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).

Tutti gli esperimenti sugli animali sono stati condotti in conformità con la guida per la cura e l&#39;uso di animali da laboratorio, pubblicato dal National Institutes of Health (NIH pubblicazione n. 85-23, rivisto 1996). Questo protocollo è stato approvato dal e in conformità con le linee guida stabilite dalla cura degli animali e del Comitato Istituzionale Usa presso National Taiwan University. 1. Chirurgia Animal <ol><li> Preparare un ago G 22 siringa ottundimento la punta di un ago su una pietra levigatura. Utilizzando pinze, plicate l&#39;ago a un angolo retto. </li><li> Prima della chirurgia, preparare gli strumenti necessari chirurgici e materiali, nonché una gabbia di recupero. Autoclavare tutti gli strumenti e forniture chirurgici prima dell&#39;uso. </li><li> Mantenere i ratti circa 200 g e tenerli sotto cicli di 12 ore luce / buio a temperatura controllata (21 ± 2 ° C), con libero accesso a cibo e acqua. Include almeno 6 topi in ciascun gruppo. Anestetizzare i topi con pentobarbital(75 mg / kg in 0,5 ml, IP) o un altro agente anestetico appropriato. Confermare la profondità dell&#39;anestesia testando il riflesso di coda. NOTA: L&#39;assenza di coda riflesso è un indicatore di un&#39;anestesia adeguata. </li><li> Inserire un topo in posizione supina su una piattaforma intervento chirurgico con una piastra elettrica per mantenere la temperatura corporea. Radere la regione addominale del ratto con un tagliatore di peli e capelli rimozione lozione per evitare contaminazioni chirurgici. Scrub l&#39;addome in modo pulito rasato con betadine o un altro reagente di pulizia prima della chirurgia. NOTA: E &#39;importante mantenere un campo sterile durante tutta la procedura. </li><li> Effettuare una incisione 2 centimetri lungo la linea mediana del ventre con un bisturi. Utilizzando normale soluzione fisiologica, mantenere l&#39;organo addominale umido durante l&#39;intervento chirurgico. Spostare organi digestivi attenzione al lato con batuffoli di cotone per esporre la vena cava inferiore che si trova nella regione peritoneale posteriore. Identificare l&#39;aorta addominale, che giace giustapposta e di solito sullasinistra della vena cava inferiore. NOTA: L&#39;aorta addominale è il vaso che pulsa nel tempo con la frequenza cardiaca. </li><li> Pierce il peritoneo con un paio di pinze per scoprire i vasi al di sotto. isolare delicatamente l&#39;aorta addominale adiacente alle arterie renali e superare un 8 cm a lunga 4-0 seta sutura sotto l&#39;aorta addominale tra le origini del diritto e delle arterie renale sinistra. </li><li> Fare un doppio nodo sciolto con la sutura; lasciare un anello di 3 mm di diametro, e posizionare l&#39;ago 22 G attenuato e piegata all&#39;interno del ciclo. Stringere il nodo intorno alla aorta e l&#39;ago, e poi rimuovere immediatamente l&#39;ago per ottenere un diametro di 0,7 millimetri costrizione. </li><li> Chiudere la cavità addominale con 6-0 materiale di sutura riassorbibile. Suturare le incisioni del muscolo o della pelle con semplici punti staccati. Per prevenire l&#39;infezione, strofinare il sito chirurgico con una tintura di iodio. </li><li> Osservare l&#39;animale accuratamente fino riprende conoscenza sufficiente, come indicato dalla libera circolazionee l&#39;assunzione di cibo. Non lasciare un animale incustodito fino a quando non ha ripreso conoscenza sufficiente a mantenere decubito sternale. Per prevenire il dolore post-operatorio, trattare il ratto con paracetamolo (300 mg / kg a 0,5 ml, ip). NOTA: gli utenti devono somministrare analgesici come approvato dalle politiche istituzionali. </li></ol> 2. tessuti e nel sangue Collection Campione <ol><li> A 10 settimane post-operatorie, pesare il ratto e anestetizzare con pentobarbital (75 mg / kg in 0,5 ml, ip). Prima dell&#39;intervento, confermare la profondità dell&#39;anestesia testando il suo riflesso di coda. Posizionare il ratto su un vassoio di metallo. </li><li> Effettuare una incisione 2 centimetri lungo la linea mediana del collo con un bisturi. Spostare i muscoli con cura con una pinza per esporre la trachea. Osservare attentamente per identificare le arterie carotidi, che sono parallele alla trachea e pulsano nel tempo con la frequenza cardiaca. </li><li> Raccogliere il sangue dalla carotide in un tubo di raccolta di sangue rivestito con EDTA. immediatamente centrifugail sangue per 15 min a 2000 xg e raccogliere il plasma. Conservare il plasma sanguigno a -80 ° C fino al momento dell&#39;uso. </li><li> Fare un cinque centimetri incisione alla regione toracica, intorno alla linea mediana del processo xifoideo. Forare il diaframma con una pinza taglienti. Utilizzando un paio di forbici, tagliare e rimuovere la gabbia toracica lungo le linee emiclaveare su entrambi i lati per esporre il cuore. Asportare cuore con cura lungo i confini cardiache e vascolari. Rimuovere il cuore delicatamente senza afferrare il tessuto. </li><li> Montare il cuore su un apparato di perfusione Langendorff modificato legando il tronco aortico all&#39;ago perfusione. Profumato cuore con tampone Krebs (contenente 110 mM NaCl, KCl 2,6 mm, 1,2 mm KH 2 PO 4, 1,2 mm MgSO 4, 25 mM NaHCO 3, e 11 mm di glucosio [pH 7.4]) per lavare il sangue. Pesare il cuore e calcolare il rapporto cuore-peso-a-corpo-peso. Fissare il cuore con 4% paraformaldeide sul ghiaccio. Ricordatevi di indossare una maschera, dal momento che il vapore paraformaldeide è toxcircuito integrato. </li></ol> 3. Tessuto Fibrosi Quantificazione <ol><li> Posizionare il tessuto cardiaco paraformaldeide-fisso su un dispositivo di sezionamento tessuto e tagliare 2 mm sezioni spesse. Posizionare le sezioni di tessuto in una cassetta embedding. Disidratare il tessuto attraverso una serie di bagni di alcool graduati (50%, 75%, 95% e 100% per 1 ora ciascuno). </li><li> Infiltrati il ​​tessuto con xilene per 1 ora e, infine, in cera per 1 ora. Posizionare il tessuto infiltrato in una cassetta embedding e incorporare con cera di paraffina. Conservare i tessuti incorporati nei blocchi di paraffina a temperatura ambiente fino microtoming. </li><li> Tagliate il tessuto incorporato in 4 sezioni micron di spessore. Place sezioni in un bagno d&#39;acqua di 45 ° C. Immergere un vetrino a bagnomaria in un angolo e avvicinare gradualmente i bordi della sezione di paraffina per consentire fissaggio parziale diapositiva. </li><li> Spostare il vetrino in e fuori del bagno per rimuovere eventuali sacche d&#39;aria sotto la sezione di tessuto e per facilitare una migliore fissaggio. t seccoscivola a 37 ° C per 1 ora e conservare a temperatura ambiente per la colorazione istologica. </li><li> Mettere il vetrino in un serbatoio. Deparaffinare il vetrino con xilene per 30 min e reidratare in sequenza alcool diluito (95%, 75% e 50% per 3 minuti ciascuno) e infine con acqua distillata. Utilizzare un&#39;adeguata soluzione rosso picrosirius per coprire completamente le sezioni di tessuto per 1 ora. Sciacquare i vetrini in una soluzione allo 0,5% di acido acetico per due modifiche, e quindi eseguire due risciacqui in alcool assoluto. </li><li> Asciugare il vetrino e montare il vetrino in resina sintetica con un vetrino. Fotografare il vetrino in un campo di luce visibile. NOTA: L&#39;area rossa nella fotografia mostra una zona positivo rosso picrosirius sotto un microscopio a 200X ingrandimento. Calcolare la percentuale della zona positivo rosso picrosirius sull&#39;intera area, che indica il grado di fibrosi 11. </li></ol> 4. Sangue Troponina Quantificazione <ol><li> Quantificare i livelli di troponina plasmaticautilizzando un saggio di immunoassorbimento enzimatico (ELISA). Analizzare ogni campione in duplicato. Carico 50 ml di plasma sanguigno e 50 ml di un cocktail di anticorpi nei pozzetti appropriati. Sigillare la piastra e incubare per 1 ora a temperatura ambiente su un agitatore a 400 rpm. NOTA: troponina cardiaca nel plasma è un marker per il danno cardiaco. </li><li> Aspirare il liquido e lavare bene ciascuno con 250 microlitri di tampone di lavaggio tre volte. Dopo l&#39;ultimo lavaggio, capovolgere la piastra e asciugare contro tovaglioli di carta puliti per rimuovere il liquido in eccesso. </li><li> Aggiungere 100 pl di substrato tetrametilbenzidina ciascun pozzetto ed incubare per 10 minuti al buio su un agitatore a 400 rpm. Aggiungere 100 ml di soluzione di stop in ogni pozzetto. Agitare la piastra su un agitatore per 1 minuto per mescolare. Registrare la densità ottica (DO) a 450 nm. NOTA: La concentrazione di troponina è proporzionale al valore 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...

L&#39;insufficienza cardiaca è una sindrome complessa clinica, i cui sintomi comprendono stanchezza, dispnea, intolleranza all&#39;esercizio, e ritenzione di liquidi nei tessuti periferici. E &#39;la principale causa di morte nei paesi sviluppati 1. Oltre a cardiomiopatia ereditaria causata da mutazioni in proteine sarcomero o canali ionici 2, disfunzione miocardica può essere causata da una varietà di condizioni mediche, tra cui l&#39;ipertensione, le malattie delle valvole cardiache, l&#39;obesità, il diabete e 3. Cambiamenti nella struttura del miocardio, conduzione elettrica, e il piombo metabolismo energetico per inadeguata capacità di pompaggio cardiaco per soddisfare le esigenze di circolazione, che alla fine si traduce in insufficienza cardiaca 3,4. Indagare i meccanismi alla base di insufficienza cardiaca, di conseguenza, è fondamentale nel campo della ricerca cardiovascolare. L&#39;identificazione dei meccanismi molecolari che portano alla progressione dell&#39;insufficienza cardiaca può eventualmente aiutare nella scoperta di nuovi bersagli terapeutici o biomarcatori utili <sup&gt; 1. E &#39;quindi importante sviluppare modelli animali di insufficienza cardiaca che condividono caratteristiche cliniche fondamentali con insufficienza cardiaca negli esseri umani 5. ipertrofia cardiaca e il rimodellamento svolge un ruolo critico nello sviluppo di insufficienza cardiaca. Cardiopatia ipertensiva è il fattore che contribuisce chiave di ipertrofia cardiaca e il rimodellamento maladaptive visto in pazienti umani 1. Per simulare queste condizioni umane, i modelli animali sono spesso stabiliti attraverso procedure chirurgiche. In particolare, l&#39;aorta addominale trasversale o può essere costretta ad aumentare la resistenza contro il ventricolo sinistro, che alla fine porta ad un sovraccarico di pressione nel cuore. Questo fenomeno provoca solitamente ipertrofia cardiaca, una compensazione fisiologica dei cardiomiociti per soddisfare la domanda funzionale del sistema cardiovascolare. Tuttavia, la domanda funzionale sostituisce i normali meccanismi compensatori fisiologici, che porta alla fibrosi cardiaca e Contraccompromissione delle mattonelle. Trasversale costrizione aortica (TAC), la chirurgia spesso comporta procedure complesse, tra cui toracotomia, la ventilazione meccanica, e la separazione del timo e del tessuto grasso dall&#39;arco aortico. Al contrario, addominale costrizione aortica richiede tecniche sperimentali semplici 6-8. L&#39;aorta addominale, tra la sinistra e la destra arterie renali, è ristretto durante l&#39;intervento chirurgico. Ipertrofia cardiaca e il rimodellamento può essere osservato parecchie settimane dopo l&#39;intervento chirurgico addominale aortico costrizione 6-8; producono cardiopatia ipertensiva robusta simile a quello generato dal trasversale chirurgia costrizione aortica 9,10. Qui, descriviamo un protocollo per condurre addominale costrizione aortica nei ratti usando un metodo efficiente, altamente riproducibile e minimamente invasiva. L&#39;aorta addominale adiacente alle arterie renali è costretto da un anello 0,72 millimetri formato da un filo di seta 4-0. Dieci settimane dopo l&#39;intervento chirurgico, l&#39;ipertrofia cardiaca e remodeling può essere osservato. Il modello di ratto di ipertrofia cardiaca costrizione indotta dell&#39;aorta addominale fornisce una piattaforma per lo studio dei meccanismi di malattia e fisiopatologia, così come lo sviluppo di potenziali terapie.

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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 settimane dopo l&#39;intervento costrizione dell&#39;aorta addominale, la patologia cardiaca risultante è stata analizzata. L&#39;istologia cardiaca è stato misurato calcolando il rapporto tra il peso del cuore al peso corporeo e rilevando la quantità di collagene nel cuore. lesione cardiaca è stata confermata attraverso la misurazione della concentrazione plasmatica di troponina cardiaca. Come mostrato in figur...

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.

Un modello di rimodellamento cardiaco Attraverso Costrizione del Aorta addominale 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 ...

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

Research

JoVE Journal

Medicine

9.9K Views.  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.

Video: 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).

La costrizione cronica del nervo sciatico e Ipersensibilità Testing Pain 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 ...

Ricerca

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.

Trapianto di fegato in ratti

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.

Crescente aortica Costrizione in ratti per la creazione di sovraccarico di pressione cardiaca Ipertrofia Modello

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.

Valutazione del ventricolo destro Struttura e funzione in modello murino di polmonare Arteria Costrizione da ecocardiografia transtoracica

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.

Visualizzazione<em&gt; Streptococcus pneumoniae</em&gt; Entro cardiache microlesioni e successive rimodellamento cardiaco

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.

Valutazione del Cardiac morfologiche e funzionali Variazioni modello murino di trasversale aortica costrizione da parte ecocardiografica 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 ...

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.

Basato su palloncino ferita per indurre iperplasia Myointimal nell'Aorta addominale di 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.

Un modello chiuso-cassa per indurre costrizione trasversale aortica in topi

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.

Formazione immagine di ultrasuono dell'Aorta toracica ed addominale in topi per determinare le dimensioni di Aneurysm

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.

Impianto di stent coronari di dimensioni umane nell'aorta addominale del ratto utilizzando un accesso trans-femorale

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