这项工作得到了村田科学基金会、铃轩纪念基金会、中谷生物医学工程测量技术促进基金会以及探索性研究（批准号21K19755，日本）和科学研究（B）（批准号23H03416，日本）的资助。

用低成本的 EEG 和毫米级线圈刺激小鼠大脑

<ol>
	<li>Ilmoniemi, R. J., 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=Neuronal+responses+to+magnetic+stimulation+reveal+cortical+reactivity+and+connectivity.">Neuronal responses to magnetic stimulation reveal cortical reactivity and connectivity.</a> Neuroreport. 8 (16), 3537-3540 (1997).</li><li>Hallett, M. <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:+a+primer.">Transcranial magnetic stimulation: a primer.</a> Neuron. 55 (2), 187-199 (2007).</li><li>Thut, G., Pascual-Leone, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Integrating+TMS+with+EEG:+How+and+what+for.">Integrating TMS with EEG: How and what for.</a> Brain Topography. 22 (4), 215-218 (2010).</li><li>Ilmoniemi, R. J., Kicic, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Methodology+for+combined+TMS+and+EEG.">Methodology for combined TMS and EEG.</a> Brain Topograpy. 22 (4), 233-248 (2010).</li><li>Daskalakis, Z. J., Farzan, F., Radhu, N., Fitzgerald, P. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Combined+transcranial+magnetic+stimulation+and+electroencephalography:+its+past,+present+and+future.">Combined transcranial magnetic stimulation and electroencephalography: its past, present and future.</a> Brain Research. 1463, 93-107 (2012).</li><li>Tremblay, S., 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=Clinical+utility+and+prospective+of+TMS-EEG.">Clinical utility and prospective of TMS-EEG.</a> Clinical Neurophysiology. 130 (5), 802-844 (2019).</li><li>Pellicciari, M. C., Veniero, D., Miniussi, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterizing+the+cortical+oscillatory+response+to+TMS+pulse.">Characterizing the cortical oscillatory response to TMS pulse.</a> Frontiers in Cellular Neuroscience. 11, 38 (2017).</li><li>Lin, Y. J., Shukla, L., Dugue, L., Valero-Cabre, A., Carrasco, M. <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+entrains+alpha+oscillatory+activity+in+occipital+cortex.">Transcranial magnetic stimulation entrains alpha oscillatory activity in occipital cortex.</a> Scientific Reports. 11 (1), 18562 (2021).</li><li>Takahashi, S., 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=Laminar+responses+in+the+auditory+cortex+using+a+multielectrode+array+substrate+for+simultaneous+stimulation+and+recording.">Laminar responses in the auditory cortex using a multielectrode array substrate for simultaneous stimulation and recording.</a> IEEJ Transactions Electrical and Electronic Engineering. 14 (2), 303-311 (2019).</li><li>Yoshikawa, T., Higuchi, H., Furukawa, R., Tateno, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Temporal+and+spatial+profiles+of+evoked+activity+induced+by+magnetic+stimulation+using+millimeter-sized+coils+in+the+mouse+auditory+cortex+in+vivo.">Temporal and spatial profiles of evoked activity induced by magnetic stimulation using millimeter-sized coils in the mouse auditory cortex in vivo.</a> Brain Research. 1796, 148092 (2022).</li><li>Tang, A. 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=Construction+and+evaluation+of+rodent-specific+rTMS+coils.">Construction and evaluation of rodent-specific rTMS coils.</a> Frontiers in Neural Circuits. 10, 47 (2016).</li><li>Li, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Controlling+annealing+and+magnetic+treatment+parameters+to+achieve+high+permeabilities+in+55+Ni-Fe+toroid+cores.">Controlling annealing and magnetic treatment parameters to achieve high permeabilities in 55 Ni-Fe toroid cores.</a> IEEE Transactions on Magnetics. 37 (4), 2315-2317 (2001).</li></ol>

作者没有什么可透露的。

这项研究涉及多点脑电图记录系统与为小动物（包括小鼠）设计的磁刺激系统相结合。构建的系统成本低，易于在生理实验室中构建，并且可以扩展其现有的测量设置。如果这些实验室以前有标准电生理实验的经验，那么从小鼠记录系统获取数据所需的外科手术非常简单。
使用这种方法的一个优点是电极放置在单个动物的头部和头皮上的良好可重复性。用于将螺旋电极分配?...

经颅磁刺激（TMS）因其非/低侵入性而成为人脑科学，临床应用和动物模型研究的有前途的工具。在TMS应用的早期阶段，人类和动物对单脉冲和配对脉冲TMS的皮质效应的测量仅限于运动皮层;易于测量的输出仅限于涉及运动皮层的运动诱发电位和感应肌电位1,2。为了扩展可以通过TMS调制测量的大脑区域，将脑电图（EEG）记录与单脉冲和配对脉冲TMS集成在一起，作为直接检查整个大脑区域的兴奋性，连通性和时空动力学的有用方法3,4,5。因此，同时将TMS和脑电图记录（TMS-EEG）应用于大脑已被用于探测人类和动物的各种浅表皮质大脑区域，以研究皮层内神经回路（见Tremblay等人6）。此外，TMS-EEG系统可用于检查其他皮质时空特征，包括信号传播到其他皮质区域和振荡活动的产生7,8。
然而，由于TMS的非侵入性，TMS在大脑中的作用机制仍然是推测性的，这限制了我们对TMS应用期间大脑功能的知识。因此，从啮齿动物到人类的动物的侵入性转化研究对于了解TMS对神经回路及其活性的影响机制至关重要。特别是，对于动物的TMS-EEG组合实验，尚未为小动物密集开发同步刺激和测量系统。因此，实验者需要根据其特定的实验要求，通过试错来构建这样的系统。此外，小鼠模型在其他 体内 动物物种模型中是有用的，因为许多转基因和品系分离的小鼠品系可作为生物资源使用。因此，对于许多神经科学研究人员来说，为小鼠构建TMS-EEG组合测量系统的便捷方法是可取的。
本研究提出了一种TMS-EEG组合方法，可用于同时刺激和记录小鼠大脑，小鼠大脑是研究中使用的主要转基因动物类型，可以在典型的神经科学实验室中轻松构建。首先，描述了使用常规螺旋电极和柔性基板的低成本脑电图记录系统，以在每个实验中可重复地分配电极阵列位置。其次，磁刺激系统是使用毫米尺寸的线圈构建的，可以在典型的实验室中轻松定制。第三，TMS-EEG组合系统记录对声音和磁刺激的神经活动。本研究中提出的方法可以揭示在小动物中产生特定疾病的机制，并且可以翻译在动物模型中获得的结果以了解相应的人类疾病。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>3D printer</td>
			<td>Zhejiang Flashforge 3D Technology Co., Ltd</td>
			<td>FFD-101</td>
			<td>The printer used for 3D-printing the donut-shaped disks</td>
		</tr>
		<tr>
			<td>ATROPINE SULFATE&#160; 0.5 mg</td>
			<td>NIPRO ES PHARMA CO., LTD.</td>
			<td>-</td>
			<td>Atropine sulfate</td>
		</tr>
		<tr>
			<td>Bipolar amplifier</td>
			<td>NF Corp.</td>
			<td>KIT61380</td>
			<td>For amplifying waveforms for coil input</td>
		</tr>
		<tr>
			<td>Butorphanol</td>
			<td>Meiji Seika Pharma 
			Co., Ltd., Tokyo, Japan</td>
			<td>-</td>
			<td>For anathesis of animals</td>
		</tr>
		<tr>
			<td>Commercial manufacturer of flexible 2D array</td>
			<td>p-ban.com Corp.</td>
			<td>-</td>
			<td>URL: https://www.p-ban.com/</td>
		</tr>
		<tr>
			<td>Computer prograom to analyze output signals</td>
			<td>Natinal Instruments</td>
			<td>NI-DAQ and&#160; NI-DAQmx Python</td>
			<td>To analyze output signals from the hall-effect sensor</td>
		</tr>
		<tr>
			<td>Connector</td>
			<td>Harwin Inc.</td>
			<td>G125-FV12005L0P</td>
			<td>For connector to conect to the measuring system</td>
		</tr>
		<tr>
			<td>Copper pad</td>
			<td>p-ban.com Corp.</td>
			<td>copper</td>
			<td>Copper pad on each substrate</td>
		</tr>
		<tr>
			<td>Copper wire</td>
			<td>Kyowa Harmonet Ltd.</td>
			<td>P644432</td>
			<td>The windings of the coil</td>
		</tr>
		<tr>
			<td>DAQ board</td>
			<td>National Instruments Corp.</td>
			<td>USB-6343</td>
			<td>For measuring the magnitic flux density of the coil</td>
		</tr>
		<tr>
			<td>Dental cement</td>
			<td>SHOFU INC.</td>
			<td>Quick Resin</td>
			<td>Self-Curing Orthodontic Resin</td>
		</tr>
		<tr>
			<td>ECoG electrode</td>
			<td>NeuroNexus Inc.</td>
			<td>HC32</td>
			<td>For reference to design of the flexible 2D array</td>
		</tr>
		<tr>
			<td>Epoxy resin</td>
			<td>Konishi Co. Ltd.</td>
			<td>#16123</td>
			<td>For coil construction</td>
		</tr>
		<tr>
			<td>Ethyl Carbamate</td>
			<td>FUJIFILM Wako Pure Chemical Corp.</td>
			<td>050-05821</td>
			<td>For urethan anesthesia</td>
		</tr>
		<tr>
			<td>Flat ribbon cable</td>
			<td>Oki Electric Cable Co., Ltd.</td>
			<td>FLEX-B2(20)-7/0.1 20028 5m</td>
			<td>For cable to connect between surface-mount connector and measuring sysytem</td>
		</tr>
		<tr>
			<td>flexible substrate</td>
			<td>p-ban.com Corp.</td>
			<td>polyimide</td>
			<td>Baseplate of flexible substrate</td>
		</tr>
		<tr>
			<td>Function generator</td>
			<td>NF Corp.</td>
			<td>WF1947</td>
			<td>For generating waveforms for coil input</td>
		</tr>
		<tr>
			<td>Hall-effect sensor</td>
			<td>Honeywell International Inc.</td>
			<td>SS94A2D</td>
			<td>For measuring the magnitic flux density of the coil</td>
		</tr>
		<tr>
			<td>IDC crimping tool</td>
			<td>Pro&#39;sKit Industries Co.</td>
			<td>6PK-214</td>
			<td>To crimp the IDC and one end of the flat ribbon cable; Flat cable connector crimping tool</td>
		</tr>
		<tr>
			<td>Instant glue</td>
			<td>Konishi Co. Ltd.</td>
			<td>#04612</td>
			<td>For coil construction</td>
		</tr>
		<tr>
			<td>Insulation-displacement connector (IDC&#160;)</td>
			<td>Uxcell Japan</td>
			<td>B07GDDG3XG</td>
			<td>2 &#215; 10 pins and a 1.27 mm pitch&#160;</td>
		</tr>
		<tr>
			<td>LCR meter</td>
			<td>NF Corp.</td>
			<td>ZM2376</td>
			<td>For measuring the AC properties of the coil</td>
		</tr>
		<tr>
			<td>Manipulator</td>
			<td>NARISHIGE Group.</td>
			<td>SM-15L</td>
			<td>For manipulating the coil</td>
		</tr>
		<tr>
			<td>Medetomidine</td>
			<td>Kobayashi Kako, Fukui, Japan</td>
			<td>-</td>
			<td>For anathesis of animals</td>
		</tr>
		<tr>
			<td>Midazolam</td>
			<td>Astellas Pharma, Tokyo, Japan</td>
			<td>-</td>
			<td>For anathesis of animals</td>
		</tr>
		<tr>
			<td>Miniature screw</td>
			<td>KOFUSEIBYO Co., Ltd.</td>
			<td>S0.6*1.5</td>
			<td>For EEG-senseing and reference electrode</td>
		</tr>
		<tr>
			<td>Mouse</td>
			<td>Japan SLC, Inc.</td>
			<td>C57BL/6J&#160;(C57BL/6JJmsSlc)</td>
			<td>Experimental animal</td>
		</tr>
		<tr>
			<td>Permalloy-45 rod</td>
			<td>The Nilaco Corp.</td>
			<td>780544</td>
			<td>The core of the coil</td>
		</tr>
		<tr>
			<td>Recording system</td>
			<td>Plexon Inc.</td>
			<td>OmniPlex</td>
			<td>For EEG data acquisition</td>
		</tr>
		<tr>
			<td>Stainless wire</td>
			<td>Wakisangyo Co., Ltd.</td>
			<td>HW-136</td>
			<td>For grasp by manipulator</td>
		</tr>
		<tr>
			<td>Stereotaxic apparatus</td>
			<td>NARISHIGE Group.</td>
			<td>SR-5M-HT</td>
			<td>To fix a mouse head</td>
		</tr>
		<tr>
			<td>Surface-mount connector</td>
			<td>Useconn Electronics Ltd.</td>
			<td>PH127-2x10MG</td>
			<td>For connector to mount on the flexible 2D array</td>
		</tr>
		<tr>
			<td>Testing equipment (LCR meter)</td>
			<td>NF Corp.</td>
			<td>ZM2372</td>
			<td>Contact check and impedance measurements</td>
		</tr>
		<tr>
			<td>White PLA filament</td>
			<td>Zhejiang Flashforge 3D Technology Co., Ltd</td>
			<td>PLA-F13</td>
			<td>The material used for 3D-printing the donut-shaped disks</td>
		</tr>
		<tr>
			<td>Xylocaine Jelly 2%</td>
			<td>Sandoz Pharma Co., Ltd.</td>
			<td>-</td>
			<td>lidocaine hydrochloride</td>
		</tr>
	</tbody>
</table>

low-cost electroencephalographic recording system combined with a millimeter-sized coil to transcranially stimulate the mouse brain in vivo

这里提出了一种低成本的脑电图（EEG）记录系统，利用毫米大小的线圈 驱动 体内小鼠大脑的经颅磁刺激（TMS）。使用传统的螺旋电极与定制的、灵活的多电极阵列基板相结合，可以从小鼠大脑进行多位点记录。此外，我们还解释了如何使用实验室中常见的低成本设备生产毫米尺寸的线圈。还介绍了制造柔性多电极阵列基板的实用程序和螺杆电极的手术植入技术，这是产生低噪声脑电信号所必需的。尽管该方法对于从任何小动物的大脑中记录是有用的，但本报告侧重于麻醉小鼠头骨中的电极实施。此外，这种方法可以很容易地扩展到清醒的小动物，它通过公共适配器 与 系留电缆连接，并在录制过程中用TMS设备固定在头部。描述了当前版本的EEG-TMS系统，其最多可以包括32个EEG通道（具有16个通道的设备作为具有较少通道的示例）和一个TMS通道设备。此外，简要报告了通过将EEG-TMS系统应用于麻醉小鼠获得的典型结果。

在麻醉的C57BL / 6J小鼠中记录的样品脑电图数据，柔性基质与螺旋电极相结合，如下所示。
作为一个典型的例子，在具有相同刺激的60项试验中，显示了响应声音刺激（8 kHz音爆，80 dB声压级[SPL]）而产生的平均EEG波形（图4A）。记录通道映射的原理图也显示在图4A的中间。来自CHS 5,7,10和12的反应是从两个颞叶的听觉皮层附近区域记录?...

Watch this Scientific Journal Video about Low-Cost Electroencephalographic Recording System Combined with a Millimeter-Sized Coil to Transcranially Stimulate the Mouse Brain In Vivo at JoVE.com

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  

提出了一种低成本的脑电图记录系统，结合毫米尺寸的线圈来驱动体内 小鼠大脑的经颅磁刺激。使用具有定制、柔性、多电极阵列基板的传统螺旋电极，可以从小鼠大脑响应经颅磁刺激进行多位点记录。

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, ...

在神经科学中，TMS和EEG记录经常被用来促进我们对大脑的理解。TMS由于其非侵入性而特别有前途。将TMS与EEG记录相结合，使研究人员能够探索不同大脑区域的兴奋性，连通性和时空动力学。该协议旨在开发用于小动物TMS-EEG实验的同步刺激测量系统。这些系统尚未得到广泛开发，迫使研究人员根据独特的实验要求创建一个试错系统。这种易于适应的方法可以在标准的神经科学实验室中建立，使其对研究人员来说更容易获得和实用。这项研究的结果将推进研究和阐明小动物特定疾病背后的机制，这是人类候选者的基本模型。此外，这些模型将提高他们对人类状况的理解，并有助于指导创造更有效的治疗和干预措施。

low-cost-electroencephalographic-recording-system-combined-with

Research

JoVE Journal

Neuroscience

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

3.1K 次观看.  Hokkaido University. 在神经科学中，TMS和EEG记录经常被用来促进我们对大脑的理解。TMS由于其非侵入性而特别有前途。将TMS与EEG记录相结合，使研究人员能够探索不同大脑区域的兴奋性，连通性和时空动力学。该协议旨在开发用于小动物TMS-EEG实验的同步刺激测量系统。这些系统尚未得到广泛开发，迫使研究人员根据独特的实验要求创建一个试错系统。这种易于适应的方法可以在标准的神经...

视频: 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  

Functional Neuroimaging Using Ultrasonic Blood-brain Barrier Disruption and Manganese-enhanced MRI

Although mice are the dominant model system for studying the genetic and molecular underpinnings of neuroscience, functional neuroimaging in mice remains technically challenging. One approach, Activation-Induced Manganese-enhanced MRI (AIM MRI), has been used successfully to map neuronal activity in rodents 1-5. In AIM MRI, Mn2+ acts a calcium analog and accumulates in depolarized neurons 6,7. Because Mn2+ shortens the T1 tissue property, regions of elevated neuronal activity will enhance in MRI. Furthermore, Mn2+ clears slowly from the activated regions; therefore, stimulation can be performed outside the magnet prior to imaging, enabling greater experimental flexibility. However, because Mn2+ does not readily cross the blood-brain barrier (BBB), the need to open the BBB has limited the use of AIM MRI, especially in mice.
One tool for opening the BBB is ultrasound. Though potentially damaging, if ultrasound is administered in combination with gas-filled microbubbles (i.e., ultrasound contrast agents), the acoustic pressure required for BBB opening is considerably lower. This combination of ultrasound and microbubbles can be used to reliably open the BBB without causing tissue damage 8-11.
Here, a method is presented for performing AIM MRI by using microbubbles and ultrasound to open the BBB. After an intravenous injection of perflutren microbubbles, an unfocused pulsed ultrasound beam is applied to the shaved mouse head for 3 minutes. For simplicity, we refer to this technique of BBB Opening with Microbubbles and UltraSound as BOMUS 12. Using BOMUS to open the BBB throughout both cerebral hemispheres, manganese is administered to the whole mouse brain. After experimental stimulation of the lightly sedated mice, AIM MRI is used to map the neuronal response.
To demonstrate this approach, herein BOMUS and AIM MRI are used to map unilateral mechanical stimulation of the vibrissae in lightly sedated mice 13. Because BOMUS can open the BBB throughout both hemispheres, the unstimulated side of the brain is used to control for nonspecific background stimulation. The resultant 3D activation map agrees well with published representations of the vibrissae regions of the barrel field cortex 14. The ultrasonic opening of the BBB is fast, noninvasive, and reversible; and thus this approach is suitable for high-throughput and/or longitudinal studies in awake mice.

A technique is described for broadly opening the blood-brain barrier in the mouse using microbubbles and ultrasound. Using this technique, manganese can be administered to the mouse brain. Because manganese is an MRI contrast agent that accumulates in depolarized neurons, this approach enables imaging of neuronal activity.

使用超声波血脑屏障破坏和锰增强MRI的脑功能成像

Although mice are the dominant model system for studying the genetic and molecular underpinnings of neuroscience, functional neuroimaging in mice remains ...

科研

神经科学

A Low Cost Setup for Behavioral Audiometry in Rodents

In auditory animal research it is crucial to have precise information about basic hearing parameters of the animal subjects that are involved in the experiments. Such parameters may be physiological response characteristics of the auditory pathway, e.g. via brainstem audiometry (BERA). But these methods allow only indirect and uncertain extrapolations about the auditory percept that corresponds to these physiological parameters. To assess the perceptual level of hearing, behavioral methods have to be used. A potential problem with the use of behavioral methods for the description of perception in animal models is the fact that most of these methods involve some kind of learning paradigm before the subjects can be behaviorally tested, e.g. animals may have to learn to press a lever in response to a sound. As these learning paradigms change perception itself 1,2 they consequently will influence any result about perception obtained with these methods and therefore have to be interpreted with caution. Exceptions are paradigms that make use of reflex responses, because here no learning paradigms have to be carried out prior to perceptual testing. One such reflex response is the acoustic startle response (ASR) that can highly reproducibly be elicited with unexpected loud sounds in na&iuml;ve animals. This ASR in turn can be influenced by preceding sounds depending on the perceptibility of this preceding stimulus: Sounds well above hearing threshold will completely inhibit the amplitude of the ASR; sounds close to threshold will only slightly inhibit the ASR. This phenomenon is called pre-pulse inhibition (PPI) 3,4, and the amount of PPI on the ASR gradually depends on the perceptibility of the pre-pulse. PPI of the ASR is therefore well suited to determine behavioral audiograms in na&iuml;ve, non-trained animals, to determine hearing impairments or even to detect possible subjective tinnitus percepts in these animals. In this paper we demonstrate the use of this method in a rodent model (cf. also ref. 5), the Mongolian gerbil (Meriones unguiculatus), which is a well know model species for startle response research within the normal human hearing range (e.g. 6).

A fast and inexpensive method for the behavioral determination of hearing parameters like hearing thresholds, hearing impairments or phantom perceptions (subjective tinnitus) is described. It uses pre-pulse inhibition of the acoustic startle response and can be easily implemented in a personal computer using a programmable AD/DA-converter and a piezo sensor.

在啮齿动物的行为测听的低成本设置

In auditory animal research it is crucial to have precise information about basic hearing parameters of the animal subjects that are involved in the ...

In vivo Optogenetic Stimulation of the Rodent Central Nervous System

The ability to probe defined neural circuits in awake, freely-moving animals with cell-type specificity, spatial precision, and high temporal resolution has been a long sought tool for neuroscientists in the systems-level search for the neural circuitry governing complex behavioral states. Optogenetics is a cutting-edge tool that is revolutionizing the field of neuroscience and represents one of the first systematic approaches to enable causal testing regarding the relation between neural signaling events and behavior. By combining optical and genetic approaches, neural signaling can be bi-directionally controlled through expression of light-sensitive ion channels (opsins) in mammalian cells. The current protocol describes delivery of specific wavelengths of light to opsin-expressing cells in deep brain structures of awake, freely-moving rodents for neural circuit modulation. Theoretical principles of light transmission as an experimental consideration are discussed in the context of performing in vivo optogenetic stimulation. The protocol details the design and construction of both simple and complex laser configurations and describes tethering strategies to permit simultaneous stimulation of multiple animals for high-throughput behavioral testing.

In vivo Optogenetic Stimulation of the Rodent Central Nervous System

Optogenetics has become a powerful tool for use in behavioral neuroscience experiments. This protocol offers a step-by-step guide to the design and set-up of laser systems, and provides a full protocol for carrying out multiple and simultaneous in vivo optogenetic stimulations compatible with most rodent behavioral testing paradigms.

在啮齿动物中枢神经系统的体内光遗传学刺激

The ability to probe defined neural circuits in awake, freely-moving animals with cell-type specificity, spatial precision, and high temporal resolution ...

The Olfactory System as a Model to Study Axonal Growth Patterns and Morphology In Vivo

The olfactory system has the unusual capacity to generate new neurons throughout the lifetime of an organism. Olfactory stem cells in the basal portion of the olfactory epithelium continuously give rise to new sensory neurons that extend their axons into the olfactory bulb, where they face the challenge to integrate into existing circuitry. Because of this particular feature, the olfactory system represents a unique opportunity to monitor axonal wiring and guidance, and to investigate synapse formation. Here we describe a procedure for in vivo labeling of sensory neurons and subsequent visualization of axons in the olfactory system of larvae of the amphibian Xenopus laevis. To stain sensory neurons in the olfactory organ we adopt the electroporation technique. In vivo electroporation is an established technique for delivering fluorophore-coupled dextrans or other macromolecules into living cells. Stained sensory neurons and their axonal processes can then be monitored in the living animal either using confocal laser-scanning or multiphoton microscopy. By reducing the number of labeled cells to few or single cells per animal, single axons can be tracked into the olfactory bulb and their morphological changes can be monitored over weeks by conducting series of in vivo time lapse imaging experiments. While the described protocol exemplifies the labeling and monitoring of olfactory sensory neurons, it can also be adopted to other cell types within the olfactory and other systems.

The Olfactory System as a Model to Study Axonal Growth Patterns and Morphology In Vivo

We describe a protocol for in vivo labeling of olfactory sensory neurons by electroporation and subsequent confocal laser-scanning or multiphoton microscopy to visualize neuronal morphology and its development over time.

嗅觉系统作为模型来研究轴突生长的模式和形态<em&gt;在体内</em

The olfactory system has the unusual capacity to generate new neurons throughout the lifetime of an organism. Olfactory stem cells in the basal portion 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 ...

Extracellular Recording of Neuronal Activity Combined with Microiontophoretic Application of Neuroactive Substances in Awake Mice

Differences in the activity of neurotransmitters and neuromodulators, and consequently different neural responses, can be found between anesthetized and awake animals. Therefore, methods allowing the manipulation of synaptic systems in awake animals are required in order to determine the contribution of synaptic inputs to neuronal processing unaffected by anesthetics. Here, we present methodology for the construction of electrodes to simultaneously record extracellular neural activity and release multiple neuroactive substances at the vicinity of the recording sites in awake mice. By combining these procedures, we performed microiontophoretic injections of gabazine to selectively block GABAA receptors in neurons of the inferior colliculus of head-restrained mice. Gabazine successfully modified neural response properties such as the frequency response area and stimulus-specific adaptation. Thus, we demonstrate that our methods are suitable for recording single-unit activity and for dissecting the role of specific neurotransmitter receptors in auditory processing.
The main limitation of the described procedure is the relatively short recording time (~3 hr), which is determined by the level of habituation of the animal to the recording sessions. On the other hand, multiple recording sessions can be performed in the same animal. The advantage of this technique over other experimental procedures used to manipulate the level of neurotransmission or neuromodulation (such as systemic injections or the use of optogenetic models), is that the drug effect is confined to the local synaptic inputs to the target neuron. In addition, the custom-manufacture of electrodes allows adjustment of specific parameters according to the neural structure and type of neuron of interest (such as the tip resistance for improving the signal-to-noise ratio of the recordings).

We present methods for the construction of electrodes to simultaneously record extracellular neural activity and release multiple neuroactive substances at the vicinity of the recording sites in awake mice. This technique allows the detailed analysis of putative local synaptic inputs to the neuron of interest.

神经元电活动外记录与神经活性物质在清醒小鼠微电泳应用相结合

Differences in the activity of neurotransmitters and neuromodulators, and consequently different neural responses, can be found between anesthetized and ...

High-density Electroencephalographic Acquisition in a Rodent Model Using Low-cost and Open-source Resources

Advanced electroencephalographic analysis techniques requiring high spatial resolution, including electrical source imaging and measures of network connectivity, are applicable to an expanding variety of questions in neuroscience. Performing these kinds of analyses in a rodent model requires higher electrode density than traditional screw electrodes can accomplish. While higher-density electroencephalographic montages for rodents exist, they are of limited availability to most researchers, are not robust enough for repeated experiments over an extended period of time, or are limited to use in anesthetized rodents.1-3 A proposed low-cost method for constructing a durable, high-count, transcranial electrode array, consisting of bilaterally implantable headpieces is investigated as a means to perform advanced electroencephalogram analyses in mice or rats.

Procedures for headpiece fabrication and surgical implantation necessary to produce high signal to noise, low-impedance electroencephalographic and electromyographic signals are presented. While the methodology is useful in both rats and mice, this manuscript focuses on the more challenging implementation for the smaller mouse skull. Freely moving mice are only tethered to cables via a common adapter during recording. One version of this electrode system that includes 26 electroencephalographic channels and 4 electromyographic channels is described below.

Instructions for the low-cost construction and surgical implantation of a chronic transcranial high-density electroencephalographic montage into mice are provided. Signal recording, extraction, and processing techniques are also described.

高密度的脑电图采集在啮齿动物模型使用低成本的开源资源

Advanced electroencephalographic analysis techniques requiring high spatial resolution, including electrical source imaging and measures of network ...

Recording Synaptic Plasticity in Acute Hippocampal Slices Maintained in a Small-volume Recycling-, Perfusion-, and Submersion-type Chamber System

Even though experiments on brain slices have been in use since 1951, problems remain that reduce the probability of achieving a stable and successful analysis of synaptic transmission modulation when performing field potential or intracellular recordings. This manuscript describes methodological aspects that might be helpful in improving experimental conditions for the maintenance of acute brain slices and for recording field excitatory postsynaptic potentials in a commercially available submersion chamber with an outflow-carbogenation unit. The outflow-carbogenation helps to stabilize the oxygen level in experiments that rely on the recycling of a small buffer reservoir to enhance the cost-efficiency of drug experiments. In addition, the manuscript presents representative experiments that examine the effects of different carbogenation modes and stimulation paradigms on the activity-dependent synaptic plasticity of synaptic transmission.

This protocol describes the stabilization of the oxygen level in a small volume of recycled buffer and methodological aspects of recording activity-dependent synaptic plasticity in submerged acute hippocampal slices.

小容积回收-灌注-浸没式室系统中维持的急性海马切片的突触可塑性记录

Even though experiments on brain slices have been in use since 1951, problems remain that reduce the probability of achieving a stable and successful ...

In vivo Positron Emission Tomography to Reveal Activity Patterns Induced by Deep Brain Stimulation in Rats

Deep brain stimulation (DBS) is an invasive neurosurgical technique based on the application of electrical pulses to brain structures involved in the patient&#39;s pathophysiology. Despite the long history of DBS, its mechanism of action and appropriate protocols remain unclear, highlighting the need for research aiming to solve these enigmas. In this sense, evaluating the in vivo effects of DBS using functional imaging techniques represents a powerful strategy to determine the impact of stimulation on brain dynamics. Here, an experimental protocol for preclinical models (Wistar rats), combined with a longitudinal study [18F]-fluorodeoxyclucose positron emission tomography (FDG-PET), to assess the acute consequences of DBS on brain metabolism is described. First, animals underwent stereotactic surgery for bilateral implantation of electrodes into the prefrontal cortex. A post-surgical computerized tomography (CT) scan of each animal was acquired to verify electrode placement. After one week of recovery, a first static FDG-PET of each operated animal without stimulation (D1) was acquired, and two days later (D2), a second FDG-PET was acquired while animals were stimulated. For that, the electrodes were connected to an isolated stimulator after administering FDG to the animals. Thus, animals were stimulated during the FDG uptake period (45 min), recording the acute effects of DBS on brain metabolism. Given the exploratory nature of this study, FDG-PET images were analyzed by a voxel-wise approach based on a paired T-test between D1 and D2 studies. Overall, the combination of DBS and imaging studies allows describing the neuromodulation consequences on neural networks, ultimately helping to unravel the conundrums surrounding DBS.

In vivo Positron Emission Tomography to Reveal Activity Patterns Induced by Deep Brain Stimulation in Rats

We describe a preclinical experimental method to evaluate metabolic neuromodulation induced by acute deep brain stimulation with in vivo FDG-PET. This manuscript includes all experimental steps, from stereotaxic surgery to the application of the stimulation treatment and the acquisition, processing, and analysis of PET images.

体内 正电子发射断层扫描揭示大鼠深部脑刺激诱导的活动模式

Deep brain stimulation (DBS) is an invasive neurosurgical technique based on the application of electrical pulses to brain structures involved in the ...

In Vivo Vascular Injury Readouts in Mouse Retina to Promote Reproducibility

Advancements in ophthalmic imaging tools offer an unprecedented level of access to researchers working with animal models of neurovascular injury. To properly leverage this greater translatability, there is a need to devise reproducible methods of drawing quantitative data from these images. Optical coherence tomography (OCT) imaging can resolve retinal histology at micrometer resolution and reveal functional differences in vascular blood flow. Here, we delineate noninvasive vascular readouts that we use to characterize pathological damage post vascular insult in an optimized mouse model of retinal vein occlusion (RVO). These readouts include live imaging analysis of retinal morphology, disorganization of retinal inner layers (DRIL) measure of capillary ischemia, and fluorescein angiography measures of retinal edema and vascular density. These techniques correspond directly to those used to examine patients with retinal disease in the clinic. Standardizing these methods enables direct and reproducible comparison of animal models with clinical phenotypes of ophthalmic disease, increasing the translational power of vascular injury models.

In Vivo Vascular Injury Readouts in Mouse Retina to Promote Reproducibility

Here, we present three data analysis protocols for fluorescein angiography (FA) and optical coherence tomography (OCT) images in the study of Retinal Vein Occlusion (RVO).

体内 小鼠视网膜血管损伤读数以促进可重复性

Advancements in ophthalmic imaging tools offer an unprecedented level of access to researchers working with animal models of neurovascular injury. To ...