Name: right ventricular systolic pressure measurements in combination with harvest of lung and immune tissue samples in mice
Uploaded: 2013-01-16T12:00:00
Description: Watch this Scientific Journal Video about Right Ventricular Systolic Pressure Measurements in Combination with Harvest of Lung and Immune Tissue Samples in Mice at JoVE.com

Эта работа финансировалась Национальным институтом здоровья 1R21HL092370-01 (GG), 1R01 HL095764-01 (GG); R01HL082694 (JW), Американской ассоциации сердца, Учредителей филиал (0855943D, GG); Stony Wold - Герберт фонда, Нью-Йорк (SHP).

1. Подготовка <ol><li> Подготовьте следующие решения и трубы (таблица 1) следующим образом: <ol><li> Хэнкс решение, не кальций, магний или индикатора с пенициллина (100 ЕД / мл) / стрептомицин (100 мкг / мл). </li><li> Фосфатным буферным раствором (PBS), 1x, ни кальций, ни магний. </li><li> Этанол, 70%, состав?...

<ol>
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J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Attenuation+of+leukocyte+recruitment+via+CXCR1/2+inhibition+stops+the+progression+of+PAH+in+mice+with+genetic+ablation+of+endothelial+BMPR-II.">Attenuation of leukocyte recruitment via CXCR1/2 inhibition stops the progression of PAH in mice with genetic ablation of endothelial BMPR-II.</a> Blood. 118, 4750-4758 (2011).</li><li>Fujita, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pulmonary+hypertension+in+TNF-alpha-overexpressing+mice+is+associated+with+decreased+VEGF+gene+expression.">Pulmonary hypertension in TNF-alpha-overexpressing mice is associated with decreased VEGF gene expression.</a> J. Appl. Physiol. 93, 2162-2170 (2002).</li><li>Motley, H. L., Cournand, A., Werko, L., Himmelstein, A., Dresdale, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Influence+of+Short+Periods+of+Induced+Acute+Anoxia+Upon+Pulmonary+Artery+Pressures+in+Man.">The Influence of Short Periods of Induced Acute Anoxia Upon Pulmonary Artery Pressures in Man.</a> Am. J. Physiol. 150, 315-320 (1947).</li><li>Liljestrand, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regulation+of+Pulmonary+Arterial+Blood+Pressure.">Regulation of Pulmonary Arterial Blood Pressure.</a> Arch. Intern. Med. 81, 162-172 (1948).</li><li>Euler, U. S. V., Liljestrand, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Observations+on+the+pulmonary+arterial+blood+pressure+in+the+cat.">Observations on the pulmonary arterial blood pressure in the cat.</a> Acta Physiol. Scand. 12, 301-320 (1946).</li><li>Van den Broeck, W., Derore, A., Simoens, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anatomy+and+nomenclature+of+murine+lymph+nodes:+Descriptive+study+and+nomenclatory+standardization+in+BALB/cAnNCrl+mice.">Anatomy and nomenclature of murine lymph nodes: Descriptive study and nomenclatory standardization in BALB/cAnNCrl mice.</a> Journal of immunological. 312, 12-19 (2006).</li><li>Rabinovitch, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Angiotensin+II+prevents+hypoxic+pulmonary+hypertension+and+vascular+changes+in+rat.">Angiotensin II prevents hypoxic pulmonary hypertension and vascular changes in rat.</a> Am. J. Physiol. 254, 500-508 (1988).</li><li>Rabinovitch, M., Gamble, W., Nadas, A. S., Miettinen, O. S., Reid, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rat+pulmonary+circulation+after+chronic+hypoxia:+hemodynamic+and+structural+features.">Rat pulmonary circulation after chronic hypoxia: hemodynamic and structural features.</a> Am. J. Physiol. 236, 818-827 (1979).</li><li>Rabinovitch, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Changes+in+pulmonary+blood+flow+affect+vascular+response+to+chronic+hypoxia+in+rats.">Changes in pulmonary blood flow affect vascular response to chronic hypoxia in rats.</a> Circ. 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Экспериментальные потока изложенные здесь обеспечивает быстрое и одновременное измерение давления правого желудочка систолическое и сбор образцов для анализа ответов в легких, сердца и иммунной системы у мышей. Процедура сочетает в себе сердце физиологии измерений, микро-рассечени?...

<table border="1"><tbody><tr><td> Имя </td><td> Компания </td><td> Номер в каталоге </td><td> Комментарии (по желанию) </td></tr><tr><td></td><td></td><td></td><td> Реагенты </td></tr><tr><td> 2-метил-2-бутанол </td><td> Sigma-Aldrich </td><td> 152463 </td><td></td></tr><tr><td> 2,2,2-Tribromoethanol </td><td> Sigma-Aldrich </td><td> T48402 </td><td></td></tr><tr><td> дезинфицирующим мылом (покрытия Spray ТБ плюс Steris) </td><td> Fisher Scientific </td><td> 1629-08 </td><td></td></tr><tr><td> Этиловый спирт, 200 Proof, абсолютный, безводный ACS / USP класс </td><td> Pharmco-AAPER </td><td> 111000200 </td><td> Разбавить до 70% дистиллированной воды </td></tr><tr><td> Раствор формальдегида </td><td> Sigma-Aldrich </td><td> F1635-500ML </td><td> Развести до 7-10% образованийldehyde концентрации в концентрации PBS в 1x PBS использованием раствора и воды </td></tr><tr><td> Хэнкс решение, не кальций, магний </td><td> Fisher Scientific </td><td> 21-022-CV </td><td></td></tr><tr><td> Октябрь </td><td> Tissue-Tek </td><td> 4583 </td><td></td></tr><tr><td> Пенициллина (10000 ЕД / мл) / стрептомицин (10.000 мг / мл) </td><td> Thermo Scientific </td><td> SV30010 </td><td></td></tr><tr><td> Фосфатным буферным раствором (PBS), ни кальций, ни магний, 1x и 10x решения </td><td> Fisher Scientific </td><td></td><td></td></tr><tr><td> Пентобарбитала натрия 26% </td><td> Fort Dodge Animal Health </td><td> NDC 0856-0471-01 </td><td></td></tr><tr><td></td><td></td><td></td><td> Лабораторное оборудование </td></tr><tr><td> Пластины 12, 24, 96 а </td><td> Сокол </td><td></td><td></td></TR&gt; <tr><td> Передача Pipet </td><td> Fisher Scientific </td><td> 13-711-9BM </td><td></td></tr><tr><td> Tube, покрытые ЭДТА </td><td> Sarstedt </td><td> 2013-08 </td><td></td></tr><tr><td> Трубы 0,65 мл и 1,7 мл микро-центрифуге </td><td> VWR </td><td></td><td></td></tr><tr><td> Трубы 12 х 75 мм, полипропилен </td><td> Fisher Scientific </td><td> 14-956-1D </td><td></td></tr><tr><td> Трубы, различных размеров, полипропилен </td><td> Fisher Scientific </td><td></td><td></td></tr><tr><td></td><td></td><td></td><td> Инструменты </td></tr><tr><td> Пинцет, Дюмон # 5 Fine </td><td> Средства изобразительных наук </td><td> 11254-20 </td><td></td></tr><tr><td> Пинцет, дополнительный штраф Грефе-0.5mm советы изогнутые </td><td> Средства изобразительных наук </td><td> 11152-10 </td><td></td></tr><tr><td> Пинцет, дополнительно Fiпе Грефе-0.5mm советы прямой </td><td> Средства изобразительных наук </td><td> 11150-10 </td><td></td></tr><tr><td> Канюля 18ga, 19 га </td><td> BD </td><td> Precision Glide иглы </td><td> Смена оптимальной длины, притупляются и за ее пределами прохрипел для создания грубой внешней поверхности </td></tr><tr><td> Ножницы, ножницы Dissector-тонкий лезвия 9см </td><td> Средства изобразительных наук </td><td> 14081-09 </td><td></td></tr><tr><td> Шовный для BAL, плетеный шовный материал шелк, 4-0 </td><td> Средства изобразительных наук </td><td> SP116 </td><td></td></tr><tr><td> Шовный для катетеризации правых отделов сердца, плетеные шелковые шва, 6-0 </td><td> Teleflex медицинских </td><td> 18020-60 </td><td></td></tr><tr><td> Шприц 1 мл </td><td> BD </td><td> 309659 </td><td></td></tr><tr><td></td><td></td><td></td><td> Оборудование </td></tr><tr><td> Усилители, PowerLab 4/30 </TD&gt; <td> ADInstrument </td><td> Модель ML866 </td><td></td></tr><tr><td> Катетер, давление F1.4 </td><td> Millar Instruments, Inc </td><td> 840-6719 </td><td></td></tr><tr><td> Препаровальная лупа </td><td> Variscope </td><td></td><td></td></tr><tr><td> Щипцы, ножницы Vannas весна-2мм лезвия </td><td> Средства изобразительных наук </td><td> 15000-00 </td><td></td></tr><tr><td> Галогенные освещенный бюро лупы </td><td> Fisher Scientific </td><td> 11-990-56 </td><td></td></tr><tr><td> Портативный компьютер </td><td> Asus </td><td> Номер модели A52F i5 процессор; 15-дюймовых </td><td></td></tr><tr><td> Источник света </td><td> Amscope </td><td> HL-250- </td><td></td></tr><tr><td> Группа контроля давления </td><td> Millar Instruments, Inc </td><td> PCU-2000 </td><td></td></tr><tr><td> Программное обеспечение, LabChart-Pro V.7 </td><td> Н.э. Вприборов </td><td></td><td></td></tr></tbody></table>

right ventricular systolic pressure measurements in combination with harvest of lung and immune tissue samples in mice

The function of the right heart is to pump blood through the lungs, thus linking right heart physiology and pulmonary vascular physiology. Inflammation is a common modifier of heart and lung function, by elaborating cellular infiltration, production of cytokines and growth factors, and by initiating remodeling processes 1.
Compared to the left ventricle, the right ventricle is a low-pressure pump that operates in a relatively narrow zone of pressure changes. Increased pulmonary artery pressures are associated with increased pressure in the lung vascular bed and pulmonary hypertension 2. Pulmonary hypertension is often associated with inflammatory lung diseases, for example chronic obstructive pulmonary disease, or autoimmune diseases 3. Because pulmonary hypertension confers a bad prognosis for quality of life and life expectancy, much research is directed towards understanding the mechanisms that might be targets for pharmaceutical intervention 4. The main challenge for the development of effective management tools for pulmonary hypertension remains the complexity of the simultaneous understanding of molecular and cellular changes in the right heart, the lungs and the immune system.
Here, we present a procedural workflow for the rapid and precise measurement of pressure changes in the right heart of mice and the simultaneous harvest of samples from heart, lungs and immune tissues. The method is based on the direct catheterization of the right ventricle via the jugular vein in close-chested mice, first developed in the late 1990s as surrogate measure of pressures in the pulmonary artery5-13. The organized team-approach facilitates a very rapid right heart catheterization technique. This makes it possible to perform the measurements in mice that spontaneously breathe room air. The organization of the work-flow in distinct work-areas reduces time delay and opens the possibility to simultaneously perform physiology experiments and harvest immune, heart and lung tissues.
The procedural workflow outlined here can be adapted for a wide variety of laboratory settings and study designs, from small, targeted experiments, to large drug screening assays. The simultaneous acquisition of cardiac physiology data that can be expanded to include echocardiography5,14-17 and harvest of heart, lung and immune tissues reduces the number of animals needed to obtain data that move the scientific knowledge basis forward. The procedural workflow presented here also provides an ideal basis for gaining knowledge of the networks that link immune, lung and heart function. The same principles outlined here can be adapted to study other or additional organs as needed.

Основным результатом для получения права кривые давления сердца достигается правильное положение правой катетер сердца. Форма кривых давления время имеет решающее значение, потому что правильное размещение катетера в правом желудочке приведет к давлению плато (рис. 4). Spiky кр...

Watch this Scientific Journal Video about Right Ventricular Systolic Pressure Measurements in Combination with Harvest of Lung and Immune Tissue Samples in Mice at JoVE.com

Right Ventricular Systolic Pressure Measurements in Combination with Harvest of Lung and Immune Tissue Samples in Mice

Конкретных и быстрых протоколов одновременно исследовать функцию правого сердца, воспаление легких, и иммунный ответ описывается как инструмент обучения. Видео и цифры описывают физиологию и микродиссекции методы в организованном командой подход, который адаптирован для использования в малых и больших размеров исследований.

Правого желудочка систолическое давление измерений в сочетании с Harvest легких и иммунной образцы тканей у мышей

The function of the right heart is to pump blood through the lungs, thus linking right heart physiology and pulmonary vascular physiology. Inflammation is ...

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Application of Consistent Massage-Like Perturbations on Mouse Calves and Monitoring the Resulting Intramuscular Pressure Changes

Применение последовательных массаж-как Пертурбации на мышителях и мониторинг результирующих внутримышечных изменений давления

Massage is generally recognized to be beneficial for relieving pain and inflammation. Although previous studies have reported anti-inflammatory effects of ...

Recurrent Escherichia coli Urinary Tract Infection Triggered by Gardnerella vaginalis Bladder Exposure in Mice

Recurrent Escherichia coli Urinary Tract Infection Triggered by Gardnerella vaginalis Bladder Exposure in Mice

Рецидивирующая инфекция мочевыводящих путей Escherichia coli , вызванная воздействием Gardnerella vaginalis на мочевой пузырь у мышей

Recurrent urinary tract infections (rUTI) caused by uropathogenic Escherichia coli (UPEC) are common and costly. Previous articles describing models of ...

Monitoring Neutrophil Elastase and Cathepsin G Activity in Human Sputum Samples

Мониторинг активности нейтрофильной эластазы и катепсина G в образцах мокроты человека

Proteases are regulators of countless physiological processes and the precise investigation of their activities remains an intriguing biomedical ...

Separation of Immune Cell Subpopulations in Peripheral Blood Samples from Children with Infectious Mononucleosis

Отделение субпопуляций иммунных клеток в образцах периферической крови у детей с инфекционным мононуклеозом

Infectious mononucleosis (IM) is an acute syndrome mostly associated with primary Epstein&#8211;Barr virus (EBV) infection. The main clinical symptoms ...

Induction of Periodontitis via a Combination of Ligature and Lipopolysaccharide Injection in a Rat Model

Induction of Periodontitis via a Combination of Ligature and Lipopolysaccharide Injection in a Rat Model

Индукция пародонтита с помощью комбинации лигатуры и инъекции липополисахарида на крысиной модели

Periodontitis (PD) is a highly prevalent, chronic immune-inflammatory disease of the periodontium, that results in a loss of gingival soft tissue, ...