Name: lateral chronic cranial window preparation enables in vivo observation following distal middle cerebral artery occlusion in mice
Uploaded: 2016-12-29T13:00:00.000+00:00
Duration: 8 min 41 s
Description: Watch this Scientific Journal Video about Lateral Chronic Cranial Window Preparation Enables In Vivo Observation Following Distal Middle Cerebral Artery Occlusion in Mice at JoVE.com

VP is a participant in the Charit&#233; Clinical Scientist Program, funded by the Charit&#233; - Universit&#228;tsmedizin Berlin and the Berlin Institute of Health. TB is an SNSF PostDoc Mobility fellow. The authors receive grant support from EinsteinStiftung/A-2012-153 to PV.

ETİK TABLOSU: Hayvan denekleri deneyler geçerliyse, Landesamt für Gesundheit und Soziales, Berlin, Almanya (G0298 / 13) ve GELİYORUM kriterlerine göre belirlenen kurallara ve yönetmeliklere uygun olarak yapıldı. 12-haftalık erkek C57BL / 6J farelerine, bu çalışmada, 10 kullanılmıştır. 1. Yanal Kronik Kranial Pencere Hazırlık <ol><li> ketamin (90 mg / kg) ve ksilazin (10 mg / kg) deri altından enjeksiyonu ile anestezi gerçekleştirin. Bir ağrı uyaran yeterli sedasyon test edin. </li><li> % 70 etanol ile cerrahi aletler ve cerrahi alan sterilize edin. </li><li> Bir kemirgen tıraş makinesi kullanarak gözlere boyundan kafa derisi saç kaldırmak. </li><li> stereotaktik çerçeve içinde başını düzelt. </li><li> dehidratasyonu önlemek için her iki göze deskpantenol göz merhemi kullanın. </li><li> tüm tüyleri ve% 74.1 etanol 3 kat ve% 10 2-propanolol ile sterilize etmek için cerrahi bölgeyi temizleyin. </li><li> Bir orta hat gerçekleştirinBir neşter kullanılarak gözlere boyun kesi. </li><li> 4 çadırlaşması dikişlerle flep span. </li><li> Temporal kas başlar noktaya bir neşter ile kapalı kazıyarak sol hemisfer dikkatlice periosta çıkarın. NOT: Bu hazırlık da cam kapak giderir diş yapıştırıcı, iyi bir yapışma alanı oluşturmak için hizmet vermektedir. </li><li> Bir mikrotur kullanarak kemik kapağının ağız kafatası inceltilerek 4 mm çapında bir fronto-parietal kranyotomi gerçekleştirin. Isı yaralanmasını önlemek için delerken tuzlu su çözeltisi uygulayın. </li><li> kanüller ile kemik flep yükseltmek ve microforceps kullanarak çıkarın. </li><li> tuzlu su çözeltisi ile dikkatlice ve yoğun yıkayınız. </li><li> O kıvama sahip ve akışkan olmayana kadar diş tutkal karıştırın (yani, tutkal artık Konuları üretmek olmamalıdır). Kraniotomi etrafındaki kemik üzerine yerleştirin. </li><li> Hazırlanan tutkal 6 mm çaplı bir cam kapak yerleştirin ve inci ile sabitleyinDiş tutkal e geri kalanı. o su geçirmez olduğundan emin olun. tutkal forseps ile test ederek sert kurutulur ve kadar bekleyin. NOT: Ek sulama kür sürecini hızlandırır. </li><li> tenting sütür çıkarın ve cilt dikişlerle yarayı kapatmak. </li><li> fare kanadını deri yukarı çekin. Bir iğne subkutan yerleştirin ve yavaşça sıvı dengesi için steril tuzlu subkutan 0.5 ml yerine. </li><li> Ameliyat sonrasında, 90 dakika süre ile ısıtıldı geri kafeste hayvanları tutmak. hayvanlar gözetimsiz bırakarak önce tamamen uyanık kadar bekleyin. hayvanlar tamamen diğer hayvanlarla bir kafes onları dönmeden önce iyileşene kadar bekleyin. </li><li> Adım 1.16 tarif edildiği gibi, yaklaşık 12 saat sonra deri altından tuz hacmi ikame tekrarlayın. </li><li> Her ameliyat (örneğin, parasetamol (10 mg / ml) ya da diğer steroidal olmayan anti-enflamatuar ilaçlar) sonra ağız boşluğu içine sonda ile veya doğrudan analjezi uygulanır. </li><li> kontrol<html> Her gün ameliyat sonrası hayvanların durumu ve her zaman bir Petri-çanak, basit yemek yapmak ve ameliyat sonrası kritik kilo kaybını önlemek için katta hayvan yiyecek püre sağlarlar. NOT: intravital mikroskopi kafatası pencere hazırlandıktan sonra ilk günü yapılabilir. </li><li> izofluran anestezi uygulayın ve bir kafa tutucu hayvan düzeltin. cilt sütür açın ve pamuk tomurcukları ve steril tuzlu su ile pencereyi temizleyin. 24 saat sonra, kafa penceresi görüntüleme sağlayan bir zaman noktası ile, beyin omurilik sıvısı ile doldurulmalıdır. Kurulan mikroskopi protokollerini 18 ile görüntüleme gerçekleştirin. </li></ol> 2. Distal MCAO NOT: MCAO prosedürü CW hazırlandıktan sonra yaklaşık 5 d yapılmalıdır. Bu vuruş kaynaklı bağışıklık reaksiyonu ile CW hazırlanması nedeniyle bağışıklık reaksiyonu girişimi en aza indirir. &quot;Src =&quot; / files / ftp_upload / 54701 / 54701fig1.jpg &quot;/&gt; Distal MCAO 1. Genel Bakış Şekil. A. Bu op önce gemiler hakkında güzel bir bakış niteliğindedir. İlk iki kutuplu temas sonrasında B. gemiler. İkinci bipolar temas sonrasında C. gemiler. D. tamamen şimdi kapalı gemiler, ilgili genel bakış. Alt büyütme ile E. Final bakış. <a href="http://ecsource.jove.com/files/ftp_upload/54701/54701fig1large.jpg" target="_blank">Bu rakamın büyük halini görmek için lütfen buraya tıklayınız.</a> <ol><li> Veteriner kadrosu (danışarak bir anestezi maskesi ve uygun bir anestezik rejimi kullanılarak fareler, uyuşturan, örneğin 1.5 ile indüksiyon - 2/3 N 2 O ve 1/3 O 2&#39;de% 1.5 izofluran üzerinden -% 2 izofluran ve 1.0 ile bakım bir buharlaştırıcı). </li><li> (Tıraş ve saç kaldırmak ve uygun bir madde ile deri ve çevredeki kürk dezenfekteörneğin,% 70 etil alkol), ve daha sonra da kurutulur. </li><li> 36.5 ° C &#39;de farelerin vücut ısısını korumak ameliyat sırasında ± 0.5 ° C. fare rektal sıcaklığına göre ısıtılmış bir geribildirim kontrollü ısıtma yastığı, şiddetle tavsiye edilir. </li><li> lateral pozisyonda hayvan yerleştirin. Anestezi maskesi burun düzeltmek ve 1.0 izofluran konsantrasyonu ayarlamak -% 1.5. </li><li> Her iki Gözleri ıslak merhem (deskpantenol) uygulayın. </li><li> CW hazırlanması için yapılan cilt kesisi kullanın. </li><li> Cildi nazikçe ayırmak ve altında zamansal kas tespit. </li><li> yüksek frekanslı jeneratör enerjisini ayarlayın (5-7 W) ve bipolar modunu kullanın. </li><li> elektrokoagülasyon forseps kullanın ve dikkatli tamamen kas çıkarmadan, bir kas flep oluşturma, kafatası zamansal kas çıkarın. </li><li> Temporal bölgenin rostral kısmında şeffaf kafatası altında MCA tespit retro orbita dorsall sinüs. MCA çatallanma tespit edilemezse, gemi en rostral belirlemeye çalışın. </li><li> sürekli ısı hasarı önlemek için sulama yaparken bir mikrotur ile MCA şube üzerinde ince kafatası. </li><li> kanüller ile kemik kaldırın ve mikroforcepslerle çıkarın. </li><li> 5 W. - 3 yüksek frekanslı jeneratör enerjisini azaltmak </li><li> yukarıdan arter yaklaşın ve yavaşça gemi kaldırmadan her iki tarafta çift kutuplu forseps ile dokunun. </li><li> damar çatallanma proksimal ve distal arter koagüle. </li><li> 30 saniye bekleyin ve sonra kan akımı kalıcı kesintiye emin olmak için hafifçe arter dokunun. Bir rekanalizasyon görülmektedir eğer elektro tekrarlayın. </li><li> Mümkünse, kemik defekti kapamak için kas ekleme 1 ya da 2 dikiş ile zamansal kas sabitleyin. </li><li> Yarayı dikin ve ısıtmalı kurtarma kutunun içine hayvan yerleştirin. hızlı uçucu ANEST sonra Genel olarak, hayvanlar kurtarmalerine. </li><li> Adım 1.16 tarif edildiği gibi hacimli ikamesi için, steril tuzlu su deri altından 0.5 mL uygulanır. </li><li> Ameliyattan sonra hayvanlar 90 dakika süre ile ısıtıldı geri kafes içinde kalmak için izin verir. hayvanlar gözetimsiz bırakarak önce bilinçli kadar bekleyin. Onlar tamamen iyileşti zaman sadece diğer hayvanlarla bir kafese onları geri. </li><li> 12 saat sonra, adım 1.16 açıklandığı gibi, ses ikame tekrarlayın. </li><li> Içme suyu (örneğin, parasetamol (10 mg / ml) ya da diğer steroidal olmayan anti-enflamatuar ilaçlar) ile ve ameliyat sonrası analjezi uygulanır. </li><li> Ameliyat sonrası her gün hayvanların sağlık durumunu kontrol edin. yeme kolaylaştırmak ve ameliyat sonrası kilo kaybı en aza indirmek için katta bir petri kabındaki püresi hayvan yiyecek sağlamak. </li></ol> 3. Sham Tedavisi <ol><li> Yukarıda da dahil olmak üzere CW maruz orta serebral ar koagüle yok hazırlık-hariç açıklanan adım 1 ve 2&#39;ye aynı bütün prosedürleri uygulayınakü. </li></ol>

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The authors have nothing to disclose.

İnme 1 dünya çapında uzun süreli sakatlık ve ölüm başlıca nedenleri arasındadır. Akut tedavi, hızlandırmak ve inme sonrası iyileşmeyi artırmak için yeni yaklaşımlar ve mekanizmaları araştıran ötesinde bir asal tıbbi gol 7 kalır. Deneysel inme araştırmacılar sıklıkla fokal serebral iskemi kemirgen modelleri kullanır. Aslında, geçici veya kalıcı MCAO uyaran modelleri hastalarda 4 fokal serebral iskemi en yaygın türlerinden biri taklit. MCA proksimal ok...

Stroke is among the principal causes of long-term disability and death worldwide1, coming second after coronary heart disease. In addition, stroke is the primary cause of long-term disability, underscoring its tremendous socioeconomic impact6-8. Beyond acute treatment, investigating new approaches and mechanisms to accelerate and enhance recovery after stroke remains a prime medical goal7. 
In the last few decades, data from experimental stroke research has contributed substantially to understanding the complex pathophysiological cascades triggered by ischemia9,10. Excitotoxicity, apoptosis, peri-infarct depolarization, and inflammation have been identified as the most relevant mediators of cell death following focal cerebral ischemia. Moreover, using animal models of cerebral ischemia, important concepts, diagnostic modalities, and therapeutic approaches have been developed and validated (e.g., "penumbra" and thrombolysis)11.
The availability of experimental stroke models, combined with non-invasive imaging modalities (e.g., magnetic resonance imaging (MRI), computed tomography, or laser speckle contrast analysis), enables the researcher to investigate hyperacute and chronic pathophysiological changes induced by the ischemic insult in a longitudinal manner12. Along with studying the spatiotemporal profile of the evolving lesion, changes resembling neuronal plasticity can be investigated and correlated to functional outcomes and histological findings. Within the last few years, further methodological advances have been made using the combination of cerebral ischemia models and in vivo microscopy via cranial windows13. These new techniques allow investigators to analyze the neurovascular unit at the cellular and molecular level, with great analytic power in the acute, subacute, and chronic phases following focal cerebral ischemia14. Moreover, in vivo microscopy imaging of microcirculatory dynamics has revealed novel aspects of cerebral microvasculature function and angioarchitecture, with significant pathophysiological relevance15-17. 
In this protocol, we present how to perform a chronic CW preparation lateral to the sagittal sinus and how to surgically induce a distal stroke underneath the window. This mouse model can be applied to sequential imaging of acute, subacute, and chronic changes following focal cerebral ischemia via epi-illuminating, confocal laser scanning, and two-photon intravital microscopy.

<table><tbody><tr><td>Binocular surgical microscope</td><td>Zeiss</td><td>Stemi 2000 C</td><td></td></tr><tr><td>Light source for microscope</td><td>Zeiss</td><td>CL 6000 LED</td><td></td></tr><tr><td>Heating pad with rectal probe</td><td>FST</td><td>21061-10</td><td></td></tr><tr><td>Stereotactic frame</td><td>Kopf</td><td>Model 930</td><td></td></tr><tr><td>Anaethesia system for isoflurane</td><td>Draeger</td><td></td><td></td></tr><tr><td>Isoflurane</td><td>Abott</td><td></td><td></td></tr><tr><td>Dumont forceps #5</td><td>FST</td><td>11251-10</td><td></td></tr><tr><td>Dumont forceps #7</td><td>FST</td><td>11271-30</td><td></td></tr><tr><td>Bipolar Forceps</td><td>Erbe</td><td>20195-501</td><td></td></tr><tr><td>Bipolar Forceps&amp;nbsp;</td><td>Erbe&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp; 20195-022</td><td></td><td></td></tr><tr><td>Microdrill</td><td>FST&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp; 18000-17&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;</td><td></td><td></td></tr><tr><td>Needle holder</td><td>FST</td><td>12010-14</td><td></td></tr><tr><td>5-0 silk suture</td><td>Feuerstein, Suprama</td><td></td><td></td></tr><tr><td>7-0 silk suture</td><td>Feuerstein,Suprama</td><td></td><td></td></tr><tr><td>8-0 silk suture</td><td>Feuerstein, Suprama</td><td></td><td></td></tr><tr><td>Veterinary Recovery Chamber</td><td>Peco Services</td><td>V1200</td><td></td></tr></tbody></table>

lateral chronic cranial window preparation enables in vivo observation following distal middle cerebral artery occlusion in mice

Fokal serebral iskemi (yani, iskemik inme) nöronal fonksiyon ciddi bir kaybı ve dolayısıyla motor ve zihinsel engelli bir dizi yol açan başlıca beyin yaralanmaya neden olabilir. Inme 1 dünya çapında uzun süreli sakatlık ve ölüm başlıca nedenleri arasında olduğu gibi yüksek prevalans, ciddi bir sağlık yükünü oluşturmaktadır. nöronal fonksiyonu geri kazanımı, çoğu durumda, sadece kısmi vardır. Şimdiye kadar, tedavi seçenekleri oldukça nedeniyle trombolitik 2,3 dar zaman penceresi, özellikle de sınırlıdır. inme kurtarma hızlandırmak için yöntemlerin belirlenmesi birinci sınıf bir tıbbi hedef olmaya devam; Ancak, bu toparlanma sürecine yetersiz mekanistik anlayışlar tarafından engellenmiştir. Deneysel inme araştırmacılar sıklıkla fokal serebral iskemi kemirgen modelleri kullanır. Akut fazda ötesinde, inme araştırma giderek serebral iskemi aşağıdaki-sub akut ve kronik fazda odaklanmıştır. Çoğu inme araştırmacılar uygulamak kalıcı veya tranfarelerde ya da sıçanlarda MCA&#39;nın SIENT oklüzyonu. Hastalarda, MCA tıkanıklıklar iskemik inme 4 en sık nedenleri arasındadır. Filament modelini kullanarak MCA proksimal oklüzyon yanı sıra, uzak MCA cerrahi tıkanması, deneysel inme araştırma 5. en sık kullanılan model muhtemelen. MCA dalı (lenticulo-striat arterlerin dallanma kadar) distalinde Tıkanıklığı tipik striatum yedek ve öncelikle neokorteks etkiler. Gemi tıkanıklığı kalıcı veya geçici olabilir. uzun dönem sonuçları açısından lezyon hacmi ve çok düşük mortalite oranları yüksek tekrarlanabilirlik bu modelin başlıca avantajları vardır. Burada, sagital sinüs kronik kafa penceresi (CW) hazırlık yanal gerçekleştirmek için, ve daha sonra nasıl bir kraniotomi yaklaşımı kullanarak pencerenin altında bir uzak inme neden cerrahi olarak nasıl gösterilmektedir. Bu yaklaşım ile iskemi sonrası akut ve kronik değişikliklerin sıralı görüntüleme için uygulanabilirKonfokal lazer tarama ve iki foton intravital mikroskopi epi-aydınlatıcı.

Zaman çizelgesi ve temsili sonuçları Şekil 2 ve 3&#39;te gösterilmiştir. Superior sagittal sinüs (Şekil 2 B, C, D), deneyimli bir cerrah tarafından gerçekleştirilen çok düşük mortalite ve morbidite oranı sonuçları için küçük bir kafatası pencere yanal ile kafa pencere hazırlama. 10 hayvanların hepsi hayatta ve tüm kronik CW bile 28 gün ameliyattan sonra, yüksek kaliteli görüntüleme için kullanılabilir. ya...

Watch this Scientific Journal Video about Lateral Chronic Cranial Window Preparation Enables In Vivo Observation Following Distal Middle Cerebral Artery Occlusion in Mice at JoVE.com

Lateral Chronic Cranial Window Preparation Enables In Vivo Observation Following Distal Middle Cerebral Artery Occlusion in Mice

distal orta serebral arter dalı (MCAO) Cerrahi tıkanması, deneysel inme araştırmalarında sıkça kullanılan bir modeldir. Bu el yazması farelerde boyuna intravital mikroskopi için bir fırsat sunuyor yanal kafatası pencere takılması, birlikte kalıcı MCAO temel tekniğini açıklar.

Yanal Kronik Kranial Pencere Hazırlık sağlar<em&gt; İn Vivo</em&gt; Farelerde Distal Orta Serebral Arter Tıkanıklığı takiben Gözlem

Focal cerebral ischemia (i.e., ischemic stroke) may cause major brain injury, leading to a severe loss of neuronal function and consequently to a host of ...

lateral-chronic-cranial-window-preparation-enables-vivo-observation

Universidad de Cartagena UDEC

Research

JoVE Journal

Neuroscience

Lateral Chronic Cranial Window Preparation Enables In Vivo Observation Following Distal Middle Cerebral Artery Occlusion in Mice

11.6K Görüntüleme.  Charité-Universitätsmedizin.  distal orta serebral arter dalı (MCAO) Cerrahi tıkanması, deneysel inme araştırmalarında sıkça kullanılan bir modeldir. Bu el yazması farelerde boyuna intravital mikroskopi için bir fırsat sunuyor yanal kafatası pencere takılması, birlikte kalıcı MCAO temel tekniğini açıklar. 

Video: Lateral Chronic Cranial Window Preparation Enables In Vivo Observation Following Distal Middle Cerebral Artery Occlusion in Mice

Intravital Microscopy of the Mouse Brain Microcirculation using a Closed Cranial Window

This experimental model was designed to assess the mouse pial microcirculation during acute and chronic, physiological and pathophysiological hemodynamic, inflammatory and metabolic conditions, using in vivo fluorescence microscopy. A closed cranial window is placed over the left parieto-occipital cortex of the mice. Local microcirculation is recorded in real time through the window using epi and fluorescence illumination, and measurements of vessels diameters and red blood cell (RBC) velocities are performed. RBC velocity is measured using real-time cross-correlation and/or fluorescent-labeled erythrocytes. Leukocyte and platelet adherence to pial vessels and assessment of perfusion and vascular leakage are made with the help of fluorescence-labeled markers such as Albumin-FITC and anti-CD45-TxR antibodies. Microcirculation can be repeatedly video-recorded over several days. We used for the first time the close window brain intravital microscopy to study the pial microcirculation to follow dynamic changes during the course of Plasmodium berghei ANKA infection in mice and show that expression of CM is associated with microcirculatory dysfunctions characterized by vasoconstriction, profound decrease in blood flow and eventually vascular collapse.

Intravital microscopy to follow temporal and spatial hemodynamic and inflammatory events in the pial microcirculation.

Kapalı Kranial Pencere kullanarak Fare Beyin Mikrosirkülasyon intravital Mikroskopi

This experimental model was designed to assess the mouse pial microcirculation during acute and chronic, physiological and pathophysiological hemodynamic, ...

Nörobilim

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 ...

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 ...

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. ...

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

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

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

Endotelin-1 Bağlı Orta Serebral Arter Tıkanıklığı sırasında Orta Serebral Arter görselleştirin bir Kranial Pencere kullanmak

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

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

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 ischemic in origin and occur in the territory of the middle cerebral artery (MCA). Currently the only Food and Drug Administration (FDA) approved drug for the treatment of this devastating disease is tissue plasminogen activator (tPA). However, tPA has a small therapeutic window for administration (3 - 6 hr), and is only effective in 4% of the patients who actually receive it. Current research focuses on understanding the pathophysiology of stroke in order to find potential therapeutic targets. Thus, reliable models are crucial, and the MCA occlusion (MCAo) model (also termed the intraluminal filament or suture model) is deemed to be the most clinically relevant surgical model of ischemic stroke, and is fairly non-invasive and easily reproducible. Typically the MCAo model is used with rodents, especially with mice due to all the genetic variations available for this species. Here we describe (and present in the video) how to successfully perform the MCAo model (with reperfusion) in mice to generate reliable and reproducible data.

In order to understand the pathophysiology of stroke, it is important to use reliable models. This paper will describe one of the most frequently used stroke models in mice, termed the middle cerebral artery occlusion (MCAo) model (also termed the intraluminal filament or suture model) with reperfusion.

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 ...

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 ...

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. ...

Yanal Kronik Kranial Pencere Hazırlık sağlar

Özet

Daha Fazla Video Keşfet

Bu videodaki bölümler

Kapalı Kranial Pencere kullanarak Fare Beyin Mikrosirkülasyon intravital Mikroskopi

Orta Serebral Arter Tıkanıklığı Fare Modeli

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

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

Endotelin-1 Bağlı Orta Serebral Arter Tıkanıklığı sırasında Orta Serebral Arter görselleştirin bir Kranial Pencere kullanmak

Farelerde Orta Serebral Arter Tıkanıklığı ve Reperfüzyon Kaynaklı İnme Cerrahi Yaklaşım

Serebral iskemi-reperfüzyon iki damar tıkanıklığı fare modeli

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

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

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