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 (图 2-3)。<ol>
<li>训练动物在视觉上被引导的抓住 (VGG; 抓住 ' 在光 ') 评估反应的 visuomotor 组分。注意: 独立于所选任务, 在训练阶段开始前至少三天内逐步限制液体摄入量。<ol>
<li>在实验课的整个时间内限制猴子的头部。</li>
			<li>在第一次疗程中, 将手对侧放在休息位置的录音室, 帮助动物到达并抓住物体, 每次尝试后给予人工奖励。</li>
			<li>在每次试验结束时, 把猴子的手放在休息的位置上。</li>
			<li>每几个试验, 松开猴子的手, 等待几秒钟观察, 如果动物自发地发起运动。</li>
			<li>当猴子到达目标时, 应用人工奖励。</li>
			<li>当获得正确的到达阶段, 帮助动物举起 (或拉扯) 对象和奖励手工。</li>
			<li>就像在1.1.1.4 和 1.1.1.5, 松开猴子的手, 等待几秒钟观察动物是否自发地启动运动。当运动正确执行时给予奖励。</li>
			<li>在手术过程中, 尽可能多地纠正到达、手部位置和手腕方向。</li>
			<li>重复以上步骤, 直到动物自动执行序列。</li>
			<li>加载自动任务。当动物在预定的时间执行到达和掌握动作时, 它会自动得到奖赏。</li>
			<li>逐渐增加对象的持有时间。</li>
			<li>介绍在物体底部投射固定点的激光。然后添加眼睛跟踪器, 以监视对象要掌握的眼睛位置。</li>
		</ol>
</li>
		<li>训练动物在记忆引导下抓取 (MGG) 来调查运动成分的反应, 不受刺激的视觉成分的影响。<ol>
<li>克制猴子的头。</li>
			<li>按照 VGG 描述的相同步骤, 确保动物在电子定义的窗口中的任务期间保持激光固定。对于这个版本的任务, 灯在固定期结束时熄灭。</li>
		</ol>
</li>
		<li>训练猴子被动固定, 以解决视觉反应和形状的选择性。<ol>
<li>克制猴子的头。</li>
			<li>使用 CRT (3D 刺激的被动固定) 或 LCD 显示器 (被动固定2D 刺激) 向猴子呈现视觉刺激。</li>
			<li>在屏幕中心显示固定点, 叠加在视觉刺激上。</li>
			<li>奖励动物在每次表现刺激之后, 逐渐增加固定期直到达到任务的标准。</li>
		</ol>
</li>
	</ol>
</li>
	<li>使用无菌工具, 窗帘和长袍进行手术。<ol>
<li>麻醉动物与氯胺酮 (15 毫克/千克, 肌肉) 和 medetomidine 盐酸 (0.01-0 04 毫升/千克肌肉), 并定期确认麻醉通过检查动物对刺激, 心率, 呼吸率和血液的反应压力。</li>
		<li>保持全身麻醉 (丙泊酚10毫克/千克/小时静脉注射), 并使用气管管来管理氧气。在麻醉下使用 lanolim 软膏防止眼部干燥。</li>
		<li>使用0.5 毫升丁丙诺啡 (静脉注射 0.3mg) 提供镇痛。在手术中心率增加的情况下, 可以进行额外的剂量治疗。</li>
		<li>用陶瓷螺钉和牙科丙烯酸植入 MRI 兼容头柱。在严格的无菌条件下执行所有的生存手术。为充分维护无菌领域, 使用一次性无菌手套, 口罩和无菌器械。</li>
		<li>以解剖学磁共振成像 (MRI) 为指导;霍斯利-克拉克坐标), 在感兴趣的地区做一个开颅手术, 并植入猴子头骨上的录音室。使用标准记录室进行单单元细胞外记录或多电极 microdrive, 用于同时记录多个神经元。</li>
		<li>手术后, 停止静脉注射丙泊酚, 直到自发呼吸恢复。不要让动物无人看管, 直到它恢复知觉并在社会群体中引入动物才完全康复。</li>
		<li>根据机构兽医的建议, 提供术后镇痛;例如使用 Meloxicam (5 毫克/毫升肌肉)。</li>
		<li>手术6周后再开始实验。这样可以更好地将头部贴在头骨上, 保证动物完全从干预中恢复。</li>
	</ol>
</li>
	<li>使用 MRI (用于单单元细胞外记录) 和计算机断层扫描 (CT; 用于多电极录音) 对记录区域进行本地化。<ol>
<li>用2% 硫酸铜溶液填充玻璃毛细血管, 并将其插入记录网格中。</li>
		<li>进行结构 MRI (切片厚度: 0.6 毫米)。</li>
	</ol>
</li>
	<li>神经活动的监测。<ol>
<li>使用阻抗为 0.8-1 MΩ的钨电极。</li>
		<li>使用23G 不锈钢导水管和液压 microdrive 将电极通过硬脑膜插入。</li>
		<li>对于穗识别, 放大和过滤300和5000赫兹之间的神经活动。</li>
		<li>对于局部场电位 (LFP) 记录, 放大并过滤1至170赫兹之间的信号。</li>
	</ol>
</li>
	<li>监视眼睛信号<ol>
<li>在动物的眼睛前调整红外线摄像头, 以获得足够的瞳孔图像和角膜反射。</li>
		<li>使用红外线摄像头在500赫兹的瞳孔位置取样。</li>
	</ol>
</li>
</ol>2. 研究背部区域的对象选择性
<ol><li>执行视觉引导抓取 (VGG)。<ol>
<li>根据研究的目标选择正确的抓取设置: 旋转木马安装或机器人安装 (图 3)。</li>
		<li>对于旋转木马安装程序, 运行 VGG 任务:<ol>
<li>让猴子把手对在完全黑暗中的静止位置的记录半球, 以启动序列。</li>
			<li>在可变时间之后 (intertrial 间隔: 2,000-3, 000 毫秒), 应用红色激光 (固定点) 在对象的基地 (距离:28 cm 从猴子的眼睛)。如果动物保持它的凝视在电子定义的固定窗口 (+/-2.5°) 为 500 ms, 照亮对象从上面与光源。</li>
			<li>在可变延迟 (300-1500 毫秒) 后, 程序的激光调光 (视觉去提示) 指示猴子从静止位置抬起手, 并达到, 抓住和保持对象的可变时间间隔 (持有时间: 300-900 毫秒)。</li>
			<li>当动物正确地执行整个序列时, 用一滴果汁奖励它。</li>
		</ol>
</li>
		<li>对机器人设置使用类似的任务序列。<ol>
<li>至于旋转木马的设置, 让猴子把手对到记录半球在完全黑暗的静止位置, 以启动序列。</li>
			<li>在可变时间之后 (intertrial 间隔: 2,000-3, 000 毫秒), 照亮对象上的 LED (固定点) (从内; 距离:28 厘米从猴子的眼睛)。同样, 如果动物保持它的凝视在电子定义的固定窗口 (+/-2.5°) 为 500 ms, 照亮对象从内部与白色光源。</li>
			<li>在可变延迟 (300-1500 毫秒) 后, 关掉 LED (视觉去提示), 指示猴子从静止位置抬起手, 到达, 抓住并按住对象的可变间隔 (保持时间: 300-900 毫秒)。</li>
			<li>当动物正确地执行整个序列时, 用一滴果汁奖励它。</li>
		</ol>
</li>
		<li>在任务期间, 量化猴子的表现, 特别注意时间。测量在走信号和手运动开始 (反应时间) 之间的时间, 以及运动和物体升力之间的距离 (抓住时间)。</li>
	</ol></li>
	<li>执行记忆引导抓取 (MGG;"抓住黑暗")。使用 MGG 任务确定神经元是 visuomotor 的还是以运动为主的。 注意: 该序列类似于 VGG 所描述的顺序, 但对象在完全黑暗中被抓住。<ol>
<li>与 VGG 任务相同的是, 让猴子把手对在完全黑暗的静止位置上的记录半球开始序列。</li>
		<li>在可变时间之后 (intertrial 间隔: 2,000-3、000毫秒), 应用红色激光/LED (固定点) 表示固定点 (在对象的底部为旋转木马设置, 在对象的中心为机器人设置; 距离: 从猴子的眼睛28厘米).如果动物保持它的凝视在电子定义的固定窗口 (+/-2.5°) 为500毫秒, 照亮对象。</li>
		<li>经过固定的时间 (400 毫秒), 关掉灯。</li>
		<li>在一个可变延迟期 (300-1500 毫秒) 后, 光偏移, 昏暗/关闭固定点 (去提示), 以指示猴子举起手和到达, 抓住, 并持有的对象 (持有时间: 300-900 毫秒)。</li>
		<li>每当动物正确地执行整个序列, 给一滴果汁作为奖励。</li>
	</ol></li>
	<li>进行被动固定。至于 VGG 任务, 根据研究的目标选择最合适的设置 (旋转木马或机器人设置)。 注意: 可以执行两种不同的被动固定任务: 现实世界物体的被动固定 (使用在旋转木马和机器人设置中的对象) 和被动固定3维/2 维图像的物体。<ol>
<li>对现实世界中的物体进行被动固定。<ol>
<li>目前的固定点 (红色激光为旋转木马设置投射在对象的底部和红色 LED 在机器人设置)。</li>
			<li>如果动物保持它的凝视在电子定义的固定窗口 (+/-2.5°) 为 500 ms, 照亮对象为 2000 ms。</li>
			<li>如果动物保持它的凝视在窗口内为1000毫秒, 奖励它与一滴果汁。</li>
		</ol>
</li>
		<li>对物体进行3维/2 维图像的被动固定。<ol>
<li>在黑色背景上呈现所有视觉刺激 (亮度为 8 cd/米2), 使用显示器 (分辨率为 1280 x 1024 像素), 配有快速衰减 P46-phosphor, 并在120赫兹 (查看距离:86 厘米) 操作。</li>
			<li>在3D 测试中, 通过在显示器 (CRT 显示器) 上交替左、右眼图像, 结合两个铁电液晶百叶窗, 呈现刺激立体感。找到这些百叶窗在猴子的眼睛前面, 操作在60赫兹和同步到显示器的垂直折回。</li>
			<li>通过在屏幕中央显示一个小正方形 (固定点; 0.2° x 0.2°) 开始试用。如果眼睛位置保持在电子定义的1°正方形窗口 (比实际对象小得多) 至少500毫秒, 则在屏幕上呈现视觉刺激, 总时间为500毫秒。</li>
			<li>当猴子保持稳定的固定, 直到刺激抵消, 奖励它一滴果汁。</li>
			<li>为了对形状选择性进行充分的研究, 在被动固定任务中, 在以下序列中运行一组2D 图像的综合测试电池。</li>
			<li>运行搜索测试。使用一组广泛的图像 (表面图像) 测试单元格的视觉选择性;图 4A), 包括在 VGG 中掌握的对象的图片。对于这个和所有后续的视觉任务, 将最强响应 (称为 "首选图像") 的图像与神经元响应较弱的第二个图像 (称为 "nonpreferred 图像") 进行比较。如果研究中的神经元也对物体的图像作出反应, 则寻找驱动细胞反应性的特定刺激成分 (轮廓测试、接受田间试验和还原试验)。</li>
			<li>运行轮廓测试。从真实物体的原始表面图像 (包含纹理、阴影和透视的2D 或3D 图像), 获得相同刺激形状 (剪影和轮廓) 的渐进简化版本;图 4B)。每个条件收集至少10个试验, 以确定神经元是否喜欢原始的表面, 剪影或轮廓从原始的形状。</li>
			<li>运行一个接受域 (RF) 测试。为了映射神经元的 RF, 在显示器上显示不同位置的物体的图像 (在此实验中, 35 位置; 3°的刺激大小), 覆盖中央视野19,20。在合理的时间内在所有可能的位置收集足够的刺激重复, 减少刺激持续时间 (闪光刺激; 刺激持续时间: 300 毫秒, intertrial 间隔: 300 毫秒)。</li>
			<li>运行还原测试。使用在 RF 中心显示的轮廓碎片进行还原测试, 以确定最小有效形状特征 (MESF)。通过裁剪主坐标轴上每个原始轮廓形状的轮廓 (图 3B), 在 Photoshop 中生成一组刺激。将 MESF 设计为最小的形状片段, 它唤起的响应至少是完整的轮廓响应的 70%, 而不是显着小于该响应8。</li>
			<li>为了更好地估计位置依赖性 (刺激位置对片段选择性的影响), 运行两个不同的测试。使用位于原始大纲形状中所占位置的碎片运行缩减测试。在形状的质心处运行还原测试。</li>
			<li>在此阶段, 使用 MESF 运行一个新的 RF 映射。</li>
		</ol>
</li>
	</ol></li>
</ol>

<ol><li>Gallese, V., Fadiga, L., Fogassi, L., Rizzolatti, G. <a target="_blank" href="https://www.ncbi.nlm.nih.gov/pubmed/8800951">Action recognition in the premotor cortex.</a> Brain. 119 (2), 593-609 (1996).</li>
<li>Fogassi, L., Gallese, V., Buccino, G., Craighero, L., Fadiga, L., Rizzolatti, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Cortical+mechanism+for+the+visual+guidance+of+hand+grasping+movements+in+the+monkey%3A+a+reversible+inactivation+study%5BTitle%5D)+AND+%22Brain%22%5BJournal%5D)">Cortical mechanism for the visual guidance of hand grasping movements in the monkey: a reversible inactivation study.</a> Brain. 124 (3), 571-586 (2001).</li>
<li>Rizzolatti, G., Camarda, R., Fogassi, L., Gentilucci, M., Luppino, G., Matelli, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Functional+organization+of+inferior+area+6+in+the+macaque+monkey.+II.+Area+F5+and+the+control+of+distal+movements%5BTitle%5D)+AND+%22Exp.+Brain+Res%22%5BJournal%5D)">Functional organization of inferior area 6 in the macaque monkey. II. Area F5 and the control of distal movements.</a> Exp. Brain Res. 71 (3), 491-507 (1988).</li>
<li>Mishkin, M., Ungerleider, L. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Contribution+of+striate+inputs+to+the+visuospatial+functions+of+parieto-preoccipital+cortex+in+monkeys%5BTitle%5D)+AND+%22Behav.+Brain+Res%22%5BJournal%5D)">Contribution of striate inputs to the visuospatial functions of parieto-preoccipital cortex in monkeys.</a> Behav. Brain Res. 6 (1), 57-77 (1982).</li>
<li>Goodale, M. A., Milner, A. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Separate+visual+pathways+for+perception+and+action%5BTitle%5D)+AND+%22Trends+Neurosci%22%5BJournal%5D)">Separate visual pathways for perception and action.</a> Trends Neurosci. 15 (1), 20-25 (1992).</li>
<li>Baumann, M. A., Fluet, M. C., Scherberger, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Context-specific+grasp+movement+representation+in+the+macaque+anterior+intraparietal+area%5BTitle%5D)+AND+%22J.+Neurosci%22%5BJournal%5D)">Context-specific grasp movement representation in the macaque anterior intraparietal area.</a> J. Neurosci. 29 (20), 6436-6438 (2009).</li>
<li>Murata, A., Gallese, V., Luppino, G., Kaseda, M., Sakata, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Selectivity+for+the+shape%2C+size%2C+and+orientation+of+objects+for+grasping+neurons+of+monkey+parietal+area+AIP%5BTitle%5D)+AND+%22J.+Neurophysiol%22%5BJournal%5D)">Selectivity for the shape, size, and orientation of objects for grasping neurons of monkey parietal area AIP.</a> J. Neurophysiol. 83 (5), 2580-2601 (2000).</li>
<li>Romero, M. C., Pani, P., Janssen, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Coding+of+shape+features+in+the+macaque+anterior+intraparietal+area%5BTitle%5D)+AND+%22J.+Neurosci%22%5BJournal%5D)">Coding of shape features in the macaque anterior intraparietal area.</a> J. Neurosci. 34 (11), 4006-4021 (2014).</li>
<li>Sakata, H., Taira, M., Kusonoki, M., Murata, A., Tanaka, Y., Tsutsui, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Neural+coding+of+3D+features+of+objects+for+hand+action+in+the+parietal+cortex+of+the+monkey%5BTitle%5D)+AND+%22Philos.+Trans.+R.+Soc.+Lond.+B.+Biol.+Sci%22%5BJournal%5D)">Neural coding of 3D features of objects for hand action in the parietal cortex of the monkey.</a> Philos. Trans. R. Soc. Lond. B. Biol. Sci. 353 (1373), 1363-1373 (1998).</li>
<li>Taira, M., Mine, S., Georgopoulos, A. P., Murata, A., Sakata, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Parietal+cortex+neurons+of+the+monkey+related+to+the+visual+guidance+of+the+hand+movement%5BTitle%5D)+AND+%22Exp+Brain+Res%22%5BJournal%5D)">Parietal cortex neurons of the monkey related to the visual guidance of the hand movement.</a> Exp Brain Res. 83 (1), 29-36 (1990).</li>
<li>Janssen, P., Scherberger, H. <a target="_blank" href="https://www.ncbi.nlm.nih.gov/pubmed/25840009">Visual guidance in control of grasping.</a> Annu. Rev. Neurosci. 8 (38), 69-86 (2015).</li>
<li>Theys, T., Pani, P., van Loon, J., Goffin, J., Janssen, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Selectivity+for+three-dimensional+contours+and+surfaces+in+the+anterior+intraparietal+area%5BTitle%5D)+AND+%22J.+Neurophysiol%22%5BJournal%5D)">Selectivity for three-dimensional contours and surfaces in the anterior intraparietal area.</a> J. Neurophysiol. 107 (3), 995-1008 (2012).</li>
<li>Goffin, J., Janssen, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Three-dimensional+shape+coding+in+grasping+circuits%3A+a+comparison+between+the+anterior+intraparietal+area+and+ventral+premotor+area+F5a%5BTitle%5D)+AND+%22J.+Cogn.+Neurosci%22%5BJournal%5D)">Three-dimensional shape coding in grasping circuits: a comparison between the anterior intraparietal area and ventral premotor area F5a.</a> J. Cogn. Neurosci. 25 (3), 352-364 (2013).</li>
<li>Raos, V., Umiltá, M. A., Murata, A., Fogassi, L., Gallese, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Functional+properties+of+grasping-related+neurons+in+the+ventral+premotor+area+F5+of+the+macaque+monkey%5BTitle%5D)+AND+%22J.+Neurophysiol%22%5BJournal%5D)">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/pubmed?term=((Simultaneous+recording+of+macaque+premotor+and+primary+motor+cortex+neuronal+populations+reveals+different+functional+contributions+to+visuomotor+grasp%5BTitle%5D)+AND+%22J.+Neurophysiol%22%5BJournal%5D)">Simultaneous recording of macaque premotor and primary motor cortex neuronal populations reveals different functional contributions to visuomotor grasp.</a> J. Neurophysiol. 98 (1), 488-501 (2007).</li>
<li>Denys, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+processing+of+visual+shape+in+the+cerebral+cortex+of+human+and+nonhuman+primates%3A+a+functional+magnetic+resonance+imaging+study%5BTitle%5D)+AND+%22J.+Neurosci%22%5BJournal%5D)">The processing of visual shape in the cerebral cortex of human and nonhuman primates: a functional magnetic resonance imaging study.</a> J. 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/pubmed?term=((Selectivity+for+three-dimensional+shape+and+grasping-related+activity+in+the+macaque+ventral+premotor+cortex%5BTitle%5D)+AND+%22J.Neurosci%22%5BJournal%5D)">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/pubmed?term=((Observing+others%3A+multiple+action+representation+in+the+frontal+lobe%5BTitle%5D)+AND+%22Science%22%5BJournal%5D)">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/pubmed?term=((Coding+of+shape+and+position+in+macaque+lateral+intraparietal+area%5BTitle%5D)+AND+%22J.+Neurosci%22%5BJournal%5D)">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/pubmed?term=((Receptive+field+properties+of+neurons+in+the+macaque+anterior+intraparietal+area%5BTitle%5D)+AND+%22J.+Neurophysiol%22%5BJournal%5D)">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="https://lirias.kuleuven.be/handle/123456789/611191">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. , Washington DC: Society for Neuroscience. Online (2017).</li>
<li>Pani, P., Theys, T., Romero, M. C., Janssen, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Grasping+execution+and+grasping+observation+activity+of+single+neurons+in+macaque+anterior+intraparietal+area%5BTitle%5D)+AND+%22J.+Cogn.+Neurosci%22%5BJournal%5D)">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/pubmed?term=((The+role+of+the+prefrontal+cortex+in+familiarity+and+recollection+processes+during+verbal+and+non-verbal+recognition+memory%5BTitle%5D)+AND+%22Neuroimage%22%5BJournal%5D)">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/pubmed?term=((Patches+of+faces-selective+cortex+in+the+macaque+frontal+lobe%5BTitle%5D)+AND+%22Nat.+Neurosci%22%5BJournal%5D)">Patches of faces-selective cortex in the macaque frontal lobe.</a> Nat. Neurosci. 11 (8), 877-879 (2008).</li>
</ol>

作者没有什么可透露的。

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/&amp;dagger;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&#039;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 ...

investigating-object-representations-macaque-dorsal-visual-stream

Research

JoVE Journal

Neuroscience

8.3K 次观看.  Katholieke Universiteit Leuven. 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。

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

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

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 nerve. In vivo recordings of ganglion cell spike trains in several animal models have revealed much of what is known about how the early visual system processes and encodes visual information. However, such recordings have been rare in one of the most common animal models, the rat, possibly owing to difficulty in detecting spikes fired by small diameter axons. The many retinal disease models involving rats motivate a need for characterizing the functional properties of ganglion cells without disturbing the eye, as with intraocular or in vitro recordings. Here, we demonstrate a method for recording ganglion cell spike trains from the optic chiasm of the anesthetized rat. We first show how to fabricate tungsten-in-glass electrodes that can pick up electrical activity from single ganglion cell axons in rat. The electrodes outperform all commercial ones that we have tried. We then illustrate our custom-designed stereotaxic system for in vivo visual neurophysiology experiments and our procedures for animal preparation and reliable and stable electrode placement in the optic chiasm.

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

Retinal ganglion cells transmit visual information from the eye to the brain with sequences of action potentials. Here, we demonstrate how to record the action potentials of single ganglion cells in vivo from anesthetized rats.

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

Automated Visual Cognitive Tasks for Recording Neural Activity Using a Floor Projection Maze

Neuropsychological tasks used in primates to investigate mechanisms of learning and memory are typically visually guided cognitive tasks. We have developed visual cognitive tasks for rats using the Floor Projection Maze1,2&#160;that are optimized for visual abilities of rats permitting stronger comparisons of experimental findings with other species.
In order to investigate neural correlates of learning and memory, we have integrated electrophysiological recordings into fully automated cognitive tasks on the Floor Projection Maze1,2. Behavioral software interfaced with an animal tracking system allows monitoring of the animal's behavior with precise control of image presentation and reward contingencies for better trained animals. Integration with an in vivo electrophysiological recording system enables examination of behavioral correlates of neural activity at selected epochs of a given cognitive task.
We describe protocols for a model system that combines automated visual presentation of information to rodents and intracranial reward with electrophysiological approaches. Our model system offers a sophisticated set of tools as a framework for other cognitive tasks to better isolate and identify specific mechanisms contributing to particular cognitive processes.

We describe protocols for training rats for chronic electrophysiological recordings in fully automated cognitive tasks on a Floor Projection Maze.

使用地面投影迷宫自动化视觉认知任务记录神经活动

Neuropsychological tasks used in primates to investigate mechanisms of learning and memory are typically visually guided cognitive tasks. We have ...

行为学

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

One of the central tasks in retinal neuroscience is to understand the circuitry of retinal neurons and how those connections are responsible for shaping the signals transmitted to the brain. Photons are detected in the retina by rod and cone photoreceptors, which convert that energy into an electrical signal, transmitting it to other retinal neurons, where it is processed and communicated to central targets in the brain via the optic nerve. Important early insights into retinal circuitry and visual processing came from the histological studies of Cajal1,2 and, later, from electrophysiological recordings of the spiking activity of retinal ganglion cells - the output cells of the retina3,4.
A detailed understanding of visual processing in the retina requires an understanding of the signaling at each step in the pathway from photoreceptor to retinal ganglion cell. However, many retinal cell types are buried deep in the tissue and therefore relatively inaccessible for electrophysiological recording. This limitation can be overcome by working with vertical slices, in which cells residing within each of the retinal layers are clearly visible and accessible for electrophysiological recording.
Here, we describe a method for making vertical sections of retinas from larval tiger salamanders (Ambystoma tigrinum). While this preparation was originally developed for recordings with sharp microelectrodes5,6, we describe a method for dual whole-cell voltage clamp recordings from photoreceptors and second-order horizontal and bipolar cells in which we manipulate the photoreceptor's membrane potential while simultaneously recording post-synaptic responses in horizontal or bipolar cells. The photoreceptors of the tiger salamander are considerably larger than those of mammalian species, making this an ideal preparation in which to undertake this technically challenging experimental approach. These experiments are described with an eye toward probing the signaling properties of the synaptic ribbon - a specialized synaptic structure found in a only a handful of neurons, including rod and cone photoreceptors, that is well suited for maintaining a high rate of tonic neurotransmitter release7,8 - and how it contributes to the unique signaling properties of this first retinal synapse.

We describe the preparation of thin retinal slices from aquatic tiger salamanders (Ambystoma tigrinum) and explain how we use these slices to study synaptic processing in the retina by obtaining dual whole-cell voltage clamp recordings from photoreceptors and second-order horizontal and bipolar cells.

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 ...

Recording Single Neurons' Action Potentials from Freely Moving Pigeons Across Three Stages of Learning

While the subject of learning has attracted immense interest from both behavioral and neural scientists, only relatively few investigators have observed single-neuron activity while animals are acquiring an operantly conditioned response, or when that response is extinguished. But even in these cases, observation periods usually encompass only a single stage of learning, i.e. acquisition or extinction, but not both (exceptions include protocols employing reversal learning; see Bingman&#160;et al.1 for an example). However, acquisition and extinction entail different learning mechanisms and are therefore expected to be accompanied by different types and/or loci of neural plasticity.
Accordingly, we developed a behavioral paradigm which institutes three stages of learning in a single behavioral session and which is well suited for the simultaneous recording of single neurons&#39; action potentials. Animals are trained on a single-interval forced choice task which requires mapping each of two possible choice responses to the presentation of different novel visual stimuli (acquisition). After having reached a predefined performance criterion, one of the two choice responses is no longer reinforced (extinction). Following a certain decrement in performance level, correct responses are reinforced again (reacquisition). By using a new set of stimuli in every session, animals can undergo the acquisition-extinction-reacquisition process repeatedly. Because all three stages of learning occur in a single behavioral session, the paradigm is ideal for the simultaneous observation of the spiking output of multiple single neurons. We use pigeons as model systems, but the task can easily be adapted to any other species capable of conditioned discrimination learning.

Learning new stimulus-response associations engages a wide range of neural processes which are ultimately reflected in changing spike output of individual neurons. Here we describe a behavioral protocol allowing for the continuous registration of single-neuron activity while animals acquire, extinguish, and reacquire a conditioned response within a single experimental session.

自由移动鸽子穿越三个阶段学习记录单个神经元&#39;动作电位

While the subject of learning has attracted immense interest from both behavioral and neural scientists, only relatively few investigators have observed ...

Simultaneous Long-term Recordings at Two Neuronal Processing Stages in Behaving Honeybees

In both mammals and insects neuronal information is processed in different higher and lower order brain centers. These centers are coupled via convergent and divergent anatomical connections including feed forward and feedback wiring. Furthermore, information of the same origin is partially sent via parallel pathways to different and sometimes into the same brain areas. To understand the evolutionary benefits as well as the computational advantages of these wiring strategies and especially their temporal dependencies on each other, it is necessary to have simultaneous access to single neurons of different tracts or neuropiles in the same preparation at high temporal resolution. Here we concentrate on honeybees by demonstrating a unique extracellular long term access to record multi unit activity at two subsequent neuropiles1, the antennal lobe (AL), the first olfactory processing stage and the mushroom body (MB), a higher order integration center involved in learning and memory formation, or two parallel neuronal tracts2 connecting the AL with the MB.&#160;The latter was chosen as an example and will be described in full. In the supporting video the construction and permanent insertion of flexible multi&#160;channel wire electrodes is demonstrated. Pairwise differential amplification of the micro&#160;wire electrode channels drastically reduces the noise and verifies that the source of the signal is closely related to the position of the electrode tip. The mechanical flexibility of the used wire electrodes allows stable invasive long&#160;term recordings over many hours up to days, which is a clear advantage compared to conventional extra&#160;and intracellular in vivo recording techniques.

Simultaneous extracellular long term recordings from two different brain neuropiles or two different anatomical tracts were established in honeybees. These recordings allow the investigation of temporal aspects of neuronal processing across different brain areas at the single neuron as well as at the ensemble level in a behaving animal.

在两个神经元处理阶段在做人蜜蜂同时长期录音

In both mammals and insects neuronal information is processed in different higher and lower order brain centers. These centers are coupled via convergent ...

Convection Enhanced Delivery of Optogenetic Adeno-associated Viral Vector to the Cortex of Rhesus Macaque Under Guidance of Online MRI Images

In non-human primate (NHP) optogenetics, infecting large cortical areas with viral vectors is often a difficult and time-consuming task. Here, we demonstrate the use of magnetic resonance (MR)-guided convection enhanced delivery (CED) of optogenetic viral vectors into primary somatosensory (S1) and motor (M1) cortices of macaques to obtain efficient, widespread cortical expression of light-sensitive ion channels. Adeno-associated viral (AAV) vectors encoding the red-shifted opsin C1V1 fused to yellow fluorescent protein (EYFP) were injected into the cortex of rhesus macaques under MR-guided CED. Three months post-infusion, epifluorescent imaging confirmed large regions of optogenetic expression (&#62;130 mm2) in M1 and S1 in two macaques. Furthermore, we were able to record reliable light-evoked electrophysiology responses from the expressing areas using micro-electrocorticographic arrays. Later histological analysis and immunostaining against the reporter revealed widespread and dense optogenetic expression in M1 and S1 corresponding to the distribution indicated by epifluorescent imaging. This technique enables us to obtain expression across large areas of the cortex within a shorter period of time with minimal damage compared to the traditional techniques and can be an optimal approach for optogenetic viral delivery in large animals such as NHPs. This approach demonstrates great potential for network-level manipulation of neural circuits with cell-type specificity in animal models evolutionarily close to humans.&#160;

Here, we demonstrate magnetic resonance (MR)-guided convection enhanced delivery (CED) of viral vectors into the cortex as an efficient and simplified approach for achieving optogenetic expression across large cortical areas in the macaque brain.

在在线 MRI 图像的指导下,对流增强将视遗传阿德诺相关病毒载体输送到 Rhesus Macaque 的皮质

In non-human primate (NHP) optogenetics, infecting large cortical areas with viral vectors is often a difficult and time-consuming task. Here, we ...

开发用于狨猴精确神经记录的 x-y 电极台

Electrophysiology of Laminar Cortical Activity in the Common Marmoset

The marmoset monkey provides an ideal model for examining laminar cortical circuits due to its smooth cortical surface, which facilitates recordings with linear arrays. The marmoset has recently grown in popularity due to its similar neural functional organization to other primates and its technical advantages for recording and imaging. However, neurophysiology in this model poses some unique challenges due to the small size and lack of gyri as anatomical landmarks. Using custom-built micro-drives, researchers can manipulate linear array placement to sub-millimeter precision and reliably record at the same retinotopically targeted location across recording days. This protocol describes the step-by-step construction of the micro-drive positioning system and the neurophysiological recording technique with silicon linear electrode arrays. With precise control of electrode placement across recording sessions, researchers can easily traverse the cortex to identify areas of interest based on their retinotopic organization and the tuning properties of the recorded neurons. Further, using this laminar array electrode system, it is possible to apply a current source density analysis (CSD) to determine the recording depth of individual neurons. This protocol also demonstrates examples of laminar recordings, including spike waveforms isolated in Kilosort, which span multiple channels on the arrays.

作者聚焦:优化 Micro-Drive Systems 以实现 Marmoset 中的精确电极定位

Custom-built micro-drives enable the sub-millimeter targeting of cortical recording sites with linear silicon arrays.

普通狨猴层流皮层活动的电生理学

The marmoset monkey provides an ideal model for examining laminar cortical circuits due to its smooth cortical surface, which facilitates recordings with ...

<meta charset="utf8"></head><body>小鼠感觉皮层发育中的神经元活动映射

In Vivo Visualization of Spontaneous Activity in Neonatal Mouse Sensory Cortex at a Single-Neuron Resolution

The mammalian brain undergoes dynamic developmental changes at both the cellular and circuit levels throughout prenatal and postnatal periods. Following the discovery of numerous genes contributing to these developmental changes, it is now known that neuronal activity also substantially modulates these processes. In the developing cerebral cortex, neurons exhibit synchronized activity patterns that are specialized to each primary sensory area. These patterns markedly differ from those observed in the mature cortex, emphasizing their role in regulating area-specific developmental processes. Deficiencies in neuronal activity during development can lead to various brain diseases. These findings highlight the need to examine the regulatory mechanisms underlying activity patterns in neuronal development. This paper summarizes a series of protocols to visualize primary sensory areas and neuronal activity in neonatal mice, to image the activity of individual neurons within the cortical subfields using two-photon microscopy in vivo, and to analyze subfield-related activity correlations. We show representative results of patchwork-like synchronous activity within individual barrels in the somatosensory cortex. We also discuss various potential applications and some limitations of this protocol.

<meta charset="utf8"></head><body>作者聚焦：从双光子显微镜和单神经元成像破译神经回路形成

Primary sensory areas in the neocortex exhibit unique spontaneous activities during development. This article describes how to visualize individual neuron activities and primary sensory areas to analyze area-specific synchronous activities in neonatal mice in vivo.

在单神经元分辨率下新生小鼠感觉皮层自发活动的体内可视化

The mammalian brain undergoes dynamic developmental changes at both the cellular and circuit levels throughout prenatal and postnatal periods. Following ...

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在单神经元分辨率下新生小鼠感觉皮层自发活动的体内可视化