Name: exploring arterial smooth muscle kv7 potassium channel function using patch clamp electrophysiology and pressure myography
Uploaded: 2012-09-14T13:00:00
Description: Watch this Scientific Journal Video about Exploring Arterial Smooth Muscle Kv7 Potassium Channel Function using Patch Clamp Electrophysiology and Pressure Myography at JoVE.com

这项工作是由国家心脏，肺和血液研究所（NIH R01-HL089564）的资助KLB和博士后奖学金由美国心脏协会（09PRE2260209）和Arthur J.施密特基金会BKM。

1。手术切除的小肠肠系膜血管商场<ol><li>麻醉300-400克SD大鼠（4％）与异氟醚吸入给药。 </li><li>执行中线剖腹手术暴露的小肠系膜。 Exteriorize通过腹部切口非常小心，以避免创伤暴露的肠管及肠系膜小肠和大肠。轻轻地扇了的肠系膜超过无菌纱布。 </li><li>外科手术切除小肠和大肠的一部分，包括盲肠（沿利润率和肠系膜上动脉/静脉）从主分支的起源仔细剪裁血管。 </li><li>切除的组织转移到烧?...

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这里所描述的方法和实验方法相当强劲，并且能够产生明确的和可重复的结果时一丝不苟的细节。良好的电生理记录和收缩/扩张动脉段是依赖于细胞和动脉血管的健康。细胞制剂可以改变每一天，甚至使用相同的协议。隔离解决方案，可用于长达2周，但如果细胞制备的质量低，随之而来的两个天应准备新的隔离解决方案。我们已经发现，一批一批，因此隔离条件（时间的培养和酶的浓度）酶活?...

<table border="1"><tbody><tr><td> 名称 </td><td> 公司 </td><td> 目录编号 </td><td>评论</td></tr><tr><td>氯化钠</td><td>西格玛</td><td> S5886 </td><td>剖析解决方案：145 沐浴液电：140 电：10内部解决方案 为心肌细胞的隔离解决方案：140 浴的压力myography解决方案：145 流明的压力myography解决方案：145 </td></tr><tr><td>氯化钾</td><td>西格玛</td><td> P5405 </td><td>剖析解决方案：4.7 沐浴液电：5.36 电：135内部解决方案 隔离解决方案肌：5.36 浴的压力myography解决方案：4.7 流明的压力myography解决方案：4.7 </td></tr><tr><td>钾EGTA </td><td>西格玛</td><td> E4378 </td><td>电：0.05内部解决方案</td></tr><tr><td> HEPES </td><td>西格玛</td><td> H9136 </td><td>沐浴液电：10 电：10内部解决方案 隔离解决方案，为心肌细胞*：10 </td></tr><tr><td>磷酸氢二钠</td><td>西格玛</td><td> S5136 </td><td>隔离解决方案，为心肌细胞*：0.34 </td></tr><tr><td>磷酸氢二钾</td><td>西格玛</td><td> P5655 </td><td>隔离解决方案，为心肌细胞*：0.44 </td></tr><tr><td>氯化镁</td><td>西格玛</td><td> M2393 </td><td>沐浴液电*：1.2 内部电：1解决方案 隔离解决方案肌：1.2 </td></tr><tr><td>氯化钙</td><td>西格玛</td><tD&gt; C7902 </td><td>沐浴液电：2 为心肌细胞的隔离解决方案：0.05 </td></tr><tr><td>磷酸钠</td><td> Fisher Scientific则</td><td> BP331-1 </td><td>剖析解决方案：1.2 浴的压力myography解决方案：1.2 流明的压力myography解决方案：1.2 </td></tr><tr><td>硫酸镁</td><td>西格玛</td><td> M2643 </td><td>剖析解决方案：1.17 浴的压力myography解决方案：1.17 流明的压力myography解决方案：1.17 </td></tr><tr><td> MOPS </td><td> Fisher Scientific则</td><td> BP308 </td><td>剖析解决方案： 浴的压力myography解决方案：3 流明的压力myography解决方案：3 </td></tr><tr><td>丙酮酸</td><td>西格玛</td><td> P4562 </td><td>剖析解决方案： 沐浴解决方案压力myography：2 流明的压力myography解决方案：2 </td></tr><tr><td> EDTA二水</td><td>研究有机物</td><td> 9572E </td><td>剖析解决方案：0.02 浴的压力myography解决方案：0.02 流明的压力myography解决方案：0.02 </td></tr><tr><td> D-葡萄糖</td><td>西格玛</td><td> G7021 </td><td>剖析解决方案： 沐浴液电：10 电：20内部解决方案 隔离解决方案，为心肌细胞*：10 浴的压力myography解决方案：5 流明的压力myography解决方案：5 </td></tr><tr><td>牛血清白蛋白</td><td>西格玛</td><td> A3912 </td><td>剖析解决方案：1％ 流明压力myography溶液：1％ </td></tr><tr><td> pH值</td><td></td><td></td><td>剖析搜索解决方案N：7.4 沐浴液电*：7.3 电：7.2的内部解决方案 隔离解决方案肌：7.2 浴的压力myography解决方案：7.4 流明的压力myography解决方案：7.4 </td></tr><tr><td>渗透压</td><td></td><td></td><td>剖析解决方案：300 沐浴液电：298 电：298内部解决方案 隔离解决方案肌：298 浴的压力myography解决方案：300 流明的压力myography解决方案：300 </td></tr><tr><td></td><td></td><td></td><td> * 11 </td></tr><tr><td></td><td></td><td></td><td> 表1中，在实验中使用的解决方案的组件。 </td></tr></tbody></table>

exploring arterial smooth muscle kv7 potassium channel function using patch clamp electrophysiology and pressure myography

电阻动脉的壁内的平滑肌细胞的收缩或松弛确定动脉直径并由此控制的血液流至容器内，并有助于全身血压。收缩过程中被限制主要是由胞内游离钙浓度（〔Ca 2 +] cyt的），它是依次由多种离子转运和频道控制。离子通道是常见的中间体，在信号转导通路激活血管活性激素对血管收缩或舒张功能。离子通道往往是有针对性的治疗药物，无论是有意（ 如钙离子通道阻断剂诱导的血管舒张和降低血压）或无意的（ 如引起不必要的心血管副作用）。 KV7（KCNQ）电压激活的钾离子通道最近被牵连的生理和治疗的重要塔尔格ETS调节平滑肌收缩。为了阐明KV7渠道在生理信号转导和治疗药物的行动中的具体作用，我们需要研究他们的活动是如何在细胞水平上调节，以及评估他们的贡献的背景下，完整的动脉。 大鼠肠系膜动脉提供了一个有用的模型系统。动脉可以很容易地解剖，结缔组织清除，并用孤立的动脉肌细胞膜片钳电生理准备，或为血管收缩剂/血管舒张反应的在相对生理条件下测量插管，加压。在这里，我们描述了这两种类型的测量方法，并提供一些例子，实验一体化设计可提供更清楚地了解这些离子通道在调节血管张力的作用。

Watch this Scientific Journal Video about Exploring Arterial Smooth Muscle Kv7 Potassium Channel Function using Patch Clamp Electrophysiology and Pressure Myography at JoVE.com

Exploring Arterial Smooth Muscle Kv7 Potassium Channel Function using Patch Clamp Electrophysiology and Pressure Myography

测量KV7（KCNQ）钾离子通道活性的孤立动脉细胞（中与测量大蟒/扩张的反应（使用压力myography），同时采用膜片钳电生理技术）可以揭示有关的重要信息的的KV7渠道作用在血管平滑肌生理和药理学。

探讨动脉平滑肌的KV7钾离子通道功能使用膜片钳电生理和压力Myography

Contraction  or relaxation of smooth muscle cells within the walls of resistance arteries  determines the artery diameter and thereby controls flow of ...

Research

JoVE Journal

Biology

Methods for Patch Clamp Capacitance Recordings from the Calyx

膜片钳从花萼电容录音方法

We demonstrate the basic techniques for presynaptic patch clamp recording at the calyx of Held, a mammalian central nervous system nerve terminal. ...

科研

生物学

Patch Clamp Recording of Ion Channels Expressed in  Xenopus Oocytes

补丁钳爪蟾卵母细胞中表达的离子通道

Since its development by Sakmann and Neher 1, 2, the patch clamp has become established as an extremely useful technique for electrophysiological ...

Pressure-polishing Pipettes for Improved Patch-clamp Recording

改进的膜片钳记录的压力抛光移液器

Pressure-polishing is a method for shaping glass pipettes for patch-clamp recording.  We first developed this method for fabricating pipettes suitable for ...

Focal Ca2+ Transient Detection in Smooth Muscle

Focal Ca2+ Transient Detection in Smooth Muscle

焦距钙 2 +</sup瞬态检测平滑肌

Ca2+ imaging of smooth muscle provides insight into cellular mechanisms that may not result in changes of membrane potential, such as the release of Ca2+ ...

Isolation of Human Umbilical Arterial Smooth Muscle Cells (HUASMC)

Isolation of Human Umbilical Arterial Smooth Muscle Cells HUASMC

人脐动脉平滑肌细胞的分离（HUASMC）

The human umbilical cord (UC) is a biological sample that can be easily obtained just after birth. This biological sample is, most of the time, discarded ...

Giant Liposome Preparation for Imaging and Patch-Clamp Electrophysiology

巨脂质体制剂成像和膜片钳电

The reconstitution of ion channels into chemically defined lipid membranes for electrophysiological recording has been a powerful technique to identify ...

Assessing Murine Resistance Artery Function Using Pressure Myography

评估抗小鼠动脉功能使用压力Myography

Pressure myograph systems are  exquisitely useful in the functional assessment of small arteries, pressurized  to a suitable transmural pressure. The  ...

One-channel Cell-attached Patch-clamp Recording

单通道细胞贴附膜片钳记录

Ion channel proteins are universal devices for fast communication across biological membranes. The temporal signature of the ionic flux they generate ...

Reconstitution of a Transmembrane Protein, the Voltage-gated Ion Channel, KvAP, into Giant Unilamellar Vesicles for Microscopy and Patch Clamp Studies

跨膜蛋白的重组，电压门控离子通道，KvAP，进入巨人单层囊的显微镜和膜片钳研究

Giant Unilamellar Vesicles (GUVs) are a popular biomimetic system for studying membrane associated phenomena. However, commonly used protocols to grow ...

Measurement of Smooth Muscle Function in the Isolated Tissue Bath-applications to Pharmacology Research

平滑肌功能中分离出来的组织浴，应用药理学研究测量

Isolated tissue bath assays are a classical pharmacological tool for evaluating concentration-response relationships in a myriad of contractile tissues.  ...