Name: automated measurements of sleep and locomotor activity in mexican cavefish
Uploaded: 2019-03-21T14:00:00
Description: Watch this Scientific Journal Video about Automated Measurements of Sleep and Locomotor Activity in Mexican Cavefish at JoVE.com

这项工作得到了 NIGMS 奖 GM127872 ack 奖、NINSD 奖105072对 ERD 和 ACK 的支持, NSF 奖对 ack 的支持。

注: 在幼虫和成虫中建立行为跟踪系统。
1. 构建幼虫睡眠系统
请注意:通过幼鱼在受精后4天通过 30 dpf a. mexicanus追踪幼虫的监测系统需要多种设备, 包括红外 (ir) 照明、丙烯酸红外光扩散器、自动照明控制 (定时器)、计算机、摄像机和二次材料, 如布线和电源控制器 (图 1a)。以下说明将说明如何建立一个系统, 以准确...

<ol>
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该协议描述了一个定制系统, 用于量化幼虫和成虫的睡眠和运动活动。紫鱼已成为研究睡眠演变的主要模型, 可用于研究睡眠调节1的遗传和神经基础。该协议中的关键步骤包括优化照明和视频质量, 以确保准确的跟踪, 这是量化睡眠所必需的。这里描述的采集和分析系统功能齐全, 许多其他商业和定制系统也是如此, 可以量化运动和行为28、29...

睡眠在生理、功能和行为水平1、2、3 上在整个动物王国高度保守。虽然哺乳动物实验室动物的睡眠通常是用脑电图来评估的, 但在小的基因间可利用的模型系统中, 电生理记录就不那么实用了, 因此睡眠通常是根据行为来衡量的,4. 与睡眠相关的行为特征在整个动物王国高度保守, 包括增加唤醒阈值、刺激可逆性和长时间的行为静止 5。这些措施可以用来描述动物的睡眠, 从线虫, 线虫, 通过人类 6 .
使用行为静止来描述睡眠需要自动跟踪软件。使用跟踪软件, 活动周期和不活动期在几天内确定, 长时间不活动被归类为睡眠 7,8。近年来, 开发了多个跟踪系统, 用于获取各种适合基因的小型模型系统中的活动数据;包括蠕虫、果蝇和鱼9,10,11。这些程序附有允许自动跟踪动物行为的软件, 包括开源免费软件和商业上可用的软件7、12、13、14.这些系统在灵活性上有所不同, 允许在许多基因修正模型中有效地筛选和表征睡眠表型。
斑马鱼睡眠的基因调查,达尼奥雷里奥, 已经导致识别了许多基因和神经回路, 调节睡眠15,16。虽然这提供了一个强大的系统, 用于调查在脊椎动物实验动物的睡眠的神经基础, 更不知道睡眠是如何演变, 以及自然变化如何有助于睡眠调节。墨西哥紫鱼,马西卡努斯(a。在睡眠、运动活动和生理节律17, 18 方面发生了巨大的差异。这些鱼作为眼睛表面鱼类存在于墨西哥和南部的得克萨斯州和墨西哥东北部的 sierra del abra 地区周围至少29个洞穴种群 19,20,21。值得注意的是, 许多行为差异, 包括睡眠不足, 似乎已经独立出现在多个空鱼群体 14,22。因此, 紫鱼为研究睡眠、生理和社会行为的收敛演变提供了一个模型。
该协议描述了一种测量a中睡眠和运动行为的系统。仙人掌幼虫和成虫。一个定制的基于红外的记录系统允许在光明和黑暗的条件下对动物进行录像。商业上可用的软件可用于测量活动, 自定义宏用于量化不活动的几个方面, 并确定睡眠时间。该协议还描述了用于跟踪水箱内多个动物活动的实验修改, 提供了检查睡眠和社会行为之间相互作用的能力。这些系统可用于测量睡眠, 生理行为, 和运动活动的其他种类, 包括斑马鱼和粘滞。

<table>
	<tbody>
		<tr>
			<td>12V power adaptor</td>
			<td>Environmental Lights</td>
			<td>24 Watt 12 VDC Power Supply</td>
			<td></td>
		</tr>
		<tr>
			<td>Acrylic dividers (adults)</td>
			<td>TAP Plastic</td>
			<td></td>
			<td>Order sheets in sizes as needed</td>
		</tr>
		<tr>
			<td>Adult infrared light power source</td>
			<td>Environmnental Lights 24 Watt 12 VDC Power Supply</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Battery pack</td>
			<td>CyberPower</td>
			<td>CP850PFCLCD</td>
			<td></td>
		</tr>
		<tr>
			<td>Camera lens (adult)</td>
			<td>Navitar Zoom 7000</td>
			<td>Zoom 7000</td>
			<td></td>
		</tr>
		<tr>
			<td>Camera lens (larval)</td>
			<td>Fujian 35mm f/1.7</td>
			<td>B01CHX7668</td>
			<td>Purchase on Amazon</td>
		</tr>
		<tr>
			<td>Camera lens adapter</td>
			<td>d</td>
			<td>1524219</td>
			<td></td>
		</tr>
		<tr>
			<td>Camera mount</td>
			<td>CowboyStudio Super Clamp</td>
			<td>B002LV7X1K</td>
			<td>Purchase on Amazon</td>
		</tr>
		<tr>
			<td>Fish tank</td>
			<td>Deep Blue Professional</td>
			<td>ADB11006</td>
			<td></td>
		</tr>
		<tr>
			<td>Heat sink (adult)</td>
			<td>M-D Building products</td>
			<td>SKU: 61085</td>
			<td>Cut to fit</td>
		</tr>
		<tr>
			<td>Heat sink (larval)</td>
			<td>M-D Building products</td>
			<td>SKU: 57000</td>
			<td>Cut to fit</td>
		</tr>
		<tr>
			<td>Infrared lights (adults)</td>
			<td>Environmental Lights Infrared 850 nm 5050 LED strip</td>
			<td>irrf850-5050-60-reel</td>
			<td>Cut to fit</td>
		</tr>
		<tr>
			<td>Infrared lights (larval)</td>
			<td>LED World</td>
			<td>B00MO9H7H4</td>
			<td>Purchase on Amazon</td>
		</tr>
		<tr>
			<td>IR-diffusing acrylic</td>
			<td>TAP Plastic</td>
			<td></td>
			<td>Order sheets in sizes as needed</td>
		</tr>
		<tr>
			<td>Laptop/computer</td>
			<td>N/A</td>
			<td>N/A</td>
			<td>Any laptop will work.</td>
		</tr>
		<tr>
			<td>LED light</td>
			<td>Chanzon 10 High Power Led Chip 3W White (6000K-6500K/600mA-700mA/DC 3V-3.4V/3 Watt)</td>
			<td>B06XKTRSP7</td>
			<td>Use with Chanzon 25pcs 1W 3W 5W LED Heat Sink (2 pin Black) Aluminum Base Plate Panel</td>
		</tr>
		<tr>
			<td>light timer</td>
			<td>Century 24 Hour Plug-in Mechanical Timer Grounded</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Plastic wall mount for IR</td>
			<td>Everbilt Plastic pegboard</td>
			<td>Model # 17961</td>
			<td></td>
		</tr>
		<tr>
			<td>Power cable</td>
			<td>BNTECHGO 22 Gauge Silicone Wire</td>
			<td>B01K4RPE0Y</td>
			<td></td>
		</tr>
		<tr>
			<td>Power source</td>
			<td>Rapid LED</td>
			<td>MOONLIGHT DRIVER (350MA)</td>
			<td></td>
		</tr>
		<tr>
			<td>Tissue culture plates</td>
			<td>Fisherbrand</td>
			<td colspan="2">12-well (FB012928) 24-well (FB012929)</td>
		</tr>
		<tr>
			<td>Tripod Ball head</td>
			<td>Demon DB-44</td>
			<td>B00TQ54CZO</td>
			<td>Purchase on Amazon</td>
		</tr>
		<tr>
			<td>USB Hardrive</td>
			<td>Seagate 3TB backup</td>
			<td>STDT3000100</td>
			<td></td>
		</tr>
		<tr>
			<td>USB Webcam</td>
			<td>Microsoft LifeCam</td>
			<td>Q2F-00014</td>
			<td>Purchase on Amazon</td>
		</tr>
		<tr>
			<td>Wall mount for camera</td>
			<td>LDR Industries 1/2&#34; Steel pipe</td>
			<td>307 12X36</td>
			<td>Mounted on wall with Flange and 90 degree pipe elbow. Could also use a tripod to hold camera.</td>
		</tr>
	</tbody>
</table>

automated measurements of sleep and locomotor activity in mexican cavefish

在整个植物区, 睡眠的特点是高度保守的行为特征, 包括提高唤醒阈值, 睡眠剥夺后反弹, 和巩固的行为不动期。墨西哥紫外鱼是研究环境扰动下性状演化的模型. mexicanus存在于眼睛表面居住的形态和多个盲孔居住种群中, 它们具有强烈的形态和行为差异。睡眠丧失发生在多个独立进化的空基鱼种群中。该方案描述了一种定量的方法, 在a. mexicanus洞穴和水面鱼类的睡眠和运动活动。一个经济高效的视频监控系统允许个人居住的幼虫或成年鱼的行为成像一周或更长时间。该系统可应用于成年后4天的鱼类。这种方法也可以用来测量社会交往对睡眠的影响, 在一个竞技场上记录多条鱼。在行为记录之后, 使用自动跟踪软件对数据进行分析, 并使用自定义脚本进行处理, 这些脚本量化多个睡眠变量, 包括持续时间、回合长度和回合数。该系统可用于测量睡眠, 生理行为, 和运动活动, 在几乎任何鱼类物种, 包括斑马鱼和粘滞。

4-30 dpf 年龄的幼虫可以可靠地记录在图 1所述的自定义构建的闭合系统中。该系统包括红外和可见光照明, 以便在光线和黑暗条件下、在各种可见光条件下进行录制 (图 1a)。然后使用跟踪软件 (图 1b, c) 对视频进行分析, 并使用自定义睡眠宏进行后处理 (请参阅补充下载)。与表面鱼?...

Watch this Scientific Journal Video about Automated Measurements of Sleep and Locomotor Activity in Mexican Cavefish at JoVE.com

Automated Measurements of Sleep and Locomotor Activity in Mexican Cavefish

该协议详细介绍了在墨西哥的运动行为和睡眠的量化方法。以前的分析扩展到衡量这些行为在社会居住的鱼。该系统可广泛应用于其他鱼类的睡眠和活动研究。

墨西哥河鱼睡眠和睡眠活动的自动测量

Across phyla, sleep is characterized by highly conserved behavioral characteristics that include elevated arousal threshold, rebound following sleep ...

该协议可用于量化任何鱼类种群的睡眠和活动，并有可能扩大用于研究睡眠调节的模型系统的数量。这种技术的优点是它的灵活性。例如，鱼可以跟踪在各种竞技场大小或单独安置或组住的实验。测量该系统睡眠的强度是高通量药物或基因筛选方法的适用性。我们应用这种方法研究两种不同种群的睡眠进化，这些洞穴鱼表现出截然不同的睡眠表型。虽然建立行为成像系统可能很困难，但我们的系统为研究幼虫和成年鱼类提供了一种经济高效的方法。为了建立系统，以准确跟踪幼虫A.mexicanus的睡眠和昼夜节律，使用0.33厘米厚的白色标志丙烯酸为幼虫跟踪系统构建平台。将录制平台放在光亮的方形散热器的顶部，在行为跟踪实验期间，含鱼竞技场将驻留在该散热器上，并在动物壳体的灯光之间放置第二个丙烯酸，以散射红外线，以实现最佳的照明和对比度。将整个幼虫跟踪台和照明装置放在封闭的塑料管内。使用旋转工具将镜头从实验合适的网络摄像头中卸下。拆下相机背面的小螺钉以拆下内部壳体，并拆下相机机身内的小螺钉以松开镜头的其余部分。使用小螺丝刀拆下切割镜头后留下的镜头壳体的任何部分。并拆下电荷耦合器件或 CCD 外壳顶部的蓝色发光二极管。接下来，将内壳体返回到其原始方向，然后再将两个外部螺钉拧回其原始位置。使用小锯将摄像机内部布线，使摄像机能够安装圆形塑料布线位，使多余的塑料平滑，直到它可以伴随镜头适配器。在不直接接触摄像机的情况下，在摄像机内部尽可能靠近 CCD 安装红外通滤波器，并拧入镜头背面的 35 毫米固定镜头适配器。然后将相机和镜头放入钻入管子顶部的孔中，该孔容纳舞台和灯光，并将 USB 连接到记录动物的计算机。要为成年鱼类构建睡眠系统，首先每隔约46厘米切割红外线条。并放置一个46由5由0.32厘米9%光通过白色标志丙烯酸板直接在每一个红外光条前，以分散红外光。然后将所有储罐放在支持后置红外照明的机架上，并使用不透明的塑料分压器和 10 升玻璃储罐创建单独的竞技场。记录墨西哥四孔鱼的运动活动，将第4至6条受精后鱼放入24孔组织板的单个井中，在6孔组织板的单个井中记录20至30天受精后。记录成人四分之四睡眠和活动在社会设置的房子最多五个成人每10升坦克有或没有分隔线。在这个具有代表性的实验中，来自三种独立洞穴鱼类种群的幼虫鱼与表层鱼相比，睡眠明显减少。这种睡眠生产在所有测试年龄是一致的，因为与表鱼相比，成年帕琼、莫利诺和蒂娜雅洞穴鱼的睡眠也显著减少。值得注意的是，社会住房有力地减少了表面鱼类的睡眠，而不影响帕乔洞穴鱼的睡眠，很可能是由于洞穴鱼群的地下室效应。记录鱼时，应注意确保相机照明最佳，以增强鱼相对于背景的信号。此协议需要一些基本命令行编码，这些代码在文本中已完全描述，并且应适合任何实验室设置。该协议已经导致许多有关睡眠演变的问题，目前正在用于药物屏幕，跟踪睡眠行为，并了解社会行为如何影响睡眠。

Research

JoVE Journal

Behavior

Mouse Short- and Long-term Locomotor Activity Analyzed by Video Tracking Software

鼠标短期和长期活动性分析视频跟踪软件

Locomotor  activity (LMA) is a simple and easily performed measurement of behavior in mice  and other rodents. Improvements in video tracking software ...

科研

行为学

Automated Visual Cognitive Tasks for Recording Neural Activity Using 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 ...

Eye Tracking, Cortisol, and a Sleep vs. Wake Consolidation Delay: Combining Methods to Uncover an Interactive Effect of Sleep and Cortisol on Memory

眼睛跟踪，皮质醇和睡眠与唤醒延迟合并：合并方法来揭开睡眠和皮质醇的交互式影响记忆

Although rises in cortisol can benefit memory consolidation, as can sleep soon after encoding, there is currently a paucity of literature as to how these ...

Behavioral and Locomotor Measurements Using an Open Field Activity Monitoring System for Skeletal Muscle Diseases

行为和运动测量使用一个开放的领域活动监控系统的骨骼肌疾病

The open field activity monitoring system comprehensively assesses locomotor and behavioral activity levels of mice. It is a useful tool for assessing ...

Use of the Open Field Maze to Measure Locomotor and Anxiety-like Behavior in Mice

使用开放式现场迷宫来衡量小鼠的运动能力和焦虑样行为

Animal models have proven to be invaluable to researchers trying to answer questions regarding the mechanisms of behavior. The Open Field Maze is one of ...

Manipulation of Epileptiform Electrocorticograms (ECoGs) and Sleep in Rats and Mice by Acupuncture

Manipulation of Epileptiform Electrocorticograms ECoGs and Sleep in Rats and Mice by Acupuncture

癫痫Electrocorticograms（ECoGs）的操作，并睡在针刺大鼠和小鼠

Ancient Chinese literature has documented that acupuncture possesses efficient therapeutic effects on epilepsy and insomnia. There is, however, little ...

Using a Split-belt Treadmill to Evaluate Generalization of Human Locomotor Adaptation

用分体式跑步机评价人类运动适应的泛化

Understanding the mechanisms underlying locomotor learning helps researchers and clinicians optimize gait retraining as part of motor rehabilitation. ...

Noninvasive, High-throughput Determination of Sleep Duration in Rodents

无创高通量测定啮齿类动物睡眠时间

Traditionally, sleep is monitored by an electroencephalogram (EEG). EEG studies in rodents require surgical implantation of the electrodes followed by a ...

A High-performance Liquid Chromatography Measurement of Kynurenine and Kynurenic Acid: Relating Biochemistry to Cognition and Sleep in Rats

高效液相色谱法测定犬尿氨和 Kynurenic 酸: 对大鼠认知和睡眠的相关生物化学研究

The kynurenine pathway (KP) of tryptophan degradation has been implicated in psychiatric disorders. Specifically, the astrocyte-derived metabolite ...

Chronic Sleep Deprivation in Mouse Pups by Means of Gentle Handling

小鼠幼犬慢性睡眠剥夺的温和处理方法

Sleep is critical for proper development and neural plasticity. Moreover, abnormal sleep patterns are characteristic of many neurodevelopmental disorders. ...