</td><td></td></tr><tr><td>7-0 silk braided surgical suture</td><td>Braintree Scientific, Braintree, MA</td><td>SUT-S103</td><td></td></tr><tr><td>0.9% sodium chloride</td><td>Morris &amp;amp; Dickson, Lake Forest, IL</td><td>0409-4888-20</td><td></td></tr><tr><td>6-0 medium MCAO suture (silicon rubber coated monofilament)</td><td>Doccol Corporation, Sharon, MA</td><td>6023PKRe</td><td></td></tr><tr><td>Sofsilk 6-0 silicone coated braided silk</td><td>Covidien, Mansfield, MA</td><td>SUT-14-1</td><td></td></tr><tr><td>Carprofen</td><td>Pfizer, New York, NY</td><td>NADA# 141-199</td><td></td></tr><tr><td>Puralube</td><td>Dechra, Norwich, UK</td><td>NDC 17033-211-38</td><td></td></tr><tr><td>Physitemp temperature controller</td><td>Harvard Apparatus, Holliston, MA</td><td>TCAT-2AC</td><td></td></tr><tr><td>Heat lamp</td><td>Harvard Apparatus, Holliston, MA</td><td>HL-1</td><td></td></tr><tr><td>Laser doppler probe</td><td>AD Instruments, Colorado Springs, CO</td><td>MSP100XP</td><td></td></tr><tr><td>24-well plates</td><td>Fisher Scientific, Waltham, MA</td><td>#353226</td><td></td></tr><tr><td>Phosphate buffered saline (PBS)</td><td>Life Technologies, Carlsbad, CA</td><td>20012-050</td><td></td></tr><tr><td>Single edge razor blades</td><td>Fisher Scientific, Waltham, MA</td><td>12-640</td><td></td></tr><tr><td>2,3,5-triphenyltetrazalium chloride (TTC)</td><td>Sigma Aldrich, St. Louis, MO</td><td>T8877-50G</td><td></td></tr><tr><td>Mouse brain matrix slicer</td><td>Braintree Scientific, Braintree, MA</td><td>BS-A 5000C</td><td></td></tr><tr><td>Water bath</td><td>VWR, Radnor, PA</td><td>#182</td><td></td></tr><tr><td>10% formalin</td><td>Sigma Aldrich, St. Louis, MO</td><td>HT501128-4L</td><td></td></tr><tr><td>ImageJ analysis software</td><td>NIH, Bethesda, MD</td><td>free download</td><td></td></tr><tr><td>Retractor</td><td>Medical Device Purchase, Newcastle, CA</td><td>MP-740</td><td></td></tr></tbody></table>

surgical approach for middle cerebral artery occlusion and reperfusion induced stroke in mice

İnme, tüm dünyada ölüm önde gelen nedenidir ve uzun vadeli yetişkin engelli temel nedenlerinden biri olmaya devam etmektedir. vuruşları yaklaşık% 87 iskemik orijinlidir ve orta serebral arter (MCA) topraklarında meydana gelir. Şu anda sadece Gıda ve İlaç Dairesi (FDA) bu yıkıcı hastalığın tedavisi için ilaç onayladı doku plazminojen aktivatörü (tPA) &#39;dir. Bununla birlikte, tPA uygulama için küçük bir terapötik penceresi olan (3-6 saat) ve aslında uygulanan hastaların% 4 etkilidir. Mevcut araştırma potansiyel terapötik hedefleri bulmak için inme patofizyolojisinin anlaşılmasında odaklanır. Böylece, güvenilir modeller çok önemlidir ve MCA oklüzyonu (MCAO) modeli iskemik inme klinik olarak en uygun cerrahi modeli olarak kabul edilir (ayrıca lümen Filament veya sütür modeli olarak adlandırılan) ve oldukça non-invaziv ve kolay tekrarlanabilir. Genellikle MCAO modeli nedeniyle özellikle fare ile, kemirgenler kullanılırBu türler için mevcut tüm genetik varyasyonları. Burada tarif (ve video mevcut) başarıyla güvenilir ve tekrarlanabilir verileri oluşturmak için farelerde (reperfüzyon) ile MCAO modelini nasıl gerçekleştirileceği.

Fareler reperfüzyon süresi ardından 30 dakika MCAO-kaynaklı beyin iskemisi (Şekil 1) uygulandı (24 saat, 1 hafta, burada sunulan, ancak reperfüzyon uzunluğunun değiştirilmesi olabilir). MCAO sırasında ölüm (yaklaşık% 2) çok az oldu. Mesaj iskemi, (ilk 24 saat içinde) ölüm oranı yaklaşık% 26 idi. Lazer Doppler flowmetre öncesi ve MCAO / reperfüzyon sonrası MCA topraklarında kan akım?...

Watch this Scientific Journal Video about Surgical Approach for Middle Cerebral Artery Occlusion and Reperfusion Induced Stroke in Mice at JoVE.com

Surgical Approach for Middle Cerebral Artery Occlusion and Reperfusion Induced Stroke in Mice

inme patofizyolojisini anlamak için, güvenilir modeller kullanmak önemlidir. Farelerde en sık kullanılan inme modellerinden birini anlatacağız Bu yazıda, orta serebral arter tıkanıklığı (MCAO) modeli olarak adlandırılan reperfüzyon ile (ayrıca lümen içi filament veya sütür modeli olarak adlandırılır).

Farelerde Orta Serebral Arter Tıkanıklığı ve Reperfüzyon Kaynaklı İnme Cerrahi Yaklaşım

Stroke is a leading cause of death worldwide and continues to be one of the major causes of long-term adult disabilities. About 87% of strokes are ...

surgical-approach-for-middle-cerebral-artery-occlusion-reperfusion

Research

JoVE Journal

Medicine

21.5K Görüntüleme.  Louisiana State University.  inme patofizyolojisini anlamak için, güvenilir modeller kullanmak önemlidir. Farelerde en sık kullanılan inme modellerinden birini anlatacağız Bu yazıda, orta serebral arter tıkanıklığı (MCAO) modeli olarak adlandırılan reperfüzyon ile (ayrıca lümen içi filament veya sütür modeli olarak adlandırılır). 

Video: Surgical Approach for Middle Cerebral Artery Occlusion and Reperfusion Induced Stroke in Mice

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

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 ...

Mouse Model of Intraluminal MCAO: Cerebral Infarct Evaluation by Cresyl Violet Staining

Stroke is the third cause of mortality and the leading cause of disability in the World. Ischemic stroke accounts for approximately 80% of all strokes. However, the thrombolytic tissue plasminogen activator (tPA) is the only treatment of acute ischemic stroke that exists. This led researchers to develop several ischemic stroke models in a variety of species. Two major types of rodent models have been developed: models of global cerebral ischemia or focal cerebral ischemia. To mimic ischemic stroke in patients, in whom approximately 80% thrombotic or embolic strokes occur in the territory of the middle cerebral artery (MCA), the intraluminal middle cerebral artery occlusion (MCAO) model is quite relevant for stroke studies. This model was first developed in rats by Koizumi et al. in 1986 1. Because of the ease of genetic manipulation in mice, these models have also been developed in this species 2-3.
	Herein, we present the transient MCA occlusion procedure in C57/Bl6 mice. Previous studies have reported that physical properties of the occluder such as tip diameter, length, shape, and flexibility are critical for the reproducibility of the infarct volume 4. Herein, a commercial silicon coated monofilaments (Doccol Corporation) have been used. Another great advantage is that this monofilament reduces the risk to induce subarachnoid hemorrhages. Using the Zeiss stereo-microscope Stemi 2000, the silicon coated monofilament was introduced into the internal carotid artery (ICA) via a cut in the external carotid artery (ECA) until the monofilament occludes the base of the MCA. Blood flow was restored 1 hour later by removal of the monofilament to mimic the restoration of blood flow after lysis of a thromboembolic clot in humans. The extent of cerebral infarct may be evaluated first by a neurologic score and by the measurement of the infarct volume. Ischemic mice were thus analyzed for their neurologic score at different post-reperfusion times. To evaluate the infarct volume, staining with 2,3,5-triphenyltetrazolium chloride (TTC) was usually performed. Herein, we used cresyl violet staining since it offers the opportunity to test many critical markers by immunohistochemistry. In this video, we report the MCAO procedure; neurological scores and the evaluation of the infarct volume by cresyl violet staining.

The intraluminal middle cerebral occlusion model in mice is herein presented. The extent of cerebral infarct is evaluated by a neurologic score and cresyl violet staining, an alternative staining to TTC, offering the great advantage to test in parallel many interest markers.

Intraluminar MCAO Mouse Model: kresil Violet Boyama tarafından Serebral İnfarktı Değerlendirilmesi

Stroke is the third  cause of mortality and the leading cause of disability in the World. Ischemic  stroke accounts for approximately 80% of all strokes. ...

Induction of Ischemic Stroke and Ischemia-reperfusion in Mice Using the Middle Artery Occlusion Technique and Visualization of Infarct Area

Cerebrovascular disease is highly prevalent in the global population and encompasses several types of conditions, including stroke. To study the impact of stroke on tissue injury and to evaluate the effectiveness of therapeutic interventions, several experimental models in a variety of species were developed. They include complete global cerebral ischemia, incomplete global ischemia, focal cerebral ischemia, and multifocal cerebral ischemia. The model described in this protocol is based on the middle cerebral artery occlusion (MCAO) and is related to the focal ischemia category. This technique produces consistent focal ischemia in a strictly defined region of the hemisphere and is less invasive than other methods. The procedure described is performed on mice, given the availability of several genetic variants and the high number of tests standardized for mice to aid in the behavioral and neurodeficit evaluation.

We describe a mouse model of stroke induced by the occlusion of the middle cerebral artery using a silicone coated suture. The protocol can be applied to induce permanent occlusion or a temporary ischemia, followed by reperfusion.

Enfarktüs Bölgesi Orta Arter Tıkanıklığı Tekniği ve Görselleştirme kullanma Fare İskemik İnme ve İskemi-reperfüzyon indüksiyon

Cerebrovascular disease is highly prevalent in the global population and encompasses several types of conditions, including stroke. To study the impact of ...

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

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

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

Orta serebral arter tıkanıklığı ortak karotid arter onarım ile reperfüzyon farelerde izin

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

Two-vessel Occlusion Mouse Model of Cerebral Ischemia-reperfusion

In this study, a middle cerebral artery (MCA) occlusion mouse model is employed to study cerebral ischemia-reperfusion. A reproducible and reliable mouse model is useful for investigating the pathophysiology of cerebral ischemia-reperfusion and determining potential therapeutic strategies for patients with stroke. Variations in the anatomy of the circle of Willis of C57BL/6 mice affects their infarct volume after cerebral-ischemia-induced injury. Studies have indicated that distal MCA occlusion (MCAO) can overcome this problem and result in a stable infarct size. In this study, we establish a two-vessel occlusion mouse model of cerebral ischemia-reperfusion through the interruption of the blood flow to the right MCA. We distally ligate the right MCA and right common carotid artery (CCA) and restore blood flow after a certain period of ischemia. This ischemia-reperfusion injury induces an infarct of stable size and a behavioral deficit. Peripheral immune cells infiltrate the ischemic brain within the 24 h infiltration period. Additionally, the neuronal loss in the cortical area is less for a longer reperfusion duration. Therefore, this two-vessel occlusion model is suitable for investigating the immune response and neuronal recovery during the reperfusion period after cerebral ischemia.

A mouse model of cerebral ischemia-reperfusion is established to investigate the pathophysiology of stroke. We distally ligate the right middle cerebral artery and right common carotid artery and restore blood flow after 10 or 40 min of ischemia.

Serebral iskemi-reperfüzyon iki damar tıkanıklığı fare modeli

In this study, a middle cerebral artery (MCA) occlusion mouse model is employed to study cerebral ischemia-reperfusion. A reproducible and reliable mouse ...

Modeling Stroke in Mice: Transient Middle Cerebral Artery Occlusion via the External Carotid Artery

Stroke is the third most common cause of mortality and the leading cause of acquired adult disability in developed countries. To date, therapeutic options are limited to a small proportion of stroke patients within the first hours after stroke. Novel therapeutic strategies are being extensively investigated, especially to prolong the therapeutic time window. These current investigations include the study of important pathophysiological pathways after stroke, such as post-stroke inflammation, angiogenesis, neuronal plasticity, and regeneration. Over the last decade, there has been increasing concern about the poor reproducibility of experimental results and scientific findings among independent research groups. To overcome the so-called &#8220;replication crisis&#8221;, detailed standardized models for all procedures are urgently needed. As an effort within the &#8220;ImmunoStroke&#8221; research consortium (https://immunostroke.de/), a standardized mouse model of transient middle cerebral artery occlusion (MCAo) is proposed. This model allows the complete restoration of the blood flow upon removal of the filament, simulating the therapeutic or spontaneous clot lysis that occurs in a large proportion of human strokes. The surgical procedure of this &#8220;filament&#8221; stroke model and tools for its functional analysis are demonstrated in the accompanying video.

Different models of middle cerebral artery occlusion (MCAo) are used in experimental stroke research. Here, an experimental stroke model of transient MCAo via the external carotid artery (ECA) is described, which aims to mimic human stroke, in which the cerebrovascular thrombus is removed due to spontaneous clot lysis or therapy.

Farelerde Modelleme İnme: Dış Karotid Arter yoluyla Geçici Orta Serebral Arter Tıkanıklığı

Stroke is the third most common cause of mortality and the leading cause of acquired adult disability in developed countries. To date, therapeutic options ...

Nörobilim

A Model for Encephalomyosynangiosis Treatment after Middle Cerebral Artery Occlusion-Induced Stroke in Mice

There is no effective treatment available for most patients suffering with ischemic stroke, making development of novel therapeutics imperative. The brain's ability to self-heal after ischemic stroke is limited by inadequate blood supply in the impacted area. Encephalomyosynangiosis (EMS) is a neurosurgical procedure that achieves angiogenesis in patients with moyamoya disease. It involves craniotomy with placement of a vascular temporalis muscle graft on the ischemic brain surface. EMS has never been studied in the setting of acute ischemic stroke in mice. The hypothesis driving this study is that EMS enhances cerebral angiogenesis at the cortical surface surrounding the muscle graft. The protocol shown here describes the procedure and provides initial data supporting the feasibility and efficacy of the EMS approach. In this protocol, after 60 min of transient middle cerebral artery occlusion (MCAo), mice were randomized to either MCAo or MCAo + EMS treatment. The EMS was performed 3-4 h after occlusion. The mice were sacrificed 7 or 21 days after MCAo or MCAo + EMS treatment. Temporalis graft viability was measured using nicotinamide adenine dinucleotide reduced-tetrazolium reductase assay. A mouse angiogenesis array quantified angiogenic and neuromodulating protein expression. Immunohistochemistry was used to visualize graft bonding with brain cortex and change in vessel density. The preliminary data here suggest that grafted muscle remained viable 21 days after EMS. Immunostaining showed successful graft implantation and increase in vessel density near the muscle graft, indicating increased angiogenesis. Data show that EMS increases fibroblast growth factor (FGF) and decreases osteopontin levels after stroke. Additionally, EMS after stroke did not increase mortality suggesting that protocol is safe and reliable. This novel procedure is effective and well-tolerated and has the potential to provide information of novel interventions for enhanced angiogenesis after acute ischemic stroke.

The protocol aims to provide methods for encephalomyosynangiosis-grafting of a vascular temporalis muscle flap on the pial surface of ischemic brain tissue-for the treatment of non-moyamoya acute ischemic stroke. The approach's efficacy in increasing angiogenesis is evaluated using a transient middle cerebral artery occlusion model in mice.

Farelerde Orta Serebral Arter Oklüzyonu Kaynaklı İnme Sonrası Ensefalomiyosinanjiyoz Tedavisi İçin Bir Model

There is no effective treatment available for most patients suffering with ischemic stroke, making development of novel therapeutics imperative. The ...

Transkraniyal Orta Serebral Arter Oklüzyon Fareleri Modelinin Kurulması

A Modified Transcranial Middle Cerebral Artery Occlusion Model to Study Stroke Outcomes in Aged Mice

In experimental stroke research, middle cerebral artery occlusion (MCAO) with an intraluminal filament is widely used to model ischemic stroke in mice. The filament MCAO model typically exhibits a massive cerebral infarction in C57Bl/6 mice that sometimes includes brain tissue in the territory supplied by the posterior cerebral artery, which is largely due to a high incidence of posterior communicating artery atresia. This phenomenon is considered a major contributor to the high mortality rate observed in C57Bl/6 mice during long-term stroke recovery after filament MCAO. Thus, many chronic stroke studies exploit distal MCAO models. However, these models usually produce infarction only in the cortex area, and consequently, the assessment of post-stroke neurologic deficits could be a challenge. This study has established a modified transcranial MCAO model in which the MCA at the trunk is partially occluded either permanently or transiently via a small cranial window. Since the occlusion location is relatively proximal to the origin of the MCA, this model generates brain damage in both the cortex and striatum. Extensive characterization of this model has demonstrated an excellent long-term survival rate, even in aged mice, as well as readily detectable neurologic deficits. Therefore, the MCAO mouse model described here represents a valuable tool for experimental stroke research.

Yazar Spotlight: İnme Araştırmalarında Yenilikçi Teknikler ve Gelecekteki Yönelimler 

This protocol demonstrates a unique mouse stroke model with a medium-sized infarct and an excellent survival rate. This model allows preclinical stroke researchers to extend the ischemia duration, use aged mice, and assess long-term functional outcomes.

Yaşlı Farelerde İnme Sonuçlarını İncelemek için Modifiye Transkraniyal Orta Serebral Arter Oklüzyon Modeli

In experimental stroke research, middle cerebral artery occlusion (MCAO) with an intraluminal filament is widely used to model ischemic stroke in mice. ...

Farelerde Orta Serebral Arter Tıkanıklığı ve Reperfüzyon Kaynaklı İnme Cerrahi Yaklaşım

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