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>在显微镜观察下，打开耳蜗顶端区域，去除血管纹和螺旋韧带使用＃11手术刀刀片的一角，一个微点挑细的镊子。 </li><li>使用镊子耳蜗mod​​iolus以Corti器，并在1mg/ml的胶原酶的L - 15在室温下5分钟。 </li><li>如果需要从耳蜗基底轮流OHCS，去除骨壳覆盖挑耳蜗基地，并单独Corti器除去了之前使用的手术刀刀片从颞骨螺旋。 </li><li> Corti器传输到录音室使用50μL汉密尔顿注射器。随后，游离细胞回流通过针。 </li></ol> 2。实验装置<ol><li>图外部的交流电场（EAEF）发生器和与图像采集系统（图1）的联系。特别是实施House耳研究所工程的核心控制电路。 </li><li>在我们的实验中所用的实验装置，由AXIOVERT 135TV倒置显微镜（蔡司，Thornwood，NY）与另一种基于LED照明系统（大功率LED系统36AD3500，光速技术，坎贝尔，加利福尼亚州），两个电子micromanipulators（Eppendorf公司"Patchman"，德国），一个电脑控制的超高速Photron Fastcam × 1024的PCI相机（Photron美国公司）在凯勒端口和一个额外的经常在三目端口CCD相机。 Fastcam相机能够捕捉到的图像在高频率（高达100,000 FPS）和高分辨率（如1024 × 1024像素，1000帧，512 × 128 10,000 FPS，384 × 96 18,000 fps的，等等）。由高速摄像机提供的图像直接观察到在电脑的显示器，而CCD相机连接到不同的显示器。基于LED的照明系统工作在两种不同的模式:低功耗模拟和高功率数字。使用低功耗的模拟模式进行了初步的所有程序（电极定位，细胞定位，重点等）。高功率照明开启相机的快门光圈快门关闭，然后关闭，促进散热。国产软件，还开发了在众议院耳研究所工程的核心，在同一台PC上运行的控制的高速摄像头，基于LED的照明系统，并在EAEF触发。在一个传统的数码照片的摄像头前端口可以根据需要为静帧。 （图2）。 </li><li>驱动电极（两个0.25毫米直径的银线提示为0.8毫米的距离）的位置使用的电子micromanipulators。电极的位置是通过视觉和显微图像监控;焦平面图像的变化表明，电极触及的试验室的底部。最初，该电场是使用外部电极校准。这种电极的措施在不同点的电势，电场产生的"地图"。如果一个单一的孤立的外毛细胞之间与应用EAEF其纵轴平行放置电极的提示，它会移动在相同的频率的电场拉长和缩短。如果细胞被置于垂直于磁场的不同类型的OHC的反应（弯曲），可以观察和调查。 （图二B） </li></ol> 3。 EAEF刺激和图像采集<ol><li>可选择四种不同的刺激协议（图3）: <ol><li>连续单频（图3）。请注意，刺激模式，频率，振幅和波型可以选择使用自制的控制软件（红色圆圈）。 </li><li>突发的单一频率（图二）。阵阵爆裂之间的差距的长度也可选择。 </li><li>线性扫描（图3 C）。最初和最终的频率是可选择的。 </li><li>多刺激（图四）。单一的频率和线性扫描，可以结合在一个单一的实验。选择相应的参数后，控制软件，配置系统，并允许操作员通过点击一个按钮启动光同步录像和细胞刺激电脑屏幕上。 </li></ol></li><li>拍摄的图像在高频率的AVI格式，作进一步的分析。 </li></ol> 4。代表性的成果<ol><li>在这部影片中，两个孤立的外毛细胞显示时，他们正在与外部的交流电场，是平行的或横向的，分别刺激的长度或曲率的变化，其纵轴。 （电影＃2）。 </li><li> OHCS能动反应分析行使用ProAnalyst软件（Xcitex公司，剑桥，马萨诸塞州）。 "特征跟踪"该软件的功能提供了帧帧（图4）两点之间的距离。在细胞缩短顶点（表皮板;红色）在选定点之间的距离和细胞（基底极;绿色）基数较小，且与细胞伸长增加。影片中展示了该软件分析帧一帧的长度变化。在图像的底部面板显示运动的轨迹。在这个例子中，总长度变化约6.5像素。 </li><li> "轮廓跟踪"ProAnalyst软件的功能，可以检测到细胞边缘和自动测量的光学部分的面积（图5）。 </li><li>聚苯乙烯微球添加到随机液，并牢牢附着到质膜（图6）。可同时选择不同的微球，该软件可以自动跟踪他们一帧一帧。这样，细胞可分为部分和每一节的蠕动独立评估。 （图6 A） </li><li>通过选择位于微球的细胞图像的外侧边缘，每个段的长度变化，并在一个段尊重的角度变化（弯曲）也可以被独立评估。 （图6B） </li></ol><img src="/files/ftp_upload/2965/2965fig1.jpg" alt="图1"/> 图1图EAEF发电机和与图像采集系统的联系。 <img src="/files/ftp_upload/2965/2965fig2.jpg" alt="图2"/> 图2:实验装置）的图片。二）在显微镜阶段，详细描绘电极和一个单一的OHC它们之间放置其纵轴平行电场的漫画。 <img src="/files/ftp_upload/2965/2965fig3.jpg" alt="图3"/> 图3。一个用户界面的配置为单频刺激国产控制软件）。选择的参数是在红色圈。 b）用户自制的控制软件的接口配置爆裂单频刺激。三）为线性扫描的刺激配置自制的控制软件的用户界面。四）控制配置多刺激国产软件的用户界面。 <img src="/files/ftp_upload/2965/2965fig4.jpg" alt="图4"/> 图4。一个OHC的单帧基地（绿色）和顶点（红色）的细胞，分别使用"功能跟踪"ProAnalyst软件的功能，选择了两个点。的单元格下方的曲线显示了电刺激所选择的点之间的距离的周期性变化。移动竖线不同的框架，可以选择个别的分析。 <img src="/files/ftp_upload/2965/2965fig5.jpg" alt="图5"/> 图5在ProAnalyst"轮廓跟踪"功能检测到细胞边缘，自动测量的光学部分的面积。 <img src="/files/ftp_upload/2965/2965fig6.jpg" alt="图6"/> 图6。一个一个孤立的OHC）拍摄的图像与聚苯乙烯微球（上），和微单独选择（下）与五个相同的图像装饰。不同的颜色分配给每个微球，和他们的位移可以单独和自动跟踪一帧一帧，分析和比较。二）任意长度的片段，可以选择聚苯乙烯微球的细胞边缘位于的定义，并在这些片段的长度变化，以及在尊重方向的一个部分的修改给别人（细胞弯曲），可自动评估框架帧图像分析软件。 电影1。豚鼠OHCS隔离。 <a href="http://www.jove.com/files/ftp_upload/2965/2965_Movie1_web.mov">点击这里观看视频</a> 电影2。OHCS平行和垂直EAEF呈现出典型的electromotility和弯曲反应。 <a href="http://www.jove.com/files/ftp_upload/2965/2965_Movie2_web.mov">点击这里观看视频</a>

<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，每个排量只有与在细胞形态，而不是与他们与外部参考帧运动的变化有关。可靠的测量横断面OHC的面积也很容易获得，它允许OHC的量的快速变化的估计。此外，更快，更灵敏的照相机和更好的图像分析软件的不断发展，保证质量的方法在不断改进。的技术，没有跨质膜电势控制，缺点是与目前所有的方法用于评估在kHz范围内的OHC蠕动共享限制。 因此，这里介绍的方法可能是一个听力研究的重要工具，能够基本OHCS"能动反应有关的细胞和分子机制提供新的和重要的线索

<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 ...

investigating-outer-hair-cell-motility-with-combination-external

Nanyang Technological University

Research

JoVE Journal

Neuroscience

13.4K 次观看.  House Ear Institute. 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。

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

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

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 display abnormalities in the expression of FMR1 - a protein required for typical, healthy neural development.3 Recent data has suggested that the loss of this protein can cause the cortex to be hyperexcitable thereby affecting overall patterns of neural plasticity.4,5
In addition, Fragile X shows a strong comorbidity with autism: in fact, 30% of children with FXS are diagnosed with autism, and 2 - 5% of autistic children suffer from FXS.6
Transcranial Magnetic Stimulation (a non-invasive neurostimulatory and neuromodulatory technique that can transiently or lastingly modulate cortical excitability via the application of localized magnetic field pulses 7,8) represents a unique method of exploring plasticity and the manifestations of FXS within affected individuals. More specifically, Theta-Burst Stimulation (TBS), a specific stimulatory protocol shown to modulate cortical plasticity for a duration up to 30 minutes after stimulation cessation in healthy populations, has already proven an efficacious tool in the exploration of abnormal plasticity.9,10 
Recent studies have shown the effects of TBS last considerably longer in individuals on the autistic spectrum - up to 90 minutes.11 This extended effect-duration suggests an underlying abnormality in the brain's natural plasticity state in autistic individuals - similar to the hyperexcitability induced by Fragile X Syndrome.
In this experiment, utilizing single-pulse motor-evoked potentials (MEPs) as our benchmark, we will explore the effects of both intermittent and continuous TBS on cortical plasticity in individuals suffering from FXS and individuals on the Autistic Spectrum.

In this article, we examine the effects of Theta-Burst TMS stimulation on cortical plasticity in individuals suffering from Fragile X syndrome and individuals on the autistic spectrum.

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

In utero electroporation (IUE) has become a powerful technique to study the development of different regions of the embryonic nervous system 1-5. To date this tool has been widely used to study the regulation of cellular proliferation, differentiation and neuronal migration especially in the developing cerebral cortex 6-8. Here we detail our protocol to electroporate in utero the cerebral cortex and the hippocampus and provide evidence that this approach can be used to study dendrites and spines in these two cerebral regions.
Visualization and manipulation of neurons in primary cultures have contributed to a better understanding of the processes involved in dendrite, spine and synapse development. However neurons growing in vitro are not exposed to all the physiological cues that can affect dendrite and/or spine formation and maintenance during normal development. Our knowledge of dendrite and spine structures in vivo in wild-type or mutant mice comes mostly from observations using the Golgi-Cox method 9. However, Golgi staining is considered to be unpredictable. Indeed, groups of nerve cells and fiber tracts are labeled randomly, with particular areas often appearing completely stained while adjacent areas are devoid of staining. Recent studies have shown that IUE of fluorescent constructs represents an attractive alternative method to study dendrites, spines as well as synapses in mutant / wild-type mice 10-11 (Figure 1A). Moreover in comparison to the generation of mouse knockouts, IUE represents a rapid approach to perform gain and loss of function studies in specific population of cells during a specific time window. In addition, IUE has been successfully used with inducible gene expression or inducible RNAi approaches to refine the temporal control over the expression of a gene or shRNA 12. These advantages of IUE have thus opened new dimensions to study the effect of gene expression/suppression on dendrites and spines not only in specific cerebral structures (Figure 1B) but also at a specific time point of development (Figure 1C).
Finally, IUE provides a useful tool to identify functional interactions between genes involved in dendrite, spine and/or synapse development. Indeed, in contrast to other gene transfer methods such as virus, it is straightforward to combine multiple RNAi or transgenes in the same population of cells. 
In summary, IUE is a powerful method that has already contributed to the characterization of molecular mechanisms underlying brain function and disease and it should also be useful in the study of dendrites and spines.

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

This article describes in detail a protocol to electroporate in utero the cerebral cortex and the hippocampus at E14.5 in mice. We also show that this is a valuable method to study dendrites and spines in these two cerebral regions.

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

In utero electroporation (IUE) has become a powerful technique to study the development of different regions of the embryonic nervous system 1-5. To date ...

Whole Cell Patch Clamp for Investigating the Mechanisms of Infrared Neural Stimulation

It has been demonstrated in recent years that pulsed, infrared laser light can be used to elicit electrical responses in neural tissue, independent of any further modification of the target tissue. Infrared neural stimulation has been reported in a variety of peripheral and sensory neural tissue in vivo, with particular interest shown in stimulation of neurons in the auditory nerve. However, while INS has been shown to work in these settings, the mechanism (or mechanisms) by which infrared light causes neural excitation is currently not well understood. The protocol presented here describes a whole cell patch clamp method designed to facilitate the investigation of infrared neural stimulation in cultured primary auditory neurons. By thoroughly characterizing the response of these cells to infrared laser illumination in vitro under controlled conditions, it may be possible to gain an improved understanding of the fundamental physical and biochemical processes underlying infrared neural stimulation.

Infrared nerve stimulation has been proposed as an alternative to electrical stimulation in a range of nerve types, including those associated with the auditory system. This protocol describes a patch clamp method for studying the mechanism of infrared nerve stimulation in a culture of primary auditory neurons.

全细胞膜片钳红外神经刺激机制的调查

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

A Human-machine-interface Integrating Low-cost Sensors with a Neuromuscular Electrical Stimulation System for Post-stroke Balance Rehabilitation

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 delivery of oxygen and nutrients. About half of the stroke survivors are left with some degree of disability. Innovative methodologies for restorative neurorehabilitation are urgently required to reduce long-term disability. The ability of the nervous system to reorganize its structure, function and connections as a response to intrinsic or extrinsic stimuli is called neuroplasticity. Neuroplasticity is involved in post-stroke functional disturbances, but also in rehabilitation. Beneficial neuroplastic changes may be facilitated with non-invasive electrotherapy, such as neuromuscular electrical stimulation (NMES) and sensory electrical stimulation (SES). NMES involves coordinated electrical stimulation of motor nerves and muscles to activate them with continuous short pulses of electrical current while SES involves stimulation of sensory nerves with electrical current resulting in sensations that vary from barely perceivable to highly unpleasant. Here, active cortical participation in rehabilitation procedures may be facilitated by driving the non-invasive electrotherapy with biosignals (electromyogram (EMG), electroencephalogram (EEG), electrooculogram (EOG)) that represent simultaneous active perception and volitional effort. To achieve this in a resource-poor setting, e.g., in low- and middle-income countries, we present a low-cost human-machine-interface (HMI) by leveraging recent advances in off-the-shelf video game sensor technology. In this paper, we discuss the open-source software interface that integrates low-cost off-the-shelf sensors for visual-auditory biofeedback with non-invasive electrotherapy to assist postural control during balance rehabilitation. We demonstrate the proof-of-concept on healthy volunteers.

A novel low-cost human-machine interface for interactive post-stroke balance rehabilitation system is presented in this article. The system integrates off-the-shelf low-cost sensors towards volitionally driven electrotherapy paradigm. The proof-of-concept software interface is demonstrated on healthy volunteers.

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 ...

Assessment of Neuromuscular Function Using Percutaneous Electrical Nerve Stimulation

Percutaneous electrical nerve stimulation is a non-invasive method commonly used to evaluate neuromuscular function from brain to muscle (supra-spinal, spinal and peripheral levels). The present protocol describes how this method can be used to stimulate the posterior tibial nerve that activates plantar flexor muscles. Percutaneous electrical nerve stimulation consists of inducing an electrical stimulus to a motor nerve to evoke a muscular response. Direct (M-wave) and/or indirect (H-reflex) electrophysiological responses can be recorded at rest using surface electromyography. Mechanical (twitch torque) responses can be quantified with a force/torque ergometer. M-wave and twitch torque reflect neuromuscular transmission and excitation-contraction coupling, whereas H-reflex provides an index of spinal excitability. EMG activity and mechanical (superimposed twitch) responses can also be recorded during maximal voluntary contractions to evaluate voluntary activation level. Percutaneous nerve stimulation provides an assessment of neuromuscular function in humans, and is highly beneficial especially for studies evaluating neuromuscular plasticity following acute (fatigue) or chronic (training/detraining) exercise.

We present a protocol to assess changes in neuromuscular function. Percutaneous electrical nerve stimulation is a non-invasive method that evokes muscular responses. Electrophysiological and mechanical properties of these responses permit the evaluation of neuromuscular function from brain to muscle (supra-spinal, spinal and peripheral levels).

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

The enteric nervous system (ENS) plays an important role in regulating gastrointestinal (GI) motility and can function independently of the central nervous system. Changes in ENS function are a major cause of GI symptoms and disease and may contribute to GI symptoms reported in neuropsychiatric disorders including autism. It is well established that isolated colon segments generate spontaneous, rhythmic contractions known as Colonic Migrating Motor Complexes (CMMCs). A procedure to analyze the enteric neural regulation of CMMCs in ex vivo preparations of mouse colon is described. The colon is dissected from the animal and flushed to remove fecal content prior to being cannulated in an organ bath. Data is acquired via a video camera positioned above the organ bath and converted to high-resolution spatiotemporal maps via an in-house software package. Using this technique, baseline contractile patterns and pharmacological effects on ENS function in colon segments can be compared over 3-4 hr. In addition, propagation length and speed of CMMCs can be recorded as well as changes in gut diameter and contraction frequency. This technique is useful for characterizing gastrointestinal motility patterns in transgenic mouse models (and in other species including rat and guinea pig). In this way, pharmacologically induced changes in CMMCs are recorded in wild type mice and in the Neuroligin-3R451C mouse model of autism. Furthermore, this technique can be applied to other regions of the GI tract including the duodenum, jejunum and ileum and at different developmental ages in mice.

This article describes a video imaging technique and high-resolution spatiotemporal mapping to identify changes in the neural regulation of colonic motility in adult mice. Subtle effects on gastrointestinal (GI) function can be detected using this approach in isolated tissue preparations to advance our understanding of GI disease.

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

Transcranial Alternating Current Stimulation (tACS) is a neuromodulatory technique able to act through sinusoidal electrical waveforms in a specific frequency and in turn modulate ongoing cortical oscillatory activity. This neurotool allows the establishment of a causal link between endogenous oscillatory activity and behavior. Most of the tACS studies have shown online effects of tACS. However, little is known about the underlying action mechanisms of this technique because of the AC-induced artifacts on Electroencephalography (EEG) signals. Here we show a unique approach to investigate online physiological frequency-specific effects of tACS of the primary motor cortex (M1) by using single pulse Transcranial Magnetic Stimulation (TMS) to probe cortical excitability changes. In our setup, the TMS coil is placed over the tACS electrode while Motor Evoked Potentials (MEPs) are collected to test the effects of the ongoing M1-tACS. So far, this approach has mainly been used to study the visual and motor systems. However, the current tACS-TMS setup can pave the way for future investigations of cognitive functions. Therefore, we provide a step-by-step manual and video guidelines for the procedure.

Transcranial Alternating Current Stimulation (tACS) allows the modulation of cortical excitability in a frequency-specific fashion. Here we show a unique approach which combines online tACS with single pulse Transcranial Magnetic Stimulation (TMS) in order to "probe" cortical excitability by means of Motor Evoked Potentials.

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

Transcranial electrical brain stimulation can modulate cortical excitability and plasticity in humans and rodents. The most common form of stimulation in humans is transcranial direct current stimulation (tDCS). Less frequently, transcranial alternating current stimulation (tACS) or transcranial random noise stimulation (tRNS), a specific form of tACS using an electrical current applied randomly within a pre-defined frequency range, is used. The increase of noninvasive electrical brain stimulation research in humans, both for experimental and clinical purposes, has yielded an increased need for basic, mechanistic, safety studies in animals. This article describes a model for transcranial electrical brain stimulation (tES) through the intact skull targeting the motor system in alert rodents. The protocol provides step-by-step instructions for the surgical set-up of a permanent epicranial electrode socket combined with an implanted counter electrode on the chest. By placing a stimulation electrode into the epicranial socket, different electrical stimulation types, comparable to tDCS, tACS, and tRNS in humans, can be delivered. Moreover, the practical steps for tES in alert rodents are introduced. The applied current density, stimulation duration, and stimulation type may be chosen depending on the experimental needs. The caveats, advantages, and disadvantages of this set-up are discussed, as well as safety and tolerability aspects.

This protocol describes a surgical set-up for a permanent epicranial electrode socket and an implanted chest electrode in rodents. By placing a second electrode into the socket, different types of transcranial electrical brain stimulation can be delivered to the motor system in alert animals through the intact skull.

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

Transcranial electrical stimulation (TES) is a powerful and relatively simple approach to diffusely influence brain activity either randomly or in a closed-loop event-triggered manner. Although many studies are focusing on the possible benefits and side-effects of TES in healthy and pathologic brains, there are still many fundamental open questions regarding the mechanism of action of the stimulation. Therefore, there is a clear need for a robust and reproducible method to test the acute and the chronic effects of TES in rodents. TES can be combined with regular behavioral, electrophysiological, and imaging techniques to investigate neuronal networks in vivo. The implantation of transcranial stimulation electrodes does not impose extra constraints on the experimental design while it offers a versatile, flexible tool to manipulate brain activity. Here we provide a detailed, step-by-step protocol to fabricate and implant transcranial stimulation electrodes to influence brain activity in a temporally constrained manner for months.

This detailed protocol describes transcranial stimulation electrode placement on the temporal bone in order to investigate the short- and long-term effects of transcranial electrical stimulation in freely moving 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

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 amplify the weak vibration of low-level sound signal. The morphology and electrophysiological property of outer hair cells (OHCs) develop in early postnatal ages. The maturation of outer hair cell may contribute to the development of the auditory system. However, the process of OHCs development is not well studied. This is partly because of the difficulty to measure their function by an electrophysiological approach. With the purpose of developing a simple method to address the above issue, here we describe a step-by-step protocol to study the function of OHCs in acutely dissociated cochlea from postnatal rats. With this method, we can evaluate the cochlear response to pure tone stimuli and examine the expression level and function of the motor protein prestin in OHCs. This method can also be used to investigate the inner hair cells (IHCs).

This protocol describes methods to record the auditory brainstem response from postnatal rat pups. To examine the functional development of outer hair cells, the experimental procedure of whole-cell patch clamp recording in isolated outer hair cells is described step-by-step.

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 ...