The authors' work was supported by a grant from Ministry of Science and Technology (MOST 103-2320-B-002-068-MY2), the National Health Research Institute (NHRI-EX104-10418SC), and National Taiwan University (NTU 104R4000).

Todos os experimentos com animais foram conduzidos de acordo com o Guia para o Cuidado e Uso de Animais de Laboratório, publicado pelos Institutos Nacionais de Saúde (NIH publicação no. 85-23, revista em 1996). Este protocolo foi aprovado e em conformidade com as diretrizes estabelecidas pelo Institutional Animal Care e do Comitê Use na Universidade Nacional de Taiwan. 1. Cirurgia animal <ol><li> Prepare um G agulha de seringa 22 por embotamento da ponta da agulha em uma pedra de afiar. Utilizando um alicate, plicate a agulha a um ângulo direito. </li><li> Antes da cirurgia, preparar os instrumentos cirúrgicos necessários e materiais, bem como uma gaiola de recuperação. Autoclave todos os instrumentos e material cirúrgico antes do uso. </li><li> Manter os ratos em torno de 200 g e mantê-los sob 12 h ciclo claro / escuro a uma temperatura controlada (21 ± 2 ° C), com livre acesso a comida e água. Incluir pelo menos 6 ratos em cada grupo. Anestesiar os ratos com pentobarbital(75 mg / kg em 0,5 ml, ip) ou um outro agente anestésico apropriado. Confirmar a profundidade da anestesia, testando o reflexo da cauda. NOTA: A ausência do reflexo da cauda é um indicador de anestesia adequada. </li><li> Coloque um rato na posição supina sobre uma plataforma de cirurgia com uma almofada de aquecimento para manter a temperatura do corpo. Raspar a região abdominal do rato com um cortador de pêlos e cabelos loção removedor para evitar contaminações cirúrgicos. Esfregar o abdômen limpa raspada com betadine ou outro reagente de limpeza antes da cirurgia. NOTA: É importante manter um campo esterilizado durante todo o procedimento. </li><li> Adicione de 2 cm da incisão ao longo da linha média do abdómen com um bisturi. Usando solução salina normal, manter o órgão abdominal úmida durante a cirurgia. Deslocar os órgãos digestivos cuidadosamente para o lado usando bolas de algodão para expor a veia cava inferior, que se situa na região peritoneal posterior. Identificar a aorta abdominal, que se encontra justaposta e, geralmente, sobre aesquerda da veia cava inferior. NOTA: A aorta abdominal é o vaso que pulsa no tempo com o ritmo cardíaco. </li><li> Furar o peritônio com um par de fórceps para descobrir os vasos abaixo. Gentilmente isolar a aorta abdominal ao lado das artérias renais e passar a 8 cm de comprimento por 4-0 sutura de seda por baixo da aorta abdominal entre as origens do direito e artérias renal esquerda. </li><li> Faça um nó duplo solto com a sutura; deixar um loop 3 mm de diâmetro e coloque a agulha 22 G embotada e dobrada dentro do loop. Apertar o nó em torno da aorta e da agulha, e em seguida, remover a agulha imediatamente para alcançar uma constrição diâmetro 0,7 mm. </li><li> Fechar a cavidade abdominal com 6-0 material de sutura absorvível. Suturar o músculo ou da pele incisões com suturas interrompidas simples. Para prevenir a infecção, esfregue o local da cirurgia com uma tintura de iodo. </li><li> Observar o animal com cuidado até que ele recupera a consciência suficiente, como indicado pela livre circulaçãoe ingestão de alimentos. Não deixe um animal sem supervisão até que tenha recuperado a consciência suficiente para manter decúbito esternal. Para prevenir a dor pós-cirurgia, o tratamento de ratos com acetaminofeno (300 mg / kg em 0,5 ml, ip). Nota: Os usuários devem administrar analgésicos conforme aprovado por políticas institucionais. </li></ol> 2. Tecidos e coleta de sangue <ol><li> No pós-operatório de 10 semanas, pesar o rato e anestesiar com pentobarbital (75 mg / kg em 0,5 ml, ip). Antes da cirurgia, confirmar a profundidade da anestesia, testando o seu reflexo cauda. Coloque o rato em uma bandeja de metal. </li><li> Adicione de 2 cm da incisão ao longo da linha média do pescoço utilizando um bisturi. Deslocar os músculos cuidadosamente com uma pinça para expor a traqueia. Observe cuidadosamente para identificar as artérias carótidas, as quais são paralelas à traqueia e vibram no tempo com o ritmo cardíaco. </li><li> Recolhe-se o sangue a partir da artéria carótida dentro de um tubo de recolha de sangue revestido com EDTA. imediatamente centrífugao sangue durante 15 minutos a 2000 xg e recolher o plasma. Armazenar o plasma do sangue a -80 ° C até ser necessário. </li><li> Adicione 5 cm de incisão na região torácica, em torno da linha média do processo xifóide. Perfurar o diafragma com uma pinça afiada. Utilizando um par de tesouras, cortar e remover a caixa torácica ao longo das linhas médio-clavicular de ambos os lados para expor o coração. Extirpar o coração cuidadosamente ao longo das fronteiras cardíacas e vasculares. Retire o coração suavemente sem agarrar o tecido. </li><li> Monte o coração em um aparelho de perfusão Langendorff modificado por amarrar o tronco da aorta para a agulha de perfusão. Perfundir o coração com tampão de Krebs (contendo NaCl 110 mM, KCl 2,6 mM, 1,2 mM de KH 2 PO 4, 1,2 mM MgSO4, 25 mM de NaHCO 3, e glucose 11 mM [pH 7,4]) para lavar o sangue. Pese o coração e calcular a relação coração de peso-para-corpo-peso. Corrigir o coração com 4% de paraformaldeído, em gelo. Lembre-se de usar uma máscara, uma vez que o vapor paraformaldeído é toxic. </li></ol> 3. tecido fibroso Quantificação <ol><li> Colocar o tecido cardíaco fixo-paraformaldeído em um dispositivo de corte de tecido e cortado 2 mm de espessura. Coloque as secções de tecido em uma cassete de incorporação. Desidratar o tecido através de uma série de banhos de álcool graduado (50%, 75%, 95%, e 100% durante 1 h cada). </li><li> Se infiltrar o tecido com xileno, durante 1 hora e finalmente em cera, durante 1 h. Coloque o tecido infiltrado numa cassete de incorporação e incorporar com cera de parafina. Armazenar os tecidos embebidos em blocos de parafina à temperatura ambiente até microtoming. </li><li> Corte o tecido embebido em 4 mm de espessura. Coloque as secções em banho-maria 45 ºC. Mergulhar uma lâmina de vidro para o banho de água a um ângulo e gradualmente se aproxima das bordas parafina secção parcial para permitir a ligação à corrediça. </li><li> Mova o controle deslizante dentro e fora do banho para remover bolsas de ar potenciais sob a secção de tecido e para facilitar uma melhor fixação. Dry tele desliza a 37 ºC durante 1 hora e armazená-los à temperatura ambiente durante coloração histológica. </li><li> Coloque o slide em um tanque. Desparafinar o diapositivo com xileno durante 30 minutos e re-hidratar-o em álcool diluído sequencialmente (95%, 75%, e 50% durante 3 min cada) e finalmente com água destilada. Use solução vermelho de picrosirius suficiente para cobrir completamente as secções de tecido, durante 1 h. Lavar as lâminas em uma solução de ácido acético 0,5% para duas alterações, e depois executar duas lavagens em álcool absoluto. </li><li> Ar seco o slide e montar o slide em resina sintética com uma lamela. Fotografar o slide em um campo de luz visível. NOTA: A área vermelha na fotografia mostra uma zona positiva vermelho picrosirius sob um microscópio com ampliação de 200X. Calcular a percentagem da zona positiva vermelho de picrosirius sobre a área total, o que indica a extensão de fibrose 11. </li></ol> 4. Sangue troponina Quantificação <ol><li> Quantificar os níveis de troponina plasmausando um ensaio de imunossorvente ligado a enzima (ELISA). Ensaiar cada amostra em duplicado. Carga de 50 ul de plasma sanguíneo e de 50 ul de uma mistura de anticorpos nos poços apropriados. Selar a placa e incubar durante 1 hora à temperatura ambiente num agitador de placas a 400 rpm. NOTA: A troponina cardíaca no plasma é um marcador de lesão cardíaca. </li><li> Aspirar o líquido e lavar cada cavidade com 250 uL de tampão de lavagem três vezes. Após a última lavagem, inverter a placa e apagá-lo contra toalhas de papel limpo para remover o líquido em excesso. </li><li> Adicionar 100 ul de substrato de tetrametilbenzidina a cada poço e incubar durante 10 min no escuro, num agitador de placas a 400 rpm. Adicionar 100 ul de solução de paragem a cada poço. Agitar a placa num agitador de placas durante 1 minuto para misturar. Grave a densidade óptica (OD) a 450 nm. NOTA: A concentração de troponina é proporcional ao valor OD. </li></ol>

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The authors have nothing to disclose.

Hypertensive heart disease, a major health problem that contributes greatly to morbidity and mortality, can lead to cardiac hypertrophy and heart failure5. The pathogenesis and progression of hypertensive heart disease in humans is complex, so an appropriate animal model is critical to investigate the underlying mechanisms and to test novel therapeutics that aim to improve cardiac structure and function5. The abdominal aortic constriction model, which simulates chronic heart disease, is an effective...

A insuficiência cardíaca é uma síndrome complexa clínica, cujos sintomas incluem fadiga, dispneia, intolerância ao exercício, e retenção de líquidos nos tecidos periféricos. É a principal causa de morte em países desenvolvidos 1. Além de cardiomiopatia hereditária causada por mutações em proteínas sarcômero ou canais iónicos 2, disfunção miocárdica pode ser causada por uma variedade de condições médicas, incluindo a hipertensão, doenças de coração valvular, obesidade, diabetes e 3. Mudanças na estrutura do miocárdio, condução elétrica, e chumbo metabolismo energético a capacidade inadequada de bombeamento cardíaco para atender às demandas do aparelho circulatório, que resulta em insuficiência cardíaca 3,4. Investigar os mecanismos subjacentes insuficiência cardíaca, portanto, é crítico no domínio da investigação cardiovascular. Identificando os mecanismos moleculares que conduzem à progressão da insuficiência cardíaca, eventualmente, pode auxiliar na identificação de novos alvos terapêuticos ou biomarcadores úteis <s-se&gt; 1. Portanto, é importante para desenvolver modelos de insuficiência cardíaca de animais que compartilham características clínicas principais com insuficiência cardíaca em humanos 5. hipertrofia e remodelação cardíaca desempenha um papel crítico no desenvolvimento de insuficiência cardíaca. Doença cardíaca hipertensiva é o fator de contribuição chave da hipertrofia cardíaca ea remodelação maladaptive visto em pacientes humanos 1. Para imitar essas condições humanas, os modelos animais são muitas vezes estabelecida através de procedimentos cirúrgicos. Em particular, a aorta abdominal transverso ou podem ser constrito para aumentar a resistência contra o ventrículo esquerdo, o que acaba por conduzir a uma sobrecarga de pressão no coração. Este fenómeno resulta geralmente em hipertrofia cardíaca, uma compensação fisiológica dos cardiomiócitos para satisfazer a exigência funcional do sistema cardiovascular. No entanto, a demanda funcional substitui os mecanismos compensatórios fisiológicas normais, levando a fibrose cardíaca e contraçãoimpairment telha. cirurgia transversal constrição aórtica (TAC) muitas vezes envolve procedimentos complicados, incluindo toracotomia, ventilação mecânica, e separação do timo e tecido adiposo do arco aórtico. Em contraste, a constrição da aorta abdominal mais simples requer técnicas experimentais 6-8. A aorta abdominal, entre a esquerda e artérias renais direita, é contraída durante a cirurgia. Hipertrofia e remodelação cardíaca pode ser observado várias semanas após a cirurgia abdominal aórtica constrição 6-8; eles produzem doença cardíaca hipertensa robusta semelhante à gerada pelo transversal cirurgia constrição da aorta 9,10. Aqui, descrevemos um protocolo para conduzir a constrição da aorta abdominal em ratos utilizando um método eficiente, altamente reprodutível e minimamente invasiva. A aorta abdominal adjacente às artérias renais é limitado por um circuito de 0,72 milímetros formado por um fio de seda 4-0. Dez semanas após a cirurgia, hipertrofia cardíaca e remodeling pode ser observada. O modelo de rato de hipertrofia cardíaca induzida por constrição da aorta abdominal fornece uma plataforma para o estudo de mecanismos da doença e fisiopatologia, bem como o desenvolvimento de potenciais terapias.

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a model of cardiac remodeling through constriction of the abdominal aorta in rats

Heart failure is one of the leading causes of death worldwide. It is a complex clinical syndromethat includes fatigue, dyspnea, exercise intolerance, and fluid retention. Changes in myocardial structure, electrical conduction, and energy metabolism develop with heart failure, leading to contractile dysfunction, increased risk of arrhythmias, and sudden death. Hypertensive heart disease is one of the key contributing factors of cardiac remodeling associated with heart failure. The most commonly-used animal model mimicking hypertensive heart disease is created via surgical interventions, such as by narrowing the aorta. Abdominal aortic constriction is a useful experimental technique to induce a pressure overload, which leads to heart failure. The surgery can be easily performed, without the need for chest opening or mechanical ventilation. Abdominal aortic constriction-induced cardiac pathology progresses gradually, making this model relevant to clinical hypertensive heart failure. Cardiac injury and remodeling can be observed 10 weeks after the surgery. The method described here provides a simple and effective approach to produce a hypertensive heart disease animal model that is suitable for studying disease mechanisms and for testing novel therapeutics.

10 semanas após a cirurgia a constrição da aorta abdominal, foi analisada a patologia cardíaca resultante. A histologia cardíaco foi medido através do cálculo da razão entre o peso do coração para o peso corporal e detectando a quantidade de colagénio no coração. lesão cardíaca foi confirmada pela medição da concentração plasmática troponina. Como mostrado na Figura 1A, o tamanho cardíaco ...

Watch this Scientific Journal Video about A Model of Cardiac Remodeling Through Constriction of the Abdominal Aorta in Rats at JoVE.com

A Model of Cardiac Remodeling Through Constriction of the Abdominal Aorta in Rats

A rat model of abdominal aortic constriction that induces cardiac hypertrophy and remodeling is described. An efficient, highly-reproducible, and minimally-invasive method is used to provide a simple yet useful platform for research in myocardial hypertrophy and dysfunction.

Um Modelo de Cardiologia remodela Através constrição da Aorta Abdominal em Ratos

Heart failure is one of the leading causes of death worldwide. It is a complex clinical syndromethat includes fatigue, dyspnea, exercise intolerance, and ...

a-model-cardiac-remodeling-through-constriction-abdominal-aorta

Research

JoVE Journal

Medicine

9.9K visualizações.  National Taiwan University. A rat model of abdominal aortic constriction that induces cardiac hypertrophy and remodeling is described. An efficient, highly-reproducible, and minimally-invasive method is used to provide a simple yet useful platform for research in myocardial hypertrophy and dysfunction.

Vídeo: A Model of Cardiac Remodeling Through Constriction of the Abdominal Aorta in Rats

Chronic Constriction of the Sciatic Nerve and Pain Hypersensitivity Testing in Rats

Chronic neuropathic pain, resulting from damage to the central or peripheral nervous system, is a prevalent and debilitating condition, affecting 7-18% of the population1,2. Symptoms include spontaneous (tingling, burning, electric-shock like) pain, dysaesthesia, paraesthesia, allodynia (pain resulting from normally non-painful stimuli) and hyperalgesia (an increased response to painful stimuli). The sensory symptoms are co-morbid with behavioural disabilities, such as insomnia and depression. To study chronic neuropathic pain several animal models mimicking peripheral nerve injury have been developed, one of the most widely used is Bennett and Xie's (1988) unilateral sciatic nerve chronic constriction injury (CCI)3 (Figure 1). Here we present a method for performing CCI and testing pain hypersensitivity.
CCI is performed under anaesthesia, with the sciatic nerve on one side exposed by making a skin incision, and cutting through the connective tissue between the gluteus superficialis and biceps femoris muscles. Four chromic gut ligatures are tied loosely around the sciatic nerve at 1 mm intervals, to just occlude but not arrest epineural blood flow. The wound is closed with sutures in the muscle and staples in the skin. The animal is then allowed to recover from surgery for 24 hrs before pain hypersensitivity testing begins.
For behavioural testing, rats are placed into the testing apparatus and are allowed to habituate to the testing procedure. The area tested is the mid-plantar surface of the hindpaw (Figure 2), which falls within the sciatic nerve distribution. Mechanical withdrawal threshold is assessed by mechanically stimulating both injured and uninjured hindpaws using an electronic dynamic plantar von Frey aesthesiometer or manual von Frey hairs4. The mechanical withdrawal threshold is the maximum pressure exerted (in grams) that triggers paw withdrawal. For measurement of thermal withdrawal latency, first described by Hargreaves et al (1988), the hindpaw is exposed to a beam of radiant heat through a transparent glass surface using a plantar analgesia meter5,6. The withdrawal latency to the heat stimulus is recorded as the time for paw withdrawal in both injured and uninjured hindpaws. Following CCI, mechanical withdrawal threshold, as well as thermal withdrawal latency in the injured paw are both significantly reduced, compared to baseline measurements and the uninjured paw (Figure 3). The CCI model of peripheral nerve injury combined with pain hypersensitivity testing provides a model system to investigate the effectiveness of potential therapeutic agents to modify chronic neuropathic pain. In our laboratory, we utilise CCI alongside thermal and mechanical sensitivity of the hindpaws to investigate the role of neuro-immune interactions in the pathogenesis and treatment of neuropathic pain.

Due to the simplicity of surgery and the robust behavioural outcome, chronic constriction of the sciatic nerve is one of the pre-eminent animal models of neuropathic pain. Within 24 hrs following surgery, pain hypersensitivity is established and can be quantitatively measured using a von Frey aesthesiometer (mechanical test) and plantar analgesia meter (thermal test).

Constrição crônica do nervo ciático e Teste de Hipersensibilidade dor em ratos

Chronic neuropathic pain, resulting from damage to the central or peripheral nervous system, is a prevalent and debilitating condition, affecting 7-18% of ...

Medicina

Orthotopic Liver Transplantation in Rats

Clinical progress in the field of liver transplantation has been largely supported by animal models1,2. Since the publication of the first orthotopic rat liver transplantation in 1979 by Kamada et al.3, this model has remained the gold standard despite various proposed alternative techniques4. Nevertheless, its broader use is limited by its steep learning curve5.
In this video paper, we show a simple and easy-to-establish revision of Kamada's two-cuff technique. The suprahepatic vena cava anastomosis is performed manually with a running suture, and the vena porta and infrahepatic vena cava anastomoses are performed utilizing a quick-linker cuff system6. Manufacturing the quick-linker kit is shown in a separate video paper.

We present an easy-to-establish revision of the classical two-cuff technique for orthotopic liver transplantation in rat.

Transplante hepático ortotópico experimental em ratos

Clinical progress in the field of liver transplantation has been largely supported by animal models1,2. Since the publication of the first orthotopic rat ...

Ascending Aortic Constriction in Rats for Creation of Pressure Overload Cardiac Hypertrophy Model

Ascending aortic constriction is the most common and successful surgical model for creating pressure overload induced cardiac hypertrophy and heart failure. Here, we describe a detailed surgical procedure for creating pressure overload and cardiac hypertrophy in rats by constriction of the ascending aorta using a small metallic clip. After anesthesia, the trachea is intubated by inserting a cannula through a half way incision made between two cartilage rings of trachea. Then a skin incision is made at the level of the second intercostal space on the left chest wall and muscle layers are cleared to locate the ascending portion of aorta. The ascending aorta is constricted to 50&#8211;60% of its original diameter by application of a small sized titanium clip. Following aortic constriction, the second and third ribs are approximated with prolene sutures. The tracheal cannula is removed once spontaneous breathing was re-established. The animal is allowed to recover on the heating pad by gradually lowering anesthesia. The intensity of pressure overload created by constriction of the ascending aorta is determined by recording the pressure gradient using trans-thoracic two dimensional Doppler-echocardiography. Overall this protocol is useful to study the remodeling events and contractile properties of the heart during the gradual onset and progression from compensated cardiac hypertrophy to heart failure stage.

We describe a stepwise procedure for creating pressure overload and left ventricular hypertrophy in Wistar rats by constriction of the ascending aorta using a small metallic clip. This model is extensively used for studying remodeling changes during cardiac hypertrophy and for identifying and evaluating strategies for regression of such changes.

Aorta ascendente constrição em ratos para a criação de sobrecarga de pressão Cardíaca Hipertrofia Modelo

Ascending aortic constriction is the most common and successful surgical model for creating pressure overload induced cardiac hypertrophy and heart ...

Assessment of Right Ventricular Structure and Function in Mouse Model of Pulmonary Artery Constriction by Transthoracic Echocardiography

Emerging clinical data support the notion that RV dysfunction is critical to the pathogenesis of cardiovascular disease and heart failure1-3. Moreover, the RV is significantly affected in pulmonary diseases such as pulmonary artery hypertension (PAH). In addition, the RV is remarkably sensitive to cardiac pathologies, including left ventricular (LV) dysfunction, valvular disease or RV infarction4. To understand the role of RV in the pathogenesis of cardiac diseases, a reliable and noninvasive method to access the RV structurally and functionally is essential.
A noninvasive trans-thoracic echocardiography (TTE) based methodology was established and validated for monitoring dynamic changes in RV structure and function in adult mice. To impose RV stress, we employed a surgical model of pulmonary artery constriction (PAC) and measured the RV response over a 7-day period using a high-frequency ultrasound microimaging system. Sham operated mice were used as controls. Images were acquired in lightly anesthetized mice at baseline (before surgery), day 0 (immediately post-surgery), day 3, and day 7 (post-surgery). Data was analyzed offline using software.
Several acoustic windows (B, M, and Color Doppler modes), which can be consistently obtained in mice, allowed for reliable and reproducible measurement of RV structure (including RV wall thickness, end-diastolic&#160;and end-systolic dimensions), and function (fractional area change, fractional shortening, PA peak velocity, and peak pressure gradient) in normal mice and following PAC.
Using this method, the pressure-gradient resulting from PAC was accurately measured in real-time using Color Doppler mode and was comparable to direct pressure measurements performed with a Millar high-fidelity microtip catheter. Taken together, these data demonstrate that RV measurements obtained from various complimentary views using echocardiography are reliable, reproducible and can provide insights regarding RV structure and function. This method will enable a better understanding of the role of RV cardiac dysfunction.

Right ventricle (RV) dysfunction is critical to the pathogenesis of cardiovascular disease, yet limited methodologies are available for its evaluation. Recent advances in ultrasound imaging provide a noninvasive and accurate option for longitudinal RV study. Herein, we detail a step-by-step echocardiographic method using a murine model of RV pressure overload.

Avaliação da Estrutura e Função do Ventrículo Direito em modelo de camundongo Pulmonary Artery constrição por Ecocardiografia transtorácica

Emerging clinical data support the notion that RV dysfunction is critical to the pathogenesis of cardiovascular disease and heart failure1-3. Moreover, ...

Visualization of Streptococcus pneumoniae within Cardiac Microlesions and Subsequent Cardiac Remodeling

During bacteremia Streptococcus pneumoniae can translocate across the vascular endothelium into the myocardium and form discrete bacteria-filled microscopic lesions (microlesions) that are remarkable due to the absence of infiltrating immune cells. Due to their release of cardiotoxic products, S. pneumoniae within microlesions are thought to contribute to the heart failure that is frequently observed during fulminate invasive pneumococcal disease in adults. Herein is demonstrated a protocol for experimental mouse infection that leads to reproducible cardiac microlesion formation within 30 hr. Instruction is provided on microlesion identification in hematoxylin &#38; eosin stained heart sections and the morphological distinctions between early and late microlesions are highlighted. Instruction is provided on a protocol for verification of S. pneumoniae within microlesions using antibodies against pneumococcal capsular polysaccharide and immunofluorescent microscopy. Last, a protocol for antibiotic intervention that rescues infected mice and for the detection and assessment of scar formation in the hearts of convalescent mice is provided. Together, these protocols will facilitate the investigation of the molecular mechanisms underlying pneumococcal cardiac invasion, cardiomyocyte death, cardiac remodeling as a result of exposure to S. pneumoniae, and the immune response to the pneumococci in the heart.

Visualization of Streptococcus pneumoniae within Cardiac Microlesions and Subsequent Cardiac Remodeling

Streptococcus pneumoniae forms discrete non-purulent microscopic lesions in the heart. Outlined is the protocol for a murine model of cardiac microlesion formation. Instruction is provided on microlesion visualization using microscopy, discrimination between early and late microlesions, and methods to detect cardiac remodeling in hearts of convalescent animals.

Visualização de<em&gt; Streptococcus pneumoniae</em&gt; Dentro Microle- cardíacas e remodelação cardíaca Subsequente

During bacteremia Streptococcus pneumoniae can translocate across the vascular endothelium into the myocardium and form discrete bacteria-filled ...

Assessment of Cardiac Morphological and Functional Changes in Mouse Model of Transverse Aortic Constriction by Echocardiographic Imaging

Transverse aortic constriction (TAC) in mice has been used as a valuable model to study mechanisms of cardiac hypertrophy and heart failure1. A reliable noninvasive method is essential to assess real-time cardiac morphological and functional changes in animal models of heart disease. Transthoracic echocardiography represents an important tool for noninvasive assessment of cardiac structure and function2. Here we used a high-resolution ultrasound imaging system to monitor myocardial remodeling and heart failure progression over time in a mouse model of TAC. B-mode, M-mode, and Doppler imaging were used to precisely assess cardiac hypertrophy, ventricular dilatation, and functional deterioration in mice following TAC. Color and pulse wave (PW) Doppler imaging was used to noninvasively measure pressure gradient across the aortic constriction created by TAC and to assess transmitral blood flow in mice. Thus transthoracic echocardiographic imaging provides comprehensive noninvasive measurements of cardiac dimensions and function in mouse models of heart disease.

The goal of this protocol is to noninvasively assess cardiac structural and functional changes in a mouse model of heart disease created by transverse aortic constriction, using B- and M-mode echocardiography and color/pulse wave Doppler imaging.

Avaliação da morfológicas e funcionais cardíacas Mudanças no Modelo rato de Transverse aórtica constrição por ecocardiográfica de imagem

Transverse aortic constriction (TAC) in mice has been used as a valuable model to study mechanisms of cardiac hypertrophy and heart failure1. A reliable ...

Balloon-based Injury to Induce Myointimal Hyperplasia in the Mouse Abdominal Aorta

The use of animal models is essential for a better understanding of MH, one major cause for arterial stenosis.In this article, we demonstrate a murine balloon denudation model, which is comparable with established vessel injury models in large animals. The aorta denudation model with balloon catheters mimics the clinical setting and leads to comparable pathobiological and physiological changes. Briefly, after performing a horizontal incision in the aorta abdominalis, a balloon catheter will be inserted into the vessel, inflated, and introduced retrogradely. Inflation of the balloon will lead to intima injury and overdistension of the vessel. After removing the catheter, the aortic incision will be closed with single stiches. The model shown in this article is reproducible, easy to perform, and can be established quickly and reliably. It is especially suitable for evaluating expensive experimental therapeutic agents, which can be applied in an economical fashion. By using different knockout-mouse strains, the impact of different genes on MH development can be assessed.

This article demonstrates a murine model to study the development of myointimal hyperplasia (MH) after aortic balloon injury.

Lesão baseada no balão para induzir a hiperplasia Myointimal da Aorta Abdominal de Mouse

The use of animal models is essential for a better understanding of MH, one major cause for arterial stenosis.In this article, we demonstrate a murine ...

A Closed-chest Model to Induce Transverse Aortic Constriction in Mice

Research on cardiac hypertrophy and heart failure is frequently based on pressure overload mouse models induced by TAC. The standard procedure is to perform a partial thoracotomy to visualize the transverse aortic arch. However, the surgical trauma caused by the thoracotomy in open-chest models changes the respiratory physiology as the ribs are dissected and left unattached after chest closure. To prevent this, we established a minimally invasive, closed chest approach via lateral thoracotomy. Herein we approach the aortic arch via the 2nd intercostal space without entering the chest cavities, leaving the mouse with a less traumatic injury to recover from. We perform this operation using standard laboratory settings for open chest TAC procedures with equal survival rates. Apart from maintaining physiological breathing patterns due to the closed chest approach, the mice seem to benefit by showing rapid recovery, as the less invasive technique appears to facilitate a fast healing process and to reduce immune response after trauma.

Here, we present a protocol of Transverse Aortic constriction (TAC) via a lateral thoracotomy. This technique is a minimally invasive, closed chest surgical procedure aiming to simulate pressure overload and heart failure in mice utilizing standard TAC laboratory settings.

Um modelo de peito fechado para induzir a constrição da aorta transversa em ratos

Research on cardiac hypertrophy and heart failure is frequently based on pressure overload mouse models induced by TAC. The standard procedure is to ...

Ultrasound Imaging of the Thoracic and Abdominal Aorta in Mice to Determine Aneurysm Dimensions

Contemporary high-resolution ultrasound instruments have sufficient resolution to facilitate the measurement of mouse aortas. These instruments have been widely used to measure aortic dimensions in mouse models of aortic aneurysms. Aortic aneurysms are defined as permanent dilations of the aorta, which occur most frequently in the ascending and abdominal regions. Sequential measurements of aortic dimensions by ultrasound are the principal approach for assessing the development and progression of aortic aneurysms in vivo. Although many reported studies used ultrasound imaging to measure aortic diameters as a primary endpoint, there are confounding factors, such as probe position and cardiac cycle, that may impact the accuracy of data acquisition, analysis, and interpretation. The purpose of this protocol is to provide a practical guide on the use of ultrasound to measure the aortic diameter in a reliable and reproducible manner. This protocol introduces the preparation of mice and instruments, the acquisition of appropriate ultrasound images, and data analysis.

Ultrasound imaging has become a common modality to determine the luminal dimensions of thoracic and abdominal aortic aneurysms in mice. This protocol describes the procedure to acquire reliable and reproducible two-dimensional ultrasound images of the ascending and abdominal aorta in mice.

Ecografia da Aorta torácica e Abdominal em ratos para determinar as dimensões de aneurisma

Contemporary high-resolution ultrasound instruments have sufficient resolution to facilitate the measurement of mouse aortas. These instruments have been ...

Implantation of Human-Sized Coronary Stents into Rat Abdominal Aorta Using a Trans-Femoral Access

Percutaneous coronary intervention (PCI), combined with the deployment of a coronary stent, represents the gold standard in interventional treatment of coronary artery disease. In-stent restenosis (ISR) is determined by an excessive proliferation of neointimal tissue within the stent and limits the long-term success of stents. A variety of animal models have been used to elucidate pathophysiological processes underlying in-stent restenosis (ISR), with the porcine coronary and the rabbit iliac artery models being the most frequently used. Murine models provide the advantages of high throughput, ease of handling and housing, reproducibility, and a broad availability of molecular markers. The apolipoprotein E deficient (apoE-/- ) mouse model has been widely used to study cardiovascular diseases. However, stents must be miniaturized to be implanted into mice, involving important changes of their mechanical and (potentially) biological properties. The use of apoE-/- rats can overcome these shortcomings as apoE-/- rats allow for the evaluation of human-sized coronary stents while at the same time providing an atherogenic phenotype. This makes them an excellent and reliable model to investigate ISR after stent implantation. Here, we describe, in detail, the implantation of commercially available human coronary stents into the abdominal aorta of rats with an apoE-/- background using a trans-femoral access.

This protocol describes the implantation of human coronary stents into the abdominal aorta of rats with an apoE-/- background using a trans-femoral access. Compared with other animal models, murine models carry the advantages of high throughput, reproducibility, ease of handling and housing, and a broad availability of molecular markers.

Implantação de stents coronários de tamanho humano na aorta abdominal de rato usando um acesso trans-femoral

Percutaneous coronary intervention (PCI), combined with the deployment of a coronary stent, represents the gold standard in interventional treatment of ...