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).

Tüm hayvan deneyleri ABD Ulusal Sağlık Enstitüleri tarafından yayınlanan Laboratuvar Hayvanları Bakım ve Kullanım için Kılavuzu (NIH yayın no. 85-23, 1996 revize) uyarınca yapılmıştır. Bu protokol ile ve Ulusal Tayvan Üniversitesi Kurumsal Hayvan Bakımı ve Kullanımı Komitesi tarafından belirlenen kurallara uygun olarak kabul edildi. 1. Hayvan Cerrahisi <ol><li> Bir honlama taş üzerinde iğne ucu köreltme ile 22 G iğne şırınga hazırlayın. pense kullanarak, bir dik açı iğne plicate. </li><li> Ameliyattan önce, gerekli cerrahi aletler ve malzemeler, yanı sıra kurtarma kafesini hazırlamak. Kullanmadan önce tüm enstrümanları ve cerrahi malzemeleri otoklavlayın. </li><li> 200 g etrafında fareler korumak ve yiyecek ve suya serbest erişimi olan kontrollü bir sıcaklıkta (21 ± 2 ° C) 12 saat aydınlık / karanlık döngüsü altında tutmak. Her bir grupta en az 6 fareleri içerir. pentobarbital anestezisi fareleriveya başka bir uygun anestezik bir madde (0.5 mi, IP olarak 75 mg / kg). kuyruk refleks testi ile anestezi derinliği onaylayın. Not: yükleme refleksinin yokluğu Yeterli anestezi bir göstergesidir. </li><li> vücut ısısını korumak için, bir ısıtma yastığı ile bir ameliyat platformunda yatar pozisyonda bir sıçan yerleştirin. Cerrahi kirlilikleri önlemek için bir hayvan saç kesme makinesi ve saç çıkarıcı losyon ile sıçan karın bölgesini tıraş. betadin veya ameliyattan önce başka temizlik reaktif ile temiz tıraşlı karın fırçalayın. NOT: işlem boyunca steril bir alan sağlamak için önemlidir. </li><li> Bir neşter ile karın orta hat boyunca 2 cm kesi olun. serum fizyolojik kullanılması, ameliyat sırasında nemli karın organı tutmak. Posterior periton bölgede yatıyor inferior vena kava maruz pamuk topları kullanarak tarafına dikkatlice sindirim organları yerinden. genellikle bitişik ve yalan abdominal aorta, tespitinferior vena kava sol. NOT: abdominal aort kalp hızı ile zaman içinde yanıp söner gemi. </li><li> altındaki damarları ortaya çıkarmak için forseps bir çift ile periton delin. Yavaşça renal arterlerin bitişik abdominal aorta izole ve sağ kökeni ve sol renal arterlerin arasında abdominal aorta altında 8 cm uzunluğunda 4-0 ipek sütür geçmektedir. </li><li> sütür ile gevşek bir çift düğüm yapmak; 3 mm çaplı döngü bırakın ve döngü içinde körleşmiş ve bükülmüş 22 G iğne yerleştirin. aort ve iğne etrafında düğüm sıkın ve ardından 0,7 mm çaplı daralma elde etmek için derhal iğne çıkarılır. </li><li> 6-0 emilebilir sütür materyali ile karın boşluğuna kapatın. Basit sütürler ile kas ya da derideki kesiler dikilmelidir. enfeksiyonu önlemek için, bir iyot tentürü ile cerrahi siteyi fırçalayın. </li><li> yeterli bilinç kavuşur kadar serbest dolaşımı ile belirtildiği gibi, dikkatli bir şekilde hayvan gözlemlemekve gıda alımı. sternal yatma korumak için yeterli bilinci yerine kadar gözetimsiz bir hayvan bırakmayın. ameliyat sonrası ağrıyı önlemek için, asetaminofen (0.5 ml, 300 mg / kg, ip) ile sıçan tedavi. NOT: kurumsal politikalar tarafından onaylanan kullanıcılar analjezikler yönetmek gerekir. </li></ol> 2. Doku ve Kan Örnek Toplama <ol><li> 10 hafta sonrası ameliyat at, sıçan tartmak ve (0.5 ml, ip 75 mg / kg), pentobarbital ile uyutmak. Ameliyattan önce, kuyruk refleks testi ile anestezi derinliği onaylayın. metal tepsi üzerinde sıçan yerleştirin. </li><li> Bir neşter kullanılarak boyun orta hat boyunca 2 cm kesi olun. trakea maruz forseps ile dikkatlice kasları yerinden. trakea paralel ve kalp hızı ile zaman içinde zonklamak karotis arterler, tanımlamak için dikkatle gözlemleyin. </li><li> EDTA ile kaplanmış bir kan toplama tüpüne karotid arterden kan toplamak. hemen santrifüj2,000 x g&#39;de 15 dakika boyunca, kan ve plazma toplama. kadar gerekli -80 ° C kan plazması saklayın. </li><li> xiphoid sürecin orta hat çevresinde, göğüs bölgesinde 5 cm kesi olun. Keskin forseps ile diyaframı delin. makas kullanılarak, kesme ve kalp maruz her iki tarafta orta klaviküler çizgisinde göğüs kafesini çıkarın. kalp ve damar sınırları boyunca dikkatle kalbi tüketim. doku kapma olmadan yavaşça kalp çıkarın. </li><li> perfüzyon iğne aort gövdesini bağlayarak modifiye Langendorff perfüzyon aparatı üzerinde kalbi monte edin. Krebs tamponu ile kalbi serpmek (110 mM NaCl, 2.6 mM KCI içeren 1.2 mM KH 2 PO 4, 1.2 mM MgSO 4, 25 mM NaHCO 3 ve 11 mM glikoz [pH 7.4]) kan yıkamak için. kalbi tartılır ve kalp-ağırlık-vücut-ağırlık oranı hesaplanır. buz üzerinde% 4 paraformaldehid ile kalbi düzeltmek. paraformaldehit buhar Zehir olduğundan, bir maske takmak unutmayınic. </li></ol> 3. Doku Fibrozis Niceleme <ol><li> Bir doku kesit cihazda paraformaldehid sabit kalp dokusu yerleştirin ve 2 mm kalınlığında kesitler kesti. Bir gömme kasetine doku bölümleri yerleştirin. dereceli alkol banyoları (% 50,% 75,% 95, ve 1 saat için her% 100), bir dizi doku kurutmak. </li><li> 1 saat boyunca mum içinde son olarak 1 st için ksilen doku infiltrasyonu ile. Bir gömme kasetine sızmış doku yerleştirin ve parafin mumu ile gömmek. microtoming kadar oda sıcaklığında parafine gömüldü dokuların saklayın. </li><li> 4 mikron kalınlığında kesitler içine gömülü doku dilimleyin. 45 ºC su banyosunda bölümleri yerleştirin. bir açıyla su banyosu içine bir cam slayt batırın ve yavaş yavaş slayt kısmi eki izin parafin kesit kenarlarını yaklaşım. </li><li> kaydırmak ve banyo dışına doku bölümünde olası hava boşluklarını kaldırmak ve daha iyi bağlanmasını kolaylaştırmak için. kuru to 1 saat 37 ºC de slaytlar ve histolojik boyama için oda sıcaklığında saklayın. </li><li> Bir tanka slayt koyun. 30 dakika boyunca ksilen slayt Deparaffinize ve son olarak da damıtılmış su içinde ve (3 dakika her biri için% 95,% 75 ve% 50) ardışık olarak seyreltilmiş alkol içinde rehidre. Tamamen 1 saat doku bölümleri kapsayacak şekilde yeterli picrosirius kırmızı çözümü kullanın. İki değişiklikler için,% 0.5 asetik asit solüsyonu içinde slaytlar durulayın, sonra mutlak alkol eden iki durulama yapmak. </li><li> slayt Hava kurutun ve bir lamel ile sentetik reçine slayt monte edin. Görünür ışık alanında slayt fotoğraf. NOT: fotoğraftaki kırmızı alan 200X büyütmede mikroskop altında bir picrosirius Kırmızı pozitif bölgeyi gösterir. Fibrozis 11 kapsamını gösteren toplam alan üzerinde picrosirius Kırmızı pozitif bölgeye yüzdesini hesaplayın. </li></ol> 4. Kan Troponin Niceleme <ol><li> plazma troponin düzeyleri ölçmekbir enzim-bağlantılı ümmünozorbent deneme (ELISA) kullanılmıştır. iki kopya halinde her bir numune deneyi. Yük, kan plazması, 50 ul uygun kuyulara bir antikor kokteyli 50 ul. plakası kapatılır ve 400 rpm&#39;de bir plaka karıştırıcısı üzerinde oda sıcaklığında 1 saat boyunca inkübe edilir. NOT: plazmada Kardiyak troponin kalp yaralanması için bir göstergedir. </li><li> sıvı aspire ve yıkama tamponu 250 ul üç kez de her yıkayın. Son yıkamadan sonra, plaka ters ve aşırı sıvı kaldırmak için temiz bir kağıt havlu karşı kurulayın. </li><li> Her kuyucuğa tetrametilbenzidin alt-tabaka 100 ul ilave edin ve 400 rpm&#39;de bir plaka karıştırıcısı üzerinde karanlıkta 10 dakika süreyle inkübe edilir. Her bir oyuğa durdurma çözeltisi 100 ul ekle. 1 dakika karıştırmak için, bir plaka karıştırıcısı üzerinde plakayı sallayın. 450 nm&#39;de optik yoğunluk (OD) kaydedin. NOT: troponin konsantrasyonu OD değeri ile orantılıdır. </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...

Kalp yetmezliği periferik dokularda bir yorgunluk, nefes darlığı dahil olduğu belirtileri, egzersiz intoleransı karmaşık klinik sendrom, ve sıvı tutulması olduğunu. Bu gelişmiş ülkelerde 1 önde gelen ölüm nedenidir. Kenara sarkomer protein ya da iyon kanalları 2 mutasyonlardan kaynaklanır miras kardiyomiyopati, miyokard disfonksiyonu hipertansiyon, kalp kapak hastalıkları, obezite ve diyabet 3 dahil olmak üzere tıbbi koşulları, çeşitli neden olabilir. Yetersiz kardiyak pompalama kapasitesine miyokard yapısı, elektrik iletim ve enerji metabolizması kurşun değişiklikler sonuçta kalp yetmezliği 3,4 sonuçlanır dolaşım taleplerini karşılamak için. kalp yetmezliği altında yatan mekanizmaları araştıran, bu nedenle, kardiyovasküler araştırma alanında kritik öneme sahiptir. kalp yetmezliği ilerlemesi için önde gelen moleküler mekanizmaları belirleme sonunda yeni tedavi hedefleri ya da yararlı biyomarkerların keşfi yardımcı olabilir <s&gt; 1 kadar. İnsanlarda 5 kalp yetmezliği olan temel klinik özellikleri paylaşan kalp yetmezliği hayvan modelleri geliştirmek için önemlidir. Kardiyak hipertrofi ve yeniden şekillenme kalp yetmezliği gelişiminde kritik bir rol oynamaktadır. Hipertansif kalp hastalığı, kalp hipertrofisi ve insan hastalarda 1 görülen uyumsuz biçimlenme anahtar bir faktör olduğunu. Bu insan koşullarını taklit etmek için, hayvan modelleri genellikle cerrahi prosedürler aracılığıyla kurulur. Özellikle, enine ya da abdominal aorta sonuçta kalbinde bir basınç yükü neden sol ventrikül karşı direncini artırmak için daraltıldığı edilebilir. Bu olgu, genellikle kardiyak hipertrofiye kardiyomiyositlerinin fizyolojik tazminat kardiyovasküler sistemin fonksiyonel talebi karşılamak için olur. Bununla birlikte, fonksiyonel talep kardiyak fibroz ve CONTRAC yol normal fizyolojik telafi edici mekanizmalar geçersizkaro bozukluğu. Enine aort daralma (TAC) cerrahi genellikle aortik arktan torakotomi, mekanik ventilasyon ve timus ayrılması ve yağ dokusu dahil karmaşık prosedürleri kapsar. Buna karşılık, abdominal aort daralma basit deneysel teknikler 6-8 gerektirir. abdominal aorta, sol ve sağ renal arter arasında, ameliyat sırasında dar olduğunu. Kardiyak hipertrofi ve yeniden şekillenme abdominal aort daralma cerrahisi 6-8 birkaç hafta sonra görülebilir; Onlar enine aort daralma cerrahisi 9,10 tarafından üretilen benzer sağlam hipertansif kalp hastalığı üretirler. Burada, etkin, yüksek tekrarlanabilir ve minimal invaziv bir yöntem kullanılarak sıçanlarda abdominal aort daralma yapmak için bir protokol açıklar. renal arterlerin bitişik abdominal aorta 4-0 ipek iplikle oluşan bir 0.72 mm döngü tarafından dar edilir. On hafta cerrahi, kalp hipertrofisi ve remodelin sonrag gözlenebilir. Abdominal aort daralma kaynaklı kalp hipertrofisi sıçan modelinde hastalık mekanizmalarını ve patofizyolojisi yanı sıra potansiyel tedavilerin gelişimini incelemek için bir platform sağlar.

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			<td style="width:119px;">ML870B2</td>
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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 hafta abdominal aort daralma cerrahisi sonrası ortaya çıkan kardiyak patoloji analiz edildi. Kalp histoloji vücut ağırlığına, kalp ağırlığına oranının hesaplanması ile ve kalbin kollajen miktarı tespit edilerek ölçülür. Kalp yaralanma plazma troponin konsantrasyonunun ölçülmesiyle doğrulandı. Şekil 1A&#39;da gösterildiği gibi, daha yüksek bir kalp ağırlık-vücut ağırlık ...

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.

Sıçanlarda Karın Aort Konstrüksiyon sayesinde Kardiyak Yaptırma Bir Model

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 Görüntüleme.  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).

Siyatik Sinir Sıçanlarda Ağrı aşırı duyarlılık Test Kronik daraltma

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

Tıp

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.

Sıçanlarda Ortotopik Karaciğer Nakli

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.

Basınç Aşırı Kardiyak hipertrofi Modeli Oluşturma Ratlarda Aort daralma, Artan

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.

Transtorasik Ekokardiyografi ile Pulmoner Arter Konstrüksiyon Fare Modeli sağ ventrikül yapı ve fonksiyonlarının değerlendirilmesi

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.

Görselleştirme<em&gt; Streptococcus pneumoniae</em&gt; Kalp Microlesions ve sonraki Kardiyak Tadilat içinde

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.

Ekokardiyografik Imaging tarafından Enine Aort Konstrüksiyon Mouse Modeli Kardiyak Morfolojik Değerlendirilmesi ve Fonksiyonel Değişiklikler

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.

Fare Abdominal aort Myointimal Hiperplazi ikna etmek için balon tabanlı yaralanma

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.

Transvers aort daralma farelerde ikna etmek için kapalı-göğüs manken

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.

Torasik ve Abdominal aort anevrizması boyutlarını belirlemek farelerde ultrason görüntüleme

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.

Trans-Femoral Erişim Kullanılarak İnsan Büyüklüğünde Koroner Stentlerin Rat Abdominal Aort'a yerleştirilmesi

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