Name: investigating outer hair cell motility with a combination of external alternating electrical field stimulation and high-speed image analysis
Uploaded: 2011-07-18T13:00:00
Description: Watch this Scientific Journal Video about Investigating Outer Hair Cell Motility with a Combination of External Alternating Electrical Field Stimulation and High-speed Image Analysis at JoVE.com

由国家卫生赠款R01DC10146/R01DC010397研究院支持的工作，NIDCD P30 DC006276研究的核心，和喜。其内容完全是作者的责任，并不一定代表美国国立卫生研究院或熹官方意见。作者宣称没有任何现有的或潜在的利益冲突。

1。隔离的OHCS <ol><li>开始收获豚鼠，小鼠或您的哺乳类动物模型的颞骨，此过程。 </li><li>接下来，打开一个锤骨钳的使用，以揭露耳蜗的颞骨，和他们沉浸在莱博维茨的L - 15。小心取出骨过剩，保持完好的骨壳。鉴于这是一个一般的程序，适用于任何哺乳类动物的颞骨，小的变化的技术处理时，可能需要从非常小的动物颞骨。泡在老动物通常是钙化，引入了新的复杂的程序。 </li><li>在显微镜观...

<ol>
	<li>Frolenkov, G. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+insights+into+the+morphogenesis+of+inner+ear+hair+cells.">Genetic insights into the morphogenesis of inner ear hair cells.</a> Nat Rev Genet. 5, 489-498 (2004).</li><li>Ashmore, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cochlear+outer+hair+cell+motility.">Cochlear outer hair cell motility.</a> Physiol Rev. 88, 173-210 (2008).</li><li>Dallos, P., Fakler, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prestin,+a+new+type+of+motor+protein.">Prestin, a new type of motor protein.</a> Nature Rev. Mol. Cell Biol. 3, 104-111 (2002).</li><li>Frolenkov, G. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cochlear+outer+hair+cell+bending+in+an+external+electrical+field.">Cochlear outer hair cell bending in an external electrical field.</a> Biophys. J. 73, 1665-1672 (1997).</li><li>Matsumoto, N., Kalinec, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extraction+of+Prestin-Dependent+and+Prestin-Independent+Components+from+Complex+Motile+Responses+in+Guinea+Pig+Outer+Hair+Cells.">Extraction of Prestin-Dependent and Prestin-Independent Components from Complex Motile Responses in Guinea Pig Outer Hair Cells.</a> Biophys J. 89, 4343-4351 (2005).</li><li>Matsumoto, N., Kalinec, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prestin-dependent+and+prestin-independent+motility+of+guinea+pig+outer+hair+cells.">Prestin-dependent and prestin-independent motility of guinea pig outer hair cells.</a> Hear Res. 208, 1-12 (2005).</li><li>Ashmore, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+remarkable+cochlear+amplifier.">The remarkable cochlear amplifier.</a> Hear Res. 266, 1-17 (2010).</li><li>Kitani, R., Kakehata, S., Kalinec, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Motile+responses+of+cochlear+outer+hair+cells+stimulated+with+an+alternating+electrical+field.">Motile responses of cochlear outer hair cells stimulated with an alternating electrical field.</a> Hearing Research. , (2011).</li><li>Dallos, P., Evans, B. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High-frequency+outer+hair+cell+motility:+corrections+and+addendum.">High-frequency outer hair cell motility: corrections and addendum.</a> Science. 268, 1420-1421 (1995).</li><li>Frank, G., Hemmert, W., Gummer, A. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Limiting+dynamics+of+high-frequency+electromechanical+transduction+in+outer+hair+cells.">Limiting dynamics of high-frequency electromechanical transduction in outer hair cells.</a> Proc. Natl. Acad. Sci. 96, 4420-4425 (1999).</li><li>Santos-Sacchi, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=On+the+frequency+limit+and+phase+of+outer+hair+cell+motility:+effects+of+the+membrane+filter.">On the frequency limit and phase of outer hair cell motility: effects of the membrane filter.</a> J. Neurosci. 12, 1906-1916 (1992).</li><li>Inou&#233;, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Video Microscopy. , (1986).</li></ol>

这里介绍的实验方法使OHC的能动反应在kHz的范围内没有任何限制细胞的运动估计。不同的刺激协议，额外的标记（微），以及细胞的方向与电场变化，使人们有可能探讨新的OHC蠕动方面与以前无法进入的细节水平。其他方法，例如那些采用光电二极管或激光多普勒vibrometry 10，需要严格控制的细 ​​胞中的位置。在这里，与此相反，所有的测量都属于同一单元格点之间进行，每个?...

<table border="1"><tbody><tr><td> 试剂名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论（可选） </td></tr><tr><td>莱博维茨的L - 15 </td><td> Gibco公司</td><td> 21083 </td><td></td></tr><tr><td>胶原酶（类型4） </td><td> Sigma - Aldrich公司</td><td> C5138 </td><td>为1mg/ml中的L - 15 </td></tr></tbody></table>

investigating outer hair cell motility with a combination of external alternating electrical field stimulation and high-speed image analysis

OHCS是圆柱形的感觉，位于尔蒂内哺乳动物内耳听觉器官的器官细胞。 “毛细胞”的名字来自它们所特有的纤毛，检测和传导声音能量1的一个关键因素根尖捆绑的。 OHCS能够改变形状拉长，缩短和弯曲，在电气，机械和化学刺激，电机响应考虑耳蜗声信号放大2的关键。 OHC的刺激诱导两个不同的能动对策：一）electromotility，又名快速蠕动，在微秒范围内的变化，电动马达蛋白在人口稠密的OHC质膜盒装的构象变化的长度，及ii）缓慢的蠕动，在造型上的变化毫秒到秒范围涉及细胞骨架重组，2，3。 OHC的弯曲与electromotility相关结果无论是从马达蛋白的不对称分布在外侧的质膜，或不对称电刺激这些马达蛋白（例如，与电场垂直于细胞长轴） 4。机械和化学刺激诱 ​​导基本上是缓慢的能动反应，即使在离子的细胞和/或他们的环境条件的变化也能刺激血浆膜嵌入式马达蛋白5， 6。由于OHC的能动反应耳蜗放大器的重要组成部分，声波频率（大约从20 Hz到20 kHz在人类的）的定性和定量分析这些能动的反应是在听觉研究7领域的一个非常重要的问题。 现在高速videocameras，基于LED的照明系统，以及先进的图像分析软件相结合的新的成像技术的发展提供了能够进行可靠的定性和定量研究的隔离OHCS能动的响应外部的交流电场（EAEF） 8。这是一个简单的和非侵入性的技术，规避大部分 9-11以前的方法的局限性。此外，基于LED的照明系统提供微不足道的热效应，对样品的极端的亮度，因为使用视频显微镜，光学分辨率比传统的光学显微镜技术12高出至少10倍。例如，与这里所描述的实验装置，改变细胞的长度约为20 nm可以经常和可靠地检测到10千赫的频率，可以进一步改善在较低的频率和本决议。 我们有信心，本实验的方法将有助于延长OHC的蠕动的基本细胞和分子机制的认识。

Watch this Scientific Journal Video about Investigating Outer Hair Cell Motility with a Combination of External Alternating Electrical Field Stimulation and High-speed Image Analysis at JoVE.com

Investigating Outer Hair Cell Motility with a Combination of External Alternating Electrical Field Stimulation and High-speed Image Analysis

调查外毛细胞（OHC）能动的反应，包括electromotility，缓慢的蠕动和弯曲，可靠的方法来描述。 OHC的蠕动刺激引起的与外部的交流电场，该方法需要高速图像记录，基于LED照明，和上一代图像分析软件的优势。

调查外毛细胞运动与外部的交流电场刺激和高速图像分析相结合

OHCs are cylindrical sensorimotor cells located in the Organ of Corti, the auditory organ inside the mammalian inner ear. The name "hair cells" derives ...

Research

JoVE Journal

Neuroscience

TMS: Using the Theta-Burst Protocol to Explore Mechanism of Plasticity in Individuals with Fragile X Syndrome and Autism

TMS：使用的Theta突发协议，以探索个人与脆性X综合征与孤独症的可塑性Mechasnism

Fragile X Syndrome (FXS), also known as Martin-Bell Syndrome, is a genetic abnormality found on the X chromosome.1,2 Individuals suffering from FXS ...

科研

神经科学

Visualization and Genetic Manipulation of Dendrites and Spines in the Mouse Cerebral Cortex and Hippocampus using In utero Electroporation

Visualization and Genetic Manipulation of Dendrites and Spines in the Mouse Cerebral Cortex and Hippocampus using In utero Electroporation

可视化和遗传操作在小鼠大脑皮层和使用海马树突和棘<em&gt;在子宫内</em&gt;电穿孔

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It has been  demonstrated in recent years that pulsed, infrared laser light can be used to  elicit electrical responses in neural tissue, independent of ...

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人机界面集成的低成本传感器，具有神经肌肉电刺激系统中风后康复平衡

A stroke is caused when an artery carrying blood from heart to an area in the brain bursts or a clot obstructs the blood flow to brain thereby preventing ...

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神经肌肉功能的应用经皮神经电刺激评估

Percutaneous electrical nerve stimulation is a non-invasive method commonly used to evaluate neuromuscular function from brain to muscle (supra-spinal, ...

Video Imaging and Spatiotemporal Maps to Analyze Gastrointestinal Motility in Mice

视频成像和时空地图来分析胃肠动力小鼠

The enteric nervous system (ENS) plays an important role in regulating gastrointestinal (GI) motility and can function independently of the central ...

Effects of Transcranial Alternating Current Stimulation on the Primary Motor Cortex by Online Combined Approach with Transcranial Magnetic Stimulation

经颅磁刺激在线联合方法对经颅交流电刺激对原发性运动皮层的影响

Transcranial Alternating Current Stimulation (tACS) is a neuromodulatory technique able to act through sinusoidal electrical waveforms in a specific ...

Transcranial Electrical Brain Stimulation in Alert Rodents

警惕啮齿动物的经颅电脑刺激

Transcranial electrical brain stimulation can modulate cortical excitability and plasticity in humans and rodents. The most common form of stimulation in ...

Chronic Transcranial Electrical Stimulation and Intracortical Recording in Rats

大鼠慢性经颅电刺激及 Intracortical 记录

Transcranial electrical stimulation (TES) is a powerful and relatively simple approach to diffusely influence brain activity either randomly or in a ...

Auditory Brainstem Response and Outer Hair Cell Whole-cell Patch Clamp Recording in Postnatal Rats

产后大鼠听觉脑干反应及外毛细胞全细胞贴片钳记录

The outer hair cell is one of the two types of sensory hair cells in the mammalian cochlea. They alter their cell length with the receptor potential to ...