Gli autori ringraziano Laurie von Melchner, Bern University Hospital, Dipartimento di Neurochirurgia, Berna, in Svizzera, per la revisione e la modifica del manoscritto e Paskus Geremia, Hospital di Boston per bambini, Boston, MA per la correzione della bozza iniziale. Apprezziamo la gestione sapiente di cura degli animali, anestesia e assistenza operativa da Daniel Mettler, DVM, Max Müller, DVM, Daniel Zalokar, e Olgica Beslac, Experimental Surgical Institute, Dipartimento di Ricerca Clinica, Università di Berna, Berna, Svizzera. Ringraziamo Michael Lensch, Responsabile Ricerca Infermiere, Dipartimento di Terapia Intensiva, Università di Berna Hospital e Università di Berna, Berna, Svizzera, per il monitoraggio dei dati in tempo reale e post-elaborazione dei parametri fisiologici. Ringraziamo Edin Nevzati, Carl Muroi, e Salome Erhardt, per la loro eccellente laboratorio di assistenza tecnica ed operativa. Questo lavoro è stato sostenuto dal Dipartimento della Intensive Care Medicine, Università di Berna Hospital e Università di Berna, Berna, Svizzera, il Dipartimento di Ricerca Clinica, Università di Berna, Berna, Svizzera, e il Fondo di ricerca del Kantonsspital Aarau, Aarau, Svizzera. Ringraziamo Elsevier, il permesso di ristampa per figure 1 e 2.

Tre mesi di età femminile conigli New Zealand che pesano 2,5-3,5 kg sono stati utilizzati per questa procedura. Lo studio è stato eseguito in conformità con il National Institutes of linee guida sanitarie per la cura e l&#39;uso di animali da esperimento e con l&#39;approvazione del Comitato del Cantone di Berna, Svizzera (approvazione N ° 105/13) Animal Care. Tutte le procedure chirurgiche sono state eseguite in condizioni sterili presso l&#39;Istituto Sperimentale Chirurgica del Dipartimento di Ricerca Clinica presso l&#39;Università di Berna Hospital di Berna, in Svizzera. Un anestesista veterinario monitorato gli animali durante l&#39;intervento chirurgico e durante il recupero. 1 Preparazione degli animali, posizionamento e succlavia Cannulazione <ol><li> Indurre anestesia generale nel coniglio con iniezione intramuscolare di ketamina (30 mg / kg; Ketalar, 50 mg / ml) e xilazina (6 mg / kg; Xylapan 20 mg / ml) e la profondità di controllo dell&#39;anestesia controllando la risposta del coniglio nocivi stimoione (ad esempio, un pizzico tep). Vedere 1.7 in caso di risposta positiva. </li><li> Abbassare le palpebre inferiori di entrambi gli occhi e applicare una piccola quantità di pomata per le palpebre per prevenire la secchezza e l&#39;ulteriore irritazione. </li><li> Cateterizzarla la vena orecchio laterale con un Gbutterfly venflon 20 (20 G catetere vascolare), fissare con nastro adesivo, e connettersi a un sacchetto di gravità contenente 0,9% di cloruro di sodio (500 ml / 24 hr) e ketamina (40 mg / kg / ora) / xilazina (4 mg / kg / ora) per endovenosa continua (iv) l&#39;anestesia. Somministrare analgesici supplementari ogni 15 min iv (Fentanyl, 1 mcg / kg). Nota: Evitare anestetici volatili del gas, che è associato con una diminuzione CPP, aumentando CBF, e la diminuzione del tasso metabolico cerebrale di ossigeno 11 anestetici endovenosi offrono più caratteristiche ideali per Neuroanestesia preservando CBF e vasocostrizione cerebrale, 12, che è di importanza superiore quando si studia cerebrale. vasospasmo. Inoltre, anche se la mortalità è aumentata in SPONTaneously respirazione animali, essa può meglio imitare la situazione umana di SAH acuta. </li><li> Fornire ossigeno (1 - 2 L / min) tramite una maschera respiratoria che permette di anidride carbonica di fine espirazione (EtCO2) monitoraggio. </li><li> Installare un elettrocardiogramma a 3 canali (ECG) .Montare tre elettrodi sottocutanei in una disposizione triangolare sul lato ventrale del coniglio; in particolare, posizionare un elettrodo sulla regione mediotoracico destra (con distanza dal campo sterile rasata per l&#39;arteria succlavia incannulamento) e due elettrodi nel basso addome distribuiti su entrambi gli arti. </li><li> Monitorare la profondità dell&#39;anestesia ogni 15 minuti durante l&#39;intervento seguendo frequenza respiratoria, frequenza cardiaca (HR) controllato dal segnale ECG, e la reazione alla stimolazione nociva. </li><li> In caso di risposta positiva alla stimolazione nocicettiva (pizzico tep), adeguare la profondità dell&#39;anestesia da ketamina bolo (6 mg / kg) e xilazina ev in bolo (0,05 mg / kg) ev e / o un bolo analgesia supplementare con Fentanyl (1 mcg / kg) iv </li><li> Fissare il coniglio in posizione supina su una piastra riscaldante corpo, inclinando la testa di 20 ° verso il basso e ruotando leggermente controlateralmente al lato su cui sarà esposta l&#39;arteria succlavia. </li><li> applicare unguento occhio e preparare l&#39;area per un intervento chirurgico con la rasatura dei capelli sopra il muscolo pettorale destro intorno al terzo medio della clavicola, e sopra il cranio frontal-, parietal- e occipitale, il collo, e sul diritto arteria femorale comune. </li><li> Disinfettare la pelle per 3 minuti con un ampio spettro di antisettico, ad es., Povidone-iodio. </li><li> Coprire il coniglio con teli sterili. Eseguire tutte le altre procedure in condizioni sterili e spesso applicano il 4% papaverina HCl e soluzione antibiotica (neomicina solfato 5 mg / ml) per via topica per prevenire il vasospasmo arterioso dalla manipolazione nave e infezioni locali. </li><li> Infiltrati il ​​muscolo pettorale con anestetici locali (lidocaina 1% massimo 6 mg / kg). Fare un incisione cutanea parasternale e preparare ilmuscolo pettorale. Utilizzando il microscopio, sezionare l&#39;arteria succlavia e fissare con una prossimale e distale legature (4-0 suture polifilamento) intorno l&#39;estremità esposta. Mantenere una legatura vicino al controllo prossimale in posizione per fissare il catetere e legare il vaso distale. </li><li> Eseguire un arteriotomia nella parete dell&#39;arteria succlavia incidendo la arteria con un microscissor curvo e cannulate succlavia retrograda con un piccolo intravasale rubinetto a 3 vie. Fissare il catetere doppio nodo legatura verso la legatura distale al fine di impedire la torsione arteriosa o piegatura della parte prossimale dell&#39;arteria e di evitare la fuoriuscita slittamento o massiccio. </li></ol> 2. Pressione del sangue e monitoraggio arterioso Sangue Gas <ol><li> Collegare il 3-way rubinetto di arresto per i) la cannula intravasale di gas del sangue arterioso (ABG) analisi, pH, PaCO2, PaO2, il bicarbonato, l&#39;eccesso di base, e SO 2, ii) il sangue arterioso invasivapressione dispositivo di misurazione, e iii) il dispositivo shunt. </li><li> Raccogliere campioni di sangue per lo stato ABG (PaCO2, PAO 2) e monitorare costantemente parametri standard cardiovascolari e respiratorie (pressione sanguigna, HR, ECG, frequenza respiratoria e di fine espirazione di CO 2) e di trasferimento dei dati tramite l&#39;interfaccia di uscita analogica ad un analogico- convertitore digitale / registratore di dati e memorizzare. NOTA: Le pressioni saranno azzerati a livello del cuore prima e dopo ogni sessione, e la calibrazione pressione analogico / digitale-convertitore e sistema di registrazione dei dati sarà fatto una volta prima che la serie si avvia. </li></ol> 3 Baseline Digital sottrazione angiografia <ol><li> Inserire un dispositivo di dimensionamento esterno (piccola sfera) su entrambi gli angoli mandibolari per calibrare l&#39;angiografia. NOTA: Questo permette di confronto esatto delle misurazioni post-hoc di basale e follow-up diametro del vaso. </li><li> Eseguire la sottrazione angiografia digitale (DSA) da retrograda intra-abolo rterial di non ionico Iopamidolo (0,6 ml / kg, 5 ml / sec per 2 sec) attraverso l&#39;arteria cannulata e lavare immediatamente la cannula dopo l&#39;iniezione del bolo di soluzione fisiologica per evitare occlusione di quest&#39;ultimo. </li><li> Ottenere immagini (7 immagini a 14 sec) del sistema vertebrobasilare utilizzando una rapida registrazione angiografia sequenziale in posizione obliqua 5 ° sinistro anteriore. </li><li> Infiltrati l&#39;area intorno alla destra arteria femorale comune con anestetici locali (lidocaina 1%, massimo 6 mg / kg). Fai una piccola incisione cutanea inguinale. Utilizzando il microscopio per la visualizzazione, sezionare l&#39;arteria femorale comune e fissare con una prossimale e distale legature (4-0 suture polifilamento) intorno l&#39;estremità esposta. </li><li> Dopo arteriotomia, incannulare l&#39;arteria femorale con una guaina 5-F. Lavare l&#39;attacco laterale della guaina con soluzione fisiologica. </li><li> Avanzare un catetere 5-F nell&#39;arteria brachiocefalica attraverso la guaina sotto fluoroscopia. Creare una mappa stradale, poi avanzare un filo guida di the sistema vertebro-basilare. Iniettare un bolo di non ionico Iopamidolo (0,6 ml / kg, 5 ml / sec a 2 sec) per DSA come descritto al punto 3.2. </li></ol> 4. rotazione alla posizione prona <ol><li> Dopo basale DSA, riposizionare il coniglio da posizione supina a quella prona. Fare attenzione a non manipolare o spostare la posizione dei cateteri intra-arteriose. </li><li> Posizionare la testa in un testa-porta ad un angolo di 30 °, orientato a testa in giù. </li></ol> 5. Cisterna Magna Puncture <ol><li> Disinfettare la pelle sopra la testa e il collo con Povidone-iodio 3 volte per 1 minuto ciascuna, e coprire l&#39;area chirurgica con teli sterili. </li><li> Inserire un 22 G x 40 mm pediatrica ago accesso spinale transcutanea nella cisterna magna, senza alcuna incisione cutanea preventiva o spostamento muscolare. </li><li> Verificare che l&#39;animale è completamente anestetizzato garantendo una mancanza di risposta toe-pinch prima di far scorrere l&#39;ago verso il basso lungo la osseo protuberanza occipitale esternafinché non viene rilevato un gap; non spingere ulteriormente l&#39;ago. </li><li> Verificare il corretto posizionamento dell&#39;ago osservando sgocciolatura spontanea del liquido cerebrospinale con la testa di coniglio inclinato verso il basso con un angolo di 20 ° per pochi minuti. </li></ol> 6 Installazione di pressione intracranica e cerebrale monitoraggio del flusso sanguigno <ol><li> Dopo la pelle linea mediana e galea incisione, inserire un piccolo divaricatore chirurgico. </li><li> Fai tre turno osteotomie (2 mm di diametro) con un microdrill ad alta velocità nella parte frontale del cranio secondo i punti di riferimento cranio esterne (Figura 1) 9, vale a dire, il bulbo olfattivo e frontale bilaterale per il posizionamento di un dispositivo neuromonitoring se necessario. Usare un righello scala millimetrata per determinare le coordinate di bava posizionamento del foro come segue: pressione intracranica (ICP) di monitoraggio in linea midpupillary, 1-2 mm dalla linea sagittale mediano; intraparenchimali sonde laser-Dopplerquattro a cinque mm anteriormente e lateralmente al bregma (Figura 1). </li><li> Visualizza la dura madre, ed eseguire l&#39;emostasi meticolosa: usare la cera ossea per l&#39;emostasi ossea in virtù della sua azione tamponamento ed eseguire l&#39;emostasi locale utilizzando la coagulazione bipolare della dura. </li><li> Posizionare il intraparenchimale pressione intracranica (ICP) tip monitor nel bulbo olfattivo di destra ad una profondità di 2 mm e quindi calibrare. </li><li> Bel posto sonde ad ago i due laser-Doppler flussimetria con un sistema di bloccaggio esterno e inserirli nei fori corrispondenti bave sia gli emisferi destro e sinistro frontali laterali al sistema ventricolare, cioè, sulla linea mediana per evitare interferenze con il liquido cerebrospinale. Posizionare una sonda ad ago a una profondità di 2,5 mm. </li><li> Dopo il posizionamento delle sonde neuromonitoring, sigillare tutti i fori bave con uno spesso tappo di cera ossea al fine di mantenere il cranio tenuta di fluido. </li><li> Misurare i parametri di base della pressione arteriosa media (MAP), ICP e il flusso ematico cerebrale (CBF) utilizzando un monitor multiparametrico e tessuti laser-Doppler Monitor perfusione sanguigna a quattro canali. </li></ol> 7 Shunt induzione <ol><li> Collegare l&#39;ago di accesso vertebrale nella cisterna magna per l&#39;arteria succlavia in precedenza cateterismo attraverso tubi monitoraggio della pressione sanguigna-riempita. Utilizzare il rubinetto a 3 vie per la misurazione della pressione arteriosa e come porta di prelievo di sangue. NOTA: La gravità di SAH dipende dalla quantità di sangue, e può essere approssimativamente stimato l&#39;ampiezza di coaguli subaracnoidea al momento del raccolto cervello 5,11. </li><li> Monitorare costantemente MAP, HR, ECG, frequenza respiratoria e di fine espirazione di CO 2 ad una frequenza di campionamento di 1 Hz da 6 min prima fino a almeno 20 min dopo SAH. </li><li> Verificare che l&#39;animale è completamente anestetizzato garantendo una mancanza di risposta pizzico punta prima di aprire il collegamento shunt tra l&#39;arteria succlavia e la cisterna magna per indurre SAH dal Tem pressionent. NOTA: Un SAH controllato può essere ottenuto chiudendo lo shunt in qualsiasi punto di tempo (ad esempio, ad un livello desiderato di ICP.). </li><li> Registrare i valori di stato stazionario per un periodo di tempo di circa 15 min. </li><li> Dopo ICP raggiunge il suo picco, tenere l&#39;ago di accesso spinale in vigore fino ICP ritorna a uno stato stazionario vicino ai valori basali. Se l&#39;altopiano ICP viene mantenuta per più di 10 secondi o se ICP diminuisce spontaneamente, chiudere lo shunt. </li><li> Rimuovere CBF sonde fine-ago e sonda ICP, tappare buchi bave con la cera ossea, rimuovere tutti i cateteri (compresi catetere succlavia, dal momento che la manipolazione del catetere con emorragia consecutiva è associata ad alta morbilità e mortalità, e aumenta il tasso di infezione), effettuare una rigorosa irrigazione ferita con solfato di neomicina e suturare la pelle. </li></ol> 8 Gestione postoperatoria <ol><li> La procedura dura circa 2 ore. A causa delle emivite di ketamina e xilazina, il tempo di recupero di animal è abbastanza breve - circa 1 ora. Gli animali sono tenuti sotto una lampada riscaldante durante il recupero. Non sono forniti altri liquidi. Durante questa prima fase di recupero post-operatorio, si applicano buprenorfina 0,02 mg / kg sc ogni 8 ore per 24 ore. </li><li> Applicare le patch fentanil transdermico a matrice rilasciando 12,5 mcg / ora nella regione del collo rasato degli animali per l&#39;analgesia efficace nei prossimi 72 hr. </li><li> Non lasciare un animale incustodito fino a quando non ha ripreso conoscenza sufficiente per mantenere decubito sternale. </li><li> Non restituire un animale che ha subito un intervento chirurgico in compagnia di altri animali fino alla completa guarigione. </li></ol> 9 Follow-up digitale sottrazione angiografia per valutare DCVS al Day 3 <ol><li> Eseguire i passaggi 1,1-3,6 come descritto sopra. </li><li> Euthanize gli animali per iniezione intra-arteriosa bolo di thiopenthal sodio (40 mg / kg) (Pentothal, Ospedalia AG, Hünenberg, Svizzera). Nei casi in cui istologia e immunohistochemistry è necessario, effettuare una perfusione-fissazione intracardiaco a temperatura ambiente ad una pressione di perfusione 100 cm H 2 O. </li></ol>

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None. The authors have no financial or commercial interest in any of the drugs, materials, or equipment used. No specific funding was received for this work. The authors are solely responsible for the design and conduct of the presented study and report no conflict of interest concerning the materials and methods used in this study or the findings specified in the paper. They confirm the adherence of ethical standards. The study was performed in accordance with the National Institutes of Health guidelines for the care and use of experimental animals and with the approval of the Animal Care Committee of the Canton of Bern, Switzerland (approval #109/07 and #107/09).

Il modello di shunt produce patologia simile a quello osservato negli esseri umani dopo SAH acuta 3,8,10. E &#39;stato suggerito che EBI può esacerbare, mantenere e persino scatenare DCVS 12, e come tale questo modello può essere di aiuto nelle indagini sia nella fase DCVS precoce e tardiva, tra cui EBI e DCVS interazioni seguenti SAH. In particolare, ripetibile in vivo DCVS tecniche di monitoraggio, compresi i DSA 13, tomografia computerizzata angiografia 14, e Dop...

Emorragia subaracnoidea aneurismatica (SAH) è uno dei più in pericolo di vita condizioni neuropatologici, spesso portando a danni neurologici permanenti o la morte 1. Precedenti ricerche si è concentrata sul vasospasmo cerebrale ritardata (DCVS) come l&#39;eziologia primaria di deficit neurologici associati con SAH 2. Tuttavia, generalmente i poveri risultati clinici di pazienti affetti da emorragia subaracnoidea dopo il trattamento del vasospasmo ha portato ad un ampliamento del centro di ricerca per includere gli effetti della lesione cerebrale precoce (EBI) dopo SAH 3. Maggiore comprensione del significato di entrambi EBI e DCVS nel contribuire a poveri risultati clinici dopo SAH è essenziale per lo sviluppo di più efficaci strategie terapeutiche. Fino ad oggi, singola e doppia iniezione di sangue autologo nella cisterna magna è stato il metodo standard per l&#39;induzione SAH per lo studio di DCVS 2-6. Anche se comunemente usato in studi precedenti,questo modello molto probabilmente non riprodurre le modifiche principali neuropatologici associati SAH indotto EBI 7. Al contrario, la perforazione endovascolare è noto per produrre gravi alterazioni fisiopatologiche acute che in parte imitano i sintomi di EBI 7. La relazione descrive un modello di coniglio romanzo SAH progettato per consentire indagine sia EBI e DCVS, consentendo caratterizzazione più accurata della patologia SAH indotta 8-10. Con la tecnica descritta, modello standard cisterna magna è adattato collegando il sistema arterioso dell&#39;arteria succlavia e la cisterna magna tramite uno shunt extracorporea. Il flusso di sangue è quindi legata alla fisiologia del coniglio e guidato dalla differenza di pressione tra il sangue arterioso e la pressione intracranica. L&#39;emorragia si arresta quando la pressione intracerebrale (ICP) è uguale pressione diastolica e il sangue nel sistema shunt coagula. Utilizzando l&#39;host &amp; #8217; s fisiologia riduce SAH induzione esaminatore-dipendenti, portando ad un modello più coerente di SAH che produce in modo affidabile sia EBI e DCVS fenotipi 3,8-10.

<table border="0" cellpadding="0" cellspacing="0" width="905">
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			<td height="34" style="height:35px;width:279px;">Name</td>
			<td style="width:236px;">Company</td>
			<td style="width:223px;">Catalog Number</td>
			<td style="width:167px;">Comments/Description</td>
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			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:279px;">operation microscope</td>
			<td style="width:236px;">Zeiss, Jena, Germany</td>
			<td colspan="2">Zeiss, OPMI-MD surgical microscope</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:279px;">surgical equipment</td>
			<td style="width:236px;">B. Braun, Germany</td>
			<td style="width:223px;">forceps medical n&#176;5, vessel sciccors 8cm, microclip 4mm</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">respirator</td>
			<td style="width:236px;">Hugo Sachs</td>
			<td style="width:223px;"></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">hair clipper</td>
			<td style="width:236px;">3M Surgical Clipper&#160;</td>
			<td style="width:223px;">&#160;Starter Kit 9667A</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">body warm plate</td>
			<td style="width:236px;">FHC</td>
			<td style="width:223px;"></td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:279px;">blood gas analyzer</td>
			<td style="width:236px;">Radiometer, Copenhagen, Denmark</td>
			<td style="width:223px;">ABL 725</td>
			<td></td>
		</tr>
		<tr height="60">
			<td height="60" style="height:60px;width:279px;">cardiac monitoring</td>
			<td style="width:236px;">Camino Multi-Parameter Monitor, Integra, Plainsboro, NJ, US</td>
			<td style="width:223px;">AP-05</td>
			<td></td>
		</tr>
		<tr height="60">
			<td height="60" style="height:60px;">software analysis</td>
			<td style="width:236px;">BIOPAC Systems, Inc., Goleta, CA, USA</td>
			<td style="width:223px;">Biopac MP100 and acqKnowledge software,version 3.8.1</td>
			<td></td>
		</tr>
		<tr height="60">
			<td height="60" style="height:60px;">software analysis</td>
			<td style="width:236px;">ImagePro Discovery, MediaCybernetics, Silver Spring, MD, USA</td>
			<td style="width:223px;">image-Pro Plus version&#160;</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">angiography apparatus</td>
			<td style="width:236px;">DFP 2000 A-Toshiba</td>
			<td style="width:223px;">MIIXR0001EAA</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">ICP monitor</td>
			<td style="width:236px;">Camino Laboratories, San Diego, CA, USA</td>
			<td style="width:223px;">ICP monitor, Model 110-4B</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">blood flow monitor</td>
			<td style="width:236px;">Oxford Optronix Ltd., Oxford, UK</td>
			<td style="width:223px;">CAL KIT microsphere solution</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">laser-Doppler flowmetry fine needle probes&#160;</td>
			<td style="width:236px;">Oxford Optronix Ltd., Oxford, UK</td>
			<td style="width:223px;">MNP110XP, 0.48 mm diameter</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">pressure tube</td>
			<td>B. Braun, Germay</td>
			<td style="width:223px;">PE 1.0 mm &#215; 2.0 mm</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:279px;">anesthesia monitor</td>
			<td>GE Medical Systems, Switzerland&#160;</td>
			<td style="width:223px;">Datex S5 Monitor</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:279px;">Material</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">20 G vascular catheter</td>
			<td style="width:236px;">Smiths Medical</td>
			<td style="width:223px;">Jelco i.v. catheter, REF 4057</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">5.5F three-lumen central venous catheter&#160;</td>
			<td style="width:236px;">Connectors, Tagelswangen, Switzerland</td>
			<td>silicone catheter STH-C040</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">22Gx40mm needle&#160;</td>
			<td style="width:236px;">Emergo Group Inc., Netherlands</td>
			<td style="width:223px;"></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">high-speed microdrill</td>
			<td style="width:236px;">Stryker, Solothurn, Switzerland</td>
			<td>5400-15&#160;</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">bone wax</td>
			<td style="width:236px;">Ethicon, Johnson &#38; Johnson,NJ, USA</td>
			<td style="width:223px;">ETHW31G</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:279px;">bipolar forceps</td>
			<td style="width:236px;">Aesculap, Inc., PA, US</td>
			<td>US349SP&#160;</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:279px;">Ketamin</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
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		<tr height="20">
			<td height="20" style="height:20px;">Xylazine</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
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		<tr height="20">
			<td height="20" style="height:20px;width:279px;">Buprenorphine</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
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		<tr height="20">
			<td height="20" style="height:20px;">Fentanyl</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">transdermal fentanyl matrix patches&#160;</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Lidocaine 1%&#160;</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">4% papaverin HCl&#160;</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Neomycin sulfate&#160;</td>
			<td style="width:236px;">Research Organics Inc., OH, USA</td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Povidone-iodine&#160;</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">0.9% sodium chloride</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:279px;">Iopamidol&#160;</td>
			<td style="width:236px;">Abott Laboratories, IL, USA</td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:279px;">3-0 resorbable suture</td>
			<td style="width:236px;">Ethicon Inc., USA</td>
			<td>VCP824G</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:279px;">5-0 non absorbable suture</td>
			<td style="width:236px;">Ethicon Inc., USA</td>
			<td>8618G</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:279px;">4-0 polyfilament sutures</td>
			<td style="width:236px;">Ethicon Inc., USA</td>
			<td style="width:223px;">VCP284G</td>
			<td></td>
		</tr>
	</tbody>
</table>

the rabbit blood-shunt model for the study of acute and late sequelae of subarachnoid hemorrhage: technical aspects

Lesione cerebrale precoce e ritardata vasospasmo cerebrale entrambi contribuiscono a risultati sfavorevoli dopo emorragia subaracnoidea (SAH). Modelli animali riproducibili e controllabili che simulano entrambe le condizioni sono al momento raro. Pertanto, sono necessari nuovi modelli, al fine di mimare condizioni fisiopatologiche umani derivanti da SAH. Questo rapporto descrive le sfumature tecniche di un sangue-shunt modello SAH coniglio che consente il controllo della pressione intracerebrale (ICP). Un shunt extracorporea è posto tra il sistema arterioso e lo spazio subaracnoideo, che consente esaminatore indipendente SAH in un cranio chiuso. Step-by-step istruzioni procedurali e le attrezzature necessarie sono descritte, così come considerazioni tecniche per produrre il modello con mortalità minima e morbilità. Dettagli importanti necessarie per la riuscita creazione chirurgica di questo modello SAH coniglio robusto, semplice e coerente ICP-controllato sono descritti.

<html> Il modello di shunt sangue di coniglio di SAH descritto in questo rapporto produce EBI nell&#39;ippocampo (Figura 2A, B), la corteccia basale (Figura 2A, B), e la vascolarizzazione cerebrale (Figura 2C) già a partire 24 ore dopo l&#39;infortunio e mostra una caratteristica distribuzione del sangue (Figura 2D) 8. Inoltre, il modello attiva da moderata a grave gradi di DCVS in tre giorni dopo SAH induzione (Figura 3) <su...

Watch this Scientific Journal Video about The Rabbit Blood-shunt Model for the Study of Acute and Late Sequelae of Subarachnoid Hemorrhage: Technical Aspects at JoVE.com

The Rabbit Blood-shunt Model for the Study of Acute and Late Sequelae of Subarachnoid Hemorrhage: Technical Aspects

The experimental intracranial pressure-controlled blood shunt subarachnoid hemorrhage (SAH) model in the rabbit combines the standard procedures &#8212; subclavian artery cannulation and transcutaneous cisterna magna puncture, which enables close mimicking of human pathophysiological conditions after SAH. We present step-by-step instructions and discuss key surgical points for successful experimental SAH creation.

Il Coniglio Blood-shunt modello per lo studio di acuta e sequele tardive di emorragia subaracnoidea: Aspetti tecnici

Early brain injury and delayed cerebral vasospasm both contribute to unfavorable outcomes after subarachnoid hemorrhage (SAH). Reproducible and ...

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13.5K Views.  University and Bern University Hospital (Inselspital). The experimental intracranial pressure-controlled blood shunt subarachnoid hemorrhage (SAH) model in the rabbit combines the standard procedures — subclavian artery cannulation and transcutaneous cisterna magna puncture, which enables close mimicking of human pathophysiological conditions after SAH. We present step-by-step instructions and discuss key surgical points for successful experimental SAH creation.

Video: The Rabbit Blood-shunt Model for the Study of Acute and Late Sequelae of Subarachnoid Hemorrhage: Technical Aspects

Microsurgical Venous Pouch Arterial-Bifurcation Aneurysms in the Rabbit Model: Technical Aspects

For ruptured human cerebral aneurysms endovascular embolization has become an equivalent alternative to aneurysm clipping.1 However, large clinical trials have shown disappointing long-term results with unacceptable high rates of aneurysm recanalization and delayed aneurysm rupture.2 To overcome these problems, animal experimental studies are crucial for the development of better endovascular devices.3-5
Several animal models in rats, rabbits, canines and swine are available.6-8 Comparisons of the different animal models showed the superiority of the rabbit model with regard to hemodynamics and comparability of the coagulation system and cost-effectiveness.9-11 
The venous pouch arterial bifurcation model in rabbits is formed by a venous pouch sutured into an artificially created true bifurcation of both common carotid arteries (CCA). The main advantage of this model are true bifurcational hemodynamics.12 The major drawbacks are the sofar high microsurgical technical demands and high morbidity and mortality rates of up to 50%.13 These limitations have resulted in less frequent use of this aneurysm model in the recent years. These shortcomings could be overcome with improved surgical procedures and modified peri- and postoperative analgetic management and anticoagulation.14-16 Our techniques reported in this paper demonstrate this optimized technique for microsurgical creation of arterial bifurcation aneurysms.

An optimized technique for the microsurgical creation of arterial bifurcation aneurysms mimicking bifurcation human cerebral aneurysms is described. A venous pouch is sutured into an artificially created true bifurcation of both common carotid arteries. Facilitated microsurgical techniques and aggressive postoperative anticoagulation and analgesia lead to minimized morbidity rates and high aneurysm patency rates.

Microchirurgia arteriosa, venosa Pouch biforcazione aneurismi nel modello Rabbit: Aspetti Tecnici

For ruptured human cerebral aneurysms endovascular embolization has become an equivalent alternative to aneurysm clipping.1 However, large clinical trials ...

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Medicina

Biomarkers in an Animal Model for Revealing Neural, Hematologic, and Behavioral Correlates of PTSD

Identification of biomarkers representing the evolution of the pathophysiology of Post Traumatic Stress Disorder (PTSD) is vitally important, not only for objective diagnosis but also for the evaluation of therapeutic efficacy and resilience to trauma. Ongoing research is directed at identifying molecular biomarkers for PTSD, including traumatic stress induced proteins, transcriptomes, genomic variances and genetic modulators, using biologic samples from subjects' blood, saliva, urine, and postmortem brain tissues. However, the correlation of these biomarker molecules in peripheral or postmortem samples to altered brain functions associated with psychiatric symptoms in PTSD remains unresolved. Here, we present an animal model of PTSD in which both peripheral blood and central brain biomarkers, as well as behavioral phenotype, can be collected and measured, thus providing the needed correlation of the central biomarkers of PTSD, which are mechanistic and pathognomonic but cannot be collected from people, with the peripheral biomarkers and behavioral phenotypes, which can.
Our animal model of PTSD employs restraint and tail shocks repeated for three continuous days - the inescapable tail-shock model (ITS) in rats. This ITS model mimics the pathophysiology of PTSD 17, 7, 4, 10. We and others have verified that the ITS model induces behavioral and neurobiological alterations similar to those found in PTSD subjects 17, 7, 10, 9. Specifically, these stressed rats exhibit (1) a delayed and exaggerated startle response appearing several days after stressor cessation, which given the compressed time scale of the rat's life compared to a humans, corresponds to the one to three months delay of symptoms in PTSD patients (DSM-IV-TR PTSD Criterian D/E 13), (2) enhanced plasma corticosterone (CORT) for several days, indicating compromise of the hypothalamopituitary axis (HPA), and (3) retarded body weight gain after stressor cessation, indicating dysfunction of metabolic regulation.
The experimental paradigms employed for this model are: (1) a learned helplessness paradigm in the rat assayed by measurement of acoustic startle response (ASR) and a charting of body mass; (2) microdissection of the rat brain into regions and nuclei; (3) enzyme-linked immunosorbent assay (ELISA) for blood levels of CORT; (4) a gene expression microarray plus related bioinformatics tools 18. This microarray, dubbed rMNChip, focuses on mitochondrial and mitochondria-related nuclear genes in the rat so as to specifically address the neuronal bioenergetics hypothesized to be involved in PTSD.

We describe a rat model of post traumatic stress disorder (PTSD) that reveals the persistent alterations in neuroendocrine function and the delayed long-term, exaggerated fear response, characteristic of PTSD patients. The animal model and methods described here are useful for correlating biomarkers in brain nuclei, which are mechanistic but cannot be measured in patients, with biomarkers in peripheral white blood cells, which can.

Biomarcatori in un modello animale per Rivelare correlati neurali, ematologiche, e comportamentali di PTSD

Identification of biomarkers representing the evolution of the pathophysiology of Post Traumatic Stress Disorder (PTSD) is vitally important, not only for ...

NADH Fluorescence Imaging of Isolated Biventricular Working Rabbit Hearts

Since its inception by Langendorff1, the isolated perfused heart remains a prominent tool for studying cardiac physiology2. However, it is not well-suited for studies of cardiac metabolism, which require the heart to perform work within the context of physiologic preload and afterload pressures. Neely introduced modifications to the Langendorff technique to establish appropriate left ventricular (LV) preload and afterload pressures3. The model is known as the isolated LV working heart model and has been used extensively to study LV performance and metabolism4-6. This model, however, does not provide a properly loaded right ventricle (RV). Demmy et al. first reported a biventricular model as a modification of the LV working heart model7, 8. They found that stroke volume, cardiac output, and pressure development improved in hearts converted from working LV mode to biventricular working mode8. A properly loaded RV also diminishes abnormal pressure gradients across the septum to improve septal function. Biventricular working hearts have been shown to maintain aortic output, pulmonary flow, mean aortic pressure, heart rate, and myocardial ATP levels for up to 3 hours8.
When studying the metabolic effects of myocardial injury, such as ischemia, it is often necessary to identify the location of the affected tissue. This can be done by imaging the fluorescence of NADH (the reduced form of nicotinamide adenine dinucleotide)9-11, a coenzyme found in large quantities in the mitochondria. NADH fluorescence (fNADH) displays a near linearly inverse relationship with local oxygen concentration12 and provides a measure of mitochondrial redox state13. fNADH imaging during hypoxic and ischemic conditions has been used as a dye-free method to identify hypoxic regions14, 15 and to monitor the progression of hypoxic conditions over time10.
The objective of the method is to monitor the mitochondrial redox state of biventricular working hearts during protocols that alter the rate of myocyte metabolism or induce hypoxia or create a combination of the two. Hearts from New Zealand white rabbits were connected to a biventricular working heart system (Hugo Sachs Elektronik) and perfused with modified Krebs-Henseleit solution16 at 37 &deg;C. Aortic, LV, pulmonary artery, and left &amp; right atrial pressures were recorded. Electrical activity was measured using a monophasic action potential electrode. To image fNADH, light from a mercury lamp was filtered (350&plusmn;25 nm) and used to illuminate the epicardium. Emitted light was filtered (460&plusmn;20 nm) and imaged using a CCD camera. Changes in the epicardial fNADH of biventricular working hearts during different pacing rates are presented. The combination of the heart model and fNADH imaging provides a new and valuable experimental tool for studying acute cardiac pathologies within the context of realistic physiological conditions.

The objective is to monitor the mitochondrial redox state of isolated hearts within the context of physiologic preload and afterload pressures. A biventricular working rabbit heart model is presented. High spatiotemporal resolution fluorescence imaging of NADH is used to monitor the mitochondrial redox state of epicardial tissue.

NADH Imaging di fluorescenza biventricolare isolato lavoro Hearts Coniglio

Since its inception by Langendorff1, the isolated perfused heart remains a prominent tool for studying cardiac physiology2. However, it is not well-suited ...

A Low Mortality Rat Model to Assess Delayed Cerebral Vasospasm After Experimental Subarachnoid Hemorrhage

Objective: To characterize and establish a reproducible model that demonstrates delayed cerebral vasospasm after aneurysmal subarachnoid hemorrhage (SAH) in rats, in order to identify the initiating events, pathophysiological changes and potential targets for treatment.
Methods: Twenty-eight male Sprague-Dawley rats (250 - 300 g) were arbitrarily assigned to one of two groups - SAH or saline control. Rat subarachnoid hemorrhage in the SAH group (n=15) was induced by double injection of autologous blood, 48 hr apart, into the cisterna magna. Similarly, normal saline (n=13) was injected into the cisterna magna of the saline control group. Rats were sacrificed on day five after the second blood injection and the brains were preserved for histological analysis. The degree of vasospasm was measured using sections of the basilar artery, by measuring the internal luminal cross sectional area using NIH Image-J software. The significance was tested using Tukey/Kramer's statistical analysis.
Results: After analysis of histological sections, basilar artery luminal cross sectional area were smaller in the SAH than in the saline group, consistent with cerebral vasospasm in the former group. In the SAH group, basilar artery internal area (.056 &mu;m &plusmn; 3) were significantly smaller from vasospasm five days after the second blood injection (seven days after the initial blood injection), compared to the saline control group with internal area (.069 &plusmn; 3; p=0.004). There were no mortalities from cerebral vasospasm.
Conclusion: The rat double SAH model induces a mild, survivable, basilar artery vasospasm that can be used to study the pathophysiological mechanisms of cerebral vasospasm in a small animal model. A low and acceptable mortality rate is a significant criterion to be satisfied for an ideal SAH animal model so that the mechanisms of vasospasm can be elucidated 7, 8. Further modifications of the model can be made to adjust for increased severity of vasospasm and neurological exams.

Aneurysmal subarachnoid hemorrhage (SAH) is bleeding that occurs into the subarachnoid space when an aneurysm ruptures. While the morbidity and mortality from this event has been on a decline due to improved treatment approaches, the risk of vasospasm after subarachnoid hemorrhage continues to be the same as it was several years ago. The importance of establishing a comprehensive and reproducible animal model to identify initiating events of cerebral vasospasm has been the focus of research since the first use of rats in an experimental vasospasm model in 1979 by Barry et al. Early work in rats demonstrated that a single injection of autologous blood into the cisterna magna led to acute (within minutes) but not delayed cerebral vasospasm 3, 6, 14. Here we characterize a low mortality SAH rat model that results in reproducible delayed vasospasm.

Un basso Rat modello di mortalità per valutare ritardata vasospasmo cerebrale dopo emorragia subaracnoidea sperimentale

Objective: To characterize and establish a reproducible model that demonstrates delayed cerebral vasospasm after aneurysmal subarachnoid hemorrhage (SAH) ...

Human Neuroendocrine Tumor Cell Lines as a Three-Dimensional Model for the Study of Human Neuroendocrine Tumor Therapy

Neuroendocrine tumors (NETs) are rare tumors, with an incidence of two per 100,&#x200A;000 individuals per year, and they account for 0.5% of all human malignancies.1 Other than surgery for the minority of patients who present with localized disease, there is little or no survival benefit of systemic therapy. Therefore, there is a great need to better understand the biology of NETs, and in particular define new therapeutic targets for patients with nonresectable or metastatic neuroendocrine tumors. 3D cell culture is becoming a popular method for drug screening due to its relevance in modeling the in vivo tumor tissue organization and microenvironment.2,3 The 3D multicellular spheroids could provide valuable information in a more timely and less expensive manner than directly proceeding from 2D cell culture experiments to animal (murine) models.
To facilitate the discovery of new therapeutics for NET patients, we have developed an in vitro 3D multicellular spheroids model using the human NET cell lines. The NET cells are plated in a non-adhesive agarose-coated 24-well plate and incubated under physiological conditions (5% CO2, 37 &deg;C) with a very slow agitation for 16-24 hr after plating. The cells form multicellular spheroids starting on the 3rd or 4th day. The spheroids become more spherical by the 6th day, at which point the drug treatments are initiated. The efficacy of the drug treatments on the NET spheroids is monitored based on the morphology, shape and size of the spheroids with a phase-contrast light microscope. The size of the spheroids is estimated automatically using a custom-developed MATLAB program based on an active contour algorithm. Further, we demonstrate a simple method to process the HistoGel embedding on these 3D spheroids, allowing the use of standard histological and immunohistochemical techniques. 
This is the first report on generating 3D spheroids using NET cell lines to examine the effect of therapeutic drugs. We have also performed histology on these 3D spheroids, and displayed an example of a single drug's effect on growth and proliferation of the NET spheroids. Our results support that the NET spheroids are valuable for further studies of NET biology and drug development.

We present a simple agarose overlay platform to grow 3D multicellular spheroids using neuroendocrine cancer cell lines. This method provides a very convenient way to examine the effect of therapeutic drugs on the neuroendocrine tumor cells. It could also help us establish human neuroendocrine tumor spheroids for cancer therapy.

Neuroendocrini umani cella linee tumorali come un modello tridimensionale per lo studio dei diritti dell&#39;uomo e terapia dei tumori neuroendocrini

Neuroendocrine tumors (NETs) are rare tumors, with an incidence of two per 100,&#x200A;000 individuals per year, and they account for 0.5% of all human ...

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

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

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

Un modello murino di impianto di stent a livello dell&#39;arteria carotidea per lo Studio della restenosi

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

Technical Aspects of the Mouse Aortocaval Fistula

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta and inferior vena cava (IVC) are exposed. The proximal infrarenal aorta and the distal aorta are dissected for clamp placement and needle puncture, respectively. Special attention is paid to avoid dissection between the aorta and the IVC. After clamping the aorta, a 25 G needle is used to puncture both walls of the aorta into the IVC. The surrounding connective tissue is used for hemostatic compression. Successful creation of the AVF will show pulsatile arterial blood flow in the IVC. Further confirmation of successful AVF can be achieved by post-operative Doppler ultrasound.

The goal is to produce an arteriovenous fistula that is simple and reproducible. This method does not use sutures or glue adhesive. Therefore the samples can be used with the least amount of foreign materials for analysis.

Aspetti tecnici della fistola aorto-cavale mouse

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta ...

A Murine Model of Subarachnoid Hemorrhage

In this video publication a standardized mouse model of subarachnoid hemorrhage (SAH) is presented. Bleeding is induced by endovascular Circle of Willis perforation (CWp) and proven by intracranial pressure (ICP) monitoring. Thereby a homogenous blood distribution in subarachnoid spaces surrounding the arterial circulation and cerebellar fissures is achieved. Animal physiology is maintained by intubation, mechanical ventilation, and continuous on-line monitoring of various physiological and cardiovascular parameters: body temperature, systemic blood pressure, heart rate, and hemoglobin saturation. Thereby the cerebral perfusion pressure can be tightly monitored resulting in a less variable volume of extravasated blood. This allows a better standardization of endovascular filament perforation in mice and makes the whole model highly reproducible. Thus it is readily available for pharmacological and pathophysiological studies in wild type and genetically altered mice.

A standardized mouse model of subarachnoid hemorrhage by intraluminal Circle of Willis perforation is described. Vessel perforation and subarachnoid bleeding are monitored by intracranial pressure monitoring. In addition various vital parameters are recorded and controlled to maintain physiologic conditions.

Un modello murino di emorragia subaracnoidea

In this video publication a standardized mouse model of subarachnoid  hemorrhage (SAH) is presented. Bleeding is induced by endovascular Circle of  Willis ...

A Murine Model of Arterial Restenosis: Technical Aspects of Femoral Wire Injury

Cardiovascular disease caused by atherosclerosis is the leading cause of death in the developed world. Narrowing of the vessel lumen, due to atherosclerotic plaque development or the rupturing of established plaques, interrupts normal blood flow leading to various morbidities such as myocardial infarction and stroke. In the clinic endovascular procedures such as angioplasty are commonly performed to reopen the lumen. However, these treatments inevitably damage the vessel wall as well as the vascular endothelium, triggering an excessive healing response and the development of a neointimal plaque that extends into the lumen causing vessel restenosis (re-narrowing). Restenosis remains a major cause of failure of endovascular treatments for atherosclerosis. Thus, preclinical animal models of restenosis are vitally important for investigating the pathophysiological mechanisms as well as translational approaches to vascular interventions. Among several murine experimental models, femoral artery wire injury is widely accepted as the most suitable for studies of post-angioplasty restenosis because it closely resembles the angioplasty procedure that injures both endothelium and vessel wall. However, many researchers have difficulty utilizing this model due to its high degree of technical difficulty. This is primarily because a metal wire needs to be inserted into the femoral artery, which is approximately three times thinner than the wire, to generate sufficient injury to induce prominent neointima. Here, we describe the essential surgical details to effectively overcome the major technical difficulties of this model. By following the presented procedures, performing the mouse femoral artery wire injury becomes easier. Once familiarized, the whole procedure can be completed within 20 min.

The mouse femoral artery wire injury model of restenosis is technically challenging. In this protocol we show the key technical details essential for successfully performing wire injury to induce consistent&#160;neointima for studies of restenosis.

Un modello murino di restenosi arteriosa: aspetti tecnici del femore Wire Injury

Cardiovascular disease caused by atherosclerosis is the leading cause of death in the developed world. Narrowing of the vessel lumen, due to ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and drugs. It is widely used in many studies on the physiological functions of vascular smooth muscle, the pathogenesis of vascular diseases such as hypertension, and the development of smooth muscle relaxant drugs. The mouse is a widely used model animal with a large pool of disease models and genetically modified strains. We introduced this method to measure the isometric contraction of mouse mesenteric artery in detail. A 1.4-mm segment of mouse mesenteric resistance artery was isolated and mounted on a myograph chamber by passing two steel wires through its lumen. After equilibration and normalization steps, the vessel segment was potentiated by a high-K+ solution twice prior to the contraction assay. As an example of the application of this method in drug development, we measured the relaxant effect of a novel natural substance, neoliensinine, isolated from a Chinese herb, embryos of the lotus seed (Nelumbo nucifera Gaertn.) on mouse mesenteric arteries. The vessel segments mounted on the myograph chamber were stimulated with a high-K+ solution. When the force tension reached a stable sustained phase, cumulative doses of neoliensinine were added to the chamber. We found that neoliensinine had a dose-dependent relaxant effect on smooth muscle contraction, thus suggesting that it bears potential activity against hypertension. In addition, as the vessel segment can survive at least 4 hours after mounting and maintain contractility induced by the high-K+ solution for many times, we suggest that the wire myograph system may be used for the time-consuming process of drug screening.

The wire myograph technique is used to investigate vascular smooth muscle functions and screen new drugs. We report a detailed protocol for measuring the isometric contractility of the mouse mesenteric artery and for screening new relaxants of vascular smooth muscle.

Misurazione di contrattilità isometrica dell'arteria mesenterica Mouse con filo Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...