Name: phenotyping mouse pulmonary function in vivo with the lung diffusing capacity
Uploaded: 2015-01-06T10:00:00.000+00:00
Duration: 7 min 13 s
Description: Watch this Scientific Journal Video about Phenotyping Mouse Pulmonary Function In Vivo with the Lung Diffusing Capacity at JoVE.com

This work was supported by NIH HL-10342

참고 : 모든 동물 프로토콜은 존스 홉킨스 대학 동물 관리 및 사용위원회에 의해 승인되었습니다. 1. 동물 준비 <ol><li> 아래의 단계 2.3에 설명 된대로 케타민과 자일 라진으로 그들을 마취에 의해, DFCO 측정을 위해 6 C57BL / 6 컨트롤 마우스를 준비합니다. </li><li> 컨트롤과 동일한 방법을 사용하여 표 1에 나타내는 다른 폐 병변을 가진 다른 모든 실험군을 준비한다. 이들 각각의 모델을 구축하기 위해 필요한 특정 세부 사항이 관련 참고 문헌에서 발견된다. 제어 마우스 및 기타 병리학 적 코호트에서 그 나이의 모든 6~12주 있습니다. </li></ol> 일산화탄소에 대한 확산 계수 2. 측정 (DFCO) <ol><li> 질소, 산소, 네온, 및 일산화탄소에 대한 피크를 측정하는 기기에 공급되는 가스 크로마토 모듈을 설정한다. 이 응용 프로그램에만 사용 네온 및 CO 데이터. 참고 :이 장비는 분자의 사용12.00 μm의 필름, 320.00 μm의 ID와 10m 길이의 캐리어 가스로 헬륨 ieve 열. 크로마토 그래피 컬럼은 0.8 ml의 부피를 가지고 있지만, 우리는 샘플 튜브의 적절한 연결을 보장하기 위해 클리어 2ml를 사용했다. </li><li> 각 실험 하루의 시작시 마우스의 샘플의 측정을하기에 앞서, 약 0.5 %의 Ne, 0.5 % CO를 함유하는 가스 혼합물 백에서 직접 2 ㎖의 샘플을 채취하고, 공기를 밸런스, 보정이 샘플을 사용 가스 크로마토 그래피. </li><li> 케타민 (90 ㎎ / ㎏)과 자일 라진 (15 ㎎ / ㎏)와 쥐를 마취하고, 반사 운동의 부재로 마취를 확인합니다. 건조 함을 방지하기 위해 눈에 수의학 연고를 적용합니다. (아주 어린 쥐에서 성인의 18 G, 20 G) 스텁 바늘 캐 뉼러와 마우스를 Tracheostomize. 참고 : DFCO 마취 후 미만 10 분에 완료하고 기계 환기, 또는 다른 절차에 앞서있다. </li><li> 6 주령 마우스에서보다 크고, 3 ㎖ 주사기를 사용하여 tO 혼합 가스 주머니로부터 기체를 인출 2,3,4-. 기관 정맥에 주사기를 연결하고 신속하게 폐를 팽창. 메트로놈을 사용하여, 9 초를 계산 한 다음 신속하게 0.8 ㎖ (호기, 공기)을 철회. </li><li> 이 철회 0.8 ml의 실내 공기와 2 ml의 공기를 내뿜어 희석, 적어도 15 초 동안 휴식 할 수 있습니다. 그 후 분석을 위해 가스 크로마토 그래피로 시료를 주입 전체. </li><li> 이 제 DFCO 샘플을 분석하는 동안, 가스 혼합물로부터 제 가방 0.8 ml의 마우스 폐 팽창하고 첫 번째 샘플이 동일한 샘플을 처리한다. 두 DFCO 측정에 대한 평균값. 주 : 0.8 ml의 부피가 매우 어린 마우스의 폐에서 측정을 수행하기에 너무 크기 때문에 생후 이주로 어린 마우스에서의 측정을 위해, 0.4 ml의 부피를 사용한다. 그것은 6주보다 오래된 쥐 0.8 ㎖의 부피를 사용하는 것이 더이며, 0.4 ml의 부피가 약간 생쥐에 필요하면,이 연구되고 코호트 모든 생쥐 일관 사용해야 함. </li><li> DFCO를 계산(CO 9 / CO의 C) / (느 9 / 네브라스카 C), C와 9 첨자가 주입 된 교정 가스의 농도가 각각 9 초 숨 유지 시간 후에 제거 가스를 참조 - 1 등. </li><li> 분석하고 일방향 ANOVA와 차이점을 비교하고 모든 코호트 마우스에서 다중 비교에 대한 보정 Tukey의 유의 수준을 평가. P &lt;0.05과 같은 중요한 가치를 고려한다. 참고 : 여기에 사용 된 마우스의 모든 여기에보고되지 않은 폐 환기, 기계, 폐 세척, 또는 조직 학적 여러 후속 측정을 포함하는 실험 연구의 일부. 상기 방법은 대조군 마우스 위에서 행해졌 모든 실험 모델에서 동일하기 때문에, 이러한 다양한 병리 모델 만 결과가 제시된다. 이 모델에 대한 추가 정보는 보조 표에 표시됩니다. </li><li> 깊은 anesthe에 의해 동물을 안락사TIC 과다 복용은 자궁 경부 전위 또는 잘린 하였다. 필요한 경우, 추가 생물학적 또는 조직 학적 처리 및 분석을 위해 죽은 쥐에서 폐 세포 및 / 또는 조직을 제거합니다. </li></ol>

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관심 아무것도 없음 충돌은 공개가 없습니다.

본 연구에서는 마우스 폐의 가스 교환 능력을 정량화하는 새로운 메트릭을 정의했다. 이 메트릭은 확산 능, 즉 폐의 주 기능을 측정하는 일반적인 임상 측정 가스를 교환 할 수있는 능력과 유사하다. 확산 능 쉽고 빠르게 마우스 및 인간 모두에서 수행 될 수있는 유일한 폐 기능의 측정이다. 마우스의 폐 질환을 검출하기위한, 주요 목적은 제어 및 실험 코호트 간의 폐 기능의 변화를 정량화하는 ?...

이제 마우스 폐 질환의 다양한 모델링하는데 사용되는 기본 동물이다. 이러한 병리를 underly 메커니즘을 연구하기 위해, 표현형 방법은 그것을 병리학 적 변화를 정량화 할 수 필요합니다. 환기 역학을 측정 마우스 많은 연구가 있지만, 이러한 측정들은 일반적으로 인간 정상적으로 이루어 폐 기능의 평가 표준과 무관하다. 이것은 인간의 질병에 마우스 모델에서의 번역 결과를 용이하게 할 수있다 마우스 및 인간 대상에서 동등한 측정을 수행 할 수있는 능력 때문에, 유감. 피험자의 가장 일반적이고 용이하게 측정 중 하나는 일산화탄소 (DLCO) 1,2 확산 능이지만,이 측정은 거의 마우스 모델에서 수행되지 않았다. 절차는 종종 복잡하거나 REQ 수 있기는 3-7보고되었다 이러한 연구에서, 부분적으로, 어떠한 후속 연구는 없었다복잡한 장비를 uire. 또 다른 방법은 민감한 마우스에서 CO 확산을 측정 할 수 있다는 장점을 갖는다 정상 상태 시스템에서 CO 재 호흡 방법을 사용하는 것이다. 그러나이 방법은 매우 복잡하고, 결과는 마우스의 환기 수준뿐만 아니라 O 2, CO 2 농도 8,9에 따라 달라질 수 있습니다. 이러한 어려움은 여러 가지 장점에도 불구하고, 마우스의 폐 병변을 감지하는 능력을 확산의 일상적인 사용을 배제 것 같다. 마우스에서의 용량의 측정과 확산 문제를 피하기 위해, 간단한 방법의 세부 사항을 마우스에서 측정하기 위해 최근 10보고되었다. 절차는 신속히 전체 영감 가스 동일한 부피를 샘플링함으로써 오염 폐포 가스 채취 어려운 문제를 제거한다. 매우 재현성있는 측정이 프로 시저의 결과, PA의 호스트에 민감 일산화탄소 (DFCO)에 대한 확산 인자로 지칭폐 표현형 thologic 변경됩니다. (CO 9 / CO의 C) / (느 9 / 네브라스카 C), C 및 9 첨자가 주입 된 교정 가스의 농도와 9 초 숨 유지 시간 후에 제거 가스를 참조 - DFCO 따라서 1과 같이 계산된다 각각. DFCO 1은 모든 CO의 완전한 흡수를 반영하고, 0은 어떠한 CO의 흡수를 반영하지으로 0과 1 사이에서 변화 무 차원 변수이다. 이 프레젠테이션에서 우리는이 확산 능 측정을하는 방법을 보여줍니다, 그것은 거의 모든 기종, 섬유증, 급성 폐 손상 및 바이러스 및 곰팡이 감염을 포함하여 기존의 마우스 폐 질환 모델의 변화를 문서화하는 방법을 사용할 수 있습니다.

<table><tbody><tr><td>Gas Chromatograph</td><td>Inficon</td><td>Micro GC Model 3000A</td><td>Agilent makes a comparable model</td></tr><tr><td>18 G Luer stub needle</td><td>Becton Dickenson</td><td></td><td>Several other possible vendors</td></tr><tr><td>3 ml plastic syringe</td><td>Becton Dickenson</td><td></td><td>Several other possible vendors</td></tr><tr><td>Polypropylene gas sample bags</td><td>SKC</td><td>1 or 2 L capacity works well</td><td>Other gas tight bags will work well</td></tr><tr><td>Gas tank, 0.3% Ne, 0.3% CO, balance air; (size ME)</td><td>Airgas, Inc</td><td>Z04 NI785ME3012</td><td>This is the standard mixture used for DLCO in humans</td></tr><tr><td>25 TCID50/mouse of influenza virus A/PR8 diluted in phosphate buffered saline.</td><td></td><td></td><td></td></tr><tr><td>Porcine pancreatic elastase</td><td>Elastin Products, Owensville, MO</td><td></td><td>5.4 U</td></tr><tr><td>Bleomycin</td><td>APP Pharmaceuticals, Schaumburg, IL</td><td></td><td>0.25 U</td></tr><tr><td>&lt;em&gt;Escherichia coli&lt;/em&gt; LPS</td><td>Sigma L2880</td><td></td><td>3 &amp;mu;g/g body weight; O55:B5</td></tr><tr><td>&lt;i&gt;Aspergillus fumigatus &lt;/i&gt; (isolate Af293) conidia were collected from mature colonies grown on potato dextrose agar.</td><td></td><td></td><td></td></tr></tbody></table>

phenotyping mouse pulmonary function in vivo with the lung diffusing capacity

이제 마우스 폐 질환의 다양한 모델링하는데 사용되는 기본 동물이다. 이러한 병리의 기초 메커니즘을 연구하기 위해, 표현형 방법은 병리학 적 변화를 정량화 할 수 필요합니다. 게다가, 마우스 모델 병진 관련성을 제공하기 위해, 이러한 측정은 쉽게 인간 및 마우스 모두에서 시험을 수행 할 수 있어야한다. 불행하게도, 본 문헌에서 폐 기능의 몇 가지 표현형 측정은 인체에 직접 응용 프로그램이 있습니다. 한 가지 예외는 일상적으로 인간에서 수행되는 측정 일산화탄소에 대한 확산 용량입니다. 본 보고서에서는 신속하고 간단하게 마우스에서이 확산 능력을 측정하는 방법을 설명한다. 절차는 1 분 가스 분석 시간 뒤에 마취 마우스의 추적 가스와 간단한 폐 인플레이션을 포함한다. 우리는 폐기종, 섬유증, 급성 폐 손상, 인플루엔자 등 여러 가지 폐 병변을 감지하는이 방법의 능력을 테스트 한진균 폐 감염뿐만 아니라, 젊은 새끼에서 모니터링 폐 성숙. 결과는 상당한 모든 폐 병변의 감소뿐만 아니라 폐 성숙 확산 용량 증가를 나타낸다. 폐 확산 능의이 측정 따라서 기존의 병리학 적 폐 모형의 대부분과 표현형의 구조적인 변화를 감지 할 수있는 능력과 폭 넓은 응용 프로그램이있는 폐 기능 검사를 제공합니다.

도 1이 퍼질러 인플루엔자 감염 모두 상당한 감소뿐만 아니라 섬유증, 폐기종에서 상당한 감소가 있었고, 급성 그룹 A, B, C, D, E, 및 F.에서 성숙한 생쥐 DFCO 측정치를 나타낸다 폐 손상 모델. 그림 2는 2~6주 쥐 나이 시간에 DFCO의 그룹 G 발달 변화를 보여줍니다. 이 때 코스 위에 폐 개발 약간이지만 상당한 증가가 있었다. 작은 팽창 체적의 사용 효과는 6 주 시점에서 아주 명?...

Watch this Scientific Journal Video about Phenotyping Mouse Pulmonary Function In Vivo with the Lung Diffusing Capacity at JoVE.com

Phenotyping Mouse Pulmonary Function In Vivo with the Lung Diffusing Capacity

We describe a means to quickly and simply measure the lung diffusing capacity in mice and show that it is sufficiently sensitive to phenotype changes in multiple common lung pathologies. This metric thus brings direct translational relevance to the mouse models, since diffusing capacity is also easily measured in humans.

표현형 마우스 폐 기능<em&gt; 생체</em&gt; 폐 확산 능력

The mouse is now the primary animal used to model a variety of lung diseases. To study the mechanisms that underlie such pathologies, phenotypic methods ...

phenotyping-mouse-pulmonary-function-vivo-with-lung-diffusing

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JoVE Journal

Biology

Phenotyping Mouse Pulmonary Function In Vivo with the Lung Diffusing Capacity

10.5K 조회수.  Johns Hopkins University Bloomberg School of Public Health. We describe a means to quickly and simply measure the lung diffusing capacity in mice and show that it is sufficiently sensitive to phenotype changes in multiple common lung pathologies. This metric thus brings direct translational relevance to the mouse models, since diffusing capacity is also easily measured in humans.

비디오: Phenotyping Mouse Pulmonary Function In Vivo with the Lung Diffusing Capacity

A Standardized Method for Measuring Internal Lung Surface Area via Mouse Pneumonectomy and Prosthesis Implantation

Pulmonary morphology, physiology, and respiratory functions change in both physiological and pathological conditions. Internal lung surface area (ISA), representing the gas-exchange capacity of the lung, is a critical criterion to assess respiratory function. However, observer bias can significantly influence measured values for lung morphological parameters. The protocol that we describe here minimizes variations during measurements of two morphological parameters used for ISA calculation: internal lung volume (ILV) and mean linear intercept (MLI). Using ISA as a morphometric and functional parameter to determine the outcome of alveolar regeneration in both pneumonectomy (PNX) and prosthesis implantation mouse models, we found that the increased ISA following PNX treatment was significantly blocked by implantation of a prosthesis into the thoracic cavity1. The ability to accurately quantify ISA is not only expected to improve the reliability and reproducibility of lung function studies in injured-induced alveolar regeneration models, but also to promote mechanistic discoveries of multiple pulmonary diseases.

Internal lung surface area (ISA) is a critical criterion for assessing lung morphology and physiology in lung diseases and injury-induced alveolar regeneration. We describe here a standardized method that can minimize the measurement bias for ISA in both lung pneumonectomy and prosthesis implantation mouse models.

마우스 폐 전 절제술 및 보철물 삽입을 통한 내시경 표면 측정을위한 표준화 된 방법

Pulmonary morphology, physiology, and respiratory functions change in both physiological and pathological conditions. Internal lung surface area (ISA), ...

연구

발생학

Automated Measurement of Pulmonary Emphysema and Small Airway Remodeling in Cigarette Smoke-exposed Mice

COPD is projected to be the third most common cause of mortality world-wide by 2020(1). Animal models of COPD are used to identify molecules that contribute to the disease process and to test the efficacy of novel therapies for COPD. Researchers use a number of models of COPD employing different species including rodents, guinea-pigs, rabbits, and dogs(2). However, the most widely-used model is that in which mice are exposed to cigarette smoke. Mice are an especially useful species in which to model COPD because their genome can readily be manipulated to generate animals that are either deficient in, or over-express individual proteins. Studies of gene-targeted mice that have been exposed to cigarette smoke have provided valuable information about the contributions of individual molecules to different lung pathologies in COPD(3-5). Most studies have focused on pathways involved in emphysema development which contributes to the airflow obstruction that is characteristic of COPD. However, small airway fibrosis also contributes significantly to airflow obstruction in human COPD patients(6), but much less is known about the pathogenesis of this lesion in smoke-exposed animals. To address this knowledge gap, this protocol quantifies both emphysema development and small airway fibrosis in smoke-exposed mice. This protocol exposes mice to CS using a whole-body exposure technique, then measures respiratory mechanics in the mice, inflates the lungs of mice to a standard pressure, and fixes the lungs in formalin. The researcher then stains the lung sections with either Gill&#8217;s stain to measure the mean alveolar chord length (as a readout of emphysema severity) or Masson&#8217;s trichrome stain to measure deposition of extracellular matrix (ECM) proteins around small airways (as a readout of small airway fibrosis). Studies of the effects of molecular pathways on both of these lung pathologies will lead to a better understanding of the pathogenesis of COPD.

The goal of this protocol is to provide automated methods to quantify chronic lung pathologies in a murine model of COPD. The protocol includes exposing mice to cigarette smoke (CS), measuring pulmonary function, inflating the lungs, and using morphometry methods to measure emphysema and small airway remodeling in mice.

담배 연기에 노출 된 마우스의 폐 폐기종 및 작은기도 리모델링 자동 측정

COPD is projected to be the third most common cause of mortality world-wide by 2020(1).  Animal models of COPD are used to identify molecules that ...

의학

Measurement of the Pressure-volume Curve in Mouse Lungs

In recent decades the mouse has become the primary animal model of a variety of lung diseases. In models of emphysema or fibrosis, the essential phenotypic changes are best assessed by measurement of the changes in lung elasticity. To best understand specific mechanisms underlying such pathologies in mice, it is essential to make functional measurements that can reflect the developing pathology. Although there are many ways to measure elasticity, the classical method is that of the total lung pressure-volume (PV) curve done over the whole range of lung volumes. This measurement has been made on adult lungs from nearly all mammalian species dating back almost 100 years, and such PV curves also played a major role in the discovery and understanding of the function of pulmonary surfactant in fetal lung development. Unfortunately, such total PV curves have not been widely reported in the mouse, despite the fact that they can provide useful information on the macroscopic effects of structural changes in the lung. Although partial PV curves measuring just the changes in lung volume are sometimes reported, without a measure of absolute volume, the nonlinear nature of the total PV curve makes these partial ones very difficult to interpret. In the present study, we describe a standardized way to measure the total PV curve. We have then tested the ability of these curves to detect changes in mouse lung structure in two common lung pathologies, emphysema and fibrosis. Results showed significant changes in several variables consistent with expected structural changes with these pathologies. This measurement of the lung PV curve in mice thus provides a straightforward means to monitor the progression of the pathophysiologic changes over time and the potential effect of therapeutic procedures.

Here we present a protocol to simply and reliably measure the lung pressure-volume curve in mice, showing that it is sufficiently sensitive to detect phenotypic parenchymal changes in two common lung pathologies, pulmonary fibrosis and emphysema. This metric provides a means to quantify the lung&#8217;s structural changes with developing pathology.

마우스 폐의 압력 볼륨 곡선의 측정

In recent decades the mouse has become the primary animal model of a variety of lung diseases. In models of emphysema or fibrosis, the essential ...

Intraperitoneal Glucose Tolerance Test, Measurement of Lung Function, and Fixation of the Lung to Study the Impact of Obesity and Impaired Metabolism on Pulmonary Outcomes

Obesity and respiratory disorders are major health problems. Obesity is becoming an emerging epidemic with an expected number of over 1 billion obese individuals worldwide by 2030, thus representing a growing socioeconomic burden. Simultaneously, obesity-related comorbidities, including diabetes as well as heart and chronic lung diseases, are continuously on the rise. Although obesity has been associated with increased risk for asthma exacerbations, worsening of respiratory symptoms, and poor control, the functional role of obesity and perturbed metabolism in the pathogenesis of chronic lung disease is often underestimated, and underlying molecular mechanisms remain elusive. This article aims to present methods to assess the effect of obesity on metabolism, as well as lung structure and function. Here, we describe three techniques for mice studies: (1) assessment of intraperitoneal glucose tolerance (ipGTT) to analyze the effect of obesity on glucose metabolism; (2) measurement of airway resistance (Res) and respiratory system compliance (Cdyn) to analyze the effect of obesity on lung function; and (3) preparation and fixation of the lung for subsequent quantitative histological assessment. Obesity-related lung diseases are probably multifactorial, stemming from systemic inflammatory and metabolic dysregulation that potentially adversely influence lung function and the response to therapy. Therefore, a standardized methodology to study molecular mechanisms and the effect of novel treatments is essential.

The incidence of obesity is rising and increases the risk of chronic lung diseases. To establish the underlying mechanisms and preventive strategies, well-defined animal models are needed. Here, we provide three methods (glucose-tolerance-test, body plethysmography, and lung fixation) to study the effect of obesity on pulmonary outcomes in mice.

복 포도 당 내성 검사, 폐 기능 측정 및 폐의 고정 비만 및 장애인된 대사 폐 결과에 미치는 영향 연구

Obesity and respiratory disorders are major health problems. Obesity is becoming an emerging epidemic with an expected number of over 1 billion obese ...

면역학 및 감염병학

Generation of a Chronic Obstructive Pulmonary Disease Model in Mice by Repeated Ozone Exposure

Chronic obstructive pulmonary disease (COPD) is characterized by persistent airflow limitation and lung parenchymal destruction. It has a very high incidence in aging populations. The current conventional therapies for COPD focus mainly on symptom-modifying drugs; thus, the development of new therapies is urgently needed. Qualified animal models of COPD could help to characterize the underlying mechanisms and can be used for new drug screening. Current COPD models, such as lipopolysaccharide (LPS) or the porcine pancreatic elastase (PPE)-induced emphysema model, generate COPD-like lesions in the lungs and airways but do not otherwise resemble the pathogenesis of human COPD. A cigarette smoke (CS)-induced model remains one of the most popular because it not only simulates COPD-like lesions in the respiratory system, but it is also based on one of the main hazardous materials that causes COPD in humans. However, the time-consuming and labor-intensive aspects of the CS-induced model dramatically limit its application in new drug screening. In this study, we successfully generated a new COPD model by exposing mice to high levels of ozone. This model demonstrated the following: 1) decreased forced expiratory volume 25, 50, and 75/forced vital capacity (FEV25/FVC, FEV50/FVC, and FEV75/FVC), indicating the deterioration of lung function; 2) enlarged lung alveoli, with lung parenchymal destruction; 3) reduced fatigue time and distance; and 4) increased inflammation. Taken together, these data demonstrate that the ozone exposure (OE) model is a reliable animal model that is similar to humans because ozone overexposure is one of the etiological factors of COPD. Additionally, it only took 6 - 8 weeks, based on our previous work, to create an OE model, whereas it requires 3 - 12 months to induce the cigarette smoke model, indicating that the OE model might be a good choice for COPD research.

This study describes the successful generation of a new chronic obstructive pulmonary disease (COPD) animal model by repeatedly exposing mice to high concentrations of ozone.

반복된 오존 노출에 의해 쥐에 만성 폐쇄성 폐 질환 모델의 생성

Chronic obstructive pulmonary disease (COPD) is characterized by persistent airflow limitation and lung parenchymal destruction. It has a very high ...

Evaluation of Respiratory System Mechanics in Mice using the Forced Oscillation Technique

The forced oscillation technique (FOT) is a powerful, integrative and translational tool permitting the experimental assessment of lung function in mice in a comprehensive, detailed, precise and reproducible manner. It provides measurements of respiratory system mechanics through the analysis of pressure and volume signals acquired in reaction to predefined, small amplitude, oscillatory airflow waveforms, which are typically applied at the subject's airway opening. The present protocol details the steps required to adequately execute forced oscillation measurements in mice using a computer-controlled piston ventilator (flexiVent; SCIREQ Inc, Montreal, Qc, Canada). The description is divided into four parts: preparatory steps, mechanical ventilation, lung function measurements, and data analysis. It also includes details of how to assess airway responsiveness to inhaled methacholine in anesthetized mice, a common application of this technique which also extends to other outcomes and various lung pathologies. Measurements obtained in na&iuml;ve mice as well as from an oxidative-stress driven model of airway damage are presented to illustrate how this tool can contribute to a better characterization and understanding of studied physiological changes or disease models as well as to applications in new research areas.

The present protocol provides a detailed step-by-step description of the procedures required to execute measurements of respiratory system mechanics as well as the assessment of airway responsiveness to inhaled methacholine in mice using the forced oscillation technique (flexiVent; SCIREQ Inc, Montreal, Qc, Canada).

강제 진동 기술을 사용하여 마우스의 호흡기 시스템 역학의 평가

The  forced oscillation technique (FOT) is a powerful, integrative and translational  tool permitting the experimental assessment of lung function in mice ...

생물학

Assessment of Respiratory Function in Conscious Mice by Double-chamber Plethysmography

Air volume changes created by a conscious subject breathing spontaneously within a body box are at the basis of plethysmography, a technique used to non-invasively assess some features of the respiratory function in humans as well as in laboratory animals. The present article focuses on the application of the double-chamber plethysmography (DCP) in small animals. It provides background information on the methodology as well as a detailed step-by-step procedure to successfully assess respiratory function in conscious, spontaneously breathing animals in a non-invasive manner. The DCP can be used to monitor the respiratory function of multiple animals in parallel, as well as to identify changes induced by aerosolized substances over a chosen time period and in a repeated manner. Experiments on control and allergic mice are used herein to demonstrate the utility of the technique, explain the associated outcome parameters, as well as to discuss the related advantages and shortcomings. Overall, the DCP provides valid and theoretically sound readouts that can be trusted to evaluate the respiratory function of conscious small animals both at baseline and after challenges with aerosolized substances.

The objective of the present article is to provide a detailed description of the recommended procedures to evaluate respiratory function in conscious mice by double-chamber plethysmography.

더블 챔버 Plethysmography 여 의식 생쥐에 호흡기 기능의 평가

Air volume changes created by a conscious subject breathing spontaneously within a body box are at the basis of plethysmography, a technique used to ...

Hyperpolarized 129Xe Lung MRI and Spectroscopy in Mechanically Ventilated Mice

Hyperpolarized (HP) xenon-129 (129Xe) is an inhaled magnetic resonance imaging (MRI) contrast agent with unique spectral and physical properties that can be exploited to quantify pulmonary physiology, including ventilation, restricted diffusion (alveolar-airspace size), and gas exchange. In humans, it has been used to evaluate disease severity and progression in a variety of pulmonary disorders and is approved for clinical use in the United States and United Kingdom. Beyond its clinical applications, the ability of 129Xe MRI to noninvasively assess pulmonary pathophysiology and provide spatially resolved information is valuable for preclinical research. Among animal models, mice are the most widely used due to the accessibility of genetically modified disease models. Here, 129Xe MRI is promising as a minimally invasive, radiation-free, and sensitive technique to longitudinally monitor lung disease progression and therapy response (e.g., in drug discovery). This technique can extend to preclinical applications by incorporating an MRI-triggered, free-breathing apparatus or mechanical ventilator to deliver gas. Here, we describe the steps and provide checklists to ensure robust data collection and analysis, including creating a thermally polarized xenon gas phantom for quality control, optimizing polarization, animal handling (sedation, intubation, ventilation, and care for mice), and protocols for ventilation, restricted diffusion, and gas exchange data. While preclinical 129Xe MRI can be applied in various animal models (e.g., rats, pigs, sheep), this protocol focuses on mice due to the challenges posed by their small anatomy, which are balanced by their affordability and the availability of many disease models.

Hyperpolarized xenon MRI can quantify regional lung microstructure (air-space dimensions) and physiology (ventilation and gas exchange) in translational research and clinical care. Although challenging, it can provide comparable pulmonary insights in preclinical studies. This protocol describes the infrastructure and procedures needed to perform routine xenon lung MRI in mice.

Hyperpolarized (HP) xenon-129 (129Xe) is an inhaled magnetic resonance imaging (MRI) contrast agent with unique spectral and physical properties that can ...

생체공학

Measuring Respiratory Function in Mice Using Unrestrained Whole-body Plethysmography

Respiratory dysfunction is one of the leading causes of morbidity and mortality in the world and the rates of mortality continue to rise. Quantitative assessment of lung function in rodent models is an important tool in the development of future therapies. Commonly used techniques for assessing respiratory function including invasive plethysmography and forced oscillation. While these techniques provide valuable information, data collection can be fraught with artefacts and experimental variability due to the need for anesthesia and/or invasive instrumentation of the animal. In contrast, unrestrained whole-body plethysmography (UWBP) offers a precise, non-invasive, quantitative way by which to analyze respiratory parameters. This technique avoids the use of anesthesia and restraints, which is common to traditional plethysmography techniques. This video will demonstrate the UWBP procedure including the equipment set up, calibration and lung function recording. It will explain how to analyze the collected data, as well as identify experimental outliers and artefacts that results from animal movement. The respiratory parameters obtained using this technique include tidal volume, minute volume, inspiratory duty cycle, inspiratory flow rate and the ratio of inspiration time to expiration time. UWBP does not rely on specialized skills and is inexpensive to perform. A key feature of UWBP, and most appealing to potential users, is the ability to perform repeated measures of lung function on the same animal.

The assessment of respiratory physiology has traditionally relied upon techniques, which require restraint or sedation of the animal. Unrestrained whole-body plethysmography, however, provides precise, non-invasive, quantitative analysis of respiratory physiology in animal models. In addition, the technique allows repeated respiratory assessment of mice allowing for longitudinal studies.

무제한 전신 혈량 측정법을 사용하여 마우스의 호흡 기능을 측정

Respiratory dysfunction is one of the leading causes of morbidity and mortality in the world and the rates of mortality continue to rise. Quantitative ...

In vivo Evaluation of Mucociliary Clearance in Mice

Respiratory motile cilia, specialized organelles of the cell, line the apical surface of epithelial cells lining the respiratory tract. By beating in a metachronal, synchronal fashion, these multiple, motile, actin-based organelles generate a cephalad fluid flow clearing the respiratory tract of inhaled pollutants and pathogens. With increasing environmental pollution, novel viral pathogens and emerging multi-drug resistant bacteria, cilia generated mucociliary clearance (MCC) is essential for maintaining lung health. MCC is also depressed in multiple congenital disorders like primary ciliary dyskinesia, cystic fibrosis as well as acquired disorders like chronic obstructive pulmonary disease. All these disorders have established, in some case multiple, mouse models. In this publication, we detail a method using a small amount of radioactivity and dual-modality SPECT/CT imaging to accurately and reproducibly measure MCC in mice in vivo. The method allows for recovery of mice after imaging, making serial measurements possible, and testing potential therapeutics longitudinally over time. The data in wild-type mice demonstrates the reproducibility of the MCC measurement as long as adequate attention to detail is paid, and the protocol strictly adhered to.

In this publication, we describe protocols for assessing airway mucociliary clearance (MCC) in mice in vivo utilizing dual-modality radionuclide imaging. This protocol is designed for a single photon emission computed tomography (SPECT) and computed tomography (CT) acquisition protocol using mouse whole body (MWB) collimators in a dual SPECT/CT system.

마우스의 점도 클리어런스의 생체 내 평가

Respiratory motile cilia, specialized organelles of the cell, line the apical surface of epithelial cells lining the respiratory tract. By beating in a ...

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