저자는 기술 지원을 크리스티안 Pahrmann 감사합니다. SS는 독일 Forschungsgemeinschaft (DFG) (SCHR992/3-2 및 SCHR992/4-2)로부터 자금을 받았다.

모든 동물 연구는 중요한 동물 관리 지침에 따라 수행되어야한다. 이전에 시작 프로토콜 동물 작업 기관의 승인을 얻습니다. 1. Myointimal 증식 모델 (A) 내막 증식증 모델의 경우, C57BL/6J (주문 번호 000664, C57BL/6J) 약 25 g의 무게 팔주 세의 나이에 마우스를 구입할 수 있습니다. 기존의 조건에서 이러한 실험을위한 주택의 마우스; 마우스 표준 마우스 우를 공급하고 임의 량의 물을 제공합니다. <ol><li> 마우스 준비 : 유도 챔버를 이용한 이소 플루 란 (2 %)와 마취 상태로 마우스를 소개합니다. <ol><li> 그 뒷면에 마우스를 놓고 마취를 보장하기 위해 안면 마스크로 코와 입을 커버, 머리 트리머를 사용하여 복부의 머리를 제거합니다. </li><li> 프로보 - 요오드, 2 사이클에서는 80 % 에탄올이어서 보편적 복부 영역을 소독. </li><li> 이 없음을 보증마취의 충분한 깊이를 모니터하기 위해 뒷다리를 곤란에 의한 반사의. </li><li> 복부 장기를 노출하는 두 단계로 리네아 알바을 따라 피부와 근육을 분리합니다. </li><li> 식염수 보습 장갑에 내장을 놓습니다. 습기를 유지하기 위해 내장 주변의 장갑을 감싸십시오. </li><li> 주의 깊게 복부 대동맥 천축 지방 조직을 제거하다. 어떤 지점의 혈관을 손상하지 않도록주의하면서 아래의 분기점에 신동맥 대동맥을 노출합니다. </li><li> 첫째, 혈액의 흐름을 차단하기 위해, 근위에 신동맥 대동맥을 미세 수술 클램프를 놓습니다. 이제, 대동맥 분기 옆에 두 번째 원심 클램프를 적용합니다. </li><li> 혈액의 흐름이 중단되면, 근위 클램프 부근 대동맥에 작은 절개 부위를 만든다. </li><li> 0.9 %의 식염수에 수분 된 풍선 팁 카테터를 삽입하고 대동맥의 말초 클램프를 향해 전진. </li><li> 내피 손상의 경우, balloo 팽창하여 대동맥을 발가 벗기다N 신중하고 천천히 근위 클램프의 방향으로 카테터를 당겨. </li><li> 두 번 삭박 기동을 반복합니다. </li><li> 카테터를 제거하고 10-0 prolene의 실행 봉합사를 이용하여 대동맥 절개를 닫습니다. </li><li> 조심스럽게 말초 클램프를 엽니 다. aortotomy 사이트에서 출혈의 경우에는, 필요에 따라, 다시 클램프를 닫 출혈을 찾아 추가 중단 바늘을 배치. </li><li> 제공된 출혈이 관찰 될 수있는 경우, 천천히 근위 클램프를 연다. </li><li> 대동맥 펄스는 절개에서 원심 볼 수 있어야합니다. 이제 내장은 현장에 다시 배치 될 수있다. </li><li> 예열 멸균 식염수를 이용하여 복강을 씻어. </li><li> 복부 벽 6-0 prolene의 실행 봉합사를 이용하여 근육 층으로 시작 닫습니다. </li><li> 다음 5-0 prolene을 수행하는 실행 봉합사의 사용에 따라 피부를 적응. </li><li> 시간 동안 마우스가 여전히 마취, 피하 인제을 통해 4 ~ 5 ㎎ / ㎏ Carprofen을 적용의 ction. </li><li> 통증 조절을 위해 마시는 물 (100 ㎖ 당 50 밀리그램의 Metamizol)에 Metamizol를 첨가하고 3 일 수술 후 계속. 복구를 통해 동물을 모니터링합니다. 참고 :이 모델의 관찰 기간은 28 일입니다. </li></ol></li></ol> 이. 죽상 경화증 모델 (B) 사주의 나이에 마우스 (주문 번호 002052, B6.129P2-Apoetm1Unc / J), 다음 4-6개월에 공급되는 동맥 경화 모델의 경우, 아포 지단백 E 결핍을 (- - / 아포)를 구입. 기존의 조건에서 이러한 실험을위한 주택의 마우스; 쥐에게 도착시 높은 지질 서부 다이어트 (할렌 연구소 TD.88137)를 공급하고 4~6개월의 기간 동안 임의 량의 물을 제공합니다. <ol><li> 마우스 준비 : <ol><li> 아포 피드 - / - 서부 다이어트와 마우스 시대 4 주에서 시작. 식이 조작 도착시 직접 시작해야합니다. </li><li> 실험 기간에 걸쳐 서양 식단을 유지한다. &lt;/ 리&gt; </ol></li></ol> 3. 분석 <ol><li> Myointimal 증식증 모델 (A) 이 모델에 대한 관심의 영역은 &quot;병변의 영역&quot;이다. 분석을위한 옵션 내강 말살을 포함, I / M 비율 (내막 / 미디어 비율), 최대 myointima 두께, myointimal 지역, 섬유증에 대한 트리 크롬 염색, 및 단핵 세포에 대한 H &amp; E 염색 (그림 1A-1C에 도시 된 바와 같이). 과학적인 문제에 따라 Picrosirius 콜라겐 빨간색, 면역 조직 화학 염색 및 공 초점 방법 등 많은 다른 염색에 적용 할 수있다. </li><li> 죽상 경화증 모델 (B) 이 모델에 대한 관심의 영역은 대동맥 판막, 대동맥 궁뿐만 아니라 하행 대동맥이다. 분석을위한 옵션 : 수단 빨간 지질에 대한 염색 및 대동맥 판막, 대동맥 궁과 하행 대동맥 (그림 1D에 도시 된 바와 같이)의 트리 크롬 염색. 이 모델은 또한 같은 다른 염색의 다양한 옵션을 산출중성 지질, Picrosirius 레드, 면역 조직 화학 염색 및 공 초점 방법 예를 들어 오일 레드 O하십시오. <ol><li> 수단 레드 염색 : 갓 수확 대동맥 조직의 수단 레드 염색을 수행합니다. <ol><li> 수확하기 전에 다이어트 6H를 제거합니다. </li><li> 유도 챔버를 이용한 이소 플루 란 (2 %)와 마취 상태로 마우스를 소개합니다. 마취의 충분한 깊이를 모니터링하는 뒷다리 곤란하여 반사의 부재를 확신한다. </li><li> 가슴을 열고; 마음의 정점을 차단하고 대동맥을 해결하기 위해 대동맥 루트를 통해 4 % PFA (3 ㎖) 다음에, 첫 생리 식염수 (3 ㎖), 좌심실을 통해 perfuse. </li><li> 그 분기점에 루트에서 대동맥을 확보. 지방 조직에게 가능한 한 많이 제거하려고합니다. </li><li> 심장과 대동맥 루트를 차단하고, 조직 검사를위한 4 % PFA에 저장합니다. </li><li> 조직학의 경우 대동맥 루트는 신중하게 AOR의 절단을 허용하는 수직으로 파라핀 블록에 배치 할 수있다3 밸브 모양의 전단지를 표시 TIC 루트 생성 슬라이드. </li><li> 대동맥 나무의 한쪽 끝을 잘라 미세 핀에 실리콘 층에 고정합니다. 길이 방향으로 대동맥을 열고 대동맥 조직을 고정합니다. </li><li> 50 % 에탄올로 조직을 씻어. 약 15 분 동안 수단 III 염색 용액으로 조직을 담가. </li><li> 초과 수단 III를 제거하기 위해 50 %의 에탄올로 가볍게 씻어. dH보다 2 O 씻어하고, 추가 분석을 위해 사진을 찍어 ​​(50 % 에탄올이 떨어져 염료를 세척 할 수 있습니다. 참고). </li></ol></li></ol></li></ol>

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저자가 공개하는 게 없다.

두 질환은 환자와 유사한 증상을 초래할 있지만, 플라크 개발의 기본 메커니즘에 따라서 치료 방법은 2 매우 다릅니다. 동맥 협착의 다른 형태의 환자에서 임상 연구 결과뿐만 아니라 플라크 개발의 기간은 기본 메커니즘에 따라 달라집니다. 환자의 임상 연구 결과에 따라, 시료 분석의 다른 방법은 5,15를 수행해야합니다. 그것은 선택된 모델 (14)에 ?...

산업화 된 국가에서 허혈성 심장 질환 사망률 1의 주요 원인이 남아있다. 관상 동맥 협착에 대한 원인은 매니 폴드이며 가장 일반적인 치료 전략이 잔존 재관류와 myointimal 증식뿐만 아니라 아테롬성 동맥 경화증을 포함한다. 분명히 내막 증식증과 죽상 경화증의 기계적인 경로를 구분 연구 모델은 동맥 플라크 개발 병인 학적 및 병태 생리 학적 과정을 조사하는 것이 필수적이다. 이 비디오에서는 myointimal 증식 또는 죽상 경화증의 개발 중 하나를 연구하는 데 두 개의 서로 다른 마우스 모델을 소개합니다. Myointimal 증식증 원래 죽상 3에 대해 기재된 &quot;응답 투 부상&quot;패러다임을 통해 형성하는 것이 가정된다. 내피 층의 기계적 중단 주변 분비 효과에 의해 강화 강렬한 리모델링에 이르게한다. 여러 디가 있습니다fferent 동물 모델은 일반적으로 myointimal 증식을 연구하는 데 사용됩니다. 일부 그룹은 쥐 4의 대동맥에서 금속 장치를 사용한다. 풍선 카테터 수행 대동맥 삭박 모델은 또한 일반적으로 실험실 쥐 5,6에서 사용된다. 실험용 쥐 (7)에서 수행 내막 증식증 모델 경동맥의 결찰을 구현하고 myointimal 8 병변을 유도한다. 우리 실험실에서, 복부 대동맥 삭박 모델 -뿐만 아니라 인간 답게 스텐트 재 협착 모델 (9) 일반적으로 사용되는 - myointimal 증식의 개발을 연구한다. 이 비디오는 적절한 실험 동물 모델을 선택하는 것이 동맥 협착의 기계적 또는 병태 생리 학적 연구에 중요한 것을 강조한다. Myointimal 증식 모델은 동맥 경화 모델을 구별해야합니다. 후자의 경우, 아포 리포 단백질 E가 결핍 (아포 - / -) 서부 다이어트와 함께 마우스는 일반적으로 atheroscl을 유도하는 데 사용됩니다에로틱 병변 10,11,12,13. 생쥐 myointimal 질병의 이러한 유형의 모두의 유도에 대한 예는 혈관 협착 (14)를 분석하기 위해 다른 모델 적응 옵션과 함께 표시되어 있습니다.

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inducing myointimal hyperplasia versus atherosclerosis in mice: an introduction of two valid models

동맥 협착의 유도에 대한 다양한 생체 실험 쥐 모델은 허혈성 심장 질환, 예를 들어 관찰, 동맥 플라크 형성 및 협착 등의 질병을 모방하기 위해 설립되었다. 두 재현성 마우스 모델 - 두 동맥 협착의 결과로하지만, 각 개발의 다른 경로를 기본는 - 여기에 소개합니다. 모델은 동맥 협착의 가장 일반적인 두 가지 원인을 나타냅니다; 즉 하나의 마우스의 각 myointimal 증식을위한 모델 및 죽상 동맥 경화증이 표시됩니다. myointimal 증식을 유도하기 위해, 복부 대동맥의 풍선 카테터의 부상이 수행된다. 죽상 경화성 플라크의 발달을 위해, 아포 - / - 서부 지방 다이어트와 함께 마우스 모델이 사용된다. 결과의 측정 및 평가를위한 다른 모델 적용 옵션이 원고에 이름과 설명되어 있습니다. 이 두 모델의 도입과 비교 INFORMAT를 제공합니다과학자가 물었다 과학적 질문에 따라 적절한 동맥 협착 모델을 선택하는 이온.

그림 1은 서로 다른 분석 옵션과 두 모델의 유도 된 질병 상태를 보여줍니다. 내막 증식증 모델의 경우, 트리 크롬 염색 대표적인 단면의 분석이 표시됩니다. 이러한 단면에서, I / M 비율, 최대 플라크 두께, 루멘에서 최대의 내막 두께, 퍼센트 내강 폐색뿐만 아니라 플라크 지역과 같은 결과를 측정하여 계산 될 수있다. 죽상 동맥 경화증 모델의 결과를 평가하기 ?...

Watch this Scientific Journal Video about Inducing Myointimal Hyperplasia Versus Atherosclerosis in Mice: An Introduction of Two Valid Models at JoVE.com

Inducing Myointimal Hyperplasia Versus Atherosclerosis in Mice: An Introduction of Two Valid Models

이 동영상은 쥐의 동맥 내막 플라크 개발의 두 가지 모델을 보여주고 myointimal 증식과 죽상 경화증의 차이를 강조한다.

쥐에서 동맥 경화 수스 Myointimal 증식을 유도 : 두 개의 유효한 모델의 소개

Various in vivo laboratory rodent models for the induction of artery stenosis have been established to mimic diseases that include arterial plaque ...

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12.0K 조회수.  University Hospital Hamburg.  이 동영상은 쥐의 동맥 내막 플라크 개발의 두 가지 모델을 보여주고 myointimal 증식과 죽상 경화증의 차이를 강조한다. 

비디오: Inducing Myointimal Hyperplasia Versus Atherosclerosis in Mice: An Introduction of Two Valid Models

Implantation of a Carotid Cuff for Triggering Shear-stress Induced Atherosclerosis in Mice

It is widely accepted that alterations in vascular shear stress trigger the expression of inflammatory genes in endothelial cells and thereby induce atherosclerosis (reviewed in 1 and 2). The role of shear stress has been extensively studied in vitro investigating the influence of flow dynamics on cultured endothelial cells 1,3,4 and in vivo in larger animals and humans 1,5,6,7,8. However, highly reproducible small animal models allowing systematic investigation of the influence of shear stress on plaque development are rare. Recently, Nam et al. 9 introduced a mouse model in which the ligation of branches of the carotid artery creates a region of low and oscillatory flow. Although this model causes endothelial dysfunction and rapid formation of atherosclerotic lesions in hyperlipidemic mice, it cannot be excluded that the observed inflammatory response is, at least in part, a consequence of endothelial and/or vessel damage due to ligation.
In order to avoid such limitations, a shear stress modifying cuff has been developed based upon calculated fluid dynamics, whose cone shaped inner lumen was selected to create defined regions of low, high and oscillatory shear stress within the common carotid artery 10. By applying this model in Apolipoprotein E (ApoE) knockout mice fed a high cholesterol western type diet, vascular lesions develop upstream and downstream from the cuff. Their phenotype is correlated with the regional flow dynamics 11 as confirmed by in vivo Magnetic Resonance Imaging (MRI) 12: Low and laminar shear stress upstream of the cuff causes the formation of extensive plaques of a more vulnerable phenotype, whereas oscillatory shear stress downstream of the cuff induces stable atherosclerotic lesions 11. In those regions of high shear stress and high laminar flow within the cuff, typically no atherosclerotic plaques are observed.
 
In conclusion, the shear stress-modifying cuff procedure is a reliable surgical approach to produce phenotypically different atherosclerotic lesions in ApoE-deficient mice.

The constricting cuff presented in this article is designed to induce atherosclerosis in the murine common carotid artery. Due to the conical shape of its inner lumen the implanted cuff generates well-defined regions of low, high and oscillatory shear stress triggering the development of atherosclerotic lesions of different inflammatory phenotypes.

마우스의 전단 응력 유발 죽상 경화증을 실행을위한 경동맥 혈압계의 주입

It is widely accepted that alterations in vascular shear stress trigger the expression of inflammatory genes in endothelial cells and thereby induce ...

연구

의학

Quantitative Analysis and Characterization of Atherosclerotic Lesions in the Murine Aortic Sinus

Atherosclerosis is a disease of the large arteries and a major underlying cause of myocardial infarction and stroke. Several different mouse models have been developed to facilitate the study of the molecular and cellular pathophysiology of this disease. In this manuscript we describe specific techniques for the quantification and characterization of atherosclerotic lesions in the murine aortic sinus and ascending aorta. The advantage of this procedure is that it provides an accurate measurement of the cross-sectional area and total volume of the lesion, which can be used to compare atherosclerotic progression across different treatment groups. This is possible through the use of the valve leaflets as an anatomical landmark, together with careful adjustment of the sectioning angle. We also describe basic staining methods that can be used to begin to characterize atherosclerotic progression. These can be further modified to investigate antigens of specific interest to the researcher. The described techniques are generally applicable to a wide variety of existing and newly created dietary and genetically-induced models of atherogenesis.

We describe procedures to quantify and characterize atherosclerotic lesions in mouse models using precision sectioning of the aortic sinus and ascending aorta combined with histochemical and immunohistochemical analysis.

쥐의 대동맥 부비동의 동맥 경화 병변의 정량 분석​​ 및 특성

Atherosclerosis is a disease of the large arteries and a major underlying cause of myocardial infarction and stroke. Several different mouse models have ...

Acute Brain Trauma in Mice Followed By Longitudinal Two-photon Imaging

Although acute brain trauma often results from head damage in different accidents and affects a substantial fraction of the population, there is no effective treatment for it yet. Limitations of currently used animal models impede understanding of the pathology mechanism. Multiphoton microscopy allows studying cells and tissues within intact animal brains longitudinally under physiological and pathological conditions. Here, we describe two models of acute brain injury studied by means of two-photon imaging of brain cell behavior under posttraumatic conditions. A selected brain region is injured with a sharp needle to produce a trauma of a controlled width and depth in the brain parenchyma. Our method uses stereotaxic prick with a syringe needle, which can be combined with simultaneous drug application. We propose that this method can be used as an advanced tool to study cellular mechanisms of pathophysiological consequences of acute trauma in mammalian brain in vivo. In this video, we combine acute brain injury with two preparations: cranial window and skull thinning. We also discuss advantages and limitations of both preparations for multisession imaging of brain regeneration after trauma.

Acute brain trauma is a severe injury that has no adequate treatment to date. Multiphoton microscopy allows studying longitudinally the process of acute brain trauma development and probing therapeutical strategies in rodents. Two models of acute brain trauma studied with in vivo two-photon imaging of brain are demonstrated in this protocol.

세로 두 광자 이미징 다음 마우스의 급성 뇌 외상

Although acute brain trauma often results from head damage in different accidents and affects a substantial fraction of the population, there is no ...

Minimal Invasive Surgical Procedure of Inducing Myocardial Infarction in Mice

Myocardial infarction still remains the main cause of death in western countries, despite considerable progress in the stent development area in the last decades. For clarification of the underlying mechanisms and the development of new therapeutic strategies, the availability of valid animal models are mandatory. Since we need new insights into pathomechanisms of cardiovascular diseases under in vivo conditions to combat myocardial infarction, the validity of the animal model is a crucial aspect. However, protection of animals are highly relevant in this context. Therefore, we establish a minimally invasive and simple model of myocardial infarction in mice, which assures a high reproducibility and survival rate of animals. Thus, this models fulfils the requirements of the 3R principle (Replacement, Refinement and Reduction) for animal experiments and assure the scientific information needed for further developing of therapeutical strategies for cardiovascular diseases.

A highly reproducible model for myocardial infarction in mice with minimal invasive manipulations is described. The model can be easily performed, resulting in a high reproducibility and survival rate. Thus, the described model will reduce the number of required animals as requested by the 3R principle (Replacement, Refinement and Reduction).

쥐 심근 경색을 유도 최소 침습 수술

Myocardial infarction still remains the main cause of death in western countries, despite considerable progress in the stent development area in the last ...

Surgical Models of Gastroesophageal Reflux with Mice

Multiple surgical procedures have been reported to induce gastroesophageal reflux in animals. Herein, we report three surgical models with mice aiming to induce reflux of gastric contents, duodenal contents or mixed contents. Surgical procedures and general principles have been described in detail. A researcher with surgical experience should be able to grasp the technique after a short period of practice. After surgery, most mice can survive and develop reflux esophagitis similar to that in humans. However, it should be noted that histological differences between mouse and human esophagus are the inherent limitations of these surgical models. If used for research on Barrett&#8217;s esophagus and adenocarcinoma, these procedures may need to be combined with genetic modifications.

This article demonstrates surgical procedures of gastroesophageal reflux with mice. These models are useful tools for research on mechanisms and treatment of gastroesophageal reflux disease and potentially Barrett&#8217;s esophagus and esophageal adenocarcinoma.

마우스와 위식도 역류의 외과 모델

Multiple surgical procedures have been reported to induce gastroesophageal reflux in animals. Herein, we report three surgical models with mice aiming to ...

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.

This article presents a rapid and simple method for administering bleomycin directly into the mouse trachea via intubation. Key advantages of this method are that it is highly reproducible, easy to master, and does not require specialized equipment or lengthy recovery times.

마우스의기도 신속 삽관 용 개선 된 방법

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

Induction of Accelerated Atherosclerosis in Mice: The "Wire-Injury" Model

Atherosclerosis is a proliferative fibro-inflammatory disease developing in the arterial wall, inducing a deficient blood flow or a lack of blood flow. Moreover, by rupture of the defective vascular wall, atherosclerosis induces occlusive thrombus formation, which represents the main cause of myocardial infarction or stroke and the most frequent cause of death. Despite the advances in the cardiovascular field, many questions remain unanswered, and additional basic research is essential to improve our understanding of the molecular mechanisms during atherosclerosis and its effects. Due to limited clinical studies, there is a need for representative animal models recreating atherosclerotic conditions such as neointima formation after stent implantation, balloon angioplasty, or endarterectomy. Since the mouse presents many advantages and is the most frequently used model for studying molecular processes, the current study proposes an invasive procedure of endothelial denudation, also known as the wire-injury model, which is representative of the human condition of neointima formation in arteries after revascularization procedures.

This study describes an invasive procedure for the induction of accelerated atherosclerosis in mice. In comparison to other methods using electric- or cryo-induced injury, mechanical-induced injury mimics the human condition of restenosis after revascularization therapies and is ideal for the study of the molecular mechanisms involved.

마우스에서 가속 죽상 경화증의 유도: "와이어 부상" 모델

Atherosclerosis is a proliferative fibro-inflammatory disease developing in the arterial wall, inducing a deficient blood flow or a lack of blood flow. ...

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.

풍선-기반 부상 마우스 복 부 대동맥에서 Myointimal 증식을 유도 하

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 ...

Murine Drinking Models in the Development of Pharmacotherapies for Alcoholism: Drinking in the Dark and Two-bottle Choice

Alcohol Use Disorder (AUD) is a major problem with more than an estimated 76 million people worldwide meeting the diagnostic criteria. Current treatments are limited to three FDA-approved medications that are largely ineffective even when combined with psychosocial intervention, as is evident by the high relapse rate. As such, the search for more novel treatments represents an important public health goal. To this end, the following protocol utilizes two simple rodent drinking models to assess the preclinical efficacy of lead anti-alcohol compounds: two-bottle choice (TBC) and drinking in the dark (DID). The former allows mice to voluntary drink in moderation while the latter induces mice to voluntary consume a large amount of alcohol in a short period that mimics binge drinking. The simple and high throughput nature of both of these paradigms allow for rapid screening of pharmacological agents or for identifying strains of mice that exhibit certain voluntary drinking behavior.

Alcohol Use Disorder (AUD) is a major national health problem and the development of more effective treatments is&#160;required to offset the needs of this patient population. To this end, the following protocol utilizes two simple rodent drinking models to assess the preclinical efficacy of lead anti-alcohol compounds.

Alcohol Use Disorder (AUD) is a major problem with more than an estimated 76 million people worldwide meeting the diagnostic criteria. Current treatments ...

Inducing Acute Lung Injury in Mice by Direct Intratracheal Lipopolysaccharide Instillation

Airway administration of lipopolysaccharide (LPS) is a common way to study pulmonary inflammation and acute lung injury (ALI) in small animal models. Various approaches have been described, such as the inhalation of aerosolized LPS as well as nasal or intratracheal instillation. The presented protocol describes a detailed step-by-step procedure to induce ALI in mice by direct intratracheal LPS instillation and perform FACS analysis of blood samples, bronchoalveolar lavage (BAL) fluid, and lung tissue. After intraperitoneal sedation, the trachea is exposed and LPS is administered via a 22 G venous catheter. A robust and reproducible inflammatory reaction with leukocyte invasion, upregulation of proinflammatory cytokines, and disruption of the alveolo-capillary barrier is induced within hours to days, depending on the LPS dosage used. Collection of blood samples, BAL fluid, and lung harvesting, as well as the processing for FACS analysis, are described in detail in the protocol. Although the use of the sterile LPS is not suitable to study pharmacologic interventions in infectious diseases, the described approach offers minimal invasiveness, simple handling, and good reproducibility to answer mechanistic immunological questions. Furthermore, dose titration as well as the use of alternative LPS preparations or mouse strains allow modulation of the clinical effects, which can exhibit different degrees of ALI severity or early vs. late onset of disease symptoms.

Presented here is a step-by-step procedure to induce acute lung injury in mice by direct intratracheal lipopolysaccharide instillation and to perform FACS analysis of blood samples, bronchoalveolar lavage fluid, and lung tissue. Minimal invasiveness, simple handling, good reproducibility, and titration of disease severity are advantages of this approach.

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Airway administration of lipopolysaccharide (LPS) is a common way to study pulmonary inflammation and acute lung injury (ALI) in small animal models. ...