Name: an open-source virtual reality system for the measurement of spatial learning in head-restrained mice
Uploaded: 2023-03-03T13:33:00
Description: Watch this Scientific Journal Video about An Open-Source Virtual Reality System for the Measurement of Spatial Learning in Head-Restrained Mice Video at JoVE.com

我们要感谢哈维实验室的诺亚·佩蒂特（Noah Pettit）在本手稿中开发协议时的讨论和建议。这项工作得到了BBRF青年研究者奖和NIMH 1R21MH122965（G.F.T.）的支持，此外还有NINDS R56NS128177（R.H.，C.L.）和NIMH R01MH068542（R.H.）。

本协议中的所有程序均已获得纽约州精神病学研究所机构动物护理和使用委员会的批准。
注意：单板计算机用于显示与在滚轮上运行头部约束的鼠标相协调的 VR 视觉环境。运动信息以串行输入的形式从 ESP32 微控制器接收，读取耦合到轮轴的旋转编码器。VR环境在Raspberry Pi GPU上使用OpenGL硬件加速渲染，该GPU利用Raspberry Pi的pi3d Python 3D包。然后，渲染环境 通过 投影仪输?...

<ol>
	<li>Lisman, 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=Viewpoints:+How+the+hippocampus+contributes+to+memory,+navigation+and+cognition.">Viewpoints: How the hippocampus contributes to memory, navigation and cognition.</a> Nature Neuroscience. 20 (11), 1434-1447 (2017).</li><li>Buzsaki, G., Moser, E. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Memory,+navigation+and+theta+rhythm+in+the+hippocampal-entorhinal+system.">Memory, navigation and theta rhythm in the hippocampal-entorhinal system.</a> Nature Neuroscience. 16 (2), 130-138 (2013).</li><li>O'Keefe, J., Dostrovsky, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+hippocampus+as+a+spatial+map.+Preliminary+evidence+from+unit+activity+in+the+freely-moving+rat.">The hippocampus as a spatial map. Preliminary evidence from unit activity in the freely-moving rat.</a> Brain Research. 34 (1), 171-175 (1971).</li><li>O'Keefe, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Place+units+in+the+hippocampus+of+the+freely+moving+rat.">Place units in the hippocampus of the freely moving rat.</a> Experimental Neurology. 51 (1), 78-109 (1976).</li><li>Fyhn, M., Molden, S., Witter, M. P., Moser, E. I., Moser, M. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spatial+representation+in+the+entorhinal+cortex.">Spatial representation in the entorhinal cortex.</a> Science. 305 (5688), 1258-1264 (2004).</li><li>Letzkus, J. 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=A+disinhibitory+microcircuit+for+associative+fear+learning+in+the+auditory+cortex.">A disinhibitory microcircuit for associative fear learning in the auditory cortex.</a> Nature. 480 (7377), 331-335 (2011).</li><li>Lacefield, C. O., Pnevmatikakis, E. A., Paninski, L., Bruno, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reinforcement+learning+recruits+somata+and+apical+dendrites+across+layers+of+primary+sensory+cortex.">Reinforcement learning recruits somata and apical dendrites across layers of primary sensory cortex.</a> Cell Reports. 26 (8), 2000-2008 (2019).</li><li>Dombeck, D. A., Harvey, C. D., Tian, L., Looger, L. L., Tank, D. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+imaging+of+hippocampal+place+cells+at+cellular+resolution+during+virtual+navigation.">Functional imaging of hippocampal place cells at cellular resolution during virtual navigation.</a> Nature Neuroscience. 13 (11), 1433-1440 (2010).</li><li>Gauthier, J. L., Tank, D. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+dedicated+population+for+reward+coding+in+the+hippocampus.">A dedicated population for reward coding in the hippocampus.</a> Neuron. 99 (1), 179-193 (2018).</li><li>Rickgauer, J. P., Deisseroth, K., Tank, D. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Simultaneous+cellular-resolution+optical+perturbation+and+imaging+of+place+cell+firing+fields.">Simultaneous cellular-resolution optical perturbation and imaging of place cell firing fields.</a> Nature Neuroscience. 17 (12), 1816-1824 (2014).</li><li>Yadav, 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=Prefrontal+feature+representations+drive+memory+recall.">Prefrontal feature representations drive memory recall.</a> Nature. 608 (7921), 153-160 (2022).</li><li>Priestley, J. B., Bowler, J. C., Rolotti, S. V., Fusi, S., Losonczy, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Signatures+of+rapid+plasticity+in+hippocampal+CA1+representations+during+novel+experiences.">Signatures of rapid plasticity in hippocampal CA1 representations during novel experiences.</a> Neuron. 110 (12), 1978-1992 (2022).</li><li>Heys, J. G., Rangarajan, K. V., Dombeck, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+functional+micro-organization+of+grid+cells+revealed+by+cellular-resolution+imaging.">The functional micro-organization of grid cells revealed by cellular-resolution imaging.</a> Neuron. 84 (5), 1079-1090 (2014).</li><li>Harvey, C. D., Collman, F., Dombeck, D. A., Tank, D. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intracellular+dynamics+of+hippocampal+place+cells+during+virtual+navigation.">Intracellular dynamics of hippocampal place cells during virtual navigation.</a> Nature. 461 (7266), 941-946 (2009).</li><li>. Harvey Lab Mouse VR Available from: <a target="_blank" href="https://github.com/Harvey/Lab/mouseVR">https://github.com/Harvey/Lab/mouseVR</a> (2021)</li><li>Pettit, N. L., Yap, E. L., Greenberg, M. E., Harvey, C. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fos+ensembles+encode+and+shape+stable+spatial+maps+in+the+hippocampus.">Fos ensembles encode and shape stable spatial maps in the hippocampus.</a> Nature. 609 (7926), 327-334 (2022).</li><li>Turi, G. 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=Vasoactive+intestinal+polypeptide-expressing+interneurons+in+the+hippocampus+support+goal-oriented+spatial+learning.">Vasoactive intestinal polypeptide-expressing interneurons in the hippocampus support goal-oriented spatial learning.</a> Neuron. 101 (6), 1150-1165 (2019).</li><li>Ulivi, A. 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=Longitudinal+two-photon+imaging+of+dorsal+hippocampal+CA1+in+live+mice.">Longitudinal two-photon imaging of dorsal hippocampal CA1 in live mice.</a> Journal of Visual Experiments. (148), e59598 (2019).</li><li>Wang, Y., Zhu, D., Liu, B., Piatkevich, K. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Craniotomy+procedure+for+visualizing+neuronal+activities+in+hippocampus+of+behaving+mice.">Craniotomy procedure for visualizing neuronal activities in hippocampus of behaving mice.</a> Journal of Visual Experiments. (173), e62266 (2021).</li><li>Tuncdemir, S. 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=Parallel+processing+of+sensory+cue+and+spatial+information+in+the+dentate+gyrus.">Parallel processing of sensory cue and spatial information in the dentate gyrus.</a> Cell Reports. 38 (3), 110257 (2022).</li><li>Dombeck, D. A., Khabbaz, A. N., Collman, F., Adelman, T. L., Tank, D. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Imaging+large-scale+neural+activity+with+cellular+resolution+in+awake,+mobile+mice.">Imaging large-scale neural activity with cellular resolution in awake, mobile mice.</a> Neuron. 56 (1), 43-57 (2007).</li><li>Guo, Z. 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=Procedures+for+behavioral+experiments+in+head-fixed+mice.">Procedures for behavioral experiments in head-fixed mice.</a> PLoS One. 9 (2), 88678 (2014).</li><li>Jordan, J. T., Gon&#231;alves, J. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Silencing+of+hippocampal+synaptic+transmission+impairs+spatial+reward+search+on+a+head-fixed+tactile+treadmill+task.">Silencing of hippocampal synaptic transmission impairs spatial reward search on a head-fixed tactile treadmill task.</a> bioRxiv. , (2021).</li><li>Urai, A. E., 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=Citric+acid+water+as+an+alternative+to+water+restriction+for+high-yield+mouse+behavior.">Citric acid water as an alternative to water restriction for high-yield mouse behavior.</a> eNeuro. 8 (1), (2021).</li><li>Saleem, A. B., Diamanti, E. M., Fournier, J., Harris, K. D., Carandini, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Coherent+encoding+of+subjective+spatial+position+in+visual+cortex+and+hippocampus.">Coherent encoding of subjective spatial position in visual cortex and hippocampus.</a> Nature. 562 (7725), 124-127 (2018).</li><li>Ravassard, 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=Multisensory+control+of+hippocampal+spatiotemporal+selectivity.">Multisensory control of hippocampal spatiotemporal selectivity.</a> Science. 340 (6138), 1342-1346 (2013).</li><li>Aghajan, Z. 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=Impaired+spatial+selectivity+and+intact+phase+precession+in+two-dimensional+virtual+reality.">Impaired spatial selectivity and intact phase precession in two-dimensional virtual reality.</a> Nature Neuroscience. 18 (1), 121-128 (2015).</li></ol>

这款面向鼠标的开源 VR 系统只有在旋转和行为 ESP32 微控制器与单板计算机之间正确建立串行连接（步骤 2）的情况下才能运行（步骤 2），这可以使用 IDE 串行监视器（步骤 2.4.5）进行确认。为了从该协议（步骤4）获得成功的行为结果，必须使小鼠习惯于该装置，并舒适地在轮子上运行以获得液体奖励（步骤4.3-4.5）。这需要足够（但不是过度）的水限制，因为在笼子里随意喝水的小鼠不会...

空间导航是一种重要的行为，动物通过这种行为将新位置的特征编码到认知地图中，用于查找可能奖励的区域并避免潜在危险的区域。空间导航背后的认知过程与记忆有着千丝万缕的联系，它们在海马体1和皮层中共享一个神经基质，在这些区域中，这些区域的神经回路整合传入的信息，形成环境和事件的认知图谱，以供以后回忆2。虽然海马体3，4 中的位置细胞和内嗅皮层5 中网格细胞的发现揭示了海马体内认知图谱是如何形成的，但关于海马体的特定神经亚型、微电路和单个亚区（齿状回和角氨区，CA3-1）如何相互作用并参与空间记忆的形成和回忆，仍然存在许多问题。
体内双光子成像一直是揭示感觉神经生理学中细胞和群体动态的有用工具6，7;然而，头枕的典型必要性限制了这种方法在检查哺乳动物空间行为方面的实用性。虚拟现实（VR）8的出现解决了这一缺点，它提供了身临其境和逼真的视觉空间环境，而头部约束的小鼠在球或跑步机上奔跑，以研究海马体8，9，10和皮层11中的空间和上下文编码。此外，将VR环境与行为小鼠一起使用，使神经科学研究人员能够通过精确控制VR环境12的元素（例如，视觉流，上下文调制）来剖析空间行为的组成部分，这在现实世界的空间学习实验中是不可能的，例如莫里斯水迷宫，巴恩斯迷宫或孔板任务。
Visual VR 环境通常在计算机的图形处理单元 （GPU） 上渲染，该单元处理快速计算在屏幕上实时建模移动 3D 环境所需的数千个多边形的负载。较大的处理要求通常需要使用带有GPU的单独PC，该GPU将视觉环境渲染到显示器，多个屏幕13或投影仪14 ，因为从跑步机，轮子或动物下方的泡沫球记录运动。因此，由此产生的用于控制、渲染和投影 VR 环境的设备相对昂贵、笨重且笨重。此外，文献中的许多此类环境都是使用专有软件实现的，这些软件既昂贵又只能在专用PC上运行。
出于这些原因，我们设计了一个开源VR系统，使用Raspberry Pi单板计算机研究头部约束小鼠的空间学习行为。这款Linux计算机既小又便宜，但包含一个用于3D渲染的GPU芯片，允许将VR环境与各种个人设置中的显示器或行为设备集成。此外，我们还开发了一个用Python编写的图形软件包“HallPassVR”，它利用单板计算机通过重新组合使用图形用户界面（GUI）选择的自定义视觉特征来渲染简单的视觉空间环境，虚拟线性轨道或走廊。它与微控制器子系统（例如 ESP32 或 Arduino）相结合，以测量运动和协调行为，例如通过提供其他形式的感官刺激或奖励来促进强化学习。该系统提供了一种廉价、灵活且易于使用的替代方法，用于在双光子成像（或其他需要头部固定的技术）期间向头部约束的小鼠提供视觉空间 VR 环境，以研究空间学习行为背后的神经回路。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>1/4 &#34; diam aluminum rod</td>
			<td>McMaster-Carr</td>
			<td>9062K26</td>
			<td>3&#34; in length for wheel axle</td>
		</tr>
		<tr>
			<td>1/4&#34;-20 cap screws, 3/4&#34; long (x2)</td>
			<td>Amazon.com</td>
			<td>B09ZNMR41V</td>
			<td>for affixing head post holders to optical posts</td>
		</tr>
		<tr>
			<td>2&#34;x7&#34; T-slotted aluminum bar (x2)</td>
			<td>8020.net</td>
			<td>1020</td>
			<td>wheel/animal mounting frame</td>
		</tr>
		<tr>
			<td>6&#34; diam, 3&#34; wide acrylic cylinder (1/8&#34; thick)</td>
			<td>Canal Plastics</td>
			<td>33210090702</td>
			<td>Running wheel (custom width cut at canalplastics.com)</td>
		</tr>
		<tr>
			<td>8-32 x 1/2&#34; socket head screws</td>
			<td>McMaster-Carr</td>
			<td>92196A194</td>
			<td>fastening head post holder to optical post&#160;</td>
		</tr>
		<tr>
			<td>Adjustable arm (14&#34;)</td>
			<td>Amazon.com</td>
			<td>B087BZGKSL</td>
			<td>to hold/adjust lick spout</td>
		</tr>
		<tr>
			<td>Analysis code (MATLAB)</td>
			<td></td>
			<td>custom written</td>
			<td>file at github.com/GergelyTuri/HallPassVR/software/Analysis code</td>
		</tr>
		<tr>
			<td>Axle mounting flange, 1/4&#34; ID</td>
			<td>Pololu</td>
			<td>1993</td>
			<td>for mounting wheel to axle</td>
		</tr>
		<tr>
			<td>Ball bearing (5/8&#34; OD, 1/4&#34; ID, x2)</td>
			<td>McMaster-Carr</td>
			<td>57155K324</td>
			<td>for mounting wheel axle to frame</td>
		</tr>
		<tr>
			<td>Behavior ESP32 code</td>
			<td></td>
			<td>custom written</td>
			<td>file at github.com/GergelyTuri/HallPassVR/software/Arduino code/Behavior board</td>
		</tr>
		<tr>
			<td>Black opaque matte acrylic sheets (1/4&#34; thick)</td>
			<td>Canal Plastics</td>
			<td>32918353422</td>
			<td>laser cut file at github.com/GergelyTuri/HallPassVR/hardware/VR screen assembly</td>
		</tr>
		<tr>
			<td>Clear acrylic sheet (1/4&#34; thick)</td>
			<td>Canal Plastics</td>
			<td>32920770574</td>
			<td>laser cut file at github.com/GergelyTuri/HallPassVR/hardware/VR wheel assembly</td>
		</tr>
		<tr>
			<td>ESP32 devKitC v4 (x2)</td>
			<td>Amazon.com</td>
			<td>B086YS4Z3F</td>
			<td>microcontroller for behavior and rotary encoder</td>
		</tr>
		<tr>
			<td>ESP32 shield</td>
			<td>OpenMaze.org</td>
			<td>OMwSmall</td>
			<td>description at www.openmaze.org (https://claylacefield.wixsite.com/openmazehome/copy-of-om2shield). ZIP gerber files at: https://github.com/claylacefield/OpenMaze/tree/master/OM_PCBs</td>
		</tr>
		<tr>
			<td>Fasteners and brackets&#160;</td>
			<td>8020.net</td>
			<td>4138, 3382,3280</td>
			<td>for wheel frame mounts</td>
		</tr>
		<tr>
			<td>goniometers</td>
			<td>Edmund Optics</td>
			<td>66-526, 66-527</td>
			<td>optional for behavior. Fine tuning head for imaging</td>
		</tr>
		<tr>
			<td>HallPassVR python code</td>
			<td></td>
			<td>custom written</td>
			<td>file at github.com/GergelyTuri/HallPassVR/software/HallPassVR</td>
		</tr>
		<tr>
			<td>Head post holder</td>
			<td></td>
			<td>custom design</td>
			<td>3D design file at github.com/GergelyTuri/HallPassVR/hardware/VR head mount/Headpost Clamp</td>
		</tr>
		<tr>
			<td>LED projector</td>
			<td>Texas Instruments</td>
			<td>DLPDLCR230NPEVM</td>
			<td>or other small LED projector</td>
		</tr>
		<tr>
			<td>Lick spout</td>
			<td>VWR</td>
			<td>20068-638</td>
			<td>(or ~16 G metal hypodermic tubing)</td>
		</tr>
		<tr>
			<td>M 2.5 x 6 set screws</td>
			<td>McMaster-Carr</td>
			<td>92015A097</td>
			<td>securing head post&#160;</td>
		</tr>
		<tr>
			<td>Matte white diffusion paper</td>
			<td>Amazon.com</td>
			<td></td>
			<td>screen material</td>
		</tr>
		<tr>
			<td>Metal headposts</td>
			<td></td>
			<td>custom design</td>
			<td>3D design file at github.com/GergelyTuri/HallPassVR/hardware/VR head mount/head post designs</td>
		</tr>
		<tr>
			<td>Miscellenous tubing and tubing adapters (1/16&#34; ID)</td>
			<td></td>
			<td></td>
			<td>for constructing the water line</td>
		</tr>
		<tr>
			<td>Optical breadboard</td>
			<td>Thorlabs</td>
			<td>as per user&#39;s requirements</td>
			<td></td>
		</tr>
		<tr>
			<td>Optical posts, 1/2&#34; diam (2x)</td>
			<td>Thorlabs</td>
			<td>TR4</td>
			<td>for head fixation setup</td>
		</tr>
		<tr>
			<td>Processing code</td>
			<td></td>
			<td>custom written</td>
			<td>file at github.com/GergelyTuri/HallPassVR/software/Processing code</td>
		</tr>
		<tr>
			<td>Raspberry Pi 4B</td>
			<td>raspberry.com, adafruit.com</td>
			<td></td>
			<td>Single-board computer for rendering of HallPassVR envir.</td>
		</tr>
		<tr>
			<td>Right angle clamp</td>
			<td>Thorlabs</td>
			<td>RA90</td>
			<td>for head fixation setup</td>
		</tr>
		<tr>
			<td>Rotary encoder (quadrature, 256 step)</td>
			<td>DigiKey</td>
			<td>ENS1J-B28-L00256L</td>
			<td>to measure wheel rotation</td>
		</tr>
		<tr>
			<td>Rotary encoder ESP32 code</td>
			<td></td>
			<td>custom written</td>
			<td>file at github.com/GergelyTuri/HallPassVR/software/Arduino code/Rotary encoder</td>
		</tr>
		<tr>
			<td>SCIGRIP 10315 acrylic cement</td>
			<td>Amazon.com</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Shaft coupler</td>
			<td>McMaster-Carr</td>
			<td>9861T426</td>
			<td>to couple rotary encoder shaft with axle</td>
		</tr>
		<tr>
			<td>Silver mirror acrylic sheets</td>
			<td>Canal Plastics</td>
			<td>32913817934</td>
			<td>laser cut file at github.com/GergelyTuri/HallPassVR/hardware/VR screen assembly</td>
		</tr>
		<tr>
			<td>Solenoid valve</td>
			<td>Parker</td>
			<td>003-0137-900</td>
			<td>to administer water rewards</td>
		</tr>
	</tbody>
</table>

an open-source virtual reality system for the measurement of spatial learning in head-restrained mice

小鼠头部约束行为实验使神经科学家能够使用高分辨率电生理和光学成像工具观察神经回路活动，同时向行为动物提供精确的感官刺激。最近，使用虚拟现实（VR）环境的人类和啮齿动物研究表明，由于对空间和上下文线索等参数的极其精确的控制，VR是揭示海马体和皮层空间学习背后的神经机制的重要工具。然而，为啮齿动物的空间行为设置虚拟环境可能成本高昂，并且需要广泛的工程和计算机编程背景。在这里，我们提出了一个基于廉价，模块化，开源硬件和软件的简单而强大的系统，使研究人员能够使用VR环境研究头部约束小鼠的空间学习。该系统使用耦合的微控制器来测量运动并提供行为刺激，而头部约束的小鼠在轮子上奔跑，与单板计算机上运行的图形软件包呈现的虚拟线性轨道环境保持一致。对分布式处理的强调使研究人员能够设计灵活的模块化系统来引发和测量小鼠的复杂空间行为，以确定哺乳动物大脑中神经回路活动与空间学习之间的联系。

这种开源的虚拟现实行为设置使我们能够量化舔行为，作为空间学习的读出，因为头部约束的小鼠在虚拟线性轨道环境中导航。将7只4个月大的两性C57BL / 6小鼠置于限制的水时间表上，并首先训练在低水平上连续舔舐，同时在没有VR的情况下在轮子上奔跑以获得随机空间奖励（“随机觅食”）。尽管当以 2 m 随机走廊模式移动到 VR 投影屏幕设置时，它们的性能最初受到影响，但在几次 VR 会话中，?...

Watch this Scientific Journal Video about An Open-Source Virtual Reality System for the Measurement of Spatial Learning in Head-Restrained Mice Video at JoVE.com

An Open-Source Virtual Reality System for the Measurement of Spatial Learning in Head-Restrained Mice Video (Video) | JoVE

在这里，我们提出了一个简化的开源硬件和软件设置，用于使用虚拟现实（VR）研究鼠标空间学习。该系统利用微控制器网络和运行易于使用的Python图形软件包的单板计算机，向在滚轮上运行的头部受限鼠标显示虚拟线性轨迹。

一种用于测量头部约束小鼠空间学习的开源虚拟现实系统

Head-restrained behavioral experiments in mice allow neuroscientists to observe neural circuit activity with high-resolution electrophysiological and ...

这种开源虚拟现实系统是研究大脑空间学习的重要工具，因为它允许研究人员使用简单的模块化电子设置向头部受限的鼠标呈现一组一致的空间刺激。该系统的优点是价格低廉、易于设置、紧凑且模块化，允许构建多个行为设置以进行训练并与现有的头部约束行为设置集成。该系统是测量空间学习和头部约束小鼠的理想选择，但是，它同样能够为其他物种和制剂的实验提供视觉虚拟现实环境，包括人类心理物理学和神经成像。演示该程序的将是Carla Diaz和Hannah Chung，我们实验室的研究助理。首先，连接旋转编码器组件和旋转 ESP32 之间的电线。旋转编码器通常有四根线，正极、GND、A 和 B.通过跳线将它们连接到 ESP32、3 个 0.3 伏、GND 25 和 26 针。连接旋转 ESP32 和行为 ESP32 之间的串行 RX/TX 线。在旋转 ESP32、串行 0 RX/TX 和行为 ESP32 的串行 2 端口之间建立简单的两线连接。连接旋转 ESP32 和单板计算机 GPIO 或直接 USB 连接之间的串行 RX/TX 线。在单板计算机 GPIO 引脚 14、15、RX/TX 和旋转 ESP32、Serial2、TX/RX 引脚 1716 之间建立两线连接。接下来，将旋转 ESP32 USB 插入单板计算机 USB 以上传初始旋转编码器代码。将 12 伏液体电磁阀连接到 OMW 小 PCB 最左侧的 ULN2803 IC 输出端，将舔口连接到 ESP32 触摸输入端。将 USB 插入单板计算机的 USB 端口，即可将新程序上传到行为 ESP32 中，以用于不同的实验范式，并使用随附的处理草图捕获行为数据。然后将 12 伏直流墙上适配器插入行为 ESP32 OMW 小 PCB 上的 2.1 毫米桶形插孔连接器，为奖励电磁阀提供电源。将单板计算机的 HDMI 二进制输出插入投影仪 HDMI 端口。这会将单板计算机GPU渲染的图形软件环境带到投影屏幕上。在单板计算机中打开终端窗口，然后导航到霍尔通道 VR 文件夹。运行指示的虚拟现实或 VR 图形用户界面或 GUI 以打开 GUI 窗口。从列表框中为轨道上的三种模式中的每种模式选择并添加四个元素，然后单击“生成”。从下拉菜单中选择地板和天花板图像，并将此示例代码的轨道长度设置为两米。如果需要，请命名此模式。单击“开始”按钮并等待 VR 窗口启动，然后再单击其他位置。图形软件环境将出现在第二个屏幕上。运行处理草图以获取并绘制行为数据移动。在处理 IDE 中打开指示的命令。将动物更改为鼠标编号变量，并将会话分钟数设置为等于行为会话的长度（以分钟为单位）。单击处理 IDE 上的“运行”按钮。检查处理绘图窗口，该窗口应显示当前鼠标在虚拟线性轨道上的位置，因为轮子随着奖励区域和舔、圈数和奖励的运行直方图旋转，每 30 秒更新一次。手动推进运行轮以模拟鼠标运行进行测试或使用测试鼠标进行初始设置。单击绘图窗口，然后按键盘上的 Q 键停止获取行为数据。当会话分钟数过后或用户按 Q 键退出时，将保存行为事件和时间的文本文件以及 PNG 中最终绘图窗口的图像。对于具有非操作奖励的随机锻造，请使用任意视觉元素的路径运行图形软件 GUI 程序。然后将行为程序上传到行为 ESP32 中，并带有多个非操作奖励来调节鼠标运行和舔舐。轻轻地将鼠标放在头部固定装置中，将舔嘴调整到鼠标嘴正前方的位置，并将鼠标滚轮定位到投影屏幕区域的中心。在处理草图中设置动物的名称，然后在处理 IDE 中按“运行”开始获取和绘制行为数据。在 20 到 30 分钟的会话中运行鼠标，直到鼠标每次会话至少运行 20 圈，并舔舐随机位置提供的奖励。对于在交替圈上使用操作奖励的随机觅食，上传交替操作者等于 1 的行为程序并训练鼠标，直到它同时舔非操作性和操作性奖励区域。对于完全操作的随机觅食，上传具有四个操作性随机奖励区域的行为程序并训练鼠标，直到它沿着轨道一致地舔舐奖励。接下来进行空间学习，运行图形软件程序，路径为黑暗的走廊，中间有一个视觉提示。然后将带有单个隐藏奖励区域的行为程序上传到行为 ESP32。让鼠标在单个隐藏奖励区和单个视觉提示 VR 走廊上运行 30 分钟的会话，并在会话期间捕获数据，如前所述。从处理草图文件夹中下载 txt 数据文件并分析行为数据，以观察空间选择性舔作为空间学习指标的出现。此处显示了使用图形软件环境进行的空间学习。通过随机觅食训练的渐进阶段，小鼠学会了在轮子上奔跑，并在低水平上沿着轨道持续舔舐，然后切换到单个隐藏的奖励位置以显示空间学习。在这项研究中，七只小鼠中的四只在两到四个会话中通过单一的视觉提示学习了隐藏的奖励任务，如它们在奖励区附近的舔舐所显示的选择性增加。此外，小鼠在会话内和会话之间学习之间都表现出实质性。第二天每圈的空间舔舐次数显示奖励区之前的舔舐增加，其他地方的舔舐减少，表明空间特定的预期舔舐的发展。使用该系统时要记住的主要事情是，只有在跑轮上加固和舒适的情况下，鼠标才会表现良好。因此，水适当地限制动物，轻柔地处理它们，并确保它们的头部约束位置最适合在观看投影屏幕时奔跑。神经科学研究人员可以将这种开源VR系统与体内成像或电生理学相结合，以研究大脑空间学习背后的神经元回路。我们认为，这种开源VR系统的简单性将使研究人员能够将该系统集成到各种神经记录设置中。对VR环境中空间刺激的精确控制将使研究人员能够检查特定神经元回路对空间学习的贡献。

Research

JoVE Journal

Neuroscience

An Open-Source Virtual Reality System for the Measurement of Spatial Learning in Head-Restrained Mice

Low-stress Route Learning Using the Lashley III Maze in Mice

Low-stress Route Learning Using the Lashley III Maze in Mice Video (Video) | JoVE

低应力路线学习，在老鼠身上使用拉什利三迷宫

Many behavior tests designed to assess learning and memory in rodents, particularly mice, rely on visual cues, food and/or water deprivation, or other ...

科研

神经科学

Human Fear Conditioning Conducted in Full Immersion 3-Dimensional Virtual Reality 

Human Fear Conditioning Conducted in Full Immersion 3-Dimensional Virtual Reality  Video (Video) | JoVE

人类的恐惧充分浸泡三维虚拟现实中进行的空调

Fear conditioning is a widely used paradigm in non-human animal research to investigate the neural mechanisms underlying fear and anxiety. A major ...

Using MazeSuite and Functional Near Infrared Spectroscopy to Study Learning in Spatial Navigation

Using MazeSuite and Functional Near Infrared Spectroscopy to Study Learning in Spatial Navigation Video (Video) | JoVE

使用近红外光谱MazeSuite和功能研究学习空间导航

MazeSuite is a complete toolset to prepare, present and analyze navigational and spatial experiments1. MazeSuite can be used to design and edit adapted ...

Optical Recording of Suprathreshold Neural Activity with Single-cell and Single-spike Resolution

Optical Recording of Suprathreshold Neural Activity with Single-cell and Single-spike Resolution Video (Video) | JoVE

阈上的神经活动的单细胞和单穗的分辨率的光学的方法记录

Signaling of information in the vertebrate central nervous system is often carried by populations of neurons rather than individual neurons. Also ...

Building An Open-source Robotic Stereotaxic Instrument

Building An Open-source Robotic Stereotaxic Instrument Video (Video) | JoVE

建设一个开放源码的机器人立体定位仪

This protocol includes the designs and software necessary to upgrade an existing stereotaxic instrument to a robotic (CNC) stereotaxic instrument for ...

In vivo Optogenetic Stimulation of the Rodent Central Nervous System

In vivo Optogenetic Stimulation of the Rodent Central Nervous System Video (Video) | JoVE

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 ...

Immunohistochemical Visualization of Hippocampal Neuron Activity After Spatial Learning in a Mouse Model of Neurodevelopmental Disorders

Immunohistochemical Visualization of Hippocampal Neuron Activity After Spatial Learning in a Mouse Model of Neurodevelopmental Disorders Video (Video) | JoVE

海马神经元活动的可视化免疫组化空间后学习神经发育障碍的小鼠模型

Induction of phosphorylated extracellular-regulated kinase (pERK) is a reliable molecular readout of learning-dependent neuronal activation. Here, we ...

Electrophysiological Method for Recording Intracellular Voltage Responses of Drosophila Photoreceptors and Interneurons to Light Stimuli In Vivo

Electrophysiological Method for Recording Intracellular Voltage Responses of Drosophila Photoreceptors and Interneurons to Light Stimuli In Vivo Video (Video) | JoVE

为记录细胞内电压响应电法<em&gt;果蝇</em&gt;光感受器和中间对光刺激<em&gt;在体内</em

Voltage responses of insect photoreceptors and visual interneurons can be accurately recorded with conventional sharp microelectrodes. The method ...

Assessing Spatial Learning and Memory in Small Squamate Reptiles

Assessing Spatial Learning and Memory in Small Squamate Reptiles Video (Video) | JoVE

评估空间学习记忆小鳞类爬行动物

Clinical research has leveraged a variety of paradigms to assess cognitive decline, commonly targeting spatial learning and memory abilities. However, ...

An Objective and Reproducible Test of Olfactory Learning and Discrimination in Mice

An Objective and Reproducible Test of Olfactory Learning and Discrimination in Mice Video (Video) | JoVE

小鼠嗅觉学习和鉴别的客观和可重复性试验

Olfaction is the predominant sensory modality in mice and influences many important behaviors, including foraging, predator detection, mating, and ...