Name: transcranial electrical brain stimulation in alert rodents
Uploaded: 2017-11-02T14:00:00.000+00:00
Duration: 10 min 8 s
Description: Watch this Scientific Journal Video about Transcranial Electrical Brain Stimulation in Alert Rodents at JoVE.com

这项工作得到了德国研究基金会 (DFG RE 2740/3-1) 的支持。我们感谢弗兰克 Huethe 和托马斯 nther 的 in-house 生产 custom-made 工商业污水附加费和 DC 刺激。

对于涉及动物的研究, 在开始实验之前必须获得相关的 (具体国家的) 批准。这里报告的所有动物实验都是按照欧盟指令 2010/63/欧盟、更新的德国动物保护法 (和 #34; Tierschutzgesetz 和 #34;) 2013年7月, 以及2013年8月更新的德国动物研究条例进行的。地方当局和 #34 批准了动物议定书; 弗赖堡和 #34 区域理事会动物实验委员会; 和 #34; 大学医学中心动物实验委员会弗莱堡和 #34;. 
 1. 准备用于外科手术的仪器和材料 <ol> <li> 请确保在 图 1 中列出的项目已经可用, 并已放置用于手术. </li>
	<li> 准备一个薄的矩形铂金板 ( 例如, 10 mm x 6 mm x 0.15 mm), 它将用作在胸前放置的计数器电极, 并在两个相反的角板上打两个小孔. </li>
	<li> 焊接一条长度为10厘米的绝缘电缆, 使用无铅锡焊料到铂盘的一个角 (无孔). </li>
	<li> 在焊接接头上应用一小滴组织-丙烯酸胶水进行隔离. </li>
</ol> 2。外科用啮齿类动物的制备 <ol> <li> 将研究编号分配给啮齿动物, 并在准备好的手术卡上注明. </li>
	<li> 称量啮齿动物, 注意手术卡上的重量。计算注射麻醉剂的剂量 ( 例如, 氯胺酮100毫克/千克体重加上嗪70毫克/千克体重的大鼠). </li>
	<li> 通过腹腔 (ip) 注射麻醉药的量来诱导麻醉剂. 
	注: 当使用吸入麻醉代替 ( 例如, 异氟醚) 时, 将啮齿类动物置于感应腔内, 在1-2 升/分钟氧气中连续流动 4%. </li>
	<li> 检查麻醉深度, 由脚趾捏反射开始5分钟后注射。如果脚趾捏反射仍然存在, 通过注射30% 的初始剂量达到延长和深化麻醉. 
	<ol> <li> 如果在实验中的任何时间点的脚趾夹紧反射返回, 应注射30% 的初始剂量的麻醉. </li>
		<li> 当使用吸入麻醉时, 寻找在感应室的啮齿动物的姿势反射损失和检查深度的麻醉由缺乏脚趾捏反射。如果反射仍然存在, 延长在麻醉室的持续时间。在整个实验中, 将异氟醚的比例调整到麻醉深度, 直到达到 1-1. 5% 异氟醚的维持浓度. </li>
		<li> 当呼吸频率下降, 喘息发生时, 降低百分比; 当啮齿动物恢复脚趾捏反射或显示自发运动时, 增加吸入麻醉的百分比. </li>
	</ol> </li> <li> 当反射不存在时, 将啮齿类动物放在实验室的长凳上或握住它的手. 
	注: 吸入麻醉时, 使用与雾化器相连的喷嘴, 提供持续减少的异氟醚流量 (现在介于2-3% 之间). </li>
	<li> 移除老鼠头上的毛发, #39 的头部, 从耳朵到耳朵, 从侧的眼睛, 到耳朵后面, 用剪刀把区域剃掉。然后把剑上的前肢和锁骨之间的区域剃掉, 把头发移到胸前. 
	注意: 保持皮肤紧绷使剃须. </li>
	<li> 用一滴眼药膏遮住老鼠的眼睛以保护角膜. </li>
	根据指定的学习编号, <li> 标记大鼠和 #39 的耳朵. 
	注: 根据研究的长度, 尾巴标记可能是充足的, 否则规范化指定是优选的. </li>
</ol> 3。手术方法: 胸电极植入术 注意: 当将计数器电极外部放置在带背心的剃须胸前时, 可以跳过此步骤. 
<ol> <li> 将啮齿动物俯卧 (在胸部) 放在手术台上. 
	注: 在吸入麻醉的情况下, 将大鼠和 #39 的鼻口置于麻醉喷嘴中, 进一步将异氟醚浓度降低到 1.5-2%. </li>
	<li> 用消毒剂喷雾或用杀菌剂浸泡过的棉签 ( 例如, 乙醇 70%) 消毒剃光头, 让其风干。重复两次. </li>
	<li> 用手术刀在一条线上从侧的眼睛切开皮肤到中耳水平. 
	注: 这允许隧道的连接电缆从植入的胸部电极的头部顶部, 也是理想的切割 DCS 电极插座的位置. </li>
	<li> 将鼠转到仰卧位, 使胸部暴露. </li>
	<li> 步骤3.2 中所述, 对胸部皮肤进行消毒. </li>
	<li> 用纸巾钳将右胸的侧皮抬高, 用小剪刀从右腋内侧剪下0.5 厘米的扣眼。然后用剪刀在颅骨方向上做一个直矢状切口. </li>
	<li> 通过 atraumatically 从左侧主要的胸肌中断开皮肤, 形成皮下囊。这样做, 通过反复打开小剪刀 (或由盐水浸泡棉签). </li>
	<li> 将其右侧的动物转到沿颈部打开的头部皮肤的左枕角的电缆通道, 然后用平衡钳穿透浅筋膜, 进入胸袋. </li>
	<li> 小心打开平衡钳, 以抓住连接到铂电极的电极电缆的末端, 而不允许尖锐的导线偏离。拉电缆通过隧道, 直到电极进入邮袋, 以焊接点为导向的啮齿动物左后肢。将啮齿动物恢复到俯卧位置. </li>
	<li> 修复铂板与无菌合成编织吸收缝合到胸筋膜在两个对立的角落孔 (4-5 节建议稳定). </li>
	<li> 类似地将电缆连接到筋膜上的一个松散的结, 形成一个轻微的循环前的组织隧道入口. </li>
	<li> 根据切口的大小将皮肤缝合3-4 皮 (同样的缝合材料可用于电极和电缆). </li>
</ol> 4。外科手术: Epicranial 的位置 <ol> <li> 将动物置于立体定向的框架中. 
	注意: 如果使用吸入麻醉, 降低麻醉的浓度到维持异氟醚的流量〜 1.5-1%, 调整到脚趾捏反射和呼吸模式. </li>
	<li> 步骤3.2 中所述的剃光头皮进行消毒. </li>
	<li> 用手术刀在一条线上从侧的眼睛切开皮肤到中耳水平. 
	注: 如果胸电极 placem执行了耳鼻喉科, 已执行步骤4.2 和 4.3. </li>
	<li> 用手术刀把骨膜 (结缔组织放在头骨上) 刮掉, 并彻底抹掉棉花掉期。固定的结缔组织在4角的切口与斗牛犬钳夹, 让他们横向挂起, 以保持手术场开放. </li>
	<li> 应用0.9% 盐水以棉签清洁骨骼表面和组织。然后用 3% H 2 O 2 清除骨骼表面。避免与组织接触。因此, 骨头被清洗得更彻底, 骨头上的轻微出血将被停止。并且, 骨膜的残差变得可见。使用中等压力的棉签将这些残余物移除. 
	注: 去除骨膜残差将增加粘附在骨上的附加物的附着力和耐久性。
	<ol> <li> 在不可阻挡的出血情况下, 使用骨钻, 并触摸它为 1-3 s, 在骨头上的轻微压力。这种机械程序将在大多数情况下停止出血没有显着加热。不要在骨头上使用电;即使是短暂的应用也会导致脑组织损伤 (电应仅用于伤口组织出血). </li>
	</ol> </li> <li> 作为固定螺钉将提高安装的粘度, 请选择一个钻头, 使其与螺钉大小匹配。在两个不同的骨板上放置两个毛刺孔前用手钻, 然后通过轻微的垂直压力应用与骨钻。避免接近所需位置的污水附加费插座, 因为它可能会妨碍在电极拧紧 ( 例如, 左主电机皮质附加费, 选择右额和后侧壁螺钉位置). </li>
	<li> 在植入的反电极的情况下, 毛刺在右后壁骨的第三个孔, 以未来固定的隧道电缆. </li>
	<li> 将塑料螺丝放置在毛刺孔和螺钉中, 直到感觉到第一个摩擦力为止。然后执行三额外的180和 #176; 螺丝转动。用镊子检查螺丝是否稳定, 如果不够紧就再加一转. 
	注: 对于成年大鼠, 这将确保硬膜外放置的螺丝, 而不损害硬脑膜或大脑 (取决于螺纹设计, 转数可能会有所不同)。使用不锈钢螺丝也应该是可行的, 因为即使在 DCS 电流密度以上的神经阈值, 螺钉放置没有扰乱病变的位置或范围以下的螺丝. </li>
	<li> 打开焊接铁和预热约5分钟风电缆退出组织隧道 occipitally 周围的右顶叶, 然后削减它, 留下约1厘米电缆背后的绕组。小心地用手术刀把电缆末端的绝缘层剥去. </li>
	<li> 用基胶水将缠绕的电缆固定在螺钉和骨骼上. </li>
	<li> 将少量无铅锡焊料应用到连接器和反电极电缆的裸线上, 并将两个 pre-soldered 部件连接在一起, 同时接触焊接尖端, 直到锡焊料熔化 (约2-3 秒)。立即卸下焊接头, 以避免电缆的过度金属加热, 从而导致随后的组织损坏. </li>
	<li> 拿起定制的污水附加费电极插座 ( 图 1B , 在红色) 与弯曲, 锯齿状尖端镊子和应用一层薄薄的基胶水到插座的底部边缘。安置在马达皮层之上并且使用4毫米直径插口, 安置中间插座点在2毫米前和2毫米侧向从 bregma。对于这个位置, 插座的内内侧边界应直接在矢状缝合处结束, 尾端边界应以 bregma 的高度结束。将插座简要地压在骨骼上 (大多数基胶水通过压力硬化). 
	注意: 将光源直接放置在插座上方可以轻松放置插座. </li>
	<li> 确保插座区域内的骨骼不含胶水 (通过检查光, 因为胶水是反光的)。在胶水溢出的情况下, 卸下插座, 用手术刀刮胶水, 并重复步骤 4.12. </li>
	<li> 插座到位后, 以后的刺激区域没有胶水, 首先用一小滴基胶水将插座的外侧边界密封到相邻的组织上, 以避免有可能导致在这个位置分流电流的流体桥。不要应用太多的胶水, 因为它可能流入刺激区域 (如果发生这种情况, 返回步骤 4.12). 
	注: 保持刺激区不粘胶是至关重要的, 减少刺激面积可能大大增加电流密度 (一个/m & #178;). </li>
	<li> 用基胶水覆盖所有螺钉. </li>
	<li> 在小型硅管或玻璃中混合双组分牙科丙烯酸水泥。一旦它变得粘性, 应用它与牙科刮刀密封的插槽的其余边界的骨骼。避免任何流动的牙科丙烯酸水泥进入刺激区. </li>
	<li> 最后覆盖整个头骨、螺钉、计数器电极电缆和插座, 并 #8531; 与牙科丙烯酸水泥的插座。确保水泥具有正确的粘度: 如果太流体, 它会流入周围的组织;如果太硬, 很难均匀地分配. </li>
	<li> 当所有的骨头被覆盖, 水泥硬化, 去掉牛头犬钳; 皮肤应该接触已建成的水泥, 这样就不需要缝合了。(如果最初的切口选择太长, 结缔组织或肌肉是可见的, 请按照步骤3.12 中的描述应用缝合线). </li>
	<li> 应用一层碘与棉签在切口皮肤和皮下注射卡洛芬 (5 毫克/千克体重溶解在 5-7. 5 毫升0.9% 盐水为痛苦治疗和液体替换). 
	注: 如果使用吸入麻醉, 现在关闭它. </li>
	<li> 将啮齿类动物放在一个变暖的盒子里, 从麻醉中恢复, 直到啮齿动物苏醒并恢复体位稳定性. 
	注意: 根据机构和 #39 的建议, 每天检查动物和 #39 的体重发展、创面状况和一般福利标准. </li>
</ol> 5。经颅电刺激程序 注意: 由于麻醉会影响工商业污水附加费的效果, 因此建议尽可能在警报啮齿动物中执行刺激。在开始实验之前, 允许啮齿动物至少恢复5天 (头部和胸部伤口愈合)。实验可以在手术后的早期时间点进行, 当使用外电极固定在背心, 因为胸部伤口是最急躁;但是, 动物需要习惯于电极背心数天的时间和干扰行为的任务可能发生. 
<ol> <li> 用0.9% 盐水填充附加费电极插座的一半, 并除去气泡. </li>
	<li> 被前阴极 tDCS 的会议, 总是检查氯化, 如果需要 (如一个闪亮的银色表面), re-chlorinate 银/氯化电极。在阳极 tDCS 之前, 从以前的刺激中去除可能过量的氯化沉积物, 以允许良好的导电性。螺丝在工商业污水附加电极螺丝盖帽 ( 图 1B , 灰色片断). 
	警告: 如果在阴极 tDCS 之间 re-chlorinate 电极, 则会导致在刺激过程中的氯化反应耗尽, 并通过电化学化学反应产生有毒的积聚。这会引起组织损伤。如果刺激持续时间少于20分钟, 在一个疗程内不需要再加氯. </li>
	<li> 将电缆连接到头上的两个接头 (用于阳极刺激, 阳极电缆连接到螺钉盖上的接头上, 用于阴极刺激, 则相反) 
	注: 当使用外部放置的计数器电极时, 用导电胶盖住计数器电极, 并将其放置在啮齿动物和 #39 的胸前。这是最简单的, 如果电极是 pre-fixed 在一个小啮齿动物背心, 这种啮齿动物可以穿在刺激. </li>
	<li> 将啮齿动物放入实验笼中, 将电缆连接到保持架上方的旋转, 允许自由运动. </li>
	<li> 打开刺激器并调整激励参数 (刺激强度、持续时间、爬坡和下一次). </li>
	<li> 当不使用具有安全关机和断开警报的商业可用刺激装置时, 在电路中包括一米来检查恒定电流流. 
	注意: 通过这种设置, 可以在性能或训练行为任务时应用刺激. </li>
	<li> 在刺激过程中检查啮齿动物的压力或不适症状. </li>
	<li> 在刺激结束后, 断开电缆, 拧开头上的电极帽, 用棉签清洁和烘干插座。如果需要, 将啮齿动物返回家庭环境或进行行为过程. </li>
</ol>

<ol>
	<li>Bikson, M., 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=Safety+of+Transcranial+Direct+Current+Stimulation:+Evidence+Based+Update+2016.">Safety of Transcranial Direct Current Stimulation: Evidence Based Update 2016.</a> Brain Stimul. 9 (5), 641-661 (2016).</li><li>Bindman, L. J., Lippold, O. C., Redfearn, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+action+of+brief+polarizing+currents+on+the+cerebral+cortex+of+the+rat+(1)+during+current+flow+and+(2)+in+the+production+of+long-lasting+after-effects.">The action of brief polarizing currents on the cerebral cortex of the rat (1) during current flow and (2) in the production of long-lasting after-effects.</a> J Physiol. 172, 369-382 (1964).</li><li>Gartside, I. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanisms+of+sustained+increases+of+firing+rate+of+neurones+in+the+rat+cerebral+cortex+after+polarization:+role+of+protein+synthesis.">Mechanisms of sustained increases of firing rate of neurones in the rat cerebral cortex after polarization: role of protein synthesis.</a> Nature. 220 (5165), 382-383 (1968).</li><li>Purpura, D. P., McMurtry, J. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intracellular+activities+and+potential+changes+during+polarization+of+motor+cortex.">Intracellular activities and potential changes during polarization of motor cortex.</a> Neurophysiol. 28 (1), 166-185 (1965).</li><li>Nitsche, M., Paulus, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Excitability+changes+induced+in+the+human+motor+cortex+by+weak+transcranial+direct+current+stimulation.">Excitability changes induced in the human motor cortex by weak transcranial direct current stimulation.</a> J Physiol. 527 (Pt 3), 633-639 (2000).</li><li>Nitsche, M. A., Paulus, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sustained+excitability+elevations+induced+by+transcranial+DC+motor+cortex+stimulation+in+humans.">Sustained excitability elevations induced by transcranial DC motor cortex stimulation in humans.</a> Neurology. 57 (10), 1899-1901 (2001).</li><li>Cambiaghi, M., 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+transcranial+direct+current+stimulation+modulates+motor+excitability+in+mice.">Brain transcranial direct current stimulation modulates motor excitability in mice.</a> Eur J Neuro. 31 (4), 704-709 (2010).</li><li>Fritsch, B., 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=Direct+current+stimulation+promotes+BDNF-dependent+synaptic+plasticity:+potential+implications+for+motor+learning.">Direct current stimulation promotes BDNF-dependent synaptic plasticity: potential implications for motor learning.</a> Neuron. 66 (2), 198-204 (2010).</li><li>Ranieri, F., 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=Modulation+of+LTP+at+rat+hippocampal+CA3-CA1+synapses+by+direct+current+stimulation.">Modulation of LTP at rat hippocampal CA3-CA1 synapses by direct current stimulation.</a> J Neurophysiol. 107 (7), 1868-1880 (2012).</li><li>Kronberg, G., Bridi, M., Abel, T., Bikson, M., Parra, L. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct+Current+Stimulation+Modulates+LTP+and+LTD:+Activity+Dependence+and+Dendritic+Effects.">Direct Current Stimulation Modulates LTP and LTD: Activity Dependence and Dendritic Effects.</a> Brain Stimul. 10 (November), 51-58 (2016).</li><li>Sun, Y., 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=Direct+current+stimulation+induces+mGluR5-dependent+neocortical+plasticity.">Direct current stimulation induces mGluR5-dependent neocortical plasticity.</a> Ann Neurol. 80 (2), 233-246 (2016).</li><li>Podda, M. V., 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=Anodal+transcranial+direct+current+stimulation+boosts+synaptic+plasticity+and+memory+in+mice+via+epigenetic+regulation+of+Bdnf+expression.">Anodal transcranial direct current stimulation boosts synaptic plasticity and memory in mice via epigenetic regulation of Bdnf expression.</a> Sci Rep. 6, 22180 (2016).</li><li>Reis, J., Fritsch, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modulation+of+motor+performance+and+motor+learning+by+transcranial+direct+current+stimulation.">Modulation of motor performance and motor learning by transcranial direct current stimulation.</a> Curr opin Neurology. 24 (6), 590-596 (2011).</li><li>Buch, E. 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=Effects+of+tDCS+on+motor+learning+and+memory+formation+a+consensus+and+critical+position+paper.">Effects of tDCS on motor learning and memory formation a consensus and critical position paper.</a> Clin Neurophysiol. 128 (4), 589-603 (2017).</li><li>Reis, J., Fischer, J. T., Prichard, G., Weiller, C., Cohen, L. G., Fritsch, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Time-+but+not+sleep-dependent+consolidation+of+tDCS-enhanced+visuomotor+skills.">Time- but not sleep-dependent consolidation of tDCS-enhanced visuomotor skills.</a> Cerebral cortex. 25 (1), 109-117 (2015).</li><li>Monai, H., 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=Calcium+imaging+reveals+glial+involvement+in+transcranial+direct+current+stimulation-induced+plasticity+in+mouse+brain.">Calcium imaging reveals glial involvement in transcranial direct current stimulation-induced plasticity in mouse brain.</a> Nature Comm. 7, 11100 (2016).</li><li>Gellner, A. -. K., Reis, J., Fritsch, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glia:+A+Neglected+Player+in+Non-invasive+Direct+Current+Brain+Stimulation.">Glia: A Neglected Player in Non-invasive Direct Current Brain Stimulation.</a> Front Cell Neurosci. 10, 188 (2016).</li><li>Takano, Y., Yokawa, T., Masuda, A., Niimi, J., Tanaka, S., Hironaka, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+rat+model+for+measuring+the+effectiveness+of+transcranial+direct+current+stimulation+using+fMRI.">A rat model for measuring the effectiveness of transcranial direct current stimulation using fMRI.</a> Neurosci Lett. 491 (1), 40-43 (2011).</li><li>Islam, N., Moriwaki, A., Hattori, Y., Hori, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anodal+polarization+induces+protein+kinase+C+gamma+(PKC+gamma)-like+immunoreactivity+in+the+rat+cerebral+cortex.">Anodal polarization induces protein kinase C gamma (PKC gamma)-like immunoreactivity in the rat cerebral cortex.</a> Neurosci Res. 21, 169-172 (1994).</li><li>Islam, N., Aftabuddin, M., Moriwaki, A., Hattori, Y., Hori, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Increase+in+the+calcium+level+following+anodal+polarization+in+the+rat+brain.">Increase in the calcium level following anodal polarization in the rat brain.</a> Brain Res. 684 (2), 206-208 (1995).</li><li>Rohan, J. G., Carhuatanta, K. A., McInturf, S. M., Miklasevich, M. K., Jankord, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modulating+Hippocampal+Plasticity+with+In+Vivo+Brain+Stimulation.">Modulating Hippocampal Plasticity with In Vivo Brain Stimulation.</a> J Neurosci. 35 (37), 12824-12832 (2015).</li><li>Wachter, D., 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=Transcranial+direct+current+stimulation+induces+polarity-specific+changes+of+cortical+blood+perfusion+in+the+rat.">Transcranial direct current stimulation induces polarity-specific changes of cortical blood perfusion in the rat.</a> Exp Neurol. 227 (2), 322-327 (2011).</li><li>Koo, H., 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=After-effects+of+anodal+transcranial+direct+current+stimulation+on+the+excitability+of+the+motor+cortex+in+rats.">After-effects of anodal transcranial direct current stimulation on the excitability of the motor cortex in rats.</a> Rest Neurol Neurosci. 34 (5), 859-868 (2016).</li><li>Liebetanz, D., 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=After-effects+of+transcranial+direct+current+stimulation+(tDCS)+on+cortical+spreading+depression.">After-effects of transcranial direct current stimulation (tDCS) on cortical spreading depression.</a> Neurosci Lett. 398 (1-2), 85-90 (2006).</li><li>Fregni, F., 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=Effects+of+transcranial+direct+current+stimulation+coupled+with+repetitive+electrical+stimulation+on+cortical+spreading+depression.">Effects of transcranial direct current stimulation coupled with repetitive electrical stimulation on cortical spreading depression.</a> Exp Neurol. 204 (1), 462-466 (2007).</li><li>Cambiaghi, M., 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=Flash+visual+evoked+potentials+in+mice+can+be+modulated+by+transcranial+direct+current+stimulation.">Flash visual evoked potentials in mice can be modulated by transcranial direct current stimulation.</a> Neurosci. 185, 161-165 (2011).</li><li>Dockery, C. A., Liebetanz, D., Birbaumer, N., Malinowska, M., Wesierska, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cumulative+benefits+of+frontal+transcranial+direct+current+stimulation+on+visuospatial+working+memory+training+and+skill+learning+in+rats.">Cumulative benefits of frontal transcranial direct current stimulation on visuospatial working memory training and skill learning in rats.</a> Neurobiol Learn Mem. 96 (3), 452-460 (2011).</li><li>Faraji, J., Gomez-Palacio-Schjetnan, A., Luczak, A., Metz, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Beyond+the+silence:+Bilateral+somatosensory+stimulation+enhances+skilled+movement+quality+and+neural+density+in+intact+behaving+rats.">Beyond the silence: Bilateral somatosensory stimulation enhances skilled movement quality and neural density in intact behaving rats.</a> Behav Brain Res. 253, 78-89 (2013).</li><li>Pikhovych, A., 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=Transcranial+Direct+Current+Stimulation+Modulates+Neurogenesis+and+Microglia+Activation+in+the+Mouse+Brain.">Transcranial Direct Current Stimulation Modulates Neurogenesis and Microglia Activation in the Mouse Brain.</a> Stem Cells In. , 1-10 (2016).</li><li>Rueger, M. A., 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=Multi-session+transcranial+direct+current+stimulation+(tDCS)+elicits+inflammatory+and+regenerative+processes+in+the+rat+brain.">Multi-session transcranial direct current stimulation (tDCS) elicits inflammatory and regenerative processes in the rat brain.</a> PloS one. 7 (8), e43776 (2012).</li><li>Liebetanz, D., Koch, R., Mayenfels, S., K&#246;nig, F., Paulus, W., Nitsche, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Safety+limits+of+cathodal+transcranial+direct+current+stimulation+in+rats.">Safety limits of cathodal transcranial direct current stimulation in rats.</a> Clinical Neurophysiol. 120 (6), 1161-1167 (2009).</li><li>Yoon, K. J., Oh, B. -. M., Kim, D. -. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+improvement+and+neuroplastic+effects+of+anodal+transcranial+direct+current+stimulation+(tDCS)+delivered+1+day+vs.+1+week+after+cerebral+ischemia+in+rats.">Functional improvement and neuroplastic effects of anodal transcranial direct current stimulation (tDCS) delivered 1 day vs. 1 week after cerebral ischemia in rats.</a> Brain Res. 1452, 61-72 (2012).</li><li>Spezia Adachi, L. N., 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=Exogenously+induced+brain+activation+regulates+neuronal+activity+by+top-down+modulation:+conceptualized+model+for+electrical+brain+stimulation.">Exogenously induced brain activation regulates neuronal activity by top-down modulation: conceptualized model for electrical brain stimulation.</a> Exp Brain Res. 233 (5), 1377-1389 (2015).</li><li>Jackson, M. P., 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=Safety+parameter+considerations+of+anodal+transcranial+Direct+Current+Stimulation+in+rats.">Safety parameter considerations of anodal transcranial Direct Current Stimulation in rats.</a> Brain, behavior, and immunity. , (2017).</li><li>Ordek, G., Groth, J. D., Sahin, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differential+effects+of+ketamine/xylazine+anesthesia+on+the+cerebral+and+cerebellar+cortical+activities+in+the+rat.">Differential effects of ketamine/xylazine anesthesia on the cerebral and cerebellar cortical activities in the rat.</a> J Neurophysiol. 109 (5), 1435-1443 (2013).</li><li>Sykes, M., 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=Differences+in+Motor+Evoked+Potentials+Induced+in+Rats+by+Transcranial+Magnetic+Stimulation+under+Two+Separate+Anesthetics:+Implications+for+Plasticity+Studies.">Differences in Motor Evoked Potentials Induced in Rats by Transcranial Magnetic Stimulation under Two Separate Anesthetics: Implications for Plasticity Studies.</a> Front Neural Circ. 10, 80 (2016).</li><li>Zhang, D. X., Levy, W. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ketamine+blocks+the+induction+of+LTP+at+the+lateral+entorhinal+cortex-dentate+gyrus+synapses.">Ketamine blocks the induction of LTP at the lateral entorhinal cortex-dentate gyrus synapses.</a> Brain Res. 593 (1), 124-127 (1992).</li></ol>

作者没有什么可透露的。

这项协议描述了典型的材料和程序步骤的外科实现永久附加费的设置, 以及随后的刺激警报啮齿动物。在准备一项啮齿动物试验的过程中, 几个方法学方面 (安全和耐受性工商业污水附加费, 结果参数) 以及概念方面 (与人类状况的可比性, 对特定大脑刺激的预期效果区域) 需要加以考虑。从方法论的角度来看, 颅内的手术设置与植入的胸部计数器电极是有利的纵向研究, 因为它允许应用在警报, 自由移...

经颅的电流对大脑的管理已被用于研究大脑功能和修改行为。最近, 应用直接电流, 或较少频繁交流电流 (可和 tRNS), 通过使用两个或更多的电极 (阳极 (s) 和阴极 (s), 非通过完整的头骨, 获得了科学和临床的兴趣。特别是, tDCS 已被用于33200多个疗程的健康主题和精神病患者和神经精神疾病, 并已成为一个安全和方便, cost-effective 床边应用, 有可能的治疗潜力以及长期行为效果1。这清楚地产生了对机械研究, 包括安全方面的需求和科学兴趣的增加。本文着重介绍最常用的刺激形式, tDCS。
跨物种, tDCS 调节皮质兴奋性和突触可塑性。兴奋性的变化已报告为极性依赖性改变自发性神经元的射击率在大鼠和猫2,3,4, 或作为电机诱发电位的变化, 在人类和小鼠 (两个在阳极以后增加了并且减少了阴极 tDCS: 人5,6;鼠标7)。阳极 DCS 增加了运动皮层或海马突触的突触效果在体外刺激或长期增强后的几个小时, 当共同与一个特定的弱突触输入或当给定的可塑性诱导刺激8,9,10,11,12。根据训练, 刺激对运动或认知训练成功的好处经常被显露, 只有当 tDCS 是共同与培训8,13,14,15。虽然这些先前的发现主要归因于神经元的功能, 但应该指出的是, 神经细胞细胞 (胶质) 也可能有助于 tDCS 的功能作用。例如, 在警报小鼠的阳极 tDCS 中, 星细胞内钙水平增加了16。同样, 阳极 tDCS 的电流密度低于阈值神经诱发剂量依赖性激活小胶质细胞17。然而, tDCS 神经元-胶质细胞相互作用的调节需要进一步的具体研究。
一起, 动物研究清楚地推进我们的理解 tDCS 的调节作用对兴奋性和可塑性。然而, 在人类 tDCS 研究发表的指数增长中, 有一个 "反向平移间隙" 可观察到, 与在体外和体内动物模型。此外, 在研究实验室 (从透皮到 epicranial 刺激) 的高变异性中执行了啮齿动物污水附加费模型, 报告的刺激程序往往不完全透明, 阻碍了可比性和可基础研究数据以及结果解释。
在这里, 我们详细描述了一项经颅脑部刺激设置的外科实施, 针对初级运动皮层, 它允许翻译的人 tDCS 条件, 同时尽量减少变异性, 并允许重复的刺激没有阻碍行为。给出了警报大鼠后续附加费的 step-by 步骤协议。讨论了安全应用于警戒啮齿动物的方法和概念方面。

<table><tbody><tr><td>Softasept N</td><td>B. Braun Melsungen AG,&lt;br/&gt; Melsungen, Deutschland</td><td>3887138</td><td>antiseptic agent</td></tr><tr><td>Ethanol 70 %</td><td>Carl Roth GmbH &amp;amp; Co. KG, Karlsruhe, Deutschland</td><td>T913.1</td><td></td></tr><tr><td>arched tip forceps</td><td>FST Fine science tools, Heidelberg, Deutschland</td><td>11071-10</td><td></td></tr><tr><td>Iris Forceps, 10cm, Straight, Serrated</td><td>World Precision Instruments, Inc, Sarasota, FL, USA, Inc, Sarasota, FL, USA</td><td>15914</td><td></td></tr><tr><td>Scalpel Handle #3, 13cm</td><td>World Precision Instruments, Inc, Sarasota, FL, USA, Inc, Sarasota, FL, USA</td><td>500236</td><td></td></tr><tr><td>Standard Scalpel Blade #10</td><td>World Precision Instruments, Inc, Sarasota, FL, USA, Inc, Sarasota, FL, USA</td><td>500239</td><td></td></tr><tr><td>Zelletten cellulose swabs</td><td>Lohmann und Rauscher, Neuwied, Deutschland</td><td>13349</td><td>5 x 4 cm&amp;nbsp;</td></tr><tr><td>Isoflurane</td><td>AbbVie Deutschland GmbH &amp;amp; Co</td><td>N01AB06</td><td></td></tr><tr><td>Iris Scissors, 11.5cm, Straight</td><td>World Precision Instruments, Inc, Sarasota, FL, USA, Inc, Sarasota, FL, USA</td><td>501758</td><td>small scissors</td></tr><tr><td>cotton swab/cotton buds</td><td>Carl Roth GmbH &amp;amp; Co. KG, Karlsruhe, Deutschland</td><td>EH12.1</td><td>Rotilabo</td></tr><tr><td>Kelly Hemostatic Forceps, 14cm, Straight</td><td>World Precision Instruments, Inc, Sarasota, FL, USA, Inc, Sarasota, FL, USA</td><td>501241</td><td>surgical clamp</td></tr><tr><td>electrode plate (platinum)</td><td>custom made</td><td>Wissenschaftliche Werkstatt Neurozentrum Uniklinik Freiburg, Deutschland</td><td>10x6 mm, 0.15 mm thickness</td></tr><tr><td>insulated copper strands (~1 mm diameter)</td><td>Reichelt elektronik GmbH &amp;amp; Co. KG, Sande, Germany</td><td>LITZE BL</td><td>electrode cable</td></tr><tr><td>Weller EC 2002 M soldering station</td><td>Weller Tools GmbH, Besigheim, Germany</td><td>EC2002M1D</td><td></td></tr><tr><td>Iso-Core EL 0,5 mm</td><td>FELDER GMBH L&amp;ouml;ttechnik, Oberhausen, Deutschland</td><td>20970510</td><td>lead free solder</td></tr><tr><td>MERSILENE Polyester Fiber Suture</td><td>Johnson &amp;amp; Johnson Medical GmbH, Ethicon Deutschland, Norderstedt, Germany</td><td>R871H</td><td>nonabsorbable braided suture, 4-0</td></tr><tr><td>Histoacryl</td><td>B. Braun Melsungen AG,&lt;br/&gt; Melsungen, Deutschland</td><td>9381104</td><td>cyanoacrylate</td></tr><tr><td>Ketamin 10%</td><td>Medistar GmbH, Germany</td><td>n/a</td><td>anesthetics</td></tr><tr><td>Rompun 2% (Xylazine)</td><td>Bayer GmbH, Germany</td><td>n/a</td><td>anesthetics</td></tr></tbody></table>

transcranial electrical brain stimulation in alert rodents

经颅电脑刺激可以调节人和啮齿动物的皮质兴奋性和可塑性。人类最常见的刺激形式是经颅直流电刺激 (tDCS)。不常使用, 经颅交变电流刺激 (可) 或经颅随机噪声刺激 (tRNS), 一种特定形式的可使用电流在预定的频率范围内随机应用。在人类的非侵入性电脑刺激研究的增加, 无论是为实验和临床目的, 已经产生了对动物的基本的, 机械的, 安全研究的增加的需要。本文介绍了一个模型, 经颅电脑刺激 (附加费), 通过完整的头骨目标的马达系统警报啮齿动物。该协议提供了 step-by 的步骤说明的外科设置一个永久性的 epicranial 电极插座结合一个植入的柜台电极在胸部。通过放置一个刺激电极到 epicranial 插座, 不同的电刺激类型, 可比较 tDCS, 可, 和 tRNS 在人, 可以交付。并介绍了警示啮齿动物的实用步骤。根据实验需要, 可选择应用电流密度、刺激持续时间和刺激类型。讨论了这种设置的注意事项、优点和缺点, 以及安全性和耐受性方面。

在警报啮齿类动物的可靠重复附加费的设置的被描述的实施可以容易地集成入机械实验、剂量反应研究或者实验包括行为任务。到目前为止, 从动物研究中使用 (无创性) 污水附加费的数据可比, 由于实验室之间的工商业污水附加费刺激的变化和刺激参数的差异 (例如,各种电流密度的应用在与人类的应用相比过高的高等级)。因此, 动物研究在工商业污水附加费方面的?...

Watch this Scientific Journal Video about Transcranial Electrical Brain Stimulation in Alert Rodents at JoVE.com

Transcranial Electrical Brain Stimulation in Alert Rodents

警惕啮齿动物的经颅电脑刺激

本协议描述了一个永久性的 epicranial 电极插座和植入的啮齿动物的胸电极的外科设置。通过把第二个电极插入插座, 不同类型的经颅电脑刺激可以通过完整的头骨传递到警报动物的马达系统。

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

transcranial-electrical-brain-stimulation-in-alert-rodents

Research

JoVE Journal

Neuroscience

10.6K 次观看.  Albert-Ludwigs-University Freiburg. 本协议描述了一个永久性的 epicranial 电极插座和植入的啮齿动物的胸电极的外科设置。通过把第二个电极插入插座, 不同类型的经颅电脑刺激可以通过完整的头骨传递到警报动物的马达系统。

视频: Transcranial Electrical Brain Stimulation in Alert Rodents

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.

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, ...

科研

行为学

A Wireless, Bidirectional Interface for In Vivo Recording and Stimulation of Neural Activity in Freely Behaving Rats

In vivo electrophysiology is a powerful technique to investigate the relationship between brain activity and behavior at a millisecond and micrometer scale. However, current methods mostly rely on tethered cable recordings or only use unidirectional systems, allowing either recording or stimulation of neural activity, but not at the same time or same target. Here, a new wireless, bidirectional device for simultaneous multichannel recording and stimulation of neural activity in freely behaving rats is described. The system operates through a single portable head stage that both transmits recorded activity and can be targeted in real-time for brain stimulation using a telemetry-based multichannel software. The head stage is equipped with a preamplifier and a rechargeable battery, allowing stable long-term recordings or stimulation for up to 1 h. Importantly, the head stage is compact, weighs 12 g (including battery) and thus has minimal impact on the animal&#180;s behavioral repertoire, making the method applicable to a broad set of behavioral tasks. Moreover, the method has the major advantage that the effect of brain stimulation on neural activity and behavior can be measured simultaneously, providing a tool to assess the causal relationships between specific brain activation patterns and behavior. This feature makes the method particularly valuable for the field of deep brain stimulation, allowing precise assessment, monitoring, and adjustment of stimulation parameters during long-term behavioral experiments. The applicability of the system has been validated using the inferior colliculus as a model structure.

A Wireless, Bidirectional Interface for In Vivo Recording and Stimulation of Neural Activity in Freely Behaving Rats

A wireless, bidirectional system for multi-channel neural recordings and stimulation in freely behaving rats is introduced. The system is light and compact, thus having minimal impact on the animal&#180;s behavioral repertoire. Moreover, this bidirectional system provides a sophisticated tool to assess causal relationships between brain activation patterns and behavior.

用于体内的无线双向接口记录和刺激自由行为大鼠的神经活动

In vivo electrophysiology is a powerful technique to investigate the relationship between brain activity and behavior at a millisecond and micrometer ...

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

Microelectrode Guided Implantation of Electrodes into the Subthalamic Nucleus of Rats for Long-term Deep Brain Stimulation

Deep brain stimulation (DBS) is a widely used and effective therapy for several neurologic disorders, such as idiopathic Parkinson&#8217;s disease, dystonia or tremor. DBS is based on the delivery of electrical stimuli to specific deep anatomic structures of the central nervous system. However, the mechanisms underlying the effect of DBS remain enigmatic. This has led to an interest in investigating the impact of DBS in animal models, especially in rats. As DBS is a long-term therapy, research should be focused on molecular-genetic changes of neural circuits that occur several weeks after DBS. Long-term DBS in rats is challenging because the rats move around in their cage, which causes problems in keeping in place the wire leading from the head of the animal to the stimulator. Furthermore, target structures for stimulation in the rat brain are small and therefore electrodes cannot easily be placed at the required position. Thus, a set-up for long-lasting stimulation of rats using platinum/iridium electrodes with an impedance of about 1 M&#937; was developed for this study. An electrode with these specifications allows for not only adequate stimulation but also recording of deep brain structures to identify the target area for DBS. In our set-up, an electrode with a plug for the wire was embedded in dental cement with four anchoring screws secured onto the skull. The wire from the plug to the stimulator was protected by a stainless-steel spring. A swivel was connected to the circuit to prevent the wire from becoming tangled. Overall, this stimulation set-up offers a high degree of free mobility for the rat and enables the head plug, as well as the wire connection between the plug and the stimulator, to retain long-lasting strength.

A method for implanting electrodes into the subthalamic nucleus (STN) of rats is described. Better localization of the STN was achieved by using a microrecording system. Furthermore, a stimulation set-up is presented that is characterized by long-lasting connections between the head of the animal and the stimulator.

微电极引导电极植入到大鼠丘脑底核的长期深部脑刺激

Deep brain stimulation (DBS) is a widely used and effective therapy for several neurologic disorders, such as idiopathic Parkinson&#8217;s disease, ...

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 ...

Transcranial Direct Current Stimulation (tDCS) in Mice

Transcranial direct current stimulation (tDCS) is a non-invasive neuromodulation technique proposed as an alternative or complementary treatment for several neuropsychiatric diseases. The biological effects of tDCS are not fully understood, which is in part explained due to the difficulty in obtaining human brain tissue. This protocol describes a tDCS mouse model that uses a chronically implanted electrode allowing the study of the long-lasting biological effects of tDCS. In this experimental model, tDCS changes the cortical gene expression and offers a prominent contribution to the understanding of the rationale for its therapeutic use.

Transcranial Direct Current Stimulation tDCS in Mice

Transcranial direct current stimulation (tDCS) is a therapeutic technique proposed to treat psychiatric diseases. An animal model is essential for understanding the specific biological alterations evoked by tDCS. This protocol describes a tDCS mouse model that uses a chronically implanted electrode.

经颅直流电刺激 (tDCS) 在小鼠中的作用

Transcranial direct current stimulation (tDCS) is a non-invasive neuromodulation technique proposed as an alternative or complementary treatment for ...

Establishment of Acute Pontine Infarction in Rats by Electrical Stimulation

Pontine infarction is the most common stroke subtype in the posterior circulation, while there lacks a rodent model mimicking pontine infarction. Provided here is a protocol for successfully establishing a rat model of acute pontine infarction. Rats weighing about 250 g are used, and a probe with an insulated sheath is injected into the pons using a stereotaxic apparatus. A lesion is produced by the electrical stimulation with a single pulse. The Longa score, Berderson score, and beam balance test are used to assess neurological deficits. Additionally, the adhesive-removal somatosensory test is used to determine sensorimotor function, and the limb placement test is used to evaluate proprioception. MRI scans are then used to assess the infarction in vivo, and TTC staining is used to confirm the infarction in vitro. Here, a successful infarction is identified that is located in the anterolateral basis of the rostral pons. In conclusion, a new method is described to establish an acute pontine infarction rat model.

Presented here is a protocol for establishing acute pontine infarction in a rat model via electrical stimulation with a single pulse.

电刺激在大鼠中建立急性庞廷梗死

Pontine infarction is the most common stroke subtype in the posterior circulation, while there lacks a rodent model mimicking pontine infarction. Provided ...

Head Implants for the Neuroimaging of Awake, Head-Fixed Rats

Anesthetics, commonly used in preclinical and fundamental scientific research, have a depressive influence on the metabolic, neuronal, and vascular functions of the brain and can adversely influence neurophysiological results. The use of awake animals for research studies is advantageous but poses the major challenge of keeping the animals calm and stationary to minimize motion artifacts throughout data acquisition. Awake imaging in smaller-sized rodents (e.g., mice) is very common but remains scant in rats as rats are bigger, stronger, and have a greater tendency to oppose movement restraints and head fixation over the long durations required for imaging. A new model of neuroimaging of awake, head-fixed rats using customized hand-sewn slings, 3D-printed head implants, head caps, and a headframe is described. The results acquired following a single trial of single-whisker stimulation suggest an increase in the intensity of the evoked functional response. The acquisition of the evoked functional response from awake, head-fixed rats is faster than that from anesthetized rats, reliable, reproducible, and can be used for repeated longitudinal studies.

A detailed new procedure for functional imaging of awake, head-fixed rats is described.

用于清醒、头部固定大鼠神经成像的头部植入物

Anesthetics, commonly used in preclinical and fundamental scientific research, have a depressive influence on the metabolic, neuronal, and vascular ...

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, ...

绘制脑血流和电场图:小鼠的电极放置

Placement of Extracranial Stimulating Electrodes and Measurement of Cerebral Blood Flow and Intracranial Electrical Fields in Anesthetized Mice

The detection of cerebral blood flow (CBF) responses to various forms of neuronal activation is critical for understanding dynamic brain function and variations in the substrate supply to the brain. This paper describes a protocol for measuring CBF responses to transcranial alternating current stimulation (tACS). Dose-response curves are estimated both from the CBF change occurring with tACS (mA) and from the intracranial electric field (mV/mm). We estimate the intracranial electrical field based on the different amplitudes measured by glass microelectrodes within each side of the brain. In this paper, we describe the experimental setup, which involves using either bilateral laser Doppler (LD) probes or laser speckle imaging (LSI) to measure the CBF; as a result, this setup requires anesthesia for the electrode placement and stability. We present a correlation between the CBF response and the current as a function of age, showing a significantly larger response at higher currents (1.5 mA and 2.0 mA) in young control animals (12-14 weeks) compared to older animals (28-32 weeks) (p &lt; 0.005 difference). We also demonstrate a significant CBF response at electrical field strengths &lt;5 mV/mm, which is an important consideration for eventual human studies. These CBF responses are also strongly influenced by the use of anesthesia compared to awake animals, the respiration control (i.e., intubated vs. spontaneous breathing), systemic factors (i.e., CO2), and local conduction within the blood vessels, which is mediated by pericytes and endothelial cells. Likewise, more detailed imaging/recording techniques may limit the field size from the entire brain to only a small region. We describe the use of extracranial electrodes for applying tACS stimulation, including both homemade and commercial electrode designs for rodents, the concurrent measurement of the CBF and intracranial electrical field using bilateral glass DC recording electrodes, and the imaging approaches. We are currently applying these techniques to implement a closed-loop format for augmenting the CBF in animal models of Alzheimer's disease and stroke.

作者聚焦:基于刺激的方法改善阿尔茨海默病模型中的脑血流 

We describe a protocol for assessing dose-response curves for extracranial stimulation in terms of brain electrical field measurements and a relevant biomarker-cerebral blood flow. Since this protocol involves invasive electrode placement into the brain, general anesthesia is needed, with spontaneous breathing preferred rather than controlled respirations.

麻醉小鼠颅外刺激电极的放置和脑血流量和颅内电场的测量

The detection of cerebral blood flow (CBF) responses to various forms of neuronal activation is critical for understanding dynamic brain function and ...