用于这些实验的拉什利三迷宫建添Bowmaster，保罗Corman，巴里Dutrow，瑞安Jabco，从宾夕法尼亚州立大学物理系机店和蒂姆Treaster。作者想感谢Shoba Belegundu女士，女士塔拉Chrzanowski，女士和亚历山德拉刘易斯协助执行在图2-4中所提出的实验。 Walter先生杰克逊在本议定书的准备提供宝贵的意见。这项工作是从美国国立精神卫生研究所（MH064756 MH077978到玛）的资金支持。内容完全是作者的责任，并不一定代表的国家精神健康或美国国​​立卫生研究院研究所的官方意见。

1。制备方法: <ol><li>拉什利三迷宫建造的有机玻璃，如图1所示。开始框，迷宫武器，和目标框是模块化的，允许清洁和可变性的迷宫。开始框，迷宫武器，和目标框的墙壁是黑色树脂玻璃构造，以消除视觉线索，从外部和内部的迷宫。迷宫武器是长45厘米，高7厘米，5厘米宽的。门通往迷宫的连续武器4厘米× 4厘米，位于迷宫的外墙11厘米。目标框长19.5厘米x 7厘米高x宽5厘米。启动框为8厘米× 9.5厘米× 7厘米。独立明确的有机玻璃块放在顶部开始框，迷宫，和目标框，让每个鼠标的可视化，因为它通过迷宫的进展，同时防止老鼠逃跑。迷宫由基地构建了一个大到足以容纳迷宫加伪家里的笼子里是作为动物的标准的家庭笼的大小相同的三件红色有机玻璃。 </li><li>为了准备伪家笼前的第一个迷宫试验，在动物的家笼中使用的是添加到个人的笼子底部有门道（2厘米× 2厘米），切成双方同类新鲜的床上用品材料。放置导线上装上这些笼底部，并单独编号，为每一个测试队列中的动物之一。这些笼子将作为个别伪家里每个动物笼测试时间表的时间。因为我们正在调查的应力的作用和/或焦虑相关的学习和记忆的行为，我们用伪家笼，以避免个别住房小鼠在自己的笼子。但是，如果这些因素不关心的首要，动物可以被单独安置在门口和这些笼子修改，可以作为成功的迷宫导航首页笼的家庭笼。 </li></ol> 2。测试: <ol><li>每一天，动物行为测试室至少一小时前移到测试，使他们能够适应环境的测试环境。 </li><li> 30分钟测试开始的时间，食物和水前拆除所有家庭的笼子。保持在行为测试室，但小鼠达到了食物和水。食物和水通常是30分钟后取出的暗相轻相变化，后发生高峰期喂养。这确保动物大多食物和水酒足饭饱前测试，然而，在此期间的测试，提供一小笔额外的原动力导航迷宫，即使是没有公开的食物或水任务完成后奖励。 </li><li>预试期已过30分钟后，用70％乙醇清洁迷宫。敞开大门，从一开始就中的领先进入迷宫武器封闭。在目标框两端的门是处于打开状态。 </li><li>将伪家庭相应的目标框年底的第一个动物笼。 </li><li>卡里的第一个鼠标迷宫，拔罐，在你的手。在开始框中鼠标和更换盖子。 </li><li>当位置观察有无梗阻的迷宫，打开启动箱门，并立即启动秒表。 </li><li>当鼠标的所有四个爪子已经离开了开始框，关闭启动箱门和停止一个计时器。 </li><li>手工记录的途径鼠标旅行，因为它的迷宫导航。一个鼠标进入一个新的区域时，所有四个爪子已经进入该地区越过迷宫（参见图1）或的手臂。 </li><li>当所有四个爪子进入目标中，领导之间的迷宫和目标框，关门停止秒的秒表。 </li><li>当鼠标已经完全进入的伪家笼，关闭门之间的目标框笼。停止第三秒表。 </li><li>从三个秒表记录时间。计数鼠标的错误。定义为一个错误是进入一个死胡同的尽头囊区的一个条目（例如，从手臂大号到K区;，图1）或穿越迷宫已经走过手臂（例如，会从ARM C臂F到图1）。虽然我们的得分鼠标的性能，因为它是发生，这是录像带或以其他方式记录的迷宫试验，为随后的进球让眼睛或自动化的软件。实验人员的自由裁量权，应选择一个合适的方法。 </li><li>返回到其家笼之前，留在1分钟的伪家笼的动物。 </li><li>重复步骤2.3至2.12的测试队列中的所有小鼠。 </li><li>当鼠标已经在队列中的最后返回到其家笼中，开始30分钟计时器。在这30分钟内结束，返回在队列中的所有小鼠的食物和水，然后返回他们的殖民地里的动物。 </li><li>连续两晚，重复此过程中，每一个测试天每只动物的试验，直到在队列中的所有老鼠都达到了定义的学习标准，直到预定的Ñ赭测试试验发生。鼠标被列为学到的迷宫时，它可以执行与0或1的错误（S），连续两个试验任务。在相同的顺序在连续试验测试小鼠。 </li></ol> 3。代表性的成果: 学习（达到标准）的天数，可以分析和比较不同群体之间在一个特定的研究（图2A和3A）。此外，一个学习型指数（学习的比例），迷宫收购的特点，可以评估在第4天测试期间（图2B和3B）。学习指数是正确的路径段数的比例总数的路径段走遍每个审判。平均学习指数应约0.5试验1，当老鼠是第一次接触时的迷宫，一个偶然的机会导航。学习指数升幅接近1和试验2和说明4之间，学习是发生。男性C57Bl/6NCr在图2中，青年（2月）和老龄化（24个月）的小鼠进行了培训长达15试验拉什利迷宫。 7.2中了解到，年轻老鼠迷宫（± 1.5）D.相比之下，老化的动物了11.7（± 2.1）d达到标准。老化C57Bl/6NCr小鼠有一个强烈的趋势，需要更长的时间来学习的迷宫（[T（23）= 1.59，P = 0.056];单尾t检验，先验假设天的学习时间较长老龄化小鼠）。当学习指数的计算方法，这两个年龄组，从试验2 4（主要效果显着的时间，P &lt;0.001），通过学习指数稳步上升。然而，有年轻的C57小鼠和旧的C57小鼠（年龄，P = 0.58的主要作用）之间的学习指数无显着差异。 图3是有代表性的研究背景应变的影响，对学习行为的研究。年轻男性C57Bl/6NCr的数据，如图所示。 2，现在比较年轻CRL的数据:的CD - 1（ICR）BR小鼠的CD - 1，莫4-5]。 CD - 1小鼠的经验教训4.5拉什利三迷宫（± 1.4）D，然而，有天的菌株之间没有显着差异，学习（图3A; [T（18）= 1.328，P = 0.20];尾t检验）。相比之下，检查的学习，而所有的动物导航一审偶然的迷宫，CD - 1小鼠学习的任务更加迅速（应变，P &lt;0.001的主要作用）的指数显示，。 CD - 1小鼠在学习指数显着增加，在临床试验中2至4 C57Bl/6NCr小鼠相比，说明了这一点。有趣的是，如果继续在达到标准之外的迷宫训练的CD - 1小鼠，他们表现出的行为表明过度训练（图4）这是在下面详细讨论。 <img src="/files/ftp_upload/1786/1786fig1.jpg" alt="图1" /> 图1:拉什利三迷宫死胡同CUL - DE - SAC的迷宫区，仅用于演示目的是在这个图中的红色。基地和武器的实际迷宫纯色（插图）。迷宫是按比例绘制（文字尺寸）。 <img src="/files/ftp_upload/1786/1786fig2.jpg" alt="图2" /> 图2:在2和24月龄的雄性C57Bl/6NCr小鼠第三拉什利迷宫性能拉什利三迷宫被用来评估年轻（2个月）和老龄化（24个月）的男性C57Bl/6NCr小鼠的学习路线。测量的参数是:（一 ）标准，鼠标运行连续两晚0错误或1个错误的迷宫所需的天数天;及（b）学习指数，正确的四个爪子手臂条目的数量作出与拉什利三迷宫测试1-4天的手臂条目总数。每组动物的数量为N = 10 2月龄和24月龄小鼠N = 15。 <img src="/files/ftp_upload/1786/1786fig3.jpg" alt="图3" /> 图3:拉什利第三年轻的男性C57Bl/6NCr和CD - 1小鼠的迷宫性能拉什利三迷宫是用来评估路线，在2个月大的C57Bl/6NCr和4-5个月大的CD - 1小鼠学习。测量的参数是:（一 ）标准，鼠标运行连续两晚0错误或1个错误的迷宫所需的天数天;及（b）学习指数，正确的四个爪子手臂条目的数量作出与拉什利三迷宫测试1-4天的手臂条目总数。每组动物的数量，N = 10 C57Bl/6NCr和N = 10，CD - 1小鼠。统计意义:* P &lt;0.05，** P &lt;0.01与C57Bl/6NCr小鼠。 <img src="/files/ftp_upload/1786/1786fig4.jpg" alt="图4" /> 图4:拉什利III在4-5个月大的雄性CD - 1小鼠的迷宫性能拉什利三迷宫 。是用来评估十个4-5月龄雄性CD - 1小鼠连续15天的学习路线。测量参数分别为:（一）每天的错误的数量，与定义到一条死胡同的尽头囊区的四个爪子入境或旅行的迷宫已经走过手臂错误;及（b）所花费的时间在每天的迷宫。该标准是达到一天是用红色箭头表示。

<ol>
	<li>Blizard, 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=Return+to+home+cage'+as+a+reward+for+maze+learning+in+young+and+old+genetically+heterogeneous+mice.">Return to home cage' as a reward for maze learning in young and old genetically heterogeneous mice.</a> Comparative Med. 56, 196-201 (2006).</li><li>Crawley, J. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> What's wrong with my mouse?: Behavioral phenotyping of transgenic and knockout mice. , (2007).</li><li>Blizard, D. A., Klein, L. C., Cohen, R., McClearn, G. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+mouse-friendly+cognitive+task+suitable+for+use+in+aging+studies.">A novel mouse-friendly cognitive task suitable for use in aging studies.</a> Behav. Genet. 33, 181-189 (2003).</li><li>Warren, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Appetitive+learning+by+old+mice.">Appetitive learning by old mice.</a> Exp. Aging Res. 12, 99-105 (1986).</li><li>Lashley, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Brain+mechanisms+and+intelligence:+A+quantitative+study+of+injuries+to+the+brain.">Brain mechanisms and intelligence: A quantitative study of injuries to the brain.</a> , (1929).</li><li>Barnes, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Memory+deficits+associated+with+senescence:+A+neurophysiological+and+behavioral+study+in+the+rat.">Memory deficits associated with senescence: A neurophysiological and behavioral study in the rat.</a> J. Comp. Physiol. Psych. 93, 74-104 (1979).</li><li>Matzel, L. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Individual+differences+in+the+expression+of+a+general+learning+ability+in+mice.">Individual differences in the expression of a general learning ability in mice.</a> J. Neurosci. 23, 6423-6433 (2003).</li><li>Brandeis, R., Brandys, Y., Yehuda, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+use+of+the+Morris+water+maze+in+the+study+of+memory+and+learning.">The use of the Morris water maze in the study of memory and learning.</a> Int. J. Neurosci. 48, 29-69 (1989).</li><li>Hepler, D. J., Wenk, G. L., Cribbs, B. L., Olton, D. S., Coyle, 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=Memory+impairments+following+basal+forebrain+lesions.">Memory impairments following basal forebrain lesions.</a> Brain Res. 346, 8-14 (1985).</li><li>Sci, B. e. h. a. v. B. r. a. i. n. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> 2, 313-313 (1979).</li></ol>

所有的行为协议是由美国宾夕法尼亚州立大学机构动物护理和使用委员会批准。有没有财务披露报告。

最常用的啮齿类动物的行为测试，以评估学习和记忆Morris水迷宫和恐惧空调2变种。这些测试依赖于视觉或听觉线索的学习和厌恶的刺激（如，水，电击）激励动物完成的任务3。在啮齿类动物老化，老化人类感官的方式随着时间的推移下降。几个背景的小鼠株失去他们的听力或白内障的发展，因为他们的年龄，死亡率23.5 h禁水后已被证明在小鼠增加，因为他们4岁。因此，?...

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<td>Lashley III maze</td>
<td>&nbsp;</td>
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low-stress route learning using the lashley iii maze in mice

许多行为测试旨在评估在啮齿类动物的学习和记忆能力，尤其是小鼠，依靠视觉线索，食品和/或水剥夺，或其他厌恶刺激来激励的任务收购。动物的年龄，感官的方式恶化。例如，许多品系小鼠发展听力赤字或白内障。在引导任务收购期间小鼠所需的感官系统的变化，目前在解释衰老动物的学习变化的潜在的混淆。此外，使用厌恶刺激，激励学习任务的动物是比较差的敏感性时，小鼠强调潜在的混杂。为了最大限度地减少干扰的影响这些类型，我们已经实施了修改后的版本拉什利三迷宫。这个迷宫路线的学习，使小鼠学习导航通过反复暴露在低应力条件，例如黑暗的阶段，没有剥夺食物/水，一个迷宫，直到他们从一开始的位置路径导航，以0比伪家笼或依赖错误（1）连续两次试验。我们归类为一个低应力的行为测试，因为它不依靠厌恶刺激，鼓励探索的迷宫和任务的学习。该仪器包括一个模块化的开始框，4臂迷宫身体，和一个目标"框中。在目标框的结束是一个伪家笼包含类似的发现在动物的笼子和迷宫测试期间，具体到每个动物的床上用品。以前已经证明，这种伪家笼提供足够的奖励激励小鼠学习导航迷宫<sup&gt; 1</sup&gt;。在这里，我们目前在评估小鼠学习记忆的年龄差异比较两个不同的背景品系小鼠的学习行为的的背景下拉什利三迷宫程序的可视化。我们希望一个有吸引力的替代涉及更紧张的学习任务和/或视觉线索的行为测试，评估老龄化的影响或压力在小鼠中的漏洞感兴趣的其他研究人员将考虑这个迷宫。

Watch this Scientific Journal Video about Low-stress Route Learning Using the Lashley III Maze in Mice at JoVE.com

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

拉什利三迷宫是一个路线的学习任务，不依赖于厌恶刺激或视觉线索。因此，这是一个极具吸引力的选项，用于评估学习和记忆，特别是在衰老小鼠或其他压力是一个考虑因素。

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 ...

low-stress-route-learning-using-the-lashley-iii-maze-in-mice

Research

JoVE Journal

Neuroscience

17.8K 次观看.  Pennsylvania State University. 拉什利三迷宫是一个路线的学习任务，不依赖于厌恶刺激或视觉线索。因此，这是一个极具吸引力的选项，用于评估学习和记忆，特别是在衰老小鼠或其他压力是一个考虑因素。

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

Morris Water Maze Test for Learning and Memory Deficits in Alzheimer&#39;s Disease Model Mice

The Morris Water Maze (MWM) was first established by neuroscientist Richard G. Morris in 1981 in order to test hippocampal-dependent learning, including acquisition of spatial memoryand long-term spatial memory 1. The MWM is a relatively simple procedure typically consisting of six day trials, the main advantage being the differentiation between the spatial (hidden-platform) and non-spatial (visible platform) conditions 2-4. In addition, the MWM testing environment reduces odor trail interference 5. This has led the task to be used extensively in the study of the neurobiology and neuropharmacology of spatial learning and memory. The MWM plays an important role in the validation of rodent models for neurocognitive disorders such as Alzheimer&#x2019;s Disease 6, 7. In this protocol we discussed the typical procedure of MWM for testing learning and memory and data analysis commonly used in Alzheimer&#x2019;s disease transgenic model mice.

Morris Water Maze Test for Learning and Memory Deficits in Alzheimer&#039;s Disease Model Mice

The Morris Water Maze is a behavioral task to test hippocampal-dependent learning and memory. It has been widely used in the study of neurobiology, neuropharmacology and neurocognitive disorders in rodent models.

在阿尔茨海默氏病模型小鼠学习和记忆障碍的Morris水迷宫测试

The Morris Water Maze (MWM) was first established by neuroscientist Richard G. Morris in 1981 in order to test hippocampal-dependent learning, including ...

科研

神经科学

T-maze Forced Alternation and Left-right Discrimination Tasks for Assessing Working and Reference Memory in Mice

Forced alternation and left-right discrimination tasks using the T-maze have been widely used to assess working and reference memory, respectively, in rodents. In our laboratory, we evaluated the two types of memory in more than 30 strains of genetically engineered mice using the automated version of this apparatus. Here, we present the modified T-maze apparatus operated by a computer with a video-tracking system and our protocols in a movie format. The T-maze apparatus consists of runways partitioned off by sliding doors that can automatically open downward, each with a start box, a T-shaped alley, two boxes with automatic pellet dispensers at one side of the box, and two L-shaped alleys. Each L-shaped alley is connected to the start box so that mice can return to the start box, which excludes the effects of experimenter handling on mouse behavior. This apparatus also has an advantage that in vivo microdialysis, in vivo electrophysiology, and optogenetics techniques can be performed during T-maze performance because the doors are designed to go down into the floor. In this movie article, we describe T-maze tasks using the automated apparatus and the T-maze performance of &alpha;-CaMKII+/- mice, which are reported to show working memory deficits in the eight-arm radial maze task. Our data indicated that &alpha;-CaMKII+/- mice showed a working memory deficit, but no impairment of reference memory, and are consistent with previous findings using the eight-arm radial maze task, which supports the validity of our protocol. In addition, our data indicate that mutants tended to exhibit reversal learning deficits, suggesting that &alpha;-CaMKII deficiency causes reduced behavioral flexibility. Thus, the T-maze test using the modified automatic apparatus is useful for assessing working and reference memory and behavioral flexibility in mice.

This article presents the protocol of T-maze tests using a modified automated apparatus for assessing the learning and memory functions in mice.

T-迷宫强迫交替和评估工作，并参考在小鼠记忆的左，右的歧视任务

Forced alternation and left-right discrimination tasks using the T-maze have been widely used to assess working and reference memory, respectively, in ...

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

Signaling of information in the vertebrate central nervous system is often carried by populations of neurons rather than individual neurons. Also propagation of suprathreshold spiking activity involves populations of neurons. Empirical studies addressing cortical function directly thus require recordings from populations of neurons with high resolution. Here we describe an optical method and a deconvolution algorithm to record neural activity from up to 100 neurons with single-cell and single-spike resolution. This method relies on detection of the transient increases in intracellular somatic calcium concentration associated with suprathreshold electrical spikes (action potentials) in cortical neurons. High temporal resolution of the optical recordings is achieved by a fast random-access scanning technique using acousto-optical deflectors (AODs)1. Two-photon excitation of the calcium-sensitive dye results in high spatial resolution in opaque brain tissue2. Reconstruction of spikes from the fluorescence calcium recordings is achieved by a maximum-likelihood method. Simultaneous electrophysiological and optical recordings indicate that our method reliably detects spikes (&gt;97% spike detection efficiency), has a low rate of false positive spike detection (&lt; 0.003 spikes/sec), and a high temporal precision (about 3 msec) 3. This optical method of spike detection can be used to record neural activity in vitro and in anesthetized animals in vivo3,4.

Understanding the function of the vertebrate central nervous system requires recordings from many neurons because cortical function arises on the level of populations of neurons. Here we describe an optical method to record suprathreshold neural activity with single-cell and single-spike resolution, dithered random-access scanning. This method records somatic fluorescence calcium signals from up to 100 neurons with high temporal resolution. A maximum-likelihood algorithm deconvolves the underlying suprathreshold neural activity from the somatic fluorescence calcium signals. This method reliably detects spikes with high detection efficiency and a low rate of false positives and can be used to study neural populations in vitro and in vivo.

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 ...

Mapping Cortical Dynamics Using Simultaneous MEG/EEG and Anatomically-constrained Minimum-norm Estimates: an Auditory Attention Example

Magneto- and electroencephalography (MEG/EEG) are neuroimaging techniques that provide a high temporal resolution particularly suitable to investigate the cortical networks involved in dynamical perceptual and cognitive tasks, such as attending to different sounds in a cocktail party. Many past studies have employed data recorded at the sensor level only, i.e., the magnetic fields or the electric potentials recorded outside and on the scalp, and have usually focused on activity that is time-locked to the stimulus presentation. This type of event-related field / potential analysis is particularly useful when there are only a small number of distinct dipolar patterns that can be isolated and identified in space and time. Alternatively, by utilizing anatomical information, these distinct field patterns can be localized as current sources on the cortex. However, for a more sustained response that may not be time-locked to a specific stimulus (e.g., in preparation for listening to one of the two simultaneously presented spoken digits based on the cued auditory feature) or may be distributed across multiple spatial locations unknown a priori, the recruitment of a distributed cortical network may not be adequately captured by using a limited number of focal sources.
Here, we describe a procedure that employs individual anatomical MRI data to establish a relationship between the sensor information and the dipole activation on the cortex through the use of minimum-norm estimates (MNE). This inverse imaging approach provides us a tool for distributed source analysis. For illustrative purposes, we will describe all procedures using FreeSurfer and MNE software, both freely available. We will summarize the MRI sequences and analysis steps required to produce a forward model that enables us to relate the expected field pattern caused by the dipoles distributed on the cortex onto the M/EEG sensors. Next, we will step through the necessary processes that facilitate us in denoising the sensor data from environmental and physiological contaminants. We will then outline the procedure for combining and mapping MEG/EEG sensor data onto the cortical space, thereby producing a family of time-series of cortical dipole activation on the brain surface (or "brain movies") related to each experimental condition. Finally, we will highlight a few statistical techniques that enable us to make scientific inference across a subject population (i.e., perform group-level analysis) based on a common cortical coordinate space.

We use magneto- and electroencephalography (MEG/EEG), combined with anatomical information captured by magnetic resonance imaging (MRI), to map the dynamics of the cortical network associated with auditory attention.

同时MEG / EEG和解剖约束的最小模估计皮质动态映射:听觉注意示例

Magneto- and electroencephalography (MEG/EEG) are neuroimaging techniques that provide a high temporal resolution particularly suitable to investigate the ...

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

Voltage responses of insect photoreceptors and visual interneurons can be accurately recorded with conventional sharp microelectrodes. The method described here enables the investigator to measure long-lasting (from minutes to hours) high-quality intracellular responses from single Drosophila R1-R6 photoreceptors and Large Monopolar Cells (LMCs) to light stimuli. Because the recording system has low noise, it can be used to study variability among individual cells in the fly eye, and how their outputs reflect the physical properties of the visual environment. We outline all key steps in performing this technique. The basic steps in constructing an appropriate electrophysiology set-up for recording, such as design and selection of the experimental equipment are described. We also explain how to prepare for recording by making appropriate (sharp) recording and (blunt) reference electrodes. Details are given on how to fix an intact fly in a bespoke fly-holder, prepare a small window in its eye and insert a recording electrode through this hole with minimal damage. We explain how to localize the center of a cell's receptive field, dark- or light-adapt the studied cell, and to record its voltage responses to dynamic light stimuli. Finally, we describe the criteria for stable normal recordings, show characteristic high-quality voltage responses of individual cells to different light stimuli, and briefly define how to quantify their signaling performance. Many aspects of the method are technically challenging and require practice and patience to master. But once learned and optimized for the investigator's experimental objectives, it grants outstanding in vivo neurophysiological data.

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

Sharp microelectrodes enable accurate electrophysiological characterization of photoreceptor and visual interneuron output in living Drosophila. Here we show how to use this method to record high-quality voltage responses of individual cells to controlled light stimulation. This method is ideal for studying neural information processing in insect compound eyes.

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 ...

Microstate and Omega Complexity Analyses of the Resting-state Electroencephalography

Microstate and omega complexity are two reference-free electroencephalography (EEG) measures that can represent the temporal and spatial complexities of EEG data and have been widely used to investigate the neural mechanisms in some brain disorders. The goal of this article is to describe the protocol underlying EEG microstate and omega complexity analyses step by step. The main advantage of these two measures is that they could eliminate the reference-dependent problem inherent to traditional spectrum analysis. In addition, microstate analysis makes good use of high time resolution of resting-state EEG, and the four obtained microstate classes could match the corresponding resting-state networks respectively. The omega complexity characterizes the spatial complexity of the whole brain or specific brain regions, which has obvious advantage compared with traditional complexity measures focusing on the signal complexity in a single channel. These two EEG measures could complement each other to investigate the brain complexity from the temporal and spatial domain respectively.

This article describes the protocol underlying electroencephalography (EEG) microstate analysis and omega complexity analysis, which are two reference-free EEG measures and highly valuable to explore the neural mechanisms of brain disorders.

静止状态脑电图的微状态和欧米茄复杂性分析

Microstate and omega complexity are two reference-free electroencephalography (EEG) measures that can represent the temporal and spatial complexities of ...

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

Olfaction is the predominant sensory modality in mice and influences many important behaviors, including foraging, predator detection, mating, and parenting. Importantly, mice can be trained to associate novel odors with specific behavioral responses to provide insight into olfactory circuit function. This protocol details the procedure for training mice on a Go/No-Go operant learning task. In this approach, mice are trained on hundreds of automated trials daily for 2&#8211;4 weeks and can then be tested on novel Go/No-Go odor pairs to assess olfactory discrimination, or be used for studies on how odor learning alters the structure or function of the olfactory circuit. Additionally, the mouse olfactory bulb (OB) features ongoing integration of adult-born neurons. Interestingly, olfactory learning increases both the survival and synaptic connections of these adult-born neurons. Therefore, this protocol can be combined with other biochemical, electrophysiological, and imaging techniques to study learning and activity-dependent factors that mediate neuronal survival and plasticity.

Here, we train mice on an associative learning task to test odor discrimination. This protocol also allows for studies on learning-induced structural changes in the brain.

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 ...

Targeting the Corticospinal Tract in Neonatal Rats with a Double-Viral Vector using Combined Brain and Spine Surgery

Successfully tackling the obstacles that constrain research on neonatal rats is important for studying the differences in outcomes seen in pediatric spinal cord injuries (SCIs) compared to adult SCIs. In addition, reliably introducing therapies into the target cells of the central nervous system (CNS) can be challenging, and inaccuracies can compromise the efficacy of the study or therapy. This protocol combines viral vector technology with a novel surgical technique&#160;to accurately introduce gene therapies into neonatal rats at postnatal day 5. Here, a virus engineered for retrograde transport (retroAAV2) of Cre is introduced at the axon terminals of corticospinal neurons in the spinal cord, where it is subsequently transported to the cell bodies. A double-floxed&#160;inverted orientation (DIO) designer receptor exclusively activated by designer drug(s) (DREADD) virus is then injected into the somatomotor cortex of the brain. This double-infection technique promotes the expression of the DREADDs only in the co-infected corticospinal tract (CST) neurons.&#160;Thus, the simultaneous co-injection of the somatomotor cortex and cervical CST terminals is a valid method for studying the chemogenetic modulation of recovery following cervical SCI models in neonatal rats.

This protocol demonstrates a novel method for applying gene therapies to subpopulations of cells in neonatal rats at postnatal ages 5-10 days by injecting an anterograde chemogenetic modifier into the somatomotor cortex and a retrogradely transportable Cre recombinase into the cervical spinal cord.

使用脑和脊柱联合手术用双病毒载体靶向新生大鼠的皮质脊髓束

Successfully tackling the obstacles that constrain research on neonatal rats is important for studying the differences in outcomes seen in pediatric ...

Shuttle Box Assay as an Associative Learning Tool for Cognitive Assessment in Learning and Memory Studies using Adult Zebrafish

Cognitive deficits, including impaired learning and memory, are a primary symptom of various developmental and age-related neurodegenerative diseases and traumatic brain injury (TBI). Zebrafish are an important neuroscience model due to their transparency during development and robust regenerative capabilities following neurotrauma. While various cognitive tests exist in zebrafish, most of the cognitive assessments that are rapid examine non-associative learning. At the same time, associative-learning assays often require multiple days or weeks. Here, we describe a rapid associative-learning test that utilizes an adverse stimulus (electric shock) and requires minimal preparation time. The shuttle box assay, presented here, is simple, ideal for novice investigators, and requires minimal equipment. We demonstrate that, following TBI, this shuttle box test reproducibly assesses cognitive deficit and recovery from young to old zebrafish. Additionally, the assay is adaptable to examine either immediate or delayed memory. We demonstrate that both a single TBI and repeated TBI events negatively affect learning and immediate memory but not delayed memory. We, therefore, conclude that the shuttle box assay reproducibly tracks the progression and recovery of cognitive impairment.

Learning and memory are potent metrics in studying either developmental, disease-dependent, or environmentally induced cognitive impairments. Most cognitive assessments require specialized equipment and extensive time commitments. However, the shuttle box assay is an associative learning tool that utilizes a conventional gel box for rapid and reliable assessment of adult zebrafish cognition.

穿梭盒测定作为使用成人斑马鱼进行学习和记忆研究认知评估的关联学习工具

Cognitive deficits, including impaired learning and memory, are a primary symptom of various developmental and age-related neurodegenerative diseases and ...

Measurement of the Directional Information Flow in fNIRS-Hyperscanning Data using the Partial Wavelet Transform Coherence Method

Social interaction is of vital importance for human beings. While the hyperscanning approach has been extensively used to study interpersonal neural synchronization (INS) during social interactions, functional near-infrared spectroscopy (fNIRS) is one of the most popular techniques for hyperscanning naturalistic social interactions because of its relatively high spatial resolution, sound anatomical localization, and exceptionally high tolerance of motion artifacts. Previous fNIRS-based hyperscanning studies usually calculate a time-lagged INS using wavelet transform coherence (WTC) to describe the direction and temporal pattern of information flow between individuals. However, the results of this method might be confounded by the autocorrelation effect of the fNIRS signal of each individual. For addressing this issue, a method termed partial wavelet transform coherence (pWTC) was introduced, which aimed to remove the autocorrelation effect and maintain the high temporal-spectrum resolution of the fNIRS signal. In this study, a simulation experiment was performed first to show the effectiveness of the pWTC in removing the impact of autocorrelation on INS. Then, step-by-step guidance was offered on the operation of the pWTC based on the fNIRS dataset from a social interaction experiment. Additionally, a comparison between the pWTC method and the traditional WTC method and that between the pWTC method and the Granger causality (GC) method was drawn. The results showed that pWTC could be used to determine the INS difference between different experimental conditions and INS's directional and temporal pattern between individuals during naturalistic social interactions. Moreover, it provides better temporal and frequency resolution than the traditional WTC and better flexibility than the GC method. Thus, pWTC is a strong candidate for inferring the direction and temporal pattern of information flow between individuals during naturalistic social interactions.

This protocol describes partial wavelet transform coherence (pWTC) for calculating the time-lagged pattern of interpersonal neural synchronization (INS) to infer the direction and temporal pattern of information flow during social interaction. The effectiveness of pWTC in removing the confounds of signal autocorrelation on INS was proved by two experiments.

使用偏小波变换相干法测量 fNIRS-超扫描数据中的定向信息流

Social interaction is of vital importance for human beings. While the hyperscanning approach has been extensively used to study interpersonal neural ...