La tecnica chirurgica è stata originariamente acquisite nel laboratorio del Dr. William D. Hill presso il Medical College of Georgia. Gli autori desiderano inoltre ringraziare il dottor David A. Rempe e Landa Prifti per l&#39;utilizzo della fotocamera dissezione. Questa ricerca è stata sostenuta da NIH NS041744, NS051279, F31 NS064700 e AHA 30815697D.

Questo protocollo è stato approvato dalla Università di Rochester comitato dedicato l&#39;uso etico degli animali nella ricerca (UCAR). Tecniche asettiche da seguire durante il protocollo. L&#39;utilizzo di guanti sterili e una maschera è necessario. Tutte le attrezzature, materiali, prodotti chimici, e gli strumenti che vengono utilizzati durante il protocollo sono descritte nella tabella 1. 1. La preparazione pre-chirurgica <ol><li> Iniettare topi per via sottocutanea con buprenorfina (0.05mg/kg) 2 h prima dell&#39;intervento, subito dopo l&#39;intervento chirurgico e poi ogni 3-5 ore durante le prime 24 ore post-operatorio. </li><li> Sterilizzare tutti gli strumenti chirurgici, spugne garza, e applicatori punta del cotone in autoclave. Mantenere gli strumenti chirurgici in etanolo al 70% durante l&#39;intervento chirurgico e aria secca a destra garza sterile prima dell&#39;uso. Spruzzare l&#39;area di lavoro con il 70% di etanolo. </li><li> Anestetizzare il mouse con un 3% isofluorane-20% miscela di ossigeno del gas utilizzando un vaporizzatore anestetico. Regolare la quantità di ossigeno con il misuratore di portata. Testare il livello di anestesia da dito o pizzicare la coda (che dovrebbero essere non risponde). Mantenere il livello di anestesia con il 2% (v / v) isofluorane. </li><li> Applicare lacrime artificiali per gli occhi del mouse e usare cautela per evitare di danneggiare l&#39;occhio durante la procedura chirurgica. Posizionare il mouse sul lato destro in posizione laterale. </li><li> Radere l&#39;area sul lato sinistro tra l&#39;occhio sinistro e la base dell&#39;orecchio sinistro con il tagliaunghie animale. Pulire l&#39;area alternando tra una soluzione betadine e il 70% di etanolo con applicatori punta di cotone e strofinare leggermente la zona. Ripetere se necessario. </li><li> Posizionare il mouse su di una piastra elettrica collegato ad una sonda rettale per mantenere la temperatura corporea a 37 ° C. Inserire la sonda rettale utilizzando olio minerale. </li><li> Posizionare il mouse sul lato destro sotto il microscopio e fissare con nastro adesivo. Tagliare una finestra in una garza e coprire un&#39;area chirurgica. </li></ol> 2. Procedura chirurgica e MCA Ligation <ol><li> Fai una incisione verticale tra l&#39;occhio sinistro e la base dell&#39;orecchio sinistro utilizzando bene le forbici diritte. Usa hemostats curva per mantenere l&#39;area chirurgica aperta. </li><li> Effettuare una incisione orizzontale sul muscolo temporale utilizzando le forbici a molla e un po &#39;separare il muscolo temporale dal cranio lentamente tirando con una pinza. </li><li> Fai una piccola craniotomia subtemporal all&#39;incrocio tra zigomatica e ossa squammoso utilizzando un ago da 18G mentre si tiene la mandibola con una pinza curva. </li><li> Esporre il MCA rimuovendo il piccoli pezzi di cranio con Rongeurs osso. Dell&#39;arcata zigomatica e contenuti orbitale non deve essere danneggiato durante questo processo. Se l&#39;essiccamento eccessivo di tessuto si verifica durante questo processo, si applicano PBS sterile con applicatori punta di cotone. </li><li> Legare la porzione distale del MCA cauterizer utilizzando un piccolo vaso. </li><li> Posizionare il muscolo temporale nella sua posizione originale e chiudere il sito di incisione chirurgica con 5-0 punti di sutura in nylon. </li><li> Interrompere l&#39;anestesia e rimuovere la sonda rettale. Iniettare il mouse con una dose 2 di buprenorfina (0.05mg/kg) per via sottocutanea. Ritorna il mouse per la gabbia che è stata mantenuta a 37 ° C con un pannello di riscaldamento. </li><li> Seguire da vicino il mouse per le prossime 24 ore per ogni disagio (riduzione dell&#39;appetito / consumo di acqua, la postura ingobbita, aumento della respirazione, pilo-eretto capelli). Iniettare il mouse con buprenorfina per via sottocutanea ogni 3-5 ore dopo l&#39;intervento fino a 24 ore. Fornire il mouse con gel di recupero durante questo periodo. </li></ol> 3. TTC colorazione e Determinazione della gittata sistolica <ol><li> Ventiquattro ore dopo l&#39;intervento profondamente anestetizzare il mouse, transcardially profumato il mouse con un 4% TTC (w / v) in tampone fosfato (PBS) per 15 min e poi con una soluzione di paraformaldeide al 4% per 10 minuti utilizzando un mini pompa peristaltica a portata media. Togliere il cervello e posizionarlo nella soluzione di paraformaldeide al 4% durante la notte. </li><li> Posizionare il cervello nel blocco di sezionamento. Tagliate a fette il cervello 1 mm di spessore utilizzando lame di rasoio. </li><li> Mettere le fette accanto ad un righello scala millimetrata. Fotografare le fette utilizzando una fotocamera digitale collegata al microscopio da dissezione. </li><li> Calcolare l&#39;area colpo sottraendo il non-infartuato area del sito ipsilaterale dalla superficie totale del sito controlaterale utilizzando immagini J Software (http://rsbweb.nih.gov/ij/). Calcolare la gittata sistolica sovrapponendo l&#39;area colpo di fette di 22. </li></ol> 4. Indicazioni per l&#39;uso J software Image <ol><li> Aprire il file immagine da analizzare in software Image J cliccando sul menu File. </li><li> Fare clic sulla scheda &#39;retta&#39; nel programma. Tracciare una linea retta tra i due margini sul righello. Fare clic su &#39;analizzare&#39; menu e seleziona &#39;scala set&#39;. Impostare la distanza nota come 1, unità di lunghezza mm. </li><li> Fare clic sulla scheda &#39;cerchio a mano libera&#39;. Disegnare un cerchio a delineare l&#39;emisfero controlaterale. Fare clic sul menu &#39;analizzare&#39; e selezionare &#39;misura&#39;. La finestra di calcolo si aprirà mostra l&#39;area del cerchio disegnato. </li><li> Disegnare un altro cerchio intorno alla emisfero esclusi ictus ipsilaterale (bianco) zona. Misurare l&#39;area, come indicato al punto 4.3. Il nuovo valore verrà aggiunto alla finestra di calcolo. La differenza tra il 1 ° e 2 ° valori rappresenta l&#39;area dell&#39;infarto, che è indicato come nella formula sottostante. </li><li> Calcolare l&#39;area corsa in ogni fetta ripetendo i passaggi 4,2-4,4. Prendere sommatoria dell&#39;area corsa calcolato in ogni sezione (ΣA n). Questo rappresenta gittata sistolica conoscere il fatto che lo spessore di ogni fetta è 1 mm. ΣA n x 1 mm (spessore di ogni fetta) volume = Corsa </li></ol> 5. Rappresentante dei risultati: Il infarti ottenuto permanente legatura MCA nel topo sono in gran parte corticale. Tuttavia, è possibile ottenere le lesioni subcorticali se l&#39;MCA è legatura prossimale al ramo lenticulostriate. I volumi di ictus dopo MCA legatura può variare da 10 millimetri a 35 mm ³ ³ 19, 23. I volumi corsa calcolata con colorazione TTC in Moyanova et al. 19 sono tra ³ ³ 10mm a 22 millimetri mentre i volumi di corsa determinata da immagini MRI a Filiano e altri 23 sono tra ³ ³ 20mm a 35mm. Il possibile motivo di queste differenze forse la posizione esatta della legatura ed i diversi metodi utilizzati per misurare i volumi di corsa. Nella Figura 1, un cervello colorato TTC 24 ore posto permanente legatura MCA in un tipo selvaggio C57/BL6 del mouse viene mostrato. Il sito di corsa è di colore bianco in apparenza (Fig. 1A, B). Figure 1A e B illustrano il sito dell&#39;infarto da diverse angolazioni. La Figura 2 mostra fette 1 mm di spessore del cervello macchiato TTC corsa da anteriore a posteriore (Fig. 2A-G). Volume dell&#39;infarto è stata calcolata 24 ore post-operatorie. Il volume dell&#39;infarto è ³ ~ 23mm. <img src="/files/ftp_upload/3039/3039fig1.jpg" alt="Figura 1"/> Figura 1. Rappresentazione del sito ictus. AB, TTC macchiato immagini del cervello intero, 24 ore dopo la legatura MCA. Area bianca rappresenta l&#39;infarto. <img src="/files/ftp_upload/3039/3039fig2.jpg" alt="Figura 2"/> Figura 2. Risultati rappresentativi. (A&gt; G), TTC macchiato 1 millimetro cervello sezioni, 24 ore dopo Mcal. Fette sono allineate da anteriore a posteriore. Area bianca rappresenta l&#39;infarto.

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Nessun conflitto di interessi dichiarati.

Il metodo di legatura permanente MCA dà volumi infarto altamente riproducibili e un aumento post-operatorio tassi di sopravvivenza rispetto ad altri metodi disponibili. La facilità e la breve durata (~ 30 min) della procedura di renderlo ancora più pratico. Il metodo è ampiamente utilizzato sia in topi e ratti. Questa tecnica richiede un intervento chirurgico invasivo sotto uno stereomicroscopio. Pertanto, l&#39;esperienza nella gestione sotto un microscopio e perfezionare una craniotomi...

<table border="1"><tbody><tr><th> NOME </th><th> SOCIETA &#39;E NUMERO CATALOGO </th></tr><tr><td> Soluzioni e prodotti chimici A. </td><td></td></tr><tr><td> Povidone-iodio soluzione, lavaggio chirurgico </td><td> Aplicare, 82-209 </td></tr><tr><td> 70% di etanolo </td><td> Koptec, V1001 </td></tr><tr><td> 2,3,5 trifeniltetrazolio cloruro (TTC) </td><td> Sigma, T8877 </td></tr><tr><td> 4% paraformaldeide </td><td> Scienze della microscopia elettronica, 159SP </td></tr><tr><td> Tampone fosfato isotonico, pH: 7,4 </td><td> Realizzato e sterilizzati in laboratorio. </td></tr><tr><td> Olio minerale </td><td> PML Microbiologicals, R6570 </td></tr><tr><td> Isofluorane </td><td> La Società Butler, 029405 </td></tr><tr><td> Buprenorfina (0.3mg/ml) </td><td> Hospira, NDC 0409-2012-32 </td></tr><tr><td> Lacrime artificiali </td><td> La Società Butler, 007312 </td></tr><tr><td> Recupero Gel </td><td> Chiaro H 2 O, 72-01-5022 </td></tr><tr><td> B. Materiali e attrezzature chirurgiche </td><td></td></tr><tr><td> C57/BL6 del mouse, 10-12 settimane di età </td><td> La colonia è stato allevato nel vivaio universitario </td></tr><tr><td> 100% bombola di ossigeno </td><td> Airgas </td></tr><tr><td> Anestetico vaporizzatore e misuratore di portata </td><td> Surgivet, Modello 100 </td></tr><tr><td> Pad di riscaldamento con sonda rettale </td><td> Strumenti Scienza bene, TR200 </td></tr><tr><td> Microscopio da dissezione </td><td> Zeiss, Stemi 2000 </td></tr><tr><td> Riscaldamento a pannelli </td><td> Petco Services Ltd. 0307-013 </td></tr><tr><td> Piccola nave Set Cauterizer </td><td> Strumenti Scienza bene, 18000-00 </td></tr><tr><td> Mini pompa peristaltica </td><td> Harvard Apparatus, MPII </td></tr><tr><td> Siringa da 1 ml </td><td> BD, 329650 </td></tr><tr><td> 18G ago </td><td> Becton Dickinson, 305196 </td></tr><tr><td> Chirurgico di sutura in nylon 5,0 </td><td> Ethilon, 698H </td></tr><tr><td> Blocco di sezionamento </td><td> Kent scientifico, 1mm </td></tr><tr><td> Singolo bordo Razor Blades </td><td> Scienze della microscopia elettronica, 71962 </td></tr><tr><td> Etichetta del nastro </td><td> Scienceware, Bel-arte Prodotti F13463-2075 </td></tr><tr><td> Garza Spugne </td><td> Kendall rezza 2252, Johnson e Johnson 6415 </td></tr><tr><td> Suggerimento cotone Applicatori </td><td> Puritana 806-WCL </td></tr><tr><td> C. Strumenti di Chirurgia </td><td></td></tr><tr><td> Forbici chirurgiche Primavera </td><td> Strumenti Scienza multa 15003-08 </td></tr><tr><td> Forcipe Microdissecting angolato </td><td> Strumenti Scienza Belle 5 / 45 </td></tr><tr><td> Pinze Microdissecting curve </td><td> Belle Scienza Strumenti 5 Ti </td></tr><tr><td> Belle forbici dritto </td><td> Strumenti Scienza multa 14568-12 </td></tr><tr><td> Osso curvo pinza ossivora </td><td> Strumenti Scienza multa 16021-14 </td></tr><tr><td> Emostatica Hartman curvo </td><td> Strumenti Scienza multa 13003-10 </td></tr></tbody></table> Tabella 1. L&#39;elenco dei materiali, strumenti e sostanze chimiche usate per la procedura di legatura MCA. Soluzioni e prodotti chimici A., B. Materiali e attrezzature chirurgiche, C. Chirurgia Strumenti.

the application of permanent middle cerebral artery ligation in the mouse

Ischemia cerebrale focale è tra il tipo più comune di ictus osservato nei pazienti. A causa della rilevanza clinica c&#39;è stato un lungo lavoro per sviluppare modelli animali adatti a studiare gli eventi che si svolgono durante l&#39;insulto ischemico. Queste tecniche sono transitori o permanenti, i modelli di ischemia focale o globale usando molti diversi modelli animali, con i roditori più comuni sono. Il metodo di legatura permanente MCA, che viene anche definito come pMCAo in letteratura è ampiamente utilizzato come modello di ischemia focale nei roditori 1-6. Questo metodo è stato originariamente descritto per i ratti da Tamura et al. nel 1981 7. In questo protocollo una craniotomia è stato utilizzato per accedere alla MCA e le regioni prossimali sono state occluse da elettrocoagulazione. Il infarti coinvolgere soprattutto le regioni corticali e striatali volte a seconda della posizione della occlusione. Questa tecnica è ormai ben consolidata e utilizzata in molti laboratori 8-13. L&#39;uso precoce di questa tecnica ha portato alla definizione e descrizione del &quot;core infarto&quot; e &quot;penombra&quot; 14-16, ed è spesso utilizzata per valutare il potenziale neuroprotettivo composti 10, 12, 13, 17. Anche se gli studi iniziali sono stati condotti nel ratto, legatura permanente MCA è stato utilizzato con successo nei topi con lievi modifiche 18-20. Questo modello produce infarti riproducibile e un aumento post-tassi di sopravvivenza. Circa l&#39;80% degli ictus ischemico negli esseri umani avvengono in zona MCA 21 e quindi questo è un modello molto importante per gli studi di ictus. Attualmente, c&#39;è una scarsità di trattamenti efficaci a disposizione dei pazienti colpiti da ictus, e quindi c&#39;è bisogno di buoni modelli per testare potenziali composti farmacologici e valutare i risultati fisiologici. Questo metodo può essere utilizzato anche per studiare i meccanismi intracellulari di risposta all&#39;ipossia in vivo. Qui, vi presentiamo l&#39;intervento legatura MCA in un mouse C57/BL6. Descriviamo la preparazione pre-chirurgica, la chirurgia legatura MCA e 2,3,5 trifeniltetrazolio cloruro (TTC) colorazione per la quantificazione dei volumi di infarto.

Watch this Scientific Journal Video about The Application Of Permanent Middle Cerebral Artery Ligation in the Mouse at JoVE.com

The Application Of Permanent Middle Cerebral Artery Ligation in the Mouse

Dell&#39;arteria cerebrale media (MCA) legatura è una tecnica per lo studio ischemia cerebrale focale in modelli animali. In questo metodo, l&#39;arteria cerebrale media è esposta da craniotomia e legato da cauterizzazione. Questo metodo dà volumi infarto altamente riproducibili e un aumento post-operatorio tassi di sopravvivenza rispetto ad altri metodi disponibili.

L&#39;applicazione di permanente legatura dell&#39;arteria cerebrale media nel mouse

Focal cerebral ischemia is among the most common type of stroke seen in patients. Due to the clinical significance there has been a prolonged effort to ...

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19.5K Views.  University of Rochester. Dell'arteria cerebrale media (MCA) legatura è una tecnica per lo studio ischemia cerebrale focale in modelli animali. In questo metodo, l'arteria cerebrale media è esposta da craniotomia e legato da cauterizzazione. Questo metodo dà volumi infarto altamente riproducibili e un aumento post-operatorio tassi di sopravvivenza rispetto ad altri metodi disponibili.

Video: The Application Of Permanent Middle Cerebral Artery Ligation in the Mouse

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.

Corsa di modellazione in Mouse - Medio occlusione dell&#39;arteria cerebrale con il Modello Filament

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

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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.

Modello murino di Mezzo occlusione dell&#39;arteria cerebrale

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 ...

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.

Utilizzando una finestra craniale per visualizzare l&#39;arteria cerebrale media Durante endotelina-1 indotta da occlusione dell&#39;arteria cerebrale media

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 ...

Optimized System for Cerebral Perfusion Monitoring in the Rat Stroke Model of Intraluminal Middle Cerebral Artery Occlusion

The translational potential of pre-clinical stroke research depends on the accuracy of experimental modeling. Cerebral perfusion monitoring in animal models of acute ischemic stroke allows to confirm successful arterial occlusion and exclude subarachnoid hemorrhage. Cerebral perfusion monitoring can also be used to study intracranial collateral circulation, which is emerging as a powerful determinant of stroke outcome and a possible therapeutic target. Despite a recognized role of Laser Doppler perfusion monitoring as part of the current guidelines for experimental cerebral ischemia, a number of technical difficulties exist that limit its widespread use. One of the major issues is obtaining a secure and prolonged attachment of a deep-penetration Laser Doppler probe to the animal skull. In this video, we show our optimized system for cerebral perfusion monitoring during transient middle cerebral artery occlusion by intraluminal filament in the rat. We developed in-house a simple method to obtain a custom made holder for twin-fibre (deep-penetration) Laser Doppler probes, which allow multi-site monitoring if needed. A continuous and prolonged monitoring of cerebral perfusion could easily be obtained over the intact skull.

Cerebral perfusion monitoring has been demonstrated to improve accuracy in ischemic stroke models. Technical difficulties often limit the use of this essential tool for cerebrovascular research. In this video, an optimized system is shown to obtain a single or multi-site hemodynamic monitoring during intraluminal middle cerebral artery occlusion in rats.

Sistema ottimizzato per il monitoraggio perfusione cerebrale nel modello Rat Stroke of intraluminale occlusione dell&#39;arteria cerebrale media

The translational potential of pre-clinical stroke research depends on the accuracy of experimental modeling. Cerebral perfusion monitoring in animal ...

Permanent Cerebral Vessel Occlusion via Double Ligature and Transection

Stroke is a leading cause of death, disability, and socioeconomic loss worldwide. The majority of all strokes result from an interruption in blood flow (ischemia) 1. Middle cerebral artery (MCA) delivers a great majority of blood to the lateral surface of the cortex 2, is the most common site of human stroke 3, and ischemia within its territory can result in extensive dysfunction or death 1,4,5. Survivors of ischemic stroke often suffer loss or disruption of motor capabilities, sensory deficits, and infarct. In an effort to capture these key characteristics of stroke, and thereby develop effective treatment, a great deal of emphasis is placed upon animal models of ischemia in MCA.
Here we present a method of permanently occluding a cortical surface blood vessel. We will present this method using an example of a relevant vessel occlusion that models the most common type, location, and outcome of human stroke, permanent middle cerebral artery occlusion (pMCAO). In this model, we surgically expose MCA in the adult rat and subsequently occlude via double ligature and transection of the vessel. This pMCAO blocks the proximal cortical branch of MCA, causing ischemia in all of MCA cortical territory, a large portion of the cortex. This method of occlusion can also be used to occlude more distal portions of cortical vessels in order to achieve more focal ischemia targeting a smaller region of cortex. The primary disadvantages of pMCAO are that the surgical procedure is somewhat invasive as a small craniotomy is required to access MCA, though this results in minimal tissue damage. The primary advantages of this model, however, are: the site of occlusion is well defined, the degree of blood flow reduction is consistent, functional and neurological impairment occurs rapidly, infarct size is consistent, and the high rate of survival allows for long-term chronic assessment.

Permanent Cerebral Vessel Occlusion via Double Ligature and Transection

We describe a highly reproducible method for the permanent occlusion of a rodent major cerebral blood vessel. This technique can be accomplished with very little peripheral damage, minimal blood loss, a high rate of long-term survival, and consistent infarct volume commensurate with the human clinical population.

Cerebrale permanente occlusione di vasi<em&gt; Via</em&gt; Doppio Legatura e resezione

Stroke is a leading cause of death, disability, and socioeconomic loss worldwide. The majority of all strokes result from an interruption in blood flow ...

Modeling Stroke in Mice: Permanent Coagulation of the Distal Middle Cerebral Artery

Stroke is the third most common cause of death and a main cause of acquired adult disability in developed countries. Only very limited therapeutical options are available for a small proportion of stroke patients in the acute phase. Current research is intensively searching for novel therapeutic strategies and is increasingly focusing on the sub-acute and chronic phase after stroke because more patients might be eligible for therapeutic interventions in a prolonged time window. These delayed mechanisms include important pathophysiological pathways such as post-stroke inflammation, angiogenesis, neuronal plasticity and regeneration. In order to analyze these mechanisms and to subsequently evaluate novel drug targets, experimental stroke models with clinical relevance, low mortality and high reproducibility are sought after. Moreover, mice are the smallest mammals in which a focal stroke lesion can be induced and for which a broad spectrum of transgenic models are available. Therefore, we describe here the mouse model of transcranial, permanent coagulation of the middle cerebral artery via electrocoagulation distal of the lenticulostriatal arteries, the so-called &#8220;coagulation model&#8221;. The resulting infarct in this model is located mainly in the cortex; the relative infarct volume in relation to brain size corresponds to the majority of human strokes. Moreover, the model fulfills the above-mentioned criteria of reproducibility and low mortality. In this video we demonstrate the surgical methods of stroke induction in the &#8220;coagulation model&#8221; and report histological and functional analysis tools.

Various murine models of middle cerebral artery occlusion (MCAO) are widely used in experimental brain research. Here, we demonstrate the model of transcranial permanent distal MCAO which produces consistent cortical infarction of a size corresponding to damage imposed by the majority of human ischemic strokes.

Corsa di modellazione in Mouse: Coagulazione permanente dell&#39;arteria cerebrale media distale

Stroke is the third most common cause of death and a main cause of acquired adult disability in developed countries. Only very limited therapeutical ...

Permanent Ligation of the Left Anterior Descending Coronary Artery in Mice: A Model of Post-myocardial Infarction Remodelling and Heart Failure

Heart failure is a syndrome in which the heart fails to pump blood at a rate commensurate with cellular oxygen requirements at rest or during stress. It is characterized by fluid retention, shortness of breath, and fatigue, in particular on exertion. Heart failure is a growing public health problem, the leading cause of hospitalization, and a major cause of mortality. Ischemic heart disease is the main cause of heart failure.
Ventricular remodelling refers to changes in structure, size, and shape of the left ventricle. This architectural remodelling of the left ventricle is induced by injury (e.g., myocardial infarction), by pressure overload (e.g., systemic arterial hypertension or aortic stenosis), or by volume overload. Since ventricular remodelling affects wall stress, it has a profound impact on cardiac function and on the development of heart failure. A model of permanent ligation of the left anterior descending coronary artery in mice is used to investigate ventricular remodelling and cardiac function post-myocardial infarction. This model is fundamentally different in terms of objectives and pathophysiological relevance compared to the model of transient ligation of the left anterior descending coronary artery. In this latter model of ischemia/reperfusion injury, the initial extent of the infarct may be modulated by factors that affect myocardial salvage following reperfusion. In contrast, the infarct area at 24 hr after permanent ligation of the left anterior descending coronary artery is fixed. Cardiac function in this model will be affected by 1) the process of infarct expansion, infarct healing, and scar formation; and 2) the concomitant development of left ventricular dilatation, cardiac hypertrophy, and ventricular remodelling.
Besides the model of permanent ligation of the left anterior descending coronary artery, the technique of invasive hemodynamic measurements in mice is presented in detail.

Heart failure is the leading cause of hospitalization and a major cause of mortality. A model of permanent ligation of the left anterior descending coronary artery in mice is applied to investigate ventricular remodelling and cardiac dysfunction post-myocardial infarction. The technique of invasive hemodynamic measurements in mice is presented.

Legatura permanente dell&#39;arteria discendente anteriore sinistra coronarica nei topi: un modello di post-infarto rimodellante miocardico e insufficienza cardiaca

Heart failure is a syndrome in which the heart fails to pump blood at a rate commensurate with cellular oxygen requirements at rest or during stress. It ...

Performing Permanent Distal Middle Cerebral with Common Carotid Artery Occlusion in Aged Rats to Study Cortical Ischemia with Sustained Disability

Stroke typically occurs in elderly people with a range of comorbidities including carotid (or other arterial) atherosclerosis, high blood pressure, obesity and diabetes. Accordingly, when evaluating therapies for stroke in animals, it is important to select a model with excellent face validity. Ischemic stroke accounts for 80% of all strokes, and the majority of these occur in the territory of the middle cerebral artery (MCA), often inducing infarcts that affect the sensorimotor cortex, causing persistent plegia or paresis on the contralateral side of the body. We demonstrate in this video a method for producing ischemic stroke in elderly rats, which causes sustained sensorimotor disability and substantial cortical infarcts. Specifically, we induce permanent distal middle cerebral artery occlusion (MCAO) in elderly female rats by using diathermy forceps to occlude a short segment of this artery. The carotid artery on the ipsilateral side to the lesion was then permanently occluded and the contralateral carotid artery was transiently occluded for 60 min. We measure the infarct size using structural T2-weighted magnetic resonance imaging (MRI) at 24 hr and 8 weeks after stroke. In this study, the mean infarct volume was 4.5% &#177; 2.0% (standard deviation) of the ipsilateral hemisphere at 24 hr (corrected for brain swelling using Gerriet&#8217;s equation, n = 5). This model is feasible and clinically relevant as it permits the induction of sustained sensorimotor deficits, which is important for the elucidation of pathophysiological mechanisms and novel treatments.

Here we present a protocol to produce permanent distal middle cerebral artery occlusion in elderly female rats with simultaneous occlusion of the carotid arteries to generate large cortical infarcts and sustained deficits. We show confirmation of the lesion size using structural MRI at 24 hr and 8 weeks after stroke.

Esecuzione permanente distale Medio cerebrale con arteria carotide comune di occlusione in ratti anziani per studiare corticale Ischemia con disabilità sostenuta

Stroke typically occurs in elderly people with a range of comorbidities including carotid (or other arterial) atherosclerosis, high blood pressure, ...

Transient Middle Cerebral Artery Occlusion Model of Neonatal Stroke in P10 Rats

A number of animal models have been used to study hypoxic-ischemic injury, traumatic injury, global hypoxia, or permanent ischemia in both the immature and mature brain. Stroke occurs commonly in the perinatal period in humans, and transient ischemia-reperfusion is the most common form of stroke in neonates. The reperfusion phase is a critical component of injury progression, which occurs over a period of days to weeks, and of the endogenous response to injury. This postnatal day 10 (p10) rat model of transient middle cerebral artery occlusion (tMCAO) creates a unilateral, non-hemorrhagic focal ischemia-reperfusion injury that can be utilized to study the mechanisms of focal injury and repair in the full-term-equivalent brain. The injury pattern that is produced by tMCAO is consistent and highly reproducible and can be confirmed with MRI or histological analyses. The severity of injury can be manipulated through changes in occlusion time and other methods that will be discussed.

Neonatal stroke is a significant cause of early brain injury requiring a translational model with consistent focal injury patterns and high reproducibility in order to enable study. This study describes the detailed surgical procedure for creating a non-hemorrhagic, unilateral focal ischemia-reperfusion injury in full-term-equivalent rodents.

Transient Medio occlusione dell&#39;arteria cerebrale Modello di Stroke neonatale nei ratti P10

A number of animal models have been used to study hypoxic-ischemic injury, traumatic injury, global hypoxia, or permanent ischemia in both the immature ...

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.

Occlusione dell'arteria cerebrale centrale consentendo di riperfusione tramite riparazione dell'arteria carotica comune nei topi

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