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

这项工作是由国家机构的健康1R21HL092370-01（GG），1R01 HL095764-01（GG）; R01HL082694（JW），美国心脏协会的创始人联属公司（0855943D，GG），石溪世界 - 赫伯特基金，纽约（SHP）。

1。准备<ol><li>准备以下的解决方案和管（ 表1）如下： <ol><li> Hanks溶液，无钙，镁或指示器，与青霉素（100单位/毫升）/链霉素（100毫克/毫升）。 </li><li>磷酸盐缓冲盐水（PBS），1个，无钙，无镁。 </li><li>乙醇，70％，500毫升。 </li><li>与PBS缓冲甲醛，7％至10％，使500毫升。 </li><li>麻醉科的解决方案： <ol><li>圣阿韦坦。小心地加入2 - 甲基-2 - 丁醇的5毫升至5克的2,2,2 - 三溴乙醇。一?...

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	<li>Price, L. C., 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=Inflammation+in+pulmonary+arterial+hypertension.">Inflammation in pulmonary arterial hypertension.</a> Chest. 141, 210-221 (2012).</li><li>Olschewski, H., 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=Cellular+pathophysiology+and+therapy+of+pulmonary+hypertension.">Cellular pathophysiology and therapy of pulmonary hypertension.</a> J. Lab. Clin. Med. 138, 367-377 (2001).</li><li>Hassoun, P. 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=Inflammation,+growth+factors,+and+pulmonary+vascular+remodeling.">Inflammation, growth factors, and pulmonary vascular remodeling.</a> J. Am. Coll. Cardiol. 54, S10-S19 (2009).</li><li>Rabinovitch, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+pathogenesis+of+pulmonary+arterial+hypertension.">Molecular pathogenesis of pulmonary arterial hypertension.</a> J. Clin. Invest. 118, 2372-2379 (2008).</li><li>Steudel, W., 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=Sustained+pulmonary+hypertension+and+right+ventricular+hypertrophy+after+chronic+hypoxia+in+mice+with+congenital+deficiency+of+nitric+oxide+synthase+3.">Sustained pulmonary hypertension and right ventricular hypertrophy after chronic hypoxia in mice with congenital deficiency of nitric oxide synthase 3.</a> J. Clin. Invest. 101, 2468-2477 (1998).</li><li>Zaidi, S. H., You, X. 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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. Res. 52, 432-441 (1983).</li><li>Kugathasan, L., 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=The+angiopietin-1-Tie2+pathway+prevents+rather+than+promotes+pulmonary+arterial+hypertension+in+transgenic+mice.">The angiopietin-1-Tie2 pathway prevents rather than promotes pulmonary arterial hypertension in transgenic mice.</a> J. Exp. Med. 206, 2221-2234 (2009).</li><li>Bearer, C., Emerson, R. K., ORiordan, M. A., Roitman, E., Shackleton, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Maternal+tobacco+smoke+exposure+and+persistent+pulmonary+hypertension+of+the+newborn.">Maternal tobacco smoke exposure and persistent pulmonary hypertension of the newborn.</a> Environ. Health Persp. , 105-202 (1997).</li><li>Graham, B. 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此处列出的实验流程，允许快速和同时测量右心室收缩压和收获的样品分析的反应在肺部，心脏和免疫系统的小鼠。该过程结合心脏生理学测量，微解剖和随后的组织为活细胞研究，组织学分析，或组学分析的组织收获。的完整过程只需不到20分钟，每鼠标。由于工作区域组织的工作流程，2-3的动物可以同时进行研究。因此，程序是适用于小的，有针对性的实验12和设置在很宽的范围内进?...

<table border="1"><tbody><tr><td> 名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论（可选） </td></tr><tr><td></td><td></td><td></td><td> 试剂 </td></tr><tr><td> 2 - 甲基-2 - 丁醇</td><td> Sigma-Aldrich公司</td><td> 152463 </td><td></td></tr><tr><td> 2,2,2 - 三溴乙醇</td><td> Sigma-Aldrich公司</td><td> T48402 </td><td></td></tr><tr><td>消毒肥皂（专题喷雾TB加STERIS） </td><td> Fisher Scientific则</td><td> 1629至1608年</td><td></td></tr><tr><td> 200的证明，乙醇，绝对无水ACS / USP级</td><td> PHARMCO-AAPER </td><td> 111000200 </td><td>用蒸馏水稀释至70％的</td></tr><tr><td>甲醛溶液</td><td> Sigma-Aldrich公司</td><td> F1635-500ML </td><td>稀释至7-10％，预计ldehyde浓度在PBS中浓度为1x使用PBS原液和水</td></tr><tr><td> Hanks溶液，无钙，镁</td><td> Fisher Scientific则</td><td> 21-022-CV </td><td></td></tr><tr><td> OCT </td><td>组织康</td><td> 4583 </td><td></td></tr><tr><td> （10,000 U / ml）的青霉素/链霉素（10000毫克/毫升）解决方案</td><td> Thermo Scientific的</td><td> SV30010 </td><td></td></tr><tr><td>磷酸盐缓冲盐水（PBS），无钙，无镁，1倍和10倍的解决方案</td><td> Fisher Scientific则</td><td></td><td></td></tr><tr><td>戊巴比妥钠26％ </td><td> Fort Dodge动物保健</td><td> NDC 0856-0471-01 </td><td></td></tr><tr><td></td><td></td><td></td><td> 医学实验 </td></tr><tr><td>板12，24，96以及</td><td>鹰</td><td></td><td></td></TR&gt; <tr><td>移液管</td><td> Fisher Scientific则</td><td> 13-711-9BM </td><td></td></tr><tr><td>管，EDTA涂层; </td><td>萨尔斯塔特</td><td> 2013-08 </td><td></td></tr><tr><td>管0.65毫升和1.7毫升的微离心</td><td> VWR </td><td></td><td></td></tr><tr><td>管12×75毫米聚丙烯</td><td> Fisher Scientific则</td><td> 14-956-1D </td><td></td></tr><tr><td>管，各种大小，聚丙烯</td><td> Fisher Scientific则</td><td></td><td></td></tr><tr><td></td><td></td><td></td><td> 仪器 </td></tr><tr><td>钳，DUMON第5精细</td><td>精细科学工具</td><td> 11254-20 </td><td></td></tr><tr><td>钳，超等格雷夫-0.5mm的技巧弯曲</td><td>精细科学工具</td><td> 11152-10 </td><td></td></tr><tr><td>钳，额外的网络连接东北格雷夫-0.5mm的提示直</td><td>精细科学工具</td><td> 11150-10 </td><td></td></tr><tr><td> 19 GA 18GA，套管</td><td> BD </td><td>精密的滑翔针</td><td>切到最佳长度，减弱外嘶哑创建一个粗糙的外表面</td></tr><tr><td>剪刀，剪刀剥离超薄刀片9厘米</td><td>精细科学工具</td><td> 14081-09 </td><td></td></tr><tr><td> BAL，编织丝线缝合，缝合4-0 </td><td>精细科学工具</td><td> SP116 </td><td></td></tr><tr><td>缝合右心导管检查，编织丝线缝合，6-0 </td><td>泰利福医疗</td><td> 18020-60 </td><td></td></tr><tr><td>注射器，1毫升</td><td> BD </td><td> 309659 </td><td></td></tr><tr><td></td><td></td><td></td><td> 设备 </td></tr><tr><td>放大器，PowerLab系统4/30 </TD&gt; <td> ADInstrument </td><td>型号ML866 </td><td></td></tr><tr><td>导管，压力F1.4 </td><td>米勒仪器公司</td><td> 840-6719 </td><td></td></tr><tr><td>解剖显微镜</td><td>彩色摄影</td><td></td><td></td></tr><tr><td>钳，Vannas春风似剪刀毫米刀片</td><td>精细科学工具</td><td> 15000-00 </td><td></td></tr><tr><td>卤素照明台放大镜</td><td> Fisher Scientific则</td><td> 11-990-56 </td><td></td></tr><tr><td>笔记本电脑</td><td>华硕</td><td>型号A52F酷睿i5处理器，15英寸</td><td></td></tr><tr><td>光源</td><td> Amscope </td><td> HL-250-A </td><td></td></tr><tr><td>压力控制单元</td><td>米勒仪器公司</td><td> PCU-2000 </td><td></td></tr><tr><td>软件，Labchart。亲V.7 </td><td> AD在剂量率仪</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

正确的心脏的功能是把血液通过肺部，从而将右心脏的生理和肺血管生理。炎症的心脏和肺的功能，是一种常见的改性剂，通过制定生产的细胞因子和生长因子，细胞浸润，并开始重塑过程1。 右心室与左心室相比，是一个低压泵工作在一个相对窄的区域的压力变化。增加与肺动脉压力增加的压力在肺血管床和肺动脉高压2。肺动脉高压往往伴有炎症性肺部疾病，如慢性阻塞性肺疾病，自身免疫性疾病3。由于肺动脉高压赋予预后不良的生活质量和寿命，大量的研究是针对了解的机制，mig的HT药物干预4的目标。有效的管理手段，为肺动脉高压的发展面临的主要挑战仍然是复杂的分子和细胞变化在权利的心，肺和免疫系统的同时了解。 在这里，我们提出了一个程序的工作流程，快速，精确的测量压力变化在正确的小鼠心脏的心脏，肺和免疫组织样本的同时收获。该方法是基于直接导管右心室，通过颈内静脉接近上身的小鼠，最早是在20世纪90年代后期，作为替代措施的压力在肺动脉5-13。组织团队的方式有利于快速右心导管检查技术。这使得能够进行测量的小鼠的自发呼吸室内空气。在不同的工作领域的组织的工作流程减少了时间延迟和打开的可能性，同时进行生理实验和收获的免疫系统，心脏和肺组织。 这里列出的程序的工作流程，可以适应各种各样的实验室设置和研究设计，从小型的，有针对性的实验，到大型药物筛选试验。同时采集心肌的生理数据，可以扩展到包括超声心动图5,14-17和收获的心脏，肺和免疫组织减少了需要获得数据的科学知识为基础向前移动的动物。这里介绍的程序的工作流程也提供了理想的基础免疫，肺和心脏功能的网络链接获取知识。这里概述的相同的原则可以适用于研究根据需要的其它的或附加的器官。

获得合适的心脏压力曲线的主要结果是通过合适的心脏导管的正确位置。右心室的导管内部的正确放置的压力时间曲线的形状是重要的，因为会导致压力高原（ 图4）。相反，高低不平的曲线，表明由呼吸或心脏跳动的运动靠在墙上的右心室导管移动。为了检测的阶段的动物的生存的潜在的问题，需要计算ΔP，和心脏的速度的标准偏差。预计这两个值都没有显着不同的治疗组之间。此...

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

一个特异，快速的协议，同时探讨合适的心脏功能，肺部炎症和免疫反应，被描述为一种学习工具。视频和数字，描述的生理和显微切割技术在组织团队的方法，是适用于用于小型到大型研究。

右心室收缩压测量组合在小鼠的肺和免疫组织样品的收获

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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Seven Steps to Stellate Cells

星状细胞的七个步骤

Hepatic stellate cells are liver-resident  cells of star-like morphology and are located in the space of Disse between  liver sinusoidal endothelial cells ...

科研

免疫与感染

Measuring Bacterial Load and Immune Responses in Mice Infected with Listeria monocytogenes

Measuring Bacterial Load and Immune Responses in Mice Infected with Listeria monocytogenes

测量细菌感染小鼠和免疫反应李斯特菌</em

Listeria monocytogenes (Listeria) is a Gram-positive facultative intracellular pathogen1. Mouse studies typically employ intravenous injection of ...

Isolation of Adipose Tissue Immune Cells

脂肪组织免疫细胞的分离

The discovery of increased macrophage infiltration in  the adipose tissue (AT) of obese rodents and humans has led to an  intensification of interest in ...

Induction of Maternal Immune Activation in Mice at Mid-gestation Stage with Viral Mimic Poly(I:C)

Induction of Maternal Immune Activation in Mice at Mid-gestation Stage with Viral Mimic PolyI:C

母体免疫激活诱导小鼠的病毒性模仿聚妊娠中期阶段（I：C）

Maternal immune activation (MIA) model is increasingly well appreciated as a rodent model for the environmental risk factor of various psychiatric ...

Flow Cytometric Analysis of Natural Killer Cell Lytic Activity in Human Whole Blood

人全血中NK细胞杀伤活性的流式细胞仪分析

NK cell cytotoxicity is a widely used measure to determine the effect of outside intervention on NK cell function. However, the accuracy and ...

Assessment of the Cytotoxic and Immunomodulatory Effects of Substances in Human Precision-cut Lung Slices

人精切肺切片中物质的细胞毒性及免疫调节作用的评价

Respiratory diseases in their broad diversity need appropriate model systems to understand the underlying mechanisms and enable development of new ...

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

在小鼠小腿上应用一致的按摩样扰动，并监测由此产生的肌肉压力变化

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

由加德纳菌阴道膀胱暴露在小鼠中引发的复发性大肠杆菌尿路感染

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

监测人类痰样中的中性粒细胞和导管素 G 活性

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

Gene Knock-in by CRISPR/Cas9 and Cell Sorting in Macrophage and T Cell Lines

基因敲击由CRISPR/Cas9和细胞排序在巨噬细胞和T细胞线

Functional genomics studies of the immune system require genetic manipulations that involve both deletion of target genes and addition of elements to ...