Os autores agradecem Laurie von Melchner, Hospital Universitário de Berna, Departamento de Neurocirurgia, Berna, Suíça, para revisão e edição do manuscrito e Paskus Jeremias, Hospital Infantil de Boston, Boston, MA pela revisão do projecto inicial. Agradecemos a gestão hábil de cuidados com os animais, anestesia e assistência operacional a partir de Daniel Mettler, DVM, Max Müller, DVM, Daniel Zalokar e Olgica Beslac, Experimental Instituto Cirúrgico, Departamento de Pesquisa Clínica da Universidade de Berna, Berna, Suíça. Agradecemos Michael Lensch, Chefe de Pesquisa Enfermeira do Departamento de Medicina de Terapia Intensiva do Hospital Universitário de Berna e da Universidade de Berna, Berna, Suíça, para o monitoramento de dados em tempo real e pós-processamento dos parâmetros fisiológicos. Agradecemos Edin Nevzati, Carl Muroi, e Salomé Erhardt, por sua excelente laboratório de assistência técnica e operativa. Este trabalho foi financiado pelo Departamento de Intensive Care Medicine, Hospital Universitário de Berna e da Universidade de Berna, Berna, Suíça, o Departamento de Pesquisa Clínica da Universidade de Berna, Berna, Suíça, e ao Fundo de Investigação do Kantonsspital Aarau, Aarau, Suíça. Agradecemos a Elsevier, a permissão para reimpressão Figuras 1 e 2.

Três mulheres meses de idade coelhos da raça Nova Zelândia, pesando 2,5-3,5 kg foram utilizados para este procedimento. O estudo foi realizado de acordo com os Institutos Nacionais de Saúde diretrizes para o cuidado e uso de animais de laboratório e com a aprovação do Comitê de Cuidados Animais do Cantão de Berna, Suíça (aprovação como 105/13). Todos os procedimentos cirúrgicos foram realizados sob condições estéreis no Instituto Cirúrgico Experimental do Departamento de Pesquisa Clínica do Hospital Universitário de Berna, em Berna, na Suíça. Um anestesista veterinário acompanhou os animais durante a cirurgia e durante a recuperação. 1 Preparação Animal, Posicionamento e artéria subclávia Canulação <ol><li> Induzir a anestesia geral, no coelho com a injecção intramuscular de cetamina (30 mg / kg; Ketalar, 50 mg / ml) e xilazina (6 mg / kg; Xylapan 20 mg / ml) e da profundidade da anestesia de controlo, verificando a resposta do coelho para nocivo stimulation (por exemplo, toe pitada). Ver 1.7, em caso de resposta positiva. </li><li> Puxe para baixo as pálpebras inferiores de ambos os olhos e aplique uma pequena quantidade de pomada nas pálpebras para evitar a secura e irritação adicional. </li><li> Cateterize a veia lateral da orelha com um Gbutterfly Venflon 20 (20 G cateter vascular), fixe com fita adesiva, e conectar-se a um saco de gravidade contendo cloreto de sódio a 0,9% (500 ml / 24h) e cetamina (40 mg / kg / hr) / xilazina (4 mg / kg / hr) para a administração intravenosa contínua (iv) a anestesia. Administrar analgésicos adicional a cada 15 min iv (fentanil, 1 mcg / kg). Nota: Evite anestésicos gases voláteis, que está associado com a diminuição da CPP, aumentando a CBF, ea diminuição da taxa metabólica cerebral de oxigênio 11 anestésicos intravenosos fornecer características mais ideais para neuroanestesia preservando CBF e vasoconstrição cerebral, 12, que é de máxima importância quando se estuda cerebral. vasoespasmo. Além disso, embora a mortalidade é aumentada em espontaneously respirando animais, pode imitar melhor a situação humana de HAS aguda. </li><li> Fornecer oxigênio (1-2 L / min) através de uma máscara respiratória que permite que o dióxido de carbono ao final da expiração (ETCO2) de monitoramento. </li><li> Instale um eletrocardiograma de 3 canais (ECG) .Colocar três eletrodos subcutâneos em um arranjo triangular no lado ventral do coelho; especificamente, colocar um eletrodo na região midthoracic direita (com a distância para o campo raspada estéril para a canulação da artéria subclávia) e dois eletrodos na parte inferior do abdômen distribuídos em ambos os membros. </li><li> Monitorização da profundidade da anestesia a cada 15 minutos durante a cirurgia, seguindo a freqüência respiratória, freqüência cardíaca (FC) monitorados a partir do sinal de ECG, e reação aos estímulos dolorosos. </li><li> Em caso de resposta positiva a um estímulo nociceptivo (dedo do pé de aperto), adaptar-se a profundidade da anestesia com cloridrato de cetamina bolus (6 mg / kg) e xilazina bólus iv (0,05 mg / kg) por via intravenosa e / ou um bolus analgesia adicional com fentanil (um mcg / kg) iv </li><li> Corrigir o coelho em decúbito dorsal sobre uma placa de aquecimento do corpo, inclinando a cabeça 20 ° para baixo e gire-o levemente para o lado contra o lado em que a artéria subclávia será exposto. </li><li> aplicar pomada e preparar a área para a cirurgia raspando o cabelo por cima do músculo peitoral direita em torno do terço médio da clavícula, e sobre o crânio frontal-, parietal- e occipital, o pescoço, e sobre a artéria femoral comum direita. </li><li> Desinfecção da pele durante 3 min com um largo espectro anti-séptico, por exemplo., Povidona-iodo. </li><li> Cubra o coelho com folhas estéreis. Executar todas procedimentos adicionais em condições estéreis e freqüentemente aplicar 4% papaverina HCl e solução de antibiótico (neomicina sulfato de 5 mg / ml) topicamente para prevenir vasoespasmo arterial por manipulação navio e infecções locais. </li><li> Infiltrar o músculo peitoral com anestésicos locais (lidocaína 1% máximo de 6 mg / kg). Faça uma incisão na pele paraesternal e preparar omúsculo peitoral. Utilizando o microscópio, dissecção da artéria subclávia e prenda com um proximal e ligadura distal (4-0 polyfilament) em torno da extremidade exposta. Manter uma ligadura perto do controle proximal no lugar para fixar o cateter e ligar o vaso distalmente. </li><li> Executar uma arteriotomia na parede da artéria subclávia por uma incisão na artéria com uma microtesoura curvo e canular a artéria subclávia retrogradamente com uma pequena intravasais torneira de 3 vias. Fixar o cateter por ligadura nó duplo para a ligadura distal, de modo a evitar a torção ou flexão arterial da parte proximal da artéria e para evitar o deslizamento ou hemorragia massiva. </li></ol> 2 Pressão Arterial e Monitoramento gasometria do sangue arterial <ol><li> Conecte o pau 3-way stop para i) a cânula intravascular para o gás de sangue arterial (ABG) analisa, pH, PaCO 2, PaO 2, bicarbonato, excesso de base, e SO 2, ii) o sangue arterial invasivadispositivo de medição de pressão, e iii) o dispositivo de derivação. </li><li> Coletar amostras de sangue para o status ABG (PaCO 2, PaO 2) e monitorar continuamente os parâmetros padrão respiratório e cardiovascular (pressão arterial, FC, ECG, frequência respiratória e expirado CO 2) e de transferência de dados através da interface de saída analógica para um analógico- conversor A / registrador de dados digital e loja. NOTA: As pressões serão zerados em níveis cardíacos antes e depois de cada sessão, e calibração de pressão do / digital conversor analógico e sistema de registro de dados será feito uma vez antes da série começar. </li></ol> 3 Base Digital Subtração Angiografia <ol><li> Coloque um dispositivo de dimensionamento externo (pequena esfera) sobre ambos os ângulos da mandíbula, a fim de calibrar o angiograma. NOTA: Isto permite a comparação exata das medições post hoc de linha de base e acompanhar o diâmetro dos vasos. </li><li> Realizar a subtração angiografia digital (DSA) por retrógrada intra-uminjecção de bolus rterial de Iopamidol não iónico (0,6 ml / kg, 5 ml / seg durante 2 segundos) através da cânula da artéria e lave a cânula imediatamente após a injecção de bolus, com solução salina, a fim de prevenir a oclusão do último. </li><li> Obtenção de imagens (7 imagens em 14 seg) do sistema vertebrobasilar utilizando uma gravação sequencial rápida angiografia em posição oblíqua 5 ° esquerda anterior. </li><li> Infiltrar-se na área ao redor da artéria femoral comum direita com anestésicos locais (lidocaína 1%, máximo de 6 mg / kg). Faça uma pequena incisão na pele inguinal. Utilizando o microscópio para visualização, dissecção da artéria femoral comum e prenda com um proximal e ligadura distal (4-0 polyfilament) em torno da extremidade exposta. </li><li> Após arteriotomia, canular a artéria femoral com uma bainha de 5-F. Lave a porta lateral da bainha com uma solução salina. </li><li> Avançar um catéter 5-F na artéria inominada pela bainha sob fluoroscopia. Criar um roteiro, então avançar um fio-guia para the sistema vertebrobasilar. Injectar um bolus de Iopamidol não iónico (0,6 ml / kg, 5 ml / seg em 2 seg) para DSA como descrito na etapa 3.2. </li></ol> 4. rotação de posição propensa <ol><li> Na sequência da linha de base DSA, reposicionar o coelho da posição supina para a posição prona. Tenha cuidado para não manipular ou mudar a posição dos cateteres intra-arteriais. </li><li> Posicione a cabeça em uma porta-cabeça em um ângulo de 30 °, cabeça orientada para baixo. </li></ol> 5. Cisterna Magna Puncture <ol><li> Desinfecção da pele por cima da cabeça e pescoço, com povidona-iodo 3 vezes durante 1 min cada, e cobrir a área cirúrgica com lençóis esterilizados. </li><li> Insira um 22 G x 40 mm pediátrica agulha de acesso espinhal transcutânea na cisterna magna, sem qualquer incisão na pele antes ou deslocamento muscular. </li><li> Confirme se o animal está totalmente anestesiado, garantindo a ausência de resposta de igual pitada antes de deslizar a agulha para baixo ao longo do ósseo protuberância occipital externaaté que um intervalo é detectado; não empurrar a agulha mais. </li><li> Confirme o correto posicionamento da agulha, observando gotejamento espontâneo do líquido cefalorraquidiano com a cabeça do coelho inclinado para baixo em um ângulo de 20 ° para poucos min. </li></ol> 6 Instalação de Pressão Intracraniana e Monitoramento do fluxo sanguíneo cerebral <ol><li> Depois de pele na linha média e Galea incisão, insira um pequeno afastador cirúrgico. </li><li> Faça três osteotomias rodada (2 mm de diâmetro) usando um microdrill alta velocidade na parte frontal do crânio de acordo com os pontos de referência externo do crânio (Figura 1) 9, ou seja, sobre o bulbo olfatório e frontal bilateral para a colocação de um dispositivo neuromonitoring se necessário. Use uma régua escala milimétrica para determinar as coordenadas para o posicionamento orifício de trepanação da seguinte forma: a pressão intracraniana (PIC) na linha midpupillary, 1-2 mm da linha sagital mediano; sondas laser Doppler intraparenquimatosos4-5 mm anterior e lateral ao bregma (Figura 1). </li><li> Visualize a dura-máter, e realizar a hemostasia meticulosa: use cera óssea na hemostasia do osso em virtude da sua acção tamponamento e hemostasia local utilizando coagulação bipolar da dura-máter. </li><li> Coloque o intraparenquimal pressão intracraniana (PIC) da ponta do monitor no bulbo olfativo direito a uma profundidade de 2 mm e, em seguida, calibrar. </li><li> Coloque os dois laser-Doppler fluxometria sondas agulha fina através de um sistema de fixação externa e inseri-los nos orifícios correspondentes Burr em ambos os hemisférios direito e esquerdo frontais laterais ao sistema ventricular, ou seja, na linha média para evitar interferência com fluido cerebrospinal. Coloque as sondas de agulha a uma profundidade de 2,5 mm. </li><li> Após a colocação das sondas neuromonitoring, vedar todos os orifícios da rebarba com um tampão de espessura de cera de osso, a fim de manter o crânio estanque. </li><li> Meça os parâmetros basais de pressão arterial média (MAP), ICP e do fluxo sanguíneo cerebral (CBF) através de monitor multiparamétrico e quatro canais tecido laser-Doppler monitor de perfusão sanguínea. </li></ol> 7 Shunt Indução <ol><li> Conecte a agulha de acesso da coluna vertebral na cisterna magna para a artéria subclávia anteriormente cateterização via tubulação de monitoramento de pressão cheia de sangue. Use a torneira de 3 vias para a medição da pressão arterial e, como porta de coleta de sangue. NOTA: A gravidade da SAH depende da quantidade de sangue, e podem ser mais ou menos estimada pela extensão dos coágulos subaracnóidea no momento da colheita do cérebro 5,11. </li><li> Monitorar continuamente PAM, FC, ECG, frequência respiratória e ao final da expiração de CO 2 a uma taxa de amostragem de 1 Hz de 6 min, antes até, pelo menos, 20 minutos após a HAS. </li><li> Confirme se o animal está totalmente anestesiado, garantindo a ausência de resposta toe pitada antes de abrir a conexão shunt entre a artéria subclávia ea cisterna magna para induzir a HAS pela gradie pressãont. NOTA: A SAH controlado pode ser conseguido fechando o shunt, em qualquer ponto de tempo (por exemplo, a um nível desejado de ICP.). </li><li> Gravar os valores de estado estacionário durante um período de tempo de aproximadamente 15 minutos. </li><li> Após ICP atinge o seu pico, manter a agulha de acesso da coluna vertebral no lugar até que o ICP retorna a um estado estacionário próximo aos valores basais. Se o planalto ICP é mantida por mais de 10 segundos ou se ICP diminui espontaneamente, fechar o shunt. </li><li> Remover CBF sondas por agulha fina e sonda ICP, tapar buracos rebarba com cera de osso, remova todos os cateteres (incluindo cateter subclávia, desde a manipulação do cateter com sangramento consecutivo está associada com alta morbidade e mortalidade, e aumenta a taxa de infecção), realizar irrigação ferida rigoroso com sulfato de neomicina e sutura da pele. </li></ol> 8 manejo pós-operatório <ol><li> O processo tem a duração de cerca de 2 horas. Devido às meias-vidas de cetamina e xilazina, o tempo de recuperação do animal é bastante curta - cerca de 1 hora. Os animais são mantidos sob uma lâmpada de aquecimento durante a recuperação. Os fluidos adicionais não são fornecidas. Durante esta fase inicial de recuperação pós-operatório, aplicar buprenorfina 0,02 mg / kg sc a cada 8 horas, durante 24 horas. </li><li> Aplique patches matriz transdérmica de fentanil liberando 12,5 mcg / h na região do pescoço raspado dos animais para analgesia eficaz nas próximas 72 horas. </li><li> Não deixe um animal sem supervisão até que ele recuperou a consciência suficiente para manter decúbito esternal. </li><li> Não devolva um animal que foi submetido a cirurgia na companhia de outros animais até que esteja totalmente recuperado. </li></ol> 9 Acompanhamento Digital subtração angiografia para avaliar as DCV são no dia 3 <ol><li> Efectuar os Passos 1,1-3,6, tal como descrito acima. </li><li> Eutanásia dos animais por injecção intra-arterial de bolus de thiopenthal de sódio (40 mg / kg) (Pentothal, Ospedalia AG, Hünenberg, Suíça). Nos casos em que a histologia e immunohistochemistry for necessário, realizar uma perfusão intracardíaca de fixação à temperatura ambiente a uma pressão de perfusão de 100 cm de 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).

O modelo de derivação produz patologia semelhante ao observado em seres humanos após SAH 3,8,10 aguda. Tem sido sugerido que pode exacerbar EBI, manter e mesmo desencadear DCV são 12, e, como tal, este modelo pode ajudar na investigação de ambas as fases precoce e tardia DCV são, incluindo interacções EBI e DCV são seguintes HAS. Em particular, repetível in vivo as DCV são técnicas de controlo, incluindo DSA 13, angiotomografia 14, e Doppler transcraniano...

Hemorragia subaracnóide aneurismática (HAS) é um dos mais fatais condições neuropatológicas, muitas vezes levando a danos neurológicos permanentes ou morte 1. Pesquisas anteriores se concentrou em vasoespasmo cerebral tardio (DCV são) como etiologia primária de déficits neurológicos associados com HAS 2. No entanto, os resultados clínicos geralmente pobres de pacientes que sofrem de HAS após o tratamento de vasoespasmo levou a uma expansão do foco de pesquisa para incluir os efeitos da lesão cerebral precoce (EBI) após SAH 3. Maior compreensão da importância de ambos EBI e as DCV são em contribuir para desfechos clínicos após SAH é essencial para o desenvolvimento de estratégias terapêuticas mais eficazes. Até agora, a injecção de sangue autólogo simples e dupla na cisterna magna tem sido o método padrão para a indução de HAS para o estudo de DCV são 2-6. Embora comumente utilizada em estudos anteriores,este modelo provavelmente não reproduz as principais alterações neuropatológicas associadas à HAS induzida EBI 7. Em contraste, a perfuração endovascular é conhecido por produzir alterações fisiopatológicas graves agudos que simulam parcialmente os sintomas da EBI 7. Este relatório descreve um modelo de coelho romance de HAS projetado para permitir a investigação de ambos EBI e as DCV são, permitindo assim a caracterização mais precisa da patologia induzida pelo SAH 8-10. Com a técnica descrita, o modelo padrão cisterna magna é adaptado ligando o sistema arterial da artéria subclávia e da cisterna magna através de um shunt extracorpórea. O fluxo de sangue está, assim, ligada à fisiologia do coelho e dirigido pela diferença de pressão entre o sangue arterial e da pressão intracraniana. O sangramento pára quando a pressão intracraniana (ICP) iguala a pressão sanguínea diastólica e o sangue no sistema de derivação do coagula. Utilizando o anfitrião &amp; #8217 s fisiologia reduz indução SAH examinador-dependente, levando a um modelo mais consistente de HAS que produz de forma confiável tanto EBI e as DCV são fenótipos 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 height="21" style="height:21px;width:279px;">Equipment</td>
			<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>
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		<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>
		</tr>
		<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>
		</tr>
		<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

Lesão cerebral precoce e vasoespasmo cerebral atrasou tanto contribuir para desfechos desfavoráveis ​​após hemorragia subaracnóidea (HSA). Modelos animais reprodutíveis e controláveis ​​que simulam as duas condições são actualmente incomum. Portanto, os novos modelos são necessários para imitar condições fisiopatológicas humanos decorrentes da HAS. Este relatório descreve as nuances técnicas de um modelo HAS-sangue shunt coelho que permite o controle da pressão intracraniana (PIC). Uma derivação extracorpórea é colocada entre o sistema arterial e o espaço subaracnóide, que permite HAS-examinador independente num crânio fechado. Passo-a-passo as instruções processuais e equipamentos necessários são descritos, bem como considerações técnicas para produzir o modelo com a mortalidade e morbidade mínima. Detalhes importantes necessários para a criação cirúrgica com sucesso deste modelo de coelho HAS controlada pelo ICP robusto, simples e consistente são descritos.

<html> O modelo de derivação de sangue de coelho de HAS descrito neste relatório produz EBI no hipocampo (Figura 2A, B), córtex basal (Figura 2A, B), e vasculatura cerebral (Figura 2C) tão cedo quanto 24 h depois da lesão e mostra uma característica distribuição de sangue (Figura 2D) 8. Além disso, o modelo provoca graus moderados a graves da DCV são no terceiro dia após a indução da SAH (Figura 3) 10....

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.

O coelho Modelo Blood-shunt para o Estudo da aguda e tardia Sequelas de hemorragia subaracnóidea: Aspectos Técnicos

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

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13.5K visualizações.  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.

Vídeo: 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.

Microcirúrgica venoso Bolsa Bifurcação Arterial-aneurismas no Modelo de Coelho: Aspectos Técnicos

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

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.

Biomarcadores em um modelo animal para Revelando correlatos neurais, hematológica e Comportamental do TEPT

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 Fluorescência de Imagem biventricular isolado de coelho

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.

Um Modelo de Rato Baixa Mortalidade para Avaliar vasoespasmo cerebral adiada após hemorragia subaracnóide Experimental

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.

Humanos Linhas tumor neuroendócrino de células como um modelo tridimensional para o estudo da terapia de tumores humanos Neuroendócrino

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.

Um modelo murino de implante de stent na artéria carótida para o Estudo da reestenose

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.

Aspectos técnicos da artocava Fístula Rato

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.

Um modelo murino de hemorragia subaracnóide

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.

Um modelo murino de Arterial Restenosis: Aspectos Técnicos do Femoral Injury Fio

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.

Medição de contratilidade isométrica da artéria mesentérica Mouse usando fio miografia

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