Name: in vitro measurements of tracheal constriction using mice
Uploaded: 2012-06-25T12:00:00
Description: Watch this Scientific Journal Video about In vitro Measurements of Tracheal Constriction Using Mice at JoVE.com

这项工作是从创新和治疗气道疾病，NINDS的补助金（NS052574），防治中心和哮喘研究桑德勒计划拨款资助。

1。设备收缩测量设备的主要部件示意图如图2A所示）。 <ol><li> 组织浴。组织沐浴在温暖的温度保持含氧的生理溶液。对小鼠气管环，我们使用循环变暖的解决方案，1熔块玻璃入口泡沫氧（95％/ 5％的O 2 / CO 2的混合物），进口和出口改变端口10毫升组织洗澡，包含了水套解决方案。水库的PSS解决方案存储不断冒泡的95％/ 5％的O 2</...

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T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Methacholine-induced+airway+hyperresponsiveness+is+dependent+on+Galphaq+signaling.">Methacholine-induced airway hyperresponsiveness is dependent on Galphaq signaling.</a> Am. J. Physiol. Lung Cell Mol. Physiol. 285, 114-120 (2003).</li><li>Sausbier, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reduced+rather+than+enhanced+cholinergic+airway+constriction+in+mice+with+ablation+of+the+large+conductance+Ca2+-activated+K++channel.">Reduced rather than enhanced cholinergic airway constriction in mice with ablation of the large conductance Ca2+-activated K+ channel.</a> Faseb. J. 21, 812-822 (2007).</li><li>Scheerens, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Long-term+topical+exposure+to+toluene+diisocyanate+in+mice+leads+to+antibody+production+and+in+vivo+airway+hyperresponsiveness+three+hours+after+intranasal+challenge.">Long-term topical exposure to toluene diisocyanate in mice leads to antibody production and in vivo airway hyperresponsiveness three hours after intranasal challenge.</a> Am. J. Respir. Crit. Care. Med. 159, 1074-1080 (1999).</li><li>Kenakin, T. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> A pharmacology primer : theory, applications, and methods. , (2009).</li><li>Semenov, I., Wang, B., Herlihy, J. T., Brenner, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=BK+Channel+{beta}1+Subunits+Regulate+Airway+Contraction+Secondary+to+M2+Muscarinic+Acetylcholine+Receptor+Mediated+Depolarization.">BK Channel {beta}1 Subunits Regulate Airway Contraction Secondary to M2 Muscarinic Acetylcholine Receptor Mediated Depolarization.</a> J. Physiol. , 1803-1817 (2011).</li><li>Moffatt, J. D., Cocks, T. M., Page, C. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Role+of+the+epithelium+and+acetylcholine+in+mediating+the+contraction+to+5-hydroxytryptamine+in+the+mouse+isolated+trachea.">Role of the epithelium and acetylcholine in mediating the contraction to 5-hydroxytryptamine in the mouse isolated trachea.</a> Br. J. Pharmacol. 141, 1159-1166 (2004).</li><li>Bachar, O., Adner, M., Uddman, R., Cardell, L. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nerve+growth+factor+enhances+cholinergic+innervation+and+contractile+response+to+electric+field+stimulation+in+a+murine+in+vitro+model+of+chronic+asthma.">Nerve growth factor enhances cholinergic innervation and contractile response to electric field stimulation in a murine in vitro model of chronic asthma.</a> Clin. Exp. Allergy. 34, 1137-1145 (2004).</li><li>Weigand, L. A., Myers, A. C., Meeker, S., Undem, B. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mast+cell-cholinergic+nerve+interaction+in+mouse+airways.">Mast cell-cholinergic nerve interaction in mouse airways.</a> J. Physiol. 587, 3355-3362 (2009).</li><li>Kao, J., Fortner, C. N., Liu, L. H., Shull, G. E., Paul, R. 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S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+"classical"+ovalbumin+challenge+model+of+asthma+in+mice.">The "classical" ovalbumin challenge model of asthma in mice.</a> Curr. Drug Targets. 9, 485-494 (2008).</li></ol>

这里介绍的协议提供了生理的准备，评估气道肌肉功能。我们一般同时操作3-4器官浴准备，但是，打包的系统，可以从供应商，允许同时测量高达8制剂（ADInstruments，世界精密仪器，和哈佛仪器）。我们利用力传感器的数量和组织器官浴相同的结果。然而，我们发现，电场刺激提供一些基于刺激电极大小，电极板之间的距离，和编制内电场的位置之间的细微差别的变异。因此，格外小心，应采取?...

<table border="1"><tbody><tr><td> 试剂名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论 </td></tr><tr><td>模拟 - 数字转换器</td><td> ADInstruments </td><td> PowerLab系统4/35 </td><td></td></tr><tr><td>卡巴（Carbamoylcholine氯化物） </td><td> Sigma-Aldrich公司</td><td> C4832 </td><td> 10 -2米的水（等分可以储存在-20°C） </td></tr><tr><td>制图软件</td><td> ADInstrtuments </td><td> LabChart </td><td></td></tr><tr><td>加热循环器</td><td>哈克</td><td>调音台轧机MM400 </td><td></td></tr><tr><td>轴测力传感器</td><td>肯特科学</td><td> TRN001 </td><td></td></tr><tr><td>刺激</td><td>草技术</td><td> S88的双输出SQU脉冲刺激</td><td></td></tr><tr><td>组织浴</td><td> WPI的</td><td> 47264 </td><td></td></tr></tbody></table>

in vitro measurements of tracheal constriction using mice

转基因和基因剔除小鼠的生理和病理生理学航空公司1,2的调查，已经强大的工具。 在体外离体气管准备tensometry已被证明是有用的气道平滑肌（ASM）的转基因小鼠的收缩反应检测。 在体外气管筹备的这些都是比较简单，提供了强有力的回应，并保留两个功能胆碱能神经末梢和肌肉反应，即使经过长时间的孵化。 气管tensometry还提供了一个功能试验，以研究各种第二信使的影响平滑肌收缩的信号转导通路。在气管的收缩主要是由副交感神经，上的ASM（ 图1），释放乙酰胆碱能神经介导。 ASM乙酰胆碱受体主要是毒蕈M2和M3 G I / O和Gq蛋白偶联受体，分别<suP&gt; 3,4,5。 M3受体引起收缩耦合GQ激活磷脂酶C，增加的IP3生产的IP3介导的钙释放和肌浆网3,6,7。 M2 / G I / O信号被认为是加强宫缩抑制腺苷酸环化酶，减少cAMP水平5,8,9,10。这些途径，构成所谓的气道平滑肌11“药物收缩偶联”。此外，通过胆碱能M2受体信号（和M3的信号调制）涉及的去极化的ASM，这反过来又激活L-型电压依赖性钙通道（ 图1）和钙离子内流（即所谓的“兴奋收缩偶联的途径” ）4,7。控制气道收缩的信号传导通路的更详细的评论，可以发现4,12。出现上述途径，小鼠和其他物种之间的保守。然而，小鼠气管不同于其他物种我N部分信号通路。最突出的是他们缺乏13,14组胺和腺苷的收缩反应，无论是在人类和其他物种5,15知名的ASM调制。 在这里，我们提出了隔离小鼠气管环的协议，并在体外测量其收缩输出。包括设备配置的描述，气管环隔离和收缩测量。例子是唤起收缩间接使用高钾刺激神经 ​​和直接由ASM的肌肉去极化，激活电压依赖性钙离子涌入（1。高K +， 图1）。此外，刺激神经 ​​单独使用电场刺激（EFS时， 图1），或直接刺激肌肉的ASM使用外源性神经递质浴（3， 图1。外生的ACH）方法。这FLExibility和便于编制呈现的离体气管环模型参与气道平滑肌收缩的信号级联的一个强大和功能检测。

Watch this Scientific Journal Video about In vitro Measurements of Tracheal Constriction Using Mice at JoVE.com

In vitro Measurements of Tracheal Constriction Using Mice

转基因小鼠已归咎于基因的生理功能极为有用。由于在一般情况下，例如，研究和呼吸道功能的研究，特别是经历了一个对小鼠模型显着的转变。在这里，我们提供的协议<em&gt;在体外</em&gt;气管收缩的研究，以评估在小鼠气道平滑肌功能。

在利用老鼠的气管收缩的体外测量

Transgenic and knockout mice have been powerful tools for the investigation of the physiology and pathophysiology of airways1,2. In vitro tensometry of ...

Research

JoVE Journal

Medicine

In vitro Measurements of Tracheal Constriction Using Mice

Chronic Constriction of the Sciatic Nerve and Pain Hypersensitivity Testing in Rats

慢性缩窄大鼠坐骨神经疼痛过敏测试

Chronic neuropathic pain, resulting from damage to the central or peripheral nervous system, is a prevalent and debilitating condition, affecting 7-18% of ...

科研

医学

Ascending Aortic Constriction in Rats for Creation of Pressure Overload Cardiac Hypertrophy Model

大鼠建立压力超负荷心肌肥厚模型在升主动脉缩窄

Ascending aortic constriction is the most common and successful surgical model for creating pressure overload induced cardiac hypertrophy and heart ...

Intravital Video Microscopy Measurements of Retinal Blood Flow in Mice

视网膜血流小鼠活体视频显微镜测量

Alterations in retinal blood flow can contribute to, or be a consequence of, ocular disease and visual dysfunction. Therefore, quantitation of altered ...

Carotid Artery Infusions for Pharmacokinetic and Pharmacodynamic Analysis of Taxanes in Mice

颈动脉输液的紫杉烷类化合物在小鼠体内的药代动力学和药效学分析

When proposing the use of a drug, drug combination, or drug delivery into a novel system, one must assess the pharmacokinetics of the drug in the study ...

Measuring Ascending Aortic Stiffness In Vivo in Mice Using Ultrasound

Measuring Ascending Aortic Stiffness In Vivo in Mice Using Ultrasound

测量升主动脉刚度<em&gt;在体内</em&gt;小鼠使用超声

We present a protocol for measuring in vivo aortic stiffness in mice using high-resolution ultrasound imaging. Aortic diameter is measured by ultrasound ...

A Model of Cardiac Remodeling Through Constriction of the Abdominal Aorta in Rats

心脏重构通过大鼠腹主动脉缩窄的模型

Heart failure is one of the leading causes of death worldwide. It is a complex clinical syndromethat includes fatigue, dyspnea, exercise intolerance, and ...

Minimally Invasive Transverse Aortic Constriction in Mice

微创横向主动脉缩窄小鼠

Minimally invasive transverse aortic constriction (MTAC) is a more desirable method for the constriction of the transverse aorta in mice than standard ...

Technique of Minimally Invasive Transverse Aortic Constriction in Mice for Induction of Left Ventricular Hypertrophy

小鼠横主动脉狭窄诱导左心室肥厚的技术研究

Transverse aortic constriction (TAC) in mice is one of the most commonly used surgical techniques for experimental investigation of pressure ...

In Vitro and In Vivo Detection of Mitophagy in Human Cells, C. Elegans, and Mice

In Vitro and In Vivo Detection of Mitophagy in Human Cells, C. Elegans, and Mice

体外和体内检测人类细胞中的 Mitophagy,线虫和小鼠

Mitochondria are the powerhouses of cells and produce cellular energy in the form of ATP. Mitochondrial dysfunction contributes to biological aging and a ...

A Closed-chest Model to Induce Transverse Aortic Constriction in Mice

诱导小鼠横向主动脉狭窄的闭合胸模型

Research on cardiac hypertrophy and heart failure is frequently based on pressure overload mouse models induced by TAC. The standard procedure is to ...