This work was supported by the Korea Research Foundation Grant funded by the Korean Government (KRF-2008-313-E00458). The authors thank Jin-Joo Lee for the technical assistance.

所有的动物使用的程序进行审查，并由首尔大学医院的机构动物护理和使用委员会批准。 1.实验装置<ol><li>动物准备<ol><li>允许雄性Sprague-Dawley大鼠1周开始实验前，以适应新的环境。 注意:虽然8周龄大鼠在本研究中所使用的，显影或成人大脑可以根据研究假设来选择。 </li></ol></li><li>吸入麻醉诱导<ol><li>诱导并用5％异氟醚分别经由室与鼻锥体溶解在40％/ 60％和25％/ 75％氧/氮，2％维持麻醉。调整麻醉深度废除踏板撤回反射到脚趾捏，确认正确的麻醉水平。 注:使用苏醒动物可以在翻译方面更好的选择，但也有困难，经颅磁刺激和t内抑制哎很容易出现过度紧张。 </li><li>监测体温用直肠探头，并通过使用一个恒温毯保持它在37℃。进行监控踏板撤回反射，温度，呼吸频率和心脏率麻醉深度。 </li></ol></li><li>切换到第静脉麻醉进行维修<ol><li>准备用酒精棉签尾巴。导尿与过渡到静脉麻醉（ 图1A）一个24号静脉导管的侧尾静脉中。负载异丙酚静脉内（1毫克/ [千克·分钟]在10分钟内，用10毫克/毫升乳剂）给动物。丙泊酚开始加载后停止异氟醚5分钟。 </li><li>保持异丙酚镇静在500的输注速率- 700微克/整个实验（千克·分钟），如在先前的研究中16。 通过鼻锥在0.8升/分钟的补充氧气。 注:麻醉丙泊酚是由inhal降低大脑皮层兴奋的潜在抑制通货膨胀剂17-19。然而，麻醉是不是在磁刺激实验强制性的，并且也可以使用惊醒梦中人动物。麻醉方法应考虑研究假设的决定。 </li><li>使用于眼部兽医药膏，以防止干燥时的麻醉下。 </li><li>应用磁刺激（见第2节），以静脉麻醉完成后转型10分钟。 </li></ol></li><li>恢复条件<ol><li>在监视过程中恢复阶段生命体征。不要离开无人看管的动物，直到它已经恢复了足够的意识，以保持胸骨斜卧。如果动物已经接受了手术，不要将其返回到公司的其他动物，直到完全康复。 注:如果在执行手术的疾病模型中，手术后疼痛的管理是必要的。然而，不需要用于此磁刺激实验疼痛管理。 </li></ol></li></ol> 2.重复经颅磁刺激<ol><li>刺激和线圈<ol><li>通过使用重复的刺激经由 25毫米8字形线圈提供双相施加剌激的刺激。定位线圈0.5厘米外侧上biauricular线的顶点的中心，并具棱的线圈45°到地面。 注意:线圈的最大磁场强度为4.0 T的磁性线圈被牢固地安装在内置的支架。 </li></ol></li><li>运动阈值<ol><li>确定在热点的运动阈值（MT），与线圈的中心位置0.5厘米外侧到顶点上biauricular线和与该表面平面上的颅盖。这是在先前的研究中20所使用的相同的方法。 注:MT定义为最小刺激强度由10个连续的刺激唤起对侧前爪5个或更多可扪及收缩。验证是否刺激主要引起对侧肌肉收缩，以保证单边刺激。 </li&gt; </ol></li><li>经颅磁刺激中的应用<ol><li>深麻醉稳定后应用rTMS治疗10分钟。将线圈在目标rTMS治疗部位的中心，距离取决于研究问题大脑皮层选择。然后，倾斜线圈，确保线圈中心，并在刺激点颅骨的表面之间的直接接触。 注意:例如，棱线圈45°到地面，以减少磁刺激的对侧皮层（ 图1B和1C）的电位直接影响。 </li><li>若使动物单边半球20分钟的rTMS的会话。使用软件控制台，提供磁刺激用低频率（1赫兹），高频率（20赫兹），或假刺激方案，并在100设置刺激强度 - 对MT的110％。 </li><li>没有休息执行1 Hz的刺激。使用软件控制台输入"1200"射击为"20"分钟）。为20赫兹的刺激，进行2秒的刺激，接着28休息秒。使用软件控制台输入"1600"射击为"20"分钟。 </li><li>对于假刺激，倾斜垂直线圈（旋转90°）到颅骨和从头部表面（ 图1D）放置线圈边缘相距2厘米。牢固地固定在线圈架的主装置;没有必要通过手工在实验过程中保持线圈。 注意:为了补偿声和其他非特异性的效果，不同的假协议应当用于不同刺激方案。例如，1赫兹假刺激可用于1Hz的磁刺激实验。 </li></ol></li><li>冷却线圈<ol><li>使用一个水冷却系统在1-和20赫兹刺激频率（ 图2），以使超过20分钟重复磁刺激。流通周围在实验过程中线圈的整个长度，以防止过热冰水，虽然线圈或刺激器的温度不被监控。 注:市售冷却鼠线圈也可以使用。 </li><li>如果可能的话，监控通过查看经颅磁刺激机的加热表上的线圈温度。注:有相关的rTMS刺激没有不良后果。然而，有，如果是金属耳识别标签的刺激线圈附近使用的潜在风险烧伤。 </li></ol></li></ol> 3.微正电子发射断层扫描<ol><li>动物准备<ol><li>进行吸入麻醉诱导和静脉麻醉进行维修（见步骤1.2.1和1.3.1）。在的MT的100-110％刺激强度施加1Hz的磁刺激的动物10分钟。 </li><li>在完成磁刺激刺激五分钟后，注入2- [F-18]溶于0.5ml生理盐水的氟脱氧葡萄糖（FDG 18）的1毫居里静脉内通过使用尾静脉导管。允许18 FDG摄取30分钟。注意:将麻醉下大鼠整个微PE时Ť实验。 </li></ol></li><li>图像分析<ol><li>使用PET扫描仪脑成像重申刺激unilaterality。重建一个3-D迭代算法图像。为了评估通过磁刺激诱导的代谢的变化，在横向脑切片21的图像识别感兴趣的（投资回报）的区域。 </li></ol></li><li>安乐死<ol><li>进行微PET成像后，安乐死的大鼠与二氧化碳预填充，而老鼠是在深麻醉室。 </li></ol></li></ol> 4.基因芯片<ol><li>安乐死<ol><li>诱导并用5％异氟醚分别经由室与鼻锥体溶解在40％/ 60％和25％/ 75％氧/氮，2％维持麻醉。深麻醉在被斩首前废除踏板撤回反射到脚趾捏的水平。 </li><li>斩首大鼠安乐死5分钟后，1赫兹的rTMS第1届。 </li></ol></li><li>组织收获<ol><li>铺陈材料和外科器械中使用的顺序，包括折叠的纸巾，骨骨钳，microscissors，更大的外科剪刀，microforcep，10号或11手术刀刀片，一个带盖10厘米玻璃培养皿装满冰和1.5毫升管中。准备一个塑料袋处置尸体的。 </li><li>使颅骨anterioposteriorly中线皮肤切口。说白了解剖软组织及周围肌肉手术剪，然后取出用咬骨钳的颅骨片。迅速从头骨解剖仔细新鲜的大脑。然后，通过使用microforceps和microscissors其放在冰上。冲洗在冰冷的生理盐水的脑组织。 </li><li>脑立即传送到干冰，并随后将其在-80℃下保存在一个管，直到进一步处理。 </li><li>解冻收获前脑组织。 </li><li>将大脑背侧，并收获从ST脑组织通过使用microforceps和microscissors imulated冰大脑皮层（围绕在初级运动皮层的热点）。把收获的组织中的1.5毫升管中。 </li></ol></li><li> RNA制备<ol><li>提取用裂解试剂22组织匀浆的总RNA。过程与DNA酶消化和清理程序。定量RNA样品与等份，并将它们存储在-80℃直至使用。 </li><li>在质量控制方面，通过变性凝胶电泳，在260 OD比评估RNA的纯度和完整性:280，并分析它们对市售的分析仪。 </li></ol></li><li>标签和净化<ol><li>放大并通过使用市售的RNA扩增试剂盒，以产生生物素化的cRNA纯化总RNA。简言之，反向转录通过用T7寡（dT）引物550纳克总RNA到cDNA的。合成和体外转录第二链cDNA，然后与生物素的NTP标记它。 </li><li>经过PURification，通过使用分光光度计量化的cDNA。 </li></ol></li><li>杂交和数据导出23 <ol><li>用于表达分析表达BeadChip芯片。杂交标记750纳克cDNA样品到各大鼠-12表达珠阵列16 - 18小时在58℃。通过使用链霉亲和的Cy3进行阵列信号的检测。 </li><li>扫描阵列，共焦扫描仪。通过使用市售的软件进行阵列数据输出处理和分析。 </li></ol></li></ol>

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C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transcranial+magnetic+stimulation+of+small+animals:+a+modeling+study+of+the+influence+of+coil+geometry,+size+and+orientation.">Transcranial magnetic stimulation of small animals: a modeling study of the influence of coil geometry, size and orientation.</a> Conf Proc IEEE Eng Med Biol Soc. 2009, 674-677 (2009).</li><li>Parthoens, J., Verhaeghe, J., Servaes, S., Miranda, A., Stroobants, S., Staelens, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Performance+Characterization+of+an+Actively+Cooled+Repetitive+Transcranial+Magnetic+Stimulation+Coil+for+the+Rat.">Performance Characterization of an Actively Cooled Repetitive Transcranial Magnetic Stimulation Coil for the Rat.</a> Neuromodulation. , (2016).</li><li>Toro, R., 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=Brain+size+and+folding+of+the+human+cerebral+cortex.">Brain size and folding of the human cerebral cortex.</a> Cereb Cortex. 18 (10), 2352-2357 (2008).</li></ol>

The authors have nothing to disclose.

<html>这项研究的主要目的是介绍单边rTMS治疗的动物模型。虽然单边刺激是人类颅磁刺激研究的最根本的特征之一，许多研究都没有小动物采用。然而，罗滕贝格等人 15记录，使用具有20毫米的外瓣直径的8字形线圈100％MT的刺激对侧的MEP，而刺激112.5％和133.3％的MT产生同侧以及对侧的MEP。这可能是因为大感应电场可影响对侧半球。因此，我们的研究是此以前的工作15,24的延伸?...

重复经颅磁刺激（rTMS治疗），用于非侵入性脑刺激和神经调节的工具，已经在各种病症的治疗被应用于如中枢性疼痛1,2，抑郁心情 ，偏头痛4，甚至中风5-7。通过线圈快速变化的电流在磁头诱导对大脑皮层和所得神经元激活电场。大脑皮层的兴奋性可以通过磁刺激，从而可以持续超过30分钟的刺激终止后进行调制。 后效应的磁刺激的建议的机制包括长时程增强/抑郁症样作用8，在离子平衡9短暂移位，和代谢改变10。此外，迪拉扎罗等 。表明，间歇THETA突发刺激影响兴奋性突触输入到锥体束神经元，无论是在刺激与对侧大脑半球11。 显著的限制，但是，已经从台上证据转化为临床情况阻碍了研究人员。首先，在以前的动物研究中，磁刺激用于全脑刺激12。全脑刺激是从人类研究9所使用的协议完全不同。另一个问题是与刺激的持续时间有关。这至少部分归因于一个事实，即一个有效的冷却系统是在过去的小线圈不可用。 近年来，精液文章已经发表建议用于克服对小动物大脑中的磁刺激实验这些困难的方式。由这些动物模型中，据透露，在大鼠脑还示出了响应于低频磁刺激13相似皮质兴奋的变化作为人类。更重要的是，经颅磁刺激的细胞和分子机制越来越蓓NG使用经颅磁刺激的动物模型研究。一个典型的例子是，一个独特的类型抑制中间神经元的被称为是间歇THETA爆发刺激14最为敏感。经颅磁刺激的啮齿类动物模型，因此，探索对颅磁刺激引起的变化的分子基础备受追捧的问题提供了新的机遇。如果经颅磁刺激的小动物模型可以更加实验室使用，它可能会大大加快，并在这方面加强研究。 我们现在描述如何将磁刺激应用于大鼠脑，将先前的工作15的延伸的单方面半球。刺激诱导的变化通过使用微正电子发射断层扫描（PET）和mRNA微阵列来研究在受刺激大脑皮质的rTMS引起的变化进行评价。

<table><tbody><tr><td>Homeothermic blanket with a rectal probe</td><td>Harvard apparatus</td><td>507222F</td><td></td></tr><tr><td>Isoflurane (Forane sol.)</td><td>Choongwae</td><td></td><td></td></tr><tr><td>Propofol (Provive Inj. 1% 20 ml)</td><td>Claris Lifesciences</td><td></td><td></td></tr><tr><td>Repetitive magnetic stimulator (Magstim Rapid&lt;sup&gt;2&lt;/sup&gt;)</td><td>Magstim Company Ltd</td><td></td><td></td></tr><tr><td>25 mm figure-of-8 coil</td><td>Magstim Company Ltd</td><td>1165-00</td><td></td></tr><tr><td>PET-CT</td><td>GE Healthcare</td><td></td><td></td></tr><tr><td>QIAzol Lysis Reagent</td><td>Qiagen</td><td>(US Patent No. 5,346,994)</td><td></td></tr><tr><td>RNeasy Lipid Tissue Mini Kit</td><td>Qiagen</td><td>74804</td><td></td></tr><tr><td>RNeasy Mini Spin Columns</td><td>Qiagen</td><td>(Mat No. 1011708)</td><td></td></tr><tr><td>Agilent 2100 Bioanalyzer</td><td>Agilent Technologies</td><td></td><td></td></tr><tr><td>Ambion Illumina RNA amplification kit</td><td>Ambion</td><td></td><td></td></tr><tr><td>Nanodrop Spectrophotometer</td><td>NanoDrop</td><td>ND-1000</td><td></td></tr><tr><td>Illumina RatRef-12 Expression BeadChip</td><td>Illumina, Inc.</td><td></td><td></td></tr><tr><td>Amersham fluorolink streptavidin-Cy3</td><td>GE Healthcare Bio-Sciences</td><td></td><td></td></tr></tbody></table>

repetitive transcranial magnetic stimulation to the unilateral hemisphere of rat brain

Previous rodent models of repetitive transcranial magnetic stimulation (rTMS) adopted whole-brain stimulation instead of unilateral hemispheric rTMS, which is unlike the protocols used for human subjects. We report a successful application of rTMS to the unilateral hemisphere of rat brain. The rTMS was delivered with a low-frequency (1 Hz), high-frequency (20 Hz), or sham stimulation protocol to one side of the brain by using a small 25-mm figure-8 coil. We placed the center of the coil 1 cm lateral to the vertex on the biauricular line and angulated the coil 45&#176; to the ground to minimize a potential direct effect of rTMS on the contralateral cortex. We also used an in-house water cooling system to enable repetitive magnetic stimulation for more than 20 min, even at a 20-Hz stimulation frequency. Increases in the transcriptions of immediate early genes (Arc, Junb, and Egr2) were greater after rTMS than after sham stimulation. After 5 consecutive days of 20-min 1-Hz rTMS, bdnf mRNA expression was significantly higher in stimulated cortex than in contralateral side. The model presented herein will elucidate the molecular mechanisms of rTMS by allowing analysis of the inter-hemispheric difference in its effect.

十五8周龄雄性SD大鼠被用于MT确定一个独立的评判间可靠性分析。使用肌肉抽搐的触诊的MT在所有可获得的大鼠和33.00±4.21％最大输出刺激（％MSO）和33.93±0.88％MSO，测量分别由两个独立的研究人员。奥特曼偏倚-0.93，以及协议的95％限为-9.13至7.26％。 在6个8周龄鼠（n =在1赫兹的rTMS 4，和n = 2的假rTMS组中）微PET试验，18 F-F...

Watch this Scientific Journal Video about Repetitive Transcranial Magnetic Stimulation to the Unilateral Hemisphere of Rat Brain at JoVE.com

Repetitive Transcranial Magnetic Stimulation to the Unilateral Hemisphere of Rat Brain

We applied repetitive transcranial magnetic stimulation (rTMS) to the unilateral hemisphere of rat brain, by placing a 25-mm figure-8 coil 1 cm lateral to the vertex on the biauricular line and angulating the coil by 45&#176;. An in-house water cooling system was used for rTMS for more than 20 min.

重复经颅磁刺激对大鼠大脑的单边半球

Previous rodent models of repetitive transcranial magnetic stimulation (rTMS) adopted whole-brain stimulation instead of unilateral hemispheric rTMS, ...

repetitive-transcranial-magnetic-stimulation-to-unilateral-hemisphere

Research

JoVE Journal

Behavior

12.5K 次观看.  Chungnam National University Hospital, Daejeon. We applied repetitive transcranial magnetic stimulation (rTMS) to the unilateral hemisphere of rat brain, by placing a 25-mm figure-8 coil 1 cm lateral to the vertex on the biauricular line and angulating the coil by 45°. An in-house water cooling system was used for rTMS for more than 20 min. 

视频: Repetitive Transcranial Magnetic Stimulation to the Unilateral Hemisphere of Rat Brain

Utilizing Transcranial Magnetic Stimulation to Study the Human Neuromuscular System

Transcranial magnetic stimulation (TMS) has been in use for more than 20 years 1, and has grown exponentially in popularity over the past decade. While the use of TMS has expanded to the study of many systems and processes during this time, the original application and perhaps one of the most common uses of TMS involves studying the physiology, plasticity and function of the human neuromuscular system. Single pulse TMS applied to the motor cortex excites pyramidal neurons transsynaptically 2 (Figure 1) and results in a measurable electromyographic response that can be used to study and evaluate the integrity and excitability of the corticospinal tract in humans 3. Additionally, recent advances in magnetic stimulation now allows for partitioning of cortical versus spinal excitability 4,5. For example, paired-pulse TMS can be used to assess intracortical facilitatory and inhibitory properties by combining a conditioning stimulus and a test stimulus at different interstimulus intervals 3,4,6-8. In this video article we will demonstrate the methodological and technical aspects of these techniques. Specifically, we will demonstrate single-pulse and paired-pulse TMS techniques as applied to the flexor carpi radialis (FCR) muscle as well as the erector spinae (ES) musculature. Our laboratory studies the FCR muscle as it is of interest to our research on the effects of wrist-hand cast immobilization on reduced muscle performance6,9, and we study the ES muscles due to these muscles clinical relevance as it relates to low back pain8. With this stated, we should note that TMS has been used to study many muscles of the hand, arm and legs, and should iterate that our demonstrations in the FCR and ES muscle groups are only selected examples of TMS being used to study the human neuromuscular system.

Transcranial magnetic stimulation (TMS) is a non-invasive tool to gain insight on the physiology and function of the human nervous system. Here, we present our TMS techniques to study cortical excitability of the upper limb and lumbar musculature.

利用经颅磁刺激，以研究人类的神经肌肉系统

Transcranial magnetic stimulation (TMS) has been in use for more than 20 years 1, and has grown exponentially in popularity over the past decade.  While ...

科研

医学

Neuronavigation-guided Repetitive Transcranial Magnetic Stimulation for Aphasia

Repetitive transcranial magnetic stimulation (rTMS) is widely used for several neurological conditions, as it has gained acknowledgement for its potential therapeutic effects. Brain excitability is non-invasively modulated by rTMS, and rTMS to the language areas has proved its potential effects on treatment of aphasia. In our protocol, we aim to artificially induce virtual aphasia in healthy subjects by inhibiting Brodmann area 44 and 45 using neuronavigational TMS (nTMS), and F3 of the International 10-20 EEG system for conventional TMS (cTMS). To measure the degree of aphasia, changes in reaction time to a picture naming task pre- and post-stimulation are measured and compare the delay in reaction time between nTMS and cTMS. Accuracy of the two TMS stimulation methods is compared by averaging the Talairach coordinates of the target and the actual stimulation. Consistency of stimulation is demonstrated by the error range from the target. The purpose of this study is to demonstrate use of nTMS and to describe the benefits and limitations of the nTMS compared to those of cTMS.

This study is designed to test the hypothesis that neuronavigational system-guided transcranial magnetic stimulation has higher accuracy for targeting the intended target as demonstrated by eliciting a greater degree of virtual aphasia in healthy subjects, measured by delay in reaction time to picture naming.

神经导航引导下重复经颅磁刺激的失语

Repetitive transcranial magnetic stimulation (rTMS) is widely used for several neurological conditions, as it has gained acknowledgement for its potential ...

行为学

Simultaneous Transcranial Alternating Current Stimulation and Functional Magnetic Resonance Imaging

Transcranial alternating current stimulation (tACS) is a promising tool for noninvasive investigation of brain oscillations. TACS employs frequency-specific stimulation of the human brain through current applied to the scalp with surface electrodes. Most current knowledge of the technique is based on behavioral studies; thus, combining the method with brain imaging holds potential to better understand the mechanisms of tACS. Because of electrical and susceptibility artifacts, combining tACS with brain imaging can be challenging, however, one brain imaging technique that is well suited to be applied simultaneously with tACS is functional magnetic resonance imaging (fMRI). In our lab, we have successfully combined tACS with simultaneous fMRI measurements to show that tACS effects are state, current, and frequency dependent, and that modulation of brain activity is not limited to the area directly below the electrodes. This article describes a safe and reliable setup for applying tACS simultaneously with visual task fMRI studies, which can lend to understanding oscillatory brain function as well as the effects of tACS on the brain.

Transcranial alternating current stimulation (tACS) is a promising tool for noninvasive investigation of brain oscillations, though its effects are not completely understood. This article describes a safe and reliable setup for applying tACS simultaneously with functional magnetic resonance imaging, which can increase understanding oscillatory brain function and effects of tACS.

同时经颅交替电流刺激和功能磁共振成像

Transcranial alternating current stimulation (tACS) is a promising tool for noninvasive investigation of brain oscillations. TACS employs ...

Utilizing Repetitive Transcranial Magnetic Stimulation to Improve Language Function in Stroke Patients with Chronic Non-fluent Aphasia

Transcranial magnetic stimulation (TMS) has been shown to significantly improve language function in patients with non-fluent aphasia1. In this experiment, we demonstrate the administration of low-frequency repetitive TMS (rTMS) to an optimal stimulation site in the right hemisphere in patients with chronic non-fluent aphasia. A battery of standardized language measures is administered in order to assess baseline performance. Patients are subsequently randomized to either receive real rTMS or initial sham stimulation. Patients in the real stimulation undergo a site-finding phase, comprised of a series of six rTMS sessions administered over five days; stimulation is delivered to a different site in the right frontal lobe during each of these sessions. Each site-finding session consists of 600 pulses of 1 Hz rTMS, preceded and followed by a picture-naming task. By comparing the degree of transient change in naming ability elicited by stimulation of candidate sites, we are able to locate the area of optimal response for each individual patient. We then administer rTMS to this site during the treatment phase. During treatment, patients undergo a total of ten days of stimulation over the span of two weeks; each session is comprised of 20 min of 1 Hz rTMS delivered at 90% resting motor threshold. Stimulation is paired with an fMRI-naming task on the first and last days of treatment. After the treatment phase is complete, the language battery obtained at baseline is repeated two and six months following stimulation in order to identify rTMS-induced changes in performance. The fMRI-naming task is also repeated two and six months following treatment. Patients who are randomized to the sham arm of the study undergo sham site-finding, sham treatment, fMRI-naming studies, and repeat language testing two months after completing sham treatment. Sham patients then cross over into the real stimulation arm, completing real site-finding, real treatment, fMRI, and two- and six-month post-stimulation language testing.

We explore the use of repetitive transcranial magnetic stimulation (rTMS) to improve language abilities in patients with chronic stroke and non-fluent aphasia. After identifying a site in the right frontal gyrus for each patient that responds optimally to stimulation, we target this site during ten days of rTMS treatment.

利用重复经颅磁刺激改善脑卒中患者的语言功能与慢性非流利性失语

Transcranial magnetic stimulation (TMS) has been shown to significantly improve language function in patients with non-fluent aphasia1. In this ...

Deep Brain Stimulation with Simultaneous fMRI in Rodents

In order to visualize the global and downstream neuronal responses to deep brain stimulation (DBS) at various targets, we have developed a protocol for using blood oxygen level dependent (BOLD) functional magnetic resonance imaging (fMRI) to image rodents with simultaneous DBS. DBS fMRI presents a number of technical challenges, including accuracy of electrode implantation, MR artifacts created by the electrode, choice of anesthesia and paralytic to minimize any neuronal effects while simultaneously eliminating animal motion, and maintenance of physiological parameters, deviation from which can confound the BOLD signal.&#160;Our laboratory has developed a set of procedures that are capable of overcoming most of these possible issues. For electrical stimulation, a homemade tungsten bipolar microelectrode is used, inserted stereotactically at the stimulation site in the anesthetized subject. In preparation for imaging, rodents are fixed on a plastic headpiece and transferred to the magnet bore. For sedation and paralysis during scanning, a cocktail of dexmedetomidine and pancuronium is continuously infused, along with a minimal dose of isoflurane; this preparation minimizes the BOLD ceiling effect of volatile anesthetics. In this example experiment, stimulation of the subthalamic nucleus (STN) produces BOLD responses which are observed primarily in ipsilateral cortical regions, centered in motor cortex. Simultaneous DBS and fMRI allows the unambiguous modulation of neural circuits dependent on stimulation location and stimulation parameters, and permits observation of neuronal modulations free of regional bias. This technique may be used to explore the downstream effects of modulating neural circuitry at nearly any brain region, with implications for both experimental and clinical DBS.

This protocol describes a standard method for simultaneous functional magnetic resonance imaging and deep brain stimulation in the rodent. The combined use of these experimental tools allows for the exploration of global downstream activity in response to electrical stimulation at virtually any brain target.

深部脑刺激与同步功能磁共振成像在鼠害

In order to visualize the global and downstream neuronal responses to deep brain stimulation (DBS) at various targets, we have developed a protocol for ...

神经科学

The Use of Magnetic Resonance Spectroscopy as a Tool for the Measurement of Bi-hemispheric Transcranial Electric Stimulation Effects on Primary Motor Cortex Metabolism

Transcranial direct current stimulation (tDCS) is a neuromodulation technique that has been increasingly used over the past decade in the treatment of neurological and psychiatric disorders such as stroke and depression. Yet, the mechanisms underlying its ability to modulate brain excitability to improve clinical symptoms remains poorly understood 33. To help improve this understanding, proton magnetic resonance spectroscopy (1H-MRS) can be used as it allows the in vivo quantification of brain metabolites such as &#947;-aminobutyric acid (GABA) and glutamate in a region-specific manner 41. In fact, a recent study demonstrated that 1H-MRS is indeed a powerful means to better understand the effects of tDCS on neurotransmitter concentration 34. This article aims to describe the complete protocol for combining tDCS (NeuroConn MR compatible stimulator) with 1H-MRS at 3 T using a MEGA-PRESS sequence. We will describe the impact of a protocol that has shown great promise for the treatment of motor dysfunctions after stroke, which consists of bilateral stimulation of primary motor cortices 27,30,31. Methodological factors to consider and possible modifications to the protocol are also discussed.

This article aims to describe a basic protocol for combining transcranial direct current stimulation (tDCS) with proton magnetic resonance spectroscopy (1H-MRS) measurements to investigate the effects of bilateral stimulation on primary motor cortex metabolism.

磁共振波谱的用途，用于对初级运动皮层新陈代谢双半球经颅电刺激效果的测量工具一

Transcranial direct current stimulation (tDCS) is a neuromodulation technique that has been increasingly used over the past decade in the treatment of ...

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 ...

Functional Magnetic Resonance Spectroscopy at 7 T in the Rat Barrel Cortex During Whisker Activation

Nuclear magnetic resonance (NMR) spectroscopy offers the opportunity to measure cerebral metabolite contents in vivo and noninvasively. Thanks to technological developments over the last decade and the increase in magnetic field strength, it is now possible to obtain good resolution spectra in vivo in the rat brain. Neuroenergetics (i.e., the study of brain metabolism) and, especially, metabolic interactions between the different cell types have attracted more and more interest in recent years. Among these metabolic interactions, the existence of a lactate shuttle between neurons and astrocytes is still debated. It is, thus, of great interest to perform functional proton magnetic resonance spectroscopy (1H-MRS) in a rat model of brain activation and monitor lactate. However, the methyl lactate peak overlaps lipid resonance peaks and is difficult to quantify. The protocol described below allows metabolic and lactate fluctuations to be monitored in an activated brain area. Cerebral activation is obtained by whisker stimulation and 1H-MRS is performed in the corresponding activated barrel cortex, whose area is detected using blood-oxygen-level-dependent functional magnetic resonance imaging (BOLD fMRI). All steps are fully described: the choice of anesthetics, coils, and sequences, achieving efficient whisker stimulation directly in the magnet, and data processing.

After checking by blood-oxygen-level-dependent functional magnetic resonance imaging (BOLD fMRI) that the corresponding somatosensory barrel field cortex area (called S1BF) is correctly activated, the main goal of this study is to quantify lactate content fluctuations in the activated rat brains by localized proton magnetic resonance spectroscopy (1H-MRS) at 7 T.

大鼠桶内皮层 7 t 的功能磁共振波谱在晶氏体活化过程中的应用

Nuclear magnetic resonance (NMR) spectroscopy offers the opportunity to measure cerebral metabolite contents in vivo and noninvasively. Thanks to ...

A Pilot Study on the Repetitive Transcranial Magnetic Stimulation of A&#946; and Tau Levels in Rhesus Monkey Cerebrospinal Fluid

Previous studies have demonstrated that a non-invasive light-flickering regime and auditory tone stimulation could affect A&#946; and tau metabolism in the brain. As a non-invasive technique, repetitive transcranial magnetic stimulation (rTMS) has been applied for the treatment of neurodegenerative disorders. This study explored the effects of rTMS on A&#946; and tau levels in rhesus monkey cerebrospinal fluid (CSF). This is a single-blind, self-controlled study. Three different frequencies (low frequency, 1 Hz; high frequencies, 20 Hz and 40 Hz) of rTMS were used to stimulate the bilateral-dorsolateral prefrontal cortex (DLPFC) of the rhesus monkey. A catheterization method was used to collect CSF. All samples were subjected to liquid chip detection to analyze CSF biomarkers (A&#946;42, A&#946;42/A&#946;40, tTau, pTau). CSF biomarker levels changed with time after stimulation by rTMS. After stimulation, the A&#946;42 level in CSF showed an upward trend at all frequencies (1 Hz, 20 Hz, and 40 Hz), with more significant differences for the high-frequencies (p &lt; 0.05) than for the low frequency. 
After high-frequency rTMS, the total Tau (tTau) level of CSF immediately increased at the post-rTMS timepoint (p &lt; 0.05) and gradually decreased by 24 h. Moreover, the results showed that the level of phosphorylated Tau (pTau) increased immediately after 40 Hz rTMS (p &lt; 0.05). The ratio of A&#946;42/A&#946;40 showed an upward trend at 1 Hz and 20 Hz (p &lt; 0.05). There was no significant difference in the tau levels with low-frequency (1 Hz) stimulation. Thus, high-frequencies (20 Hz and 40 Hz) of rTMS may have positive effects on A&#946; and tau levels in rhesus monkey CSF, while low-frequency (1 Hz) rTMS can only affect A&#946; levels.

Here, we describe the procedure for a pilot study to explore the effect of repetitive transcranial magnetic stimulation with different frequencies (1 Hz/20 Hz/40 Hz) on A&#946; and tau metabolism in rhesus monkey cerebrospinal fluid.

恒河猴脑脊液中Aβ和Tau水平重复经颅磁刺激的试点研究

Previous studies have demonstrated that a non-invasive light-flickering regime and auditory tone stimulation could affect A&#946; and tau metabolism in ...

用低成本的 EEG 和毫米级线圈刺激小鼠大脑

Low-Cost Electroencephalographic Recording System Combined with a Millimeter-Sized Coil to Transcranially Stimulate the Mouse Brain In Vivo

A low-cost electroencephalographic (EEG) recording system is proposed here to drive transcranial magnetic stimulation (TMS) of the mouse brain in vivo, utilizing a millimeter-sized coil. Using conventional screw electrodes combined with a custom-made, flexible, multielectrode array substrate, multi-site recording can be carried out from the mouse brain. In addition, we explain how a millimeter-sized coil is produced using low-cost equipment usually found in laboratories. Practical procedures for fabricating the flexible multielectrode array substrate and the surgical implantation technique for screw electrodes are also presented, which are necessary to produce low-noise EEG signals. Although the methodology is useful for recording from the brain of any small animal, the present report focuses on electrode implementation in an anesthetized mouse skull. Furthermore, this method can be easily extended to an awake small animal that is connected with tethered cables via a common adapter and fixed with a TMS device to the head during recording.The present version of the EEG-TMS system, which can include a maximum of 32 EEG channels (a device with 16 channels is presented as an example with fewer channels) and one TMS channel device, is described. Additionally, typical results obtained by the application of the EEG-TMS system to anesthetized mice are briefly reported.

作者聚焦: 低成本脑电图记录系统结合毫米级线圈经颅刺激小鼠体内大脑  

A low-cost electroencephalographic recording system combined with a millimeter-sized coil is proposed to drive transcranial magnetic stimulation of the mouse brain in vivo. Using conventional screw electrodes with a custom-made, flexible, multielectrode array substrate, multi-site recording can be carried out from the mouse brain in response to transcranial magnetic stimulation.

低成本脑电图记录系统结合毫米级线圈，在体内经颅刺激小鼠大脑

A low-cost electroencephalographic (EEG) recording system is proposed here to drive transcranial magnetic stimulation (TMS) of the mouse brain in vivo, ...