Name: investigating object representations in the macaque dorsal visual stream using single-unit recordings
Uploaded: 2018-08-01T15:00:00
Description: Watch this Scientific Journal Video about Investigating Object Representations in the Macaque Dorsal Visual Stream Using Single-unit Recordings at JoVE.com

我们感谢 Puttemans、马克 de Paep、萨拉 de Pril、Wouter Depuydt、阿斯特丽德何曼思、Piet Kayenbergh、Gerrit Meulemans、克利斯朵夫 Ulens 和 Stijn Verstraeten 的技术和行政援助。

所有技术程序都是按照国家卫生研究院关于实验室动物的护理和使用指南和欧盟指令 2010/63/欧盟的规定进行的, 并得到了鲁汶的伦理委员会的批准。
1. 清醒行为猴细胞外记录的一般方法
<ol><li>训练动物执行你的具体研究问题所需的视觉和马达任务。确保动物能够在同一记录会话期间灵活地切换任务, 以便广泛地测试神经元, 并更好地了解驱动神经响应的特征 (?...

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A., Murata, A., Fogassi, L., Gallese, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+properties+of+grasping-related+neurons+in+the+ventral+premotor+area+F5+of+the+macaque+monkey.">Functional properties of grasping-related neurons in the ventral premotor area F5 of the macaque monkey.</a> J. Neurophysiol. 95 (2), 709-729 (2006).</li><li>Umilta, M. A., Brochier, T., Spinks, R. L., Lemon, R. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Simultaneous+recording+of+macaque+premotor+and+primary+motor+cortex+neuronal+populations+reveals+different+functional+contributions+to+visuomotor+grasp.">Simultaneous recording of macaque premotor and primary motor cortex neuronal populations reveals different functional contributions to visuomotor grasp.</a> J. 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Neurosci. 24 (10), 2551-2565 (2004).</li><li>Theys, T., Pani, P., van Loon, J., Goffin, J., Janssen, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Selectivity+for+three-dimensional+shape+and+grasping-related+activity+in+the+macaque+ventral+premotor+cortex.">Selectivity for three-dimensional shape and grasping-related activity in the macaque ventral premotor cortex.</a> J.Neurosci. 32 (35), 12038-12050 (2012).</li><li>Nelissen, K., Luppino, G., Vanduffel, W., Rizzolatti, G., Orban, 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=Observing+others:+multiple+action+representation+in+the+frontal+lobe.">Observing others: multiple action representation in the frontal lobe.</a> Science. 310 (5746), 332-336 (2005).</li><li>Janssen, P., Srivastava, S., Ombelet, S., Orban, 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=Coding+of+shape+and+position+in+macaque+lateral+intraparietal+area.">Coding of shape and position in macaque lateral intraparietal area.</a> J. Neurosci. 28 (26), 6679-6690 (2008).</li><li>Romero, M. C., Janssen, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Receptive+field+properties+of+neurons+in+the+macaque+anterior+intraparietal+area.">Receptive field properties of neurons in the macaque anterior intraparietal area.</a> J. Neurophysiol. 115 (3), 1542-1555 (2016).</li><li>Decramer, T., Premereur, E., Theys, T., Janssen, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multi-electrode+recordings+in+the+macaque+frontal+cortex+reveal+common+processing+of+eye-,+arm-+and+hand+movements.+Program+No.+495.15/GG14.">Multi-electrode recordings in the macaque frontal cortex reveal common processing of eye-, arm- and hand movements. Program No. 495.15/GG14.</a> Neuroscience Meeting Planner. , (2017).</li><li>Pani, P., Theys, T., Romero, M. C., Janssen, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Grasping+execution+and+grasping+observation+activity+of+single+neurons+in+macaque+anterior+intraparietal+area.">Grasping execution and grasping observation activity of single neurons in macaque anterior intraparietal area.</a> J. Cogn. Neurosci. 26 (10), 2342-2355 (2014).</li><li>Turriziani, P., Smirni, D., Oliveri, M., Semenza, C., Cipolotti, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+the+prefrontal+cortex+in+familiarity+and+recollection+processes+during+verbal+and+non-verbal+recognition+memory.">The role of the prefrontal cortex in familiarity and recollection processes during verbal and non-verbal recognition memory.</a> Neuroimage. 52 (1), 469-480 (2008).</li><li>Tsao, D. Y., Schweers, N., Moeller, S., Freiwald, W. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Patches+of+faces-selective+cortex+in+the+macaque+frontal+lobe.">Patches of faces-selective cortex in the macaque frontal lobe.</a> Nat. Neurosci. 11 (8), 877-879 (2008).</li></ol>

对背流进行研究的综合方法需要仔细选择行为任务和视觉测试: 视觉和抓取范式可以根据区域的特定属性组合或单独使用。
在这篇文章中, 我们提供的例子, 神经活动记录在 F5p 和在响应一个子集的视觉和运动任务, 但非常相似的反应可以观察到其他前沿领域, 如45B 区和 F5a。
我们提出了两个实验装置来研究物体在抓取过程中的神经表现。用垂直旋转传送带 ...

人类和非人类灵长类动物拥有执行复杂运动动作的能力, 包括物体抓取。为了成功地完成这些任务, 我们的大脑需要完成将内部对象属性转换为马达命令。这种转变依赖于位于顶叶和腹侧前运动神经皮层1,2,3 (图 1) 的背皮层区域的复杂网络。
从猴子和人类的病变研究4,5, 我们知道, 背部视觉流-起源于初级视觉皮层和定向到后顶叶皮层-参与了空间视觉和运动规划行动。然而, 大多数的背流区域并不专门用于处理独特的类型。例如, 前 intraparietal 区 (本) 是背视流的终末期区域之一, 它包含了各种神经元, 不仅在抓取6、7、8期间, 而且在视觉检查对象7,8,9,10。
类似于 F5, 位于腹前运动神经皮层 (PMv) 的神经元, 在视觉固定和对象抓取过程中也有反应, 这对于将视觉信息转化为运动动作11是很重要的。这个区域的前部 (界别分组 F5a) 包含有选择性地响应三维 (3D, 视差定义) 图像12,13, 而位于凸性 (F5c) 中的界别分组包含神经元特征为镜子属性1,3, 射击时, 动物执行或观察行动。最后, 后 F5 区 (F5p) 是一个与手相关的领域, 有很大比例的 visuomotor 神经元响应观察和掌握3D 对象14,15。F5, 面积 45B, 位于弧形沟的下支, 也可能涉及两个形状处理16,17和抓取18。
在顶叶和额叶皮层检测对象选择性是很有挑战性的, 因为很难确定这些神经元对哪些功能做出反应, 以及这些神经元的接受领域是什么。例如, 如果一个神经元响应一个板块, 而不是一个锥, 这些物体的特征是驱动这种选择性: 2D 轮廓, 3D 结构, 深度方向, 或多种不同特征的组合？为了确定在对象固定和抓取过程中响应的神经元的关键对象特征, 需要使用对象的图像和相同图像的缩小版本来进行各种视觉测试。
在 F5 和小鼠的神经元中, 有相当一部分不仅对物体的视觉呈现做出反应, 而且当动物在黑暗中抓住这个物体时 (即在没有视觉信息的情况下)。这种神经元可能不会对无法掌握的物体的图像作出反应。因此, 视觉和运动成分的反应是紧密相连的, 这使得它很难研究神经元对象表示在这些地区。由于 visuomotor 神经元只能用真实世界的物体来测试, 所以我们需要一个灵活的系统, 在不同的位置在视觉领域和不同的方向呈现不同的物体, 如果我们想确定哪些特征对这些都是重要的。神经元。后者只能通过能够在视觉空间的不同位置呈现不同物体的机器人来实现。
本文旨在为研究顶额神经元的研究者提供一个实验指南。在以下各节中, 我们将提供实验室中用于分析醒猴猴 (猕猴猕猴) 抓取和视觉对象反应的一般协议。

<table>
	<tbody>
		<tr>
			<td>Grasping robot</td>
			<td>GIBAS Universal Robots</td>
			<td>UR-6-85-5-A</td>
			<td>Robot arm equipped with a gripper</td>
		</tr>
		<tr>
			<td>Carousel motor</td>
			<td>Siboni</td>
			<td>RD066/&#8224;20 MV6, 35x23 F02</td>
			<td>Motor to be implemented in a custom-made vertical carousel. It allows the rotation of the carousel.</td>
		</tr>
		<tr>
			<td>Eye tracker</td>
			<td>SR Research</td>
			<td>EyeLink II</td>
			<td>Infrared camera system sampling at 500 Hz</td>
		</tr>
		<tr>
			<td>Filter</td>
			<td>Wavetek Rockland</td>
			<td>852</td>
			<td>Electronic filters perform a variety of signal-processing functions with the purpose of removing a signal&#39;s unwanted frequency components.</td>
		</tr>
		<tr>
			<td>Preamplifier</td>
			<td>BAK ELECTRONICS, INC.</td>
			<td>A-1</td>
			<td>The Model A-1 allows to reduce input capacity and noise pickup and allows to test impedance for metal micro-electrodes</td>
		</tr>
		<tr>
			<td>Electrodes</td>
			<td>FHC</td>
			<td>UEWLEESE*N4G</td>
			<td>Metal microelectrodes (* = Impedance, to be chosen by the researcher)</td>
		</tr>
		<tr>
			<td>CRT monitor</td>
			<td>Vision Research Graphics</td>
			<td>M21L-67S01</td>
			<td>The CRT monitor is equipped with a fast-decay P46-phosphor operating at 120 Hz</td>
		</tr>
		<tr>
			<td>Ferroelectric liquid crystal shutters</td>
			<td>Display Tech</td>
			<td>FLC Shutter Panel; LV2500P-OEM</td>
			<td>The shutters operate at 60 Hz in front of the monkeys and are synchronized to the vertical retrace of the monitor</td>
		</tr>
	</tbody>
</table>

investigating object representations in the macaque dorsal visual stream using single-unit recordings

以前的研究表明, 猕猴大脑顶额区的神经元可以高度选择性地进行现实世界中的物体、视差定义的曲面以及真实世界物体 (无差异) 的图像, 类似于在腹视流中描述。此外, 顶锋区域被认为可以将视觉对象信息转换成适当的马达输出, 如抓取时手的预整形。为了更好地描述 visuomotor 变换所涉及的皮层网络中的对象选择性, 我们提供了一个测试电池, 目的是分析顶额区域神经元的视觉对象选择性。

图 5绘制了一个示例神经元的响应, 从区域 F5p 测试了四个对象: 两个不同的形状-一个球体和一个板块-显示两个不同的大小 (6 和3厘米)。这个特殊的神经元不仅响应大球体 (最佳刺激; 左上板), 而且还对大板 (左下板)。相比之下, 对较小对象的响应较弱 (右上角和下面板)。
图 6显示了?...

Watch this Scientific Journal Video about Investigating Object Representations in the Macaque Dorsal Visual Stream Using Single-unit Recordings at JoVE.com

Investigating Object Representations in the Macaque Dorsal Visual Stream Using Single-unit Recordings

本文给出了 visuomotor 变换中顶-额神经元的对象选择性分析的详细协议。

用单单元记录法研究猕猴背视流中的对象表示

Previous studies have shown that neurons in parieto-frontal areas of the macaque brain can be highly selective for real-world objects, disparity-defined ...

This method can help answering key questions in visual and motor systems neuroscience. For example, how do neurons encode objects and and object parts? And how do neural representations change from visual to motor?The main advantage of this method is that neurons can be studied with real-world objects during reaching and grasping, and with images of objects and object parts presented on a display. Demonstrating this procedure will be Maria Romero, a postdoc, and Irene Caprara, a PhD student from my laboratory. Begin with the test subject situated in the testing chair in front of the object-grasping setup.Adjust an infrared camera in front of the subject's eyes to obtain an adequate image of the pupil and corneal reflex. To run the visually-guided grasping task using a carousel setup, first allow the macaque to place the hand contralateral to the recorded hemisphere in the resting position in complete darkness to initiate the sequence. After a variable intertrial interval of 2, 000 to 3, 000 milliseconds, apply a red laser as a fixation point at the base of the object.If the animal maintains its gaze inside an electronically-defined fixation window for 500 milliseconds, illuminate the object from above with a light source. Then, after a variable delay of 300 to 1, 500 milliseconds, program a dimming of the laser as the visual go cue. Instruct the macaque to lift the hand from the resting position and reach, grasp, and hold the object for a randomized variable interval of 300 to 900 milliseconds.Whenever the macaque performs the whole sequence correctly, reward it with a drop of juice. As for the carousel setup, let the macaque place the hand contralateral to the recorded hemisphere in the resting position in complete darkness to initiate the sequence. After 2, 000 to 3, 000 milliseconds, illuminate the LED on the object.Again, if the animal maintains its gaze inside an electronically-defined fixation window, illuminate the object with a white light source. After the delay, switch off the LED as the visual go cue. Again, the monkey lifts the hand from the resting position and reaches, grasps, and holds the object.Reward the monkey with a drop of juice after each correct sequence. Continue the training with additional objects. During the task, the software should measure the reaction time between the go signal and the onset of the hand movement.And the grasping time between the start of the movement and lifting of the object. For the 2D tasks, use a standard LCD monitor. Project all visual stimuli on a black background.For the 3D tests, locate two ferroelectric liquid crystal shutters in front of the monkey's eyes. For the 3D test, locate two ferroelectric liquid crystal shutters in front of the monkey's eyes. Present the stimuli stereoscopically by alternating the left and right eye images on a cathode ray tube monitor equipped with a fast-decay P46 phosphor, while operating the shutters at 60 hertz with synchronization to the vertical retrace of the monitor.Start the trial by presenting a small square in the center of the screen as the fixation point. If the eye position remains within an electronically-defined one-degree-square window for at least 500 milliseconds, present the visual stimulus on the screen for a total time of 500 milliseconds. When the monkey maintains a stable fixation until the stimulus offset, reward it with a drop of juice.To study shape selectivity, run multiple tests with 2D images during the passive fixation task beginning with a search test. Test the visual selectivity of the cell using a wide set of images, including pictures of the object that was grasped in the VGG. For this and all subsequent visual tasks, compare the image evoking the strongest response, termed preferred image, to a second image to which the neuron is responding weakly, termed nonpreferred image.Next, run a contour test. From the original surface images of real objects, obtain progressively simplified versions of the same stimulus shape. Collect at least 10 trials per condition.To determine whether the neuron prefers the original surface, the silhouette, or the outline from the original shape. Then run a receptive field test. Present the images of objects at different positions on a display, covering the central visual field.A total of 35 positions at three degrees are used here. Finally, run a reduction test with contour fragments presented at the center of the receptive field to identify the minimum effective shape feature. Determine the minimum effective shape feature as the smallest shape fragment evoking a response that is at least 70%of the intact outline response.This image shows the responses of an example neuron recorded from area F5p, tested with a sphere and a plate, shown in two different sizes. This particular neuron responded not only to the large sphere optimal stimulus, but also to the large plate. In comparison, the response to the smaller objects was weaker.The following images show results from an example neuron recorded in the anterior intraparietal area, and tested during both the visually-guided grasping task and passive fixation. This neuron was responsive during grasping. The neuron was also responsive to the visual presentation of 2D images of objects, including pictures of the objects used in the grasping task.The preferred was not the object to be grasped, but another 2D picture with which the animal had no previous grasping experience. An example of the responses obtained in the reduction test is shown here. This example neuron responded to the smallest fragments in the test.Once mastered, this technique can be done in less than 90 minutes, if it's performed correctly. After watching this video, you should have a good understanding of how to test neurons in the dorsal visual stream with objects and images of objects to determine the critical features that drive the responses.

Research

JoVE Journal

Neuroscience

Single-unit In vivo Recordings from the Optic Chiasm of Rat

Single-unit In vivo Recordings from the Optic Chiasm of Rat

单机在体内</em从大鼠视交叉>录音

Information about the visual world is transmitted to the brain in sequences of action potentials in retinal ganglion cell axons that make up the optic ...

科研

神经科学

Transretinal ERG Recordings from Mouse Retina: Rod and Cone Photoresponses

transretinal ERG的录音，从小鼠视网膜杆和锥光反应

There are two distinct classes of image-forming photoreceptors in the vertebrate retina: rods and cones. Rods are able to detect single photons of light ...

Retrograde Fluorescent Labeling Allows for Targeted Extracellular Single-unit Recording from Identified Neurons In vivo

Retrograde Fluorescent Labeling Allows for Targeted Extracellular Single-unit Recording from Identified Neurons In vivo

逆行荧光染料标记允许有针对性的外单细胞记录技术鉴定神经元<em&gt;在体内</em

The overall goal of this method is to record single-unit responses from an identified population of neurons. In vivo electrophysiological recordings from ...

Dual Electrophysiological Recordings of Synaptically-evoked Astroglial and Neuronal Responses in Acute Hippocampal Slices

双突触诱发的星形胶质细胞和神经元的反应，在急性海马脑片电生理记录

Astrocytes form together with neurons tripartite synapses, where they integrate and modulate neuronal activity. Indeed, astrocytes sense neuronal inputs ...

Simultaneous Whole-cell Recordings from Photoreceptors and Second-order Neurons in an Amphibian Retinal Slice Preparation

同时光感受器和二阶神经元在两栖类动物的视网膜切片制备全细胞记录

One of the central tasks in retinal neuroscience  is to understand the circuitry of retinal neurons and how those connections are  responsible for shaping ...

A Procedure for Implanting Organized Arrays of Microwires for Single-unit Recordings in Awake, Behaving Animals

一个程序植入微丝的有组织阵列的单机录音清醒，举止动物

In vivo electrophysiological recordings in the awake, behaving animal provide a powerful method for understanding neural signaling at the single-cell ...

A Guide to In vivo Single-unit Recording from Optogenetically Identified Cortical Inhibitory Interneurons

A Guide to In vivo Single-unit Recording from Optogenetically Identified Cortical Inhibitory Interneurons

指南<em&gt;在体内</em&gt;单单元从Optogenetically鉴定皮质抑制性录像

A major challenge in neurophysiology has been to characterize the response properties and function of the numerous inhibitory cell types in the cerebral ...

Simultaneous ex vivo Functional Testing of Two Retinas by in vivo Electroretinogram System

Simultaneous ex vivo Functional Testing of Two Retinas by in vivo Electroretinogram System

同时<em&gt;体外</em两个由视网膜&gt;功能测试<em&gt;在体内</em&gt;电图系统

An In vivo electroretinogram (ERG) signal is composed of several overlapping components originating from different retinal cell types, as well as noise ...

In Vitro Modeling of Cancerous Neural Invasion: The Dorsal Root Ganglion Model

In Vitro Modeling of Cancerous Neural Invasion: The Dorsal Root Ganglion Model

在癌神经侵袭的体外模型：背根神经节型号

One way that solid tumors disseminate is through neural invasion. This route is well-known in cancers of the head and neck, prostate, and pancreas. These ...

Measuring Connectivity in the Primary Visual Pathway in Human Albinism Using Diffusion Tensor Imaging and Tractography

在人类白化病的初级视途径弥散张量成像和测量束成像连通

In albinism, the number of ipsilaterally projecting retinal ganglion cells (RGCs) is significantly reduced. The retina and optic chiasm have been proposed ...