저자는 트리 크롬 염색 및 분석에 대한 도움말은 워싱턴 대학의 조직학의 브라이언 존슨과 이미징 코어 감사합니다. 이 작품은 NIH 보조금 HL098067 및 HL089455에 의해 지원되었다.

미국과 삼목 시나이 의료 센터의 대학 기관 동물 관리 및 사용위원회 (IACUC)는 이러한 연구에 필요한 동물 작업을 승인했다. 1. 준비 <ol><li> 무딘 엔드 집게와 억압자를 통해 오토 클레이브를 모두 소독. </li><li> 생물 안전 캐비닛을 사용하여 동결 건조 분말 PBS에서 BLM의 작업 주식을 준비합니다. 혼합조차 보장 35 kHz에서 10 분 동안 초음파 처리 용액. 참고 : μL (30) 사이에 45의 총 부피는 큰 볼륨과 피펫의 로우 엔드에 변화하고, 질식을 방지하는 것이 좋습니다. </li><li> 약 1 절차 자체에 대한 평방 미터뿐만 아니라 전 시술 후 모두 케이지에 대한 지정된 위치를 포함하는 깨끗한 작업 공간을 준비합니다. </li><li> 베이스와 underl를 통해 실험실 테이프의 2 또는 3 스트립을 마련하여 즉시 연구원 앞의 벤치에 프로 시저 보드의 기본 수정벤치 잉. 보드를 만드는 방법에 대한 자세한 사양은 그림 1을 참조하십시오. </li><li> 절차 기판의 두 개의 위치 사이의 나사 크기 4.0 봉합 실의 단일 길이 넥타이. </li><li> 제거하고 1 ㎖의 세 주사기에서 플런저를 폐기하고, 기밀 밀봉을 형성하여 배럴의 상부에 PBS 60 μL를 증착함으로써 임시 폐활량계를 생성한다. 느슨하게 주사기 중 하나에 카테터의 허브를 안전하고 보드의 한쪽에 놓습니다. </li><li> 1 ML의 주사기에 공기의 흡인 300 μL와 보드의 한쪽에 놓습니다. </li><li> 한쪽으로 테이프 길이와 장소에 약 6 인치의 추가 조각을 잘라. 이 단계 2.4 보드 동물을 보호하는 데 사용된다. </li><li> 이소 플루 란 실을 설정합니다. 노출 챔버와 통관 진공 모두에서 해당 포트에 O (2), 이소 플루 란, 진공을 연결합니다. 또는에서 마취를 관리 이소 플루 란 호환 생물학적안전 캐비닛. </li></ol> 2. 삽관 <ol><li> 이 의식을 잃고 호흡이 적절한 속도로 감속 할 때까지 실에서 이소 플루 란과 마우스를 마취. 전형적인 노출 3-4 분 동안 4 % 이소 플루 란 2 % O 2를 포함하고, 최적의 결과는 마취 2-2.5 분이다. 이 한 숨을 2 초마다의 호흡 속도에 해당한다. </li><li> 한쪽으로 피펫과 장소에 BLM 30, 45 μl의 사이에 설정하는 진정 작용, 흡인 기다리는 동안. </li><li> 준비가되면, 절차 플랫폼의 위치에 장착 나사 스레드에서 상부 앞니에 의해 마우스를 진정 중단. 동물의 배부 플랫폼 표면에 평평하게 놓여 있는지 확인합니다. </li><li> 환기를 제한하지 않도록주의이기 때문에, 그냥 진동판 위에 느슨하게 흉강의 하단 (꼬리) 부분에서 테이프의 조각을 놓습니다. 위치는 PROC 동안 적절한 정렬을 유지하기 위해 꽉 충분해야한다edure하지만 호흡을 제한 너무 꽉 없습니다. </li><li> 80 % 내지 100 %의 강도로 조명을 켜고 명치 부근의 피부 표면 1~2cm,되도록 거위의 방향. 주기적으로 마우스를 손상하지 않도록 열을 위해 거위의 끝을 확인합니다. </li><li> 플랫폼 뒤에 ​​서, 혀의 위치를​​ 살균, 무딘 엔드 집게를 사용합니다. 아래 앞니, 부드럽게 그립을 방지하고 구강 밖으로 혀를 그릴 조심. </li><li> 나머지 손을 사용하여, 억압 물을 넣고 구강의 바닥에 평평하게 혀를 사용. 집게를 해제하지만 다음 두 단계를위한 장소에 억압자 둡니다. </li><li> 기관지 그것이 mainstem 기관지의 수준에 도달 할 때까지 명치의 레벨로부터 근위 거위 안내하여 볼 수 있도록 빛의 방향. 주 : 기관지 용이 호흡 작용에 의해 구별 될 수 있으며, WHI채널은 강도가 변동하는 방출되는 빛을 발생합니다. 정확하게 위치 할 때,이 구조는 구강 자체 최소한의 주변 조명과 빛의 중앙에 위치한 핀과 같은 축 평면에서 식별 할 수 있습니다. </li><li> 각도가 직선 루멘, 액 적을 함유 연결된 주사기, 기관의 천연 경로를 따르며, 22-G 카테터 팁을 낮추도록 주사기. PBS 거품이 상승하고 성공적인 위치에 따라 각각의 호흡과 가을에 시작됩니다. 참고 :이 작업은 깊은 진정 작용의 결과로 몇 초 정도 지연 될 수 있습니다. </li><li> 추가 5mm에서 카테터 피드. 혀를 제거 억압자. </li><li> 부드럽게 주사기를 제거, 반대 손으로 주사기를 이동하고, 허브를 파지. </li><li> 카테터 허브의 내부의 중심으로 BLM 30 45 μL 사이 금고는 제 2 주사기를 연결하고 허브에 공기의 300 μL 분주. </li><li> 두 번째 교체제 PBS의 기포를 함유하는 주사기. 거품 상승과 절차가 성공적으로 수행 된 경우 떨어질 것입니다. </li></ol> 3. 후 절차 케어 <ol><li> 카테터 및 테이프를 제거하고 의식을 회복 할 때까지 건조 따뜻한 장소에서 동물을 배치 - 일반적으로 몇 분 내에. </li></ol>

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The authors have nothing to disclose.

에어로졸 인해 제한 시약 가용성, 안전성, 또는 비용이 비현실적 경우에, 직접 기관 행정부는 폐에 외인성 에이전트의 전달을위한 우수한 방법이다 16 Transtracheal의 점안이 광범위하게이 작업을 수행하는 데 사용되었습니다.; 그러나, 모든 외과 적 개입, 그것은 또한 그것 절차 자체에 의한 합병증의 가능성을 수행하고, 반드시 제가 주입된다. (13) 이러한 이유로, 삽관...

일부 해부학 적 생리 학적 차이에도 불구하고, 1 뮤린 모델 사육 관점에서 인간 생물학 및 질환 발병. 2 모델링 헤아릴 수, 마우스, 취급하기 쉬운 낮은 사육 시간 가속 수명을 가지며, 상대적으로 저렴 계속 집입니다. 다양한 유전자 변형 전략 (예., 조건부 녹아웃 리포터 마우스 계통 추적 방법 등)뿐만 아니라, 가능한 시약의 다양한 (예., 항체, 재조합 단백질 억제제의 개발과 함께, 등), 마우스는 인간의 질병 및 항상성 과정을 밝히기 위해서 필수적인 모델 척추 생물되었다. (3) 마우스는 인간의 4 ALI는 외상, 부상 또는 패혈증으로 인해 발생할 수 있습니다. 급성 폐 손상 (ALI) 및 폐 섬유증을 포함한 폐 조건을 공부에 특히 유용되었습니다 상피을 특징으로하고내피 누출 (예., 부종), 염증 및 초기 섬유화. 많은 환자에서 ALI 종종 인한 호흡 부전으로 사망 섬유증 결과의 중증 급성 호흡 곤란 증후군 (ARDS)으로 진행한다. 5,6- 폐 섬유증은 세포 외 기질의 과잉 증착 특징 진보적 인 심각한 병리학 , 특히. 나는 장애인 폐 기능을 선도, 콜라겐 블레오 마이신 (BLM)의 7,8 관리를 입력 한 실험 동물에 ALI 및 섬유화를 유도하기 위해 가장 널리 사용되는 최고의 특성화 된 모델입니다. (9) 설치류에 폐 섬유증을 BLM 유발하지만 않습니다 완전히 인간의 섬유 성 표현형은,이 모델 (10) 마우스 연구는 발병과 질병의 진행에 영향을 미치는 여러 중요한 요인의 발견을 주도 요점을 되풀이하지. (11) BLM 유도 섬유화 뒤에 정확한 메커니즘 (들)를 알 수 있지만, 개시 부상전도기도 및 폐포, 특히 제 1 형 pneumocytes 라이닝 상피 세포의 접촉에 의존하는 DNA 가닥 나누기에서 발생하는 것으로 생각된다. (12) BLM과 폐 상피 세포 사이의 직접 접촉에 대한 필요성은 강력한 전달 경로의 중요성을 강조 그리고 이러한 우려는 재조합 단백질, 항체, siRNA를, 바이러스, 박테리아, 미립자 등을 포함한 말초기도를 대상으로 치료의 넓은 범위에 밀접한 있습니다. 인두 흡인 (OPA)는 이러한 목적 13 널리 사용되었지만, OPA의 주요 단점은 전달 제의 일부가되어 투여 용량으로 부정확 선도 위장관으로 삼킬 수 있다는 것이다. 널리 사용되는 또 다른 방법은 직접 호흡기에 제의 기관지 점적 및 노출 강한 마취 기관 절개술을 포함 transtracheal 점안이다. (14) 그러나, 그러한 수도절차 인해 invasivity에 바람직하지 않을 수 있지만, 또한, 시간 소모적 인 훈련 공정한 비트가 필요하며으로 제제의 직접 투여를 포함한다. 15,16 여러 프로토콜들이 개발되었다 호흡기 잠재적 손상을 일으키는 수술 개입 없이도기도가 16,17,18,19,20 그러나 이러한 방법은 확장 된 것을 포함 복구 시간 강력한 마취제로 인한 고가 장비의 사용 (예., 이경 / 후두경 시판 절차 보드, 광섬유 투여 량에 관한 와이어 등), 구강 조작 과량 불확실성. 이 논문은 연구자가 신속, 저렴하고 안정적​​으로 주변 조직에 잔류 손상의 제한으로 위험 쥐의 폐에 시약을 주입 할 수 있습니다 삽관을 통한 행정의 비교적 쉬운 방법을 설명합니다.

<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.

삽관 마우스는 체중 감소와 고통에 대해 매일 모니터링, 2.5 % 2,2,2- tribromoethanol의 복강 내 주사를 통해 4, 10 또는 제 17 일 후에 희생되었다. 기관지 폐포 세척액 (BAL)는 PBS 세 세척에 수집 된 21 곳 바와 같이 , 오른쪽 폐는 10 % 포르말린에 고정 된 파라핀, 워싱턴 조직학 및 이미징 코어 (22)의 대학에 의해 메이슨의 트리 크롬 염색 하였다. <p cla...

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An Improved Method for Rapid Intubation of the Trachea in Mice

마우스의기도 신속 삽관 용 개선 된 방법

이 문서에서는 삽관을 통해 마우스 기관에 직접 블레오 마이신 관리를위한 신속하고 간단한 방법을 제공합니다. 이러한 방법의 주요 장점은 마스터 높은 재현성 쉽고, 특수 장비 나 긴 회복 시간을 요구하지 않는다는 점이다.

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

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25.1K 조회수.  University of Washington.  이 문서에서는 삽관을 통해 마우스 기관에 직접 블레오 마이신 관리를위한 신속하고 간단한 방법을 제공합니다. 이러한 방법의 주요 장점은 마스터 높은 재현성 쉽고, 특수 장비 나 긴 회복 시간을 요구하지 않는다는 점이다. 

비디오: 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.

마우스의 급성 폐 상해를 유발하는 압력 제어 환기

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

연구

의학

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.

마우스의 기관 내 삽관<em&gt;를 통해</em이경 사용&gt; 직접 후두경

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.

마우스 폐 삽관 법의 간단한 방법

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.

비 침습적 기관 내 삽관 마우스 폐의 병리학 및 생리학을 연구하는

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.

에어웨이 저항 측정 및 마우스에 Bronchoalveolar 세척 유체 샘플링에 대한 가역, 비침습 방법

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

생물학

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.

마우스에 가로 대동맥 수축 유도를 폐쇄 가슴 모델

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.

Orotracheal 삽관 및 환기 폐 허혈 재관류 수술의 마우스 모델

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

면역학 및 감염병학

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.

생체 내 마우스 열등올리브의 칼슘 이미징

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 ...

신경과학

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.

마우스에서 반복된 Orotracheal 삽관

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>Non-survival Rat 모델을 위한 기관절개술 기반 기관내 삽관 프로토콜

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>Non-survival 응용 프로그램을 위한 쥐의 기관절개술을 통한 기관내 삽관 및 후속 흉곽절개술

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.

Non-survival 응용 프로그램을 위한 쥐의 기관절개술을 통한 기관내 삽관 및 후속 흉곽절개술

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