在自由运动大鼠的反应中, 皮质局部电位和电皮质图谱的同时记录
Summary
我们开发了一种技术, 同时记录电皮质成像和局部场电位, 以响应来自自由移动的大鼠的诱发电位激光刺激。这项技术有助于在介观和宏观层面建立电皮质信号的直接关系, 这有利于大脑中的噪声信息处理的研究。
Abstract
选择性激活无噪声神经末梢的激光热脉冲引起的电皮质反应, 在许多动物和人类研究中被广泛用于研究诱发电位信息的皮质处理。这些激光唤起的脑电位 (lep) 由几个瞬态反应组成, 这些响应是随着激光刺激的发生而被时间锁定的。然而, lep 响应的功能特性在很大程度上仍然是未知的, 因为缺乏一种采样技术, 可以同时记录皮层表面的神经活动 (即电皮质图 [ecog] 和头皮脑电图 [头皮脑电图]) 和大脑内部 (即, 局部场电位 [lfp])。为了解决这个问题, 我们在这里提出了一个使用自由移动的老鼠的动物协议。该协议由三个主要程序组成: (1) 动物准备和手术程序; (2) 同时记录 ecog 和 lfp 对诱发电位激光刺激的反应; (3) 数据分析和特征提取。具体而言, 在3d 打印保护壳的帮助下, 将植入大鼠头骨上的 ecog 和 lfp 电极牢固地固定在一起。在数据收集过程中, 当动物处于自发静止时, 在老鼠的前爪上通过腔底的缝隙传递激光脉冲。持续的白噪声被播放, 以避免激活听觉系统的激光产生的超声波。因此, 只有有选择地记录的是噪声反应。使用标准的分析程序 (例如带通滤波、划时代提取和基线校正) 提取刺激相关的大脑反应, 我们得到的结果表明, 具有高信噪比的 lep 是同时记录从 ecog 和 lfp 电极。这种方法使同时记录 ecog 和 lfp 活动成为可能, 从而在介观和宏观层面提供了一座电皮质信号的桥梁, 从而促进了对噪声信息处理的调查。在大脑中。
Introduction
脑电图是一种记录大脑中数千个神经元同步活动所产生的电势和振荡大脑活动的技术。它被广泛用于许多基础研究和临床应用1,2。例如, 脑电图对强烈激光热脉冲 (即lep) 的反应被广泛用于研究诱发感觉输入3,4, 5的外围和中央处理。在人类中, lep 主要由三个不同的偏转组成: 早期成分 (n1) 是体细胞组织的, 可能反映初级体感感皮层 (s1)6的活动, 以及中心的晚期成分 (n2 和 p2)分布, 更有可能反映二次体感皮层, 脑岛, 和前扣带回皮层的活动 7,8。在以前的研究 9,10, 我们证明, 使用 ecog (一种颅内脑电图) 从直接放置在大脑暴露表面的电极中取样的大鼠 lep 也由三个不同的偏转 (即, 由睡眠动力学组织的 n1 和集中分布的 n2 和 p2)。大鼠 lep 成分的极性、顺序和地形类似于人类 lep11.然而, 由于头皮脑电图和硬膜下脑电图记录的空间分辨率有限, 13 的脑电图源分析技术不准确, 神经活动对 lep 成分的详细贡献是很多争论。例如, 目前尚不清楚 s1 是否以及在多大程度上有助于激光刺激引起的皮质反应 (n1) 的早期部分.
与宏观层面的记录技术不同, 使用微线阵列在立体定向装置和微驱动器14、15 的辅助下进行直接颅内记录, 可以测量神经活动 (例如, lfp)) 的特定区域。lfp 主要反映局部神经元群16的抑制或兴奋突触后电位的总和.由于 lfp 采样神经活动反映了记录电极周围数百微米内发生的神经元过程, 这种记录技术被广泛用于研究大脑中的介观水平的信息处理。然而, 它只关注大脑活动的精确局部变化, 无法回答来自多个区域的信号如何整合的问题 (例如, lep 组件如何在多个大脑区域集成)。
值得注意的是, 同时记录自由移动大鼠的 ecog 和皮质 lfp, 可以促进宏观和中观层面的皮质信息处理调查。此外, 这种方法提供了一个极好的机会, 以调查预定义的大脑区域的神经活动在多大程度上对 lep 的贡献。事实上, 以前的几项研究已经评估了尖峰、皮质 lfp 和 ecog 信号17、18之间的一致性, 并证明了与 eeg 电极相邻的 lfp19、20对与刺激相关的大脑反应的形成。然而, 现有的技术通常用于记录麻醉动物的大脑反应, 因为缺乏保护壳, 以防止电极被碰撞损坏。换句话说, 在自由运动的大鼠中, 仍然缺乏在介观 (皮质 lfp) 和宏观 (eeg 和 ecog) 水平上建立电皮质信号桥梁的技术。
为了解决这个问题, 我们开发了一种技术, 可以同时记录自由移动的老鼠大脑多个区域的 ecog 和皮质 lfp。这项技术有助于在介观和宏观层面建立电皮质信号的直接关系, 从而促进大脑中噪声信息处理的研究。
Protocol
Representative Results
Discussion
在本研究中, 我们描述了一种技术, 同时记录 ecog 和皮质 lfp 反应引起的噪声激光刺激从自由移动的大鼠。结果表明, 在 ecog 和 lfp 信号中, 激光刺激发生后, lep 响应可以清晰检测到。同时记录 ecog 和皮质 lfp 信号将使科学家能够研究它们之间的关系, 以便更好地了解神经元活动对 lep 组件的贡献。
应注意拟议技术中的五个关键步骤。首先, 重要的是要确保颅骨的表面是干净和干燥…
Disclosures
The authors have nothing to disclose.
Acknowledgements
这项工作得到了中国科学院 “十三五” 信息化计划 (XXH13506) 中国科学院心理健康重点实验室、心理学研究所、国家自然科学基金 (31611141 和 31671141) 的支持,和中国科学院心理学研究所科学基金会项目 (y6cx021008)。
Materials
| Male Sprague-Drawley rats | Vital River | ||
| Isoflurane | RWD Life Science | ||
| Small animal isoflurane anaesthetic system | RWD Life Science | Including the anesthesia gas mask for rats | |
| Stereotaxic apparatus | RWD Life Science | ||
| The apparatus with combined ECoG and LFP electrodes | The apparatus is home-made, which assembles the ECoG and depth wire electrodes to a connector module | ||
| 3D-printed protective shell | The texture of shell is polylactic, and the shell is home-made and contains three parts: a base, a wall and a cap. The wall is covered by copper tapers to construct as a Faraday cage | ||
| Tungsten wires (diameter: 50 mm) | California Fine Wires Company | The electrodes for cortical LFP recording | |
| Stainless steel screws (diameter: 0.6 mm) |
The electrodes for ECoG recording | ||
| Electric cranial drill | RWD Life Science | ||
| Drill bit (diameter: 0.5 mm) | RWD Life Science | The drill is used for drilling the holes of ECoG screws | |
| Drill bit (diameter: 0.2 mm) | RWD Life Science | The drill is used for drilling the holes of depth wires | |
| Dental arylic powder | SNC dental | ||
| Dental arylic liquid | SNC dental | ||
| Paraffin | Fisher Scientific | The mixture is used for seal the craniotomy to ensure the following movement of micro-wire arrays | |
| Mineral Oil | Fisher Scientific | ||
| Electrocoagulator | Bovie medical Corporation | ||
| RHD2132 Amplifier Boards | Intan Technologies | A 32-channel headstage | |
| RHD2000 systerm | Intan Technologies | The data acquisition systerm | |
| Infrared neodymium yttrium aluminum perovskite (Nd:YAP) laser generator | Electronical Engineering | ||
| Matlab R2016b | The MathWorks |
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