Yazarlar trikrom boyama ve analizi ile yardım için Washington Üniversitesi Histoloji Brian Johnson ve Görüntüleme Çekirdek teşekkür ederiz. Bu çalışma NIH hibe HL098067 ve HL089455 tarafından desteklenmiştir.

Washington ve Cedars-Sinai Tıp Merkezi Üniversitesi Kurumsal Hayvan Bakım ve Kullanım Komiteleri (IACUC) bu çalışmalar için gerekli hayvan çalışmaları onayladı. 1. Hazırlık <ol><li> Künt uç forseps ve basacağı yoluyla otoklav hem sterilize. </li><li> biyolojik güvenlik kabini kullanarak, liyofilize toz PBS içinde BLM bir çalışma stok hazırlamak. Karıştırma daha emin olmak için 35 kHz&#39;de 10 dakika süre ile çözelti sonikasyon. Not: ul 30 ile 45 toplam hacmi oldukça büyük hacimli pipet düşük ucunda varyasyon ve boğulma önlemek için tavsiye edilir. </li><li> Yaklaşık 1 prosedürü kendisi için m 2, yanı sıra işlem öncesi ve sonrası her iki kafesleri için belirlenen yerleri içeren temiz bir çalışma alanı hazırlayın. </li><li> taban ve Underl genelinde laboratuvar bant 2 ya da 3 şerit koyarak hemen araştırmacı önünde tezgah prosedür kurulu tabanını Fixtezgah ying. Bir tahta oluşturma konusunda daha fazla özellikleri için Bkz: Şekil 1. </li><li> prosedür kurulu iki konumlandırma vida arasındaki büyüklük 4.0 dikiş ipliğinin tek uzunluğunu kravat. </li><li> kaldırılması ve üç 1 ml&#39;lik şırınga pistonu atarak ve hava geçirmez bir mühür formu her varil üst içine PBS 60 ul yatırarak derme çatma bir spirometre oluşturun. gevşek şırınga birine kateter göbeğini Güvenli ve yönetim kurulu bir tarafa koyun. </li><li> 1 ml şırınganın içine havanın aspire 300 ul ve yönetim kurulu bir tarafa koyun. </li><li> bir tarafa bant uzunluğu ve yerinde yaklaşık 6 inç ek bir parça kesin. Bu aşamada 2.4 kuruluna hayvan sabitlemek için kullanılır. </li><li> Bir izofluran odasını kurmak. Pozlama odasına ve boşluk vakum hem de uygun portlara O 2, izofluran ve vakum takın. Alternatif olarak, anestezik yönetmek bir izofluran uyumlu biyolojikgüvenlik kabini. </li></ol> 2. Entübasyon <ol><li> o bilincini kaybeder ve solunum uygun bir orana yavaşlatır kadar odasında izofluran ile fare anestezisi. Tipik bir pozlama 3-4 dakika süreyle% 4 izofluran ve% 2 O 2 içerir ve ideal bir sonuç sedasyon 2 ila 2,5 dk. Bu 1 nefes her 2 sn bir solunum oranına karşı gelmektedir. </li><li> bir tarafa bir pipet ve yerine BLM 30 ve 45 ul arasında ayarlamak için sedasyon, aspirat için beklerken. </li><li> Hazır olduğunda, işlem platformu konumlandırma vida bağlı akıştan üst kesici dişlerin tarafından sedasyon fare askıya. Hayvanın sırtı platformu yüzeye karşı düz durduğundan emin olun. </li><li> havalandırma kısıtlamak için dikkatli olmak, sadece diyafram üstünde, gevşek göğüs boşluğunun alt (kaudal) kısmı boyunca bir parça bant yerleştirin. Yerleşim proc sırasında düzgün hizalama korumak için sıkı yeterli olmalıdıredure, ancak solunumu kısıtlar o kadar sıkı değil. </li><li> 80 ile% ve% 100 yoğunluğuna aydınlatıcı açın ve solar plexus yakın cilt yüzeyinden 1 2 cm, böylece deveboynu yönlendirmek. Periyodik olarak fare zarar vermez emin olmak için ısı deveboynu ucu kontrol edin. </li><li> platformun arkasında duran, dil bulmak için steril, künt uç forseps kullanabilir. Alt kesici dişler, yavaşça kavrama önlemek ve ağız boşluğunun dışında dilini çekmek için dikkatli olmak. </li><li> Kalan elinizi kullanarak, basacağı yerleştirin ve ağız boşluğunun zemin karşı dilini düzleştirmek için kullanabilirsiniz. forseps bırakın, ancak sonraki iki aşama için yerinde basacağı bırakın. </li><li> trakea o Ana kök bronşların seviyesine ulaşıncaya kadar güneş pleksus seviyesinden proksimal deveboynu rehberlik ederek görünür böylece ışığı yönlendirmek. Not: trakea kolayca solunum aksiyonu ile ayırt edilebilir, WHIch şiddeti dalgalanmaya yayılan ışığı neden olur. Doğru yerleştirilmiş, bu yapının ağız boşluğunda kendisi minimal ortam ışığı ile ışık merkezi bir pim gibi eksenel düzlemde ayırt olacaktır. </li><li> Açı düz lümenine, damlacık içeren ekli şırınga ile, trakea doğal yolu izler ve 22-G kateter ucu düşürmek, böylece şırınga. PBS kabarcık yükselişi ve başarılı yerleştirme üzerine her nefeste düşmeye başlayacaktır. Not: Bu eylem, derin sedasyon sonucunda birkaç saniye kadar gecikebilir. </li><li> ek 5 mm kateter besleyin. dil çıkarın depressor. </li><li> yavaşça şırınga kaldırmak, ters el şırınga Shift ve göbek sürükleyici. </li><li> Kateter hub iç merkezi haline BLM 30 ve 45 ul arasındaki mevduat, ikinci şırınga takın ve göbek içine hava 300 ul dağıtmak. </li><li> ikinci değiştirinİlk PBS balonu içeren şırınga. kabarcık yükselişi ve işlemi başarıyla gerçekleştirildi olup olmadığını düşmeye devam edecektir. </li></ol> 3. Post-prosedürel Bakımı <ol><li> kateter ve bandı çıkarın ve bilinci yerine gelene kadar bir kuru ılık bir yerde hayvan yerleştirin - tipik birkaç dakika içinde. </li></ol>

<ol>
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The authors have nothing to disclose.

Aerosol haline getirme sınırlı olması nedeniyle reaktif temin edilebilirliği, emniyeti ve maliyet için pratik örneklerde, doğrudan nefes borusu idaresi akciğerlere dışsal ajanların uygulanması için üstün bir yöntemdir 16 Transtrakeal instilasyon yaygın Bunu gerçekleştirmek için kullanılmaktadır.; Ancak, tüm cerrahi müdahale ile olduğu gibi, o da onunla prosedürün kendisinden kaynaklanan komplikasyonlar için potansiyel taşır ve mutlaka ajan telkin ediliyor. 13 ...

Bazı anatomik ve fizyolojik farklılıklar olmasına rağmen, 1 fare modelleri hayvancılık açısından bakıldığında, insan biyolojisi ve hastalık patogenezi. 2 modelleme için çok değerli, fareler, kullanımı kolay olan, düşük üreme zamanı, hızlandırılmış ömrü var ve nispeten ucuzdur devam eve. Farklı genetik suşları ve stratejiler (örn., Koşullu knock-out raportör fareler, soy izleme yaklaşımlar, vs.), ve bunun yanı sıra reaktifler geniş (örn., Antikorlar, rekombinant proteinler, inhibitörlerin geliştirilmesi ile, vb), fareler, insan homeostasis ve hastalık süreçlerini ortaya çıkarmak için önemli bir örnek omurgalı organizma haline gelmiştir. 3 Fareler insanlarda 4 ALİ travma, yaralanma veya sepsis neden olabilir. Akut akciğer hasarı (ALI) ve pulmoner fibrozis dahil olmak üzere akciğer koşulları, çalışmak için özellikle değerli olmuştur ve epitel ile karakterizedir veendotel sızıntı (yani., ödem), enflamasyon ve oluşmakta olan fibroz. Birçok hastada, ALI genellikle solunum yetmezliği nedeni ile fibroz ve ölümle sonuçlanan onun ağır formu, akut solunum sıkıntısı sendromu (ARDS), ilerler. 5,6 pulmoner fibrozis ekstraselüler matriksin aşırı birikmesi ile karakterize ilerleyici, ölümcül patolojidir en önemlisi. Ben engelli akciğer fonksiyonu yol açan kolajen bleomisin (BLM) 7,8 Yönetim yazın deney hayvanlarında ALI ve fibroz uyarmak için en yaygın kullanılan ve en iyi karakterize modeldir. 9 kemirgenlerde pulmoner fibrozis BLM kaynaklı olsa da yapar tamamen insan fibrotik fenotipleri, bu model ile 10 fare çalışmaları başlangıcı ve hastalığın ilerlemesini etkileyen birçok önemli faktörlerden keşfine yol açmıştır özetlemek değil. 11 BLM kaynaklı fibrogenezisin arkasında kesin mekanizması (ler) bilinmemekle birlikte, başlangıç ​​hasarınıniletken hava yolları ve alveolleri ve özellikle de tip 1 pnömositler döşeyen epitel hücrelerinde kontak bağımlı DNA zincir kırıkları kaynaklandığı düşünülmektedir. 12 BLM ve pulmoner epitel arasında doğrudan temas ihtiyacı sağlam bir dağıtım yolunun önemini vurgulamaktadır ve bu endişeleri de rekombinant proteinlerin, antikorların, siRNA, virüs, bakteri, partikül, ve daha fazlasını içeren distal hava hedeflenen tedavilerin geniş bir yelpazede, germane vardır. Orofaringeal aspirasyon (OPA), yaygın olarak, bu amaçla 13 kullanılmaktadır, ancak OPA önemli bir eksiklik teslim maddenin bir bölümü bu şekilde uygulanan dozun belirginsizlik yol mide bağırsak Yutulan edilebilir olmasıdır. Yaygın olarak kullanılan bir yaklaşım, doğrudan solunum yollarına bir ajanın trakea ve damlatma maruz güçlü anestezi altında trakeostomi gerektirir transtrakeal damlatma vardır. 14 Ancak, sadece bu tür olabilirBir prosedür nedeniyle invaziv olması istenmeyen olabilir, ama o da zaman tüketen bir eğitim adil biraz gerektirir ve içine ajanlar direkt uygulanmasını içerir. 15,16 Çeşitli protokoller geliştirilmiştir solunum yolu güçlü bir yaralanmaya neden cerrahi müdahaleye gerek kalmadan trakea, 16,17,18,19,20 ancak bu yöntemler geniş bir alanını içermektedir kurtarma süreleri güçlü anestezik neden, pahalı ekipman kullanımı (yani., otoskop / laringoskop, ticari olarak mevcut prosedür panoları, fiber-optik doz ile ilgili olarak tel ve benzeri), ağız boşluğu içinde manipülasyon aşırı ve belirsizlik. Bu çalışma, bir araştırmacının hızla ucuz ve güvenilir bir şekilde çevre dokulara bakiye hasar sınırlı risk ile murin akciğer içine bir reaktif aşılamak sağlar entübasyon yoluyla uygulama nispeten kolay bir yöntem tarif etmektedir.

<table><tbody><tr><td>Bleomycin For Injection, 30 units/vial</td><td>APP Pharmaceuticals, LLC</td><td>103720</td><td>For best results, BLM should be suspended in PBS, aliquoted, and stored as single use lyophilzed aliquots</td></tr><tr><td>Blunt End Forceps</td><td></td><td></td><td></td></tr><tr><td>Tongue Depressor (i.e. bent Valleylab Blade Electrode, 2.4&quot;)&amp;nbsp;</td><td>Covidien</td><td>E1551G</td><td>Before use, create a 45 degree bend 1.5 cm&amp;nbsp; from the blade tip. A suitable depressor can also be created from any metal implement of similar dimensions.&amp;nbsp;</td></tr><tr><td>Exel Safelet Catheter 22G X 1&quot;</td><td>Exel International</td><td>26746</td><td></td></tr><tr><td>1 ml Slip-tip Disposable Tuberculin Syringe (200/sp, 1600/ca)</td><td>BD</td><td>309659</td><td></td></tr><tr><td>0.2 ml Pipettor and Filter Tips</td><td></td><td></td><td></td></tr><tr><td>Fiber-Lite Illuminator. Model 181-1: Model 180 mated with Standard Dual Gooseneck illuminator</td><td>Dolan Jenner Industries, Inc.</td><td>181-1</td><td>Lower output LED illuminators are not recommended as they fail to suficiently illuminate the trachea.</td></tr><tr><td>Intubation Board</td><td></td><td></td><td>See Diagram 1.</td></tr><tr><td>Colored Label Tape: 0.5&amp;nbsp;in. Wide</td><td>Fisherbrand</td><td>15-901-15A</td><td></td></tr><tr><td>Oxygen&amp;nbsp;</td><td></td><td></td><td></td></tr><tr><td>Phosphate-Buffered Saline, 1X</td><td>Corning</td><td>21-040-CV</td><td>Product should be sterile</td></tr><tr><td>Non-Sterile Silk Black Braided Suture Spool, 91.4 m, Size 4-0</td><td>Harvard Apparatus</td><td>517698</td><td></td></tr><tr><td>Table Top Anesthesia Machine Isoflurane</td><td>Highland Medical Equipment</td><td></td><td>http://www.highlandmedical.net/</td></tr><tr><td>Slide Top Induction Mouse Isoflurane Chamber</td><td>MIP / Anesthesia Technologies</td><td>AS-01-0530-SM</td><td></td></tr><tr><td>FORANE (isoflurane, USP) Liquid For Inhalation 100 mL&amp;nbsp;</td><td>Baxter</td><td>1001936040</td><td></td></tr><tr><td>&amp;nbsp;Nanozoomer Digital Pathology system</td><td>Hamamatsu</td><td></td><td></td></tr><tr><td>IgM ELISA Quantification Kit&amp;nbsp;</td><td>Bethyl Laboratories</td><td>E90-101</td><td></td></tr></tbody></table>

an improved method for rapid intubation of the trachea in mice

Despite some anatomical and physiological differences, mouse models continue to be an essential tool for studying human lung disease. Bleomycin toxicity is a commonly used model to study both acute lung injury and fibrosis, and multiple methods have been developed for administering bleomycin (and other toxic agents) into the lungs. However, many of these approaches, such as transtracheal instillation, have inherent drawbacks, including the need for strong anesthetics and survival surgery. This paper reports a quick, reproducible method of intratracheal intubation that involves mild inhaled anesthesia, visualization of the trachea, and the use of a surrogate spirometer to confirm exposure. As a proof of concept, 8-12 week old C57BL/6 mice were administered either 2.0 U/kg of bleomycin or an equivalent volume of PBS, and both damage and fibrotic endpoints were measured post-exposure. This procedure allows researchers to treat a large cohort of mice in a relatively short period with little expense and minimal post-procedure care.

Entübe farenin kilo kaybı ve sıkıntı için günlük olarak izlenmiş ve% 2.5 2,2,2-tribromoetanol intraperitonal enjeksiyon yoluyla 4, 10 ya da 17 gün sonra feda edilmiştir. Bronkoalveoler lavaj (BAL) PBS üç yıkamadan içinde toplandı 21 başka bir yerde tarif edildiği gibi ve sağ akciğer,% 10 formalin sabit parafine gömülü ve Washington Histoloji ve Görüntüleme Çekirdek 22 Üniversitesi tarafından Masson trikrom ile boyandı....

Watch this Scientific Journal Video about An Improved Method for Rapid Intubation of the Trachea in Mice at JoVE.com

An Improved Method for Rapid Intubation of the Trachea in Mice

Farelerde Trakea hızlı intübasyon için geliştirilmiş bir yöntem

Bu makalede, entübasyon yoluyla fare trakea içine doğrudan bleomisin yönetmek için hızlı ve basit bir yöntem sunar. Bu yöntemin en önemli avantajları usta son derece tekrarlanabilir kolay ve özel ekipman ya da uzun iyileşme süreleri gerektirmez vardır.

Despite some anatomical and physiological differences, mouse models continue to be an essential tool for studying human lung disease. Bleomycin toxicity ...

an-improved-method-for-rapid-intubation-of-the-trachea-in-mice

Research

JoVE Journal

Medicine

25.1K Görüntüleme.  University of Washington.  Bu makalede, entübasyon yoluyla fare trakea içine doğrudan bleomisin yönetmek için hızlı ve basit bir yöntem sunar. Bu yöntemin en önemli avantajları usta son derece tekrarlanabilir kolay ve özel ekipman ya da uzun iyileşme süreleri gerektirmez vardır. 

Video: An Improved Method for Rapid Intubation of the Trachea in Mice

Pressure Controlled Ventilation to Induce Acute Lung Injury in Mice

Murine models are extensively used to investigate acute injuries of different organs systems (1-34). Acute lung injury (ALI), which occurs with prolonged mechanical ventilation, contributes to morbidity and mortality of critical illness, and studies on novel genetic or pharmacological targets are areas of intense investigation (1-3, 5, 8, 26, 30, 33-36). ALI is defined by the acute onset of the disease, which leads to non-cardiac pulmonary edema and subsequent impairment of pulmonary gas exchange (36). We have developed a murine model of ALI by using a pressure-controlled ventilation to induce ventilator-induced lung injury (2). For this purpose, C57BL/6 mice are anesthetized and a tracheotomy is performed followed by induction of ALI via mechanical ventilation. Mice are ventilated in a pressure-controlled setting with an inspiratory peak pressure of 45 mbar over 1 - 3 hours. As outcome parameters, pulmonary edema (wet-to-dry ratio), bronchoalveolar fluid albumin content, bronchoalveolar fluid and pulmonary tissue myeloperoxidase content and pulmonary gas exchange are assessed (2). Using this technique we could show that it sufficiently induces acute lung inflammation and can distinguish between different treatment groups or genotypes (1-3, 5). Therefore this technique may be helpful for researchers who pursue molecular mechanisms involved in ALI using a genetic approach in mice with gene-targeted deletion.

A murine model for ventilator induced lung injury is an important tool to study an acute lung injury in vivo. Here, we report an easy applicable in situ model for acute lung injury using high-pressure mechanical ventilation to induce acute failure of the lung.

Farelerde Akut Akciğer Hasarı tetikleyebilecek Basınç Kontrollü Havalandırma

Murine models are extensively used to investigate acute injuries of different organs systems (1-34). Acute lung injury (ALI), which occurs with prolonged ...

Tıp

Endotracheal Intubation in Mice via Direct Laryngoscopy Using an Otoscope

Mice, both wildtype and transgenic, are the principal mammalian model in biomedical research currently. Intubation and mechanical ventilation are necessary for whole animal experiments that require surgery under deep anesthesia or measurements of lung function. Tracheostomy has been the standard for intubating the airway in these mice to allow mechanical ventilation. Orotracheal intubation has been reported but has not been successfully used in many studies because of the substantial technical difficulty or a requirement for highly specialized and expensive equipment. Here we report a technique of direct laryngoscopy using an otoscope fitted with a 2.0 mm speculum and using a 20 G&#160;intravenous catheter as an endotracheal tube. We have used this technique extensively and reliably to intubate and conduct accurate assessments of lung function in mice. This technique has proven safe, with essentially no animal loss in experienced hands. Moreover, this technique can be used for repeated studies of mice in chronic models.

Endotracheal Intubation in Mice via Direct Laryngoscopy Using an Otoscope

We have developed a simple, reliable, and relatively inexpensive method for endotracheal intubation in mice via direct laryngoscopy using an otoscope with a 2.0 mm speculum. This technique is atraumatic and can be used for repeated measurements in chronic experiments. We find it superior to tracheostomy or previously reported nonsurgical techniques.

Farelerde Endotrakeal Entübasyon<em&gt; Via</emBir Otoskop Kullanma&gt; Doğrudan Larengoskopi

Mice, both wildtype and transgenic, are the principal mammalian model in biomedical research currently. Intubation and mechanical ventilation are ...

A Simple Method of Mouse Lung Intubation

A simple procedure to intubate mice for pulmonary function measurements would have several advantages in longitudinal studies with limited numbers or expensive animal. One of the reasons that this is not done more routinely is that it is relatively difficult, despite there being several published studies that describe ways to achieve it. In this paper we demonstrate a procedure that eliminates one of the major hurdles associated with this intubation, that of visualizing the trachea during the entire time of intubation. The approach uses a 0.5 mm fiberoptic light source that serves as an introducer to direct the intubation cannula into the mouse trachea. We show that it is possible to use this procedure to measure lung mechanics in individual mice over a time course of at least several weeks. The technique can be set up with relatively little expense and expertise, and it can be routinely accomplished with relatively little training. This should make it possible for any laboratory to routinely carry out this intubation, thereby allowing longitudinal studies in individual mice, thereby minimizing the number of mice needed and increasing the statistical power by using each mouse as its own control.

This paper describes a striaghforward and efficient method of intubating mice for pulmonary function measurements or pulmonary instillation, that allows the mice to recover and be studied at later times. The procedure involves an inexpensive fiberoptic light source that directly illuminates the trachea.

Fare Akciğer Entübasyonun Basit Bir Yöntem

A  simple procedure to intubate mice for pulmonary function measurements would  have several advantages in longitudinal studies with limited numbers or  ...

Noninvasive Intratracheal Intubation to Study the Pathology and Physiology of Mouse Lung

The use of a model that mimics the condition of lung diseases in humans is critical for studying the pathophysiology and/or etiology of a particular disease and for developing therapeutic intervention. With the increasing availability of knockout and transgenic derivatives, together with a vast amount of genetic information, mice provide one of the best models to study the molecular mechanisms underlying the pathology and physiology of lung diseases. Inhalation, intranasal instillation, intratracheal instillation, and intratracheal intubation are the most widely used techniques by a number of investigators to administer materials of interest to mouse lungs. There are pros and cons for each technique depending on the goals of a study. Here a noninvasive intratracheal intubation method that can directly deliver exogenous materials to mouse lungs is presented. This technique was applied to administer bleomycin to mouse lungs as a model to study pulmonary fibrosis.

The use of a model that mimics the condition of lung diseases in humans is critical for studying the pathophysiology and/or etiology of a particular disease and for developing therapeutic intervention. Here a noninvasive intratracheal intubation method that can directly deliver exogenous materials to mouse lungs is presented.

Noninvaziv Entübasyon Fare Akciğer Patolojisi ve Fizyolojisi Eğitim için

The use of a model that mimics the condition of lung diseases in humans is critical for studying the pathophysiology and/or etiology of a particular ...

A Reversible, Non-invasive Method for Airway Resistance Measurements and Bronchoalveolar Lavage Fluid Sampling in Mice

Airway hyperreactivity (AHR) measurements and bronchoalveolar lavage (BAL) fluid sampling are essential to experimental asthma models, but repeated procedures to obtain such measurements in the same animal are generally not feasible.  Here, we demonstrate protocols for obtaining from mice repeated measurements of AHR and bronchoalveolar lavage fluid samples.  Mice were challenged intranasally seven times over 14 days with a potent allergen or sham treated.  Prior to the initial challenge, and within 24 hours following each intranasal challenge, the same animals were anesthetized, orally intubated and mechanically ventilated.  AHR, assessed by comparing dose response curves of respiratory system resistance (RRS) induced by increasing intravenous doses of acetylcholine (Ach) chloride between sham and allergen-challenged animals, were determined.  Afterwards, and via the same intubation, the left lung was lavaged so that differential enumeration of airway cells could be performed.  These studies reveal that repeated measurements of AHR and BAL fluid collection are possible from the same animals and that maximal airway hyperresponsiveness and airway eosinophilia are achieved within 7-10 days of initiating allergen challenge.  This novel technique significantly reduces the number of mice required for longitudinal experimentation and is applicable to diverse rodent species, disease models and airway physiology instruments.  

Repeated measurements of rodent respiratory physiology and sampling of airway inflammatory cells are desirable, but generally not feasible. Here we describe a repeatable method for orally intubating mice that permits repeated measurements of airway hyperreactivity and sampling of airway inflammatory cells.

Havayolu Direnci Ölçümleri ve Fare Bronkoalveolar Lavaj Sıvısı Örnekleme için, geri dönüşümlü, Non-invaziv Yöntemi

Airway hyperreactivity (AHR) measurements and bronchoalveolar lavage (BAL) fluid sampling are essential to experimental asthma models, but repeated ...

Biyoloji

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 ...

A Mouse Model of Orotracheal Intubation and Ventilated Lung Ischemia Reperfusion Surgery

Ischemia reperfusion (IR) injury frequently results from processes that involve a transient period of interrupted blood flow. In the lung, isolated IR permits the experimental study of this specific process with continued alveolar ventilation, thereby avoiding the compounding injurious processes of hypoxia and atelectasis. In the clinical context, lung ischemia reperfusion injury (also known as lung IRI or LIRI) is caused by numerous processes, including but not limited to pulmonary embolism, resuscitated hemorrhagic trauma, and lung transplantation. There are currently limited effective treatment options for LIRI. Here, we present a reversible surgical model of lung IR involving first orotracheal intubation followed by unilateral left lung ischemia and reperfusion with preserved alveolar ventilation or gas exchange. Mice undergo a left thoracotomy, through which the left pulmonary artery is exposed, visualized, isolated, and compressed using a reversible slipknot. The surgical incision is then closed during the ischemic period, and the animal is awakened and extubated. With the mouse spontaneously breathing, reperfusion is established by releasing the slipknot around the pulmonary artery. This clinically relevant survival model permits the evaluation of lung IR injury, the resolution phase, downstream effects on lung function, as well as two-hit models involving experimental pneumonia. While technically challenging, this model can be mastered over the course of a few weeks to months with an eventual survival or success rate of 80%-90%.

A mouse surgical model to create left lung ischemia reperfusion (IR) injury while maintaining ventilation and avoiding hypoxia.

Orotrakeal Entübasyon ve Ventilasyonlu Akciğer İskemisi Reperfüzyon Cerrahisinin Bir Fare Modeli

Ischemia reperfusion (IR) injury frequently results from processes that involve a transient period of interrupted blood flow. In the lung, isolated IR ...

İmmünoloji ve Enfeksiyon

In vivo Calcium Imaging in Mouse Inferior Olive

Inferior olive (IO), a nucleus in the ventral medulla, is the only source of climbing fibers that form one of the two input pathways entering the cerebellum. IO has long been proposed to be crucial for motor control and its activity is currently considered to be at the center of many hypotheses of both motor and cognitive functions of the cerebellum. While its physiology and function have been relatively well studied on single-cell level in vitro, presently there are no reports on the organization of the IO network activity in living animals. This is largely due to the extremely challenging anatomical location of the IO, making it difficult to subject to conventional fluorescent imaging methods, where an optic path must be created through the entire brain located dorsally to the region of interest.
Here we describe an alternative method for obtaining state-of-the-art -level calcium imaging data from the IO network. The method takes advantage of the extreme ventral location of the IO and involves a surgical procedure for inserting a gradient-refractive index (GRIN) lens through the neck viscera to come into contact with the ventral surface of the calcium sensor GCaMP6s-expressing IO in anesthetized mice. A representative calcium imaging recording is shown to demonstrate the feasibility to record IO neuron activity after the surgery. While this is a non-survival surgery and the recordings must be conducted under anesthesia, it avoids damage to life-critical brainstem nuclei and allows conducting large variety of experiments investigating spatiotemporal activity patterns and input integration in the IO. This procedure with modifications could be used for recordings in other, adjacent regions of the ventral brainstem.

In vivo Calcium Imaging in Mouse Inferior Olive

We present a protocol to expose the brainstem of adult mouse from the ventral side. By using a gradient-refractive index lens with a miniature microscope, calcium imaging can be used to examine the activity of inferior olive neural somata in vivo.

In vivo Fare Kalitesiz Zeytinde Kalsiyum Görüntüleme

Inferior olive (IO), a nucleus in the ventral medulla, is the only source of climbing fibers that form one of the two input pathways entering the ...

Nörobilim

Repeated Orotracheal Intubation in Mice

The literature describes several methods for mouse intubation that either require visualization of the glottis through the oral cavity or incision in the ventral neck for direct confirmation of cannula placement in the trachea. The relative difficulty or the tissue trauma induced to the subject by such procedures can be an impediment to an investigator&#8217;s ability to perform longitudinal studies. This article illustrates a technique in which physical manipulation of the mouse following the use of a depilatory to remove hair from the ventral neck permits transcutaneous visualization of the trachea for orotracheal intubation regardless of degree of skin pigmentation. This method is innocuous to the subject and easily achieved with a limited understanding of murine anatomy. This refined approach facilitates repeated intubation, which may be necessary for monitoring progression of disease or instillation of treatments. Using this method may result in a reduction of the number of animals and technical skill required to measure lung function in mouse models of respiratory disease.

The goal of this article is to describe a refined method of intubation of the laboratory mouse. The method is noninvasive and, therefore, ideal for studies that require serial monitoring of respiratory function and/or instillation of treatments into the lung.

Farelerde Tekrarlanan Orotracheal Entübasyon

The literature describes several methods for mouse intubation that either require visualization of the glottis through the oral cavity or incision in the ...

<meta charset="utf8"></head><body>Hayatta Kalamayan Sıçan Modelleri için Trakeotomi Tabanlı Endotrakeal Entübasyon Protokolü

Endotracheal Intubation via Tracheotomy and Subsequent Thoracotomy in Rats for Non-Survival Applications

Endotracheal intubation and subsequent ventilation are often basic requirements for translational research in rat models for various interventions that require controlled or high ventilation pressures or access to the thoracic cavity and organs. Conventional endoorotracheal intubation using the anatomically existing route through the mouth is well suited for survival experiments. However, this procedure poses some challenges, including generally higher levels of the required experience and technical skill, more advanced equipment, and greater time effort with relevant intubation failure rates and complications such as tracheal perforation, temporary systemic hypooxygenation, and relevant aerial leakage.
This manuscript, therefore, presents a detailed step-by-step protocol for endotracheal intubation through tracheotomy in non-survival rat models when guaranteed intubation success, constant oxygenation levels, high ventilation pressures, or open thoracotomy are required.
The protocol emphasizes the importance of meticulous surgical technique to ensure consistent and reliable outcomes, especially for researchers who are inexperienced or lack routine in the technique of endoorotracheal intubation via direct laryngoscopy. This procedure is, therefore, expected to minimize animal suffering and unnecessary animal losses.

<meta charset="utf8"></head><body>Hayatta kalamayan uygulamalar için sıçanlarda trakeotomi ile endotrakeal entübasyon ve ardından torakotomi

Here we introduce a standardized procedure for endotracheal intubation via tracheotomy followed by thoracotomy in rats, aimed at enhancing the precision and reproducibility of non-survival applications requiring invasive ventilation and exposure of thoracic organs in in vivo rat models.

Hayatta kalamayan uygulamalar için sıçanlarda trakeotomi ile endotrakeal entübasyon ve ardından torakotomi

Endotracheal intubation and subsequent ventilation are often basic requirements for translational research in rat models for various interventions that ...