Name: assaying locomotor activity to study circadian rhythms and sleep parameters in drosophila
Uploaded: 2010-09-28T17:30:00
Duration: 18 min 8 s
Description: Watch this Scientific Journal Video about Assaying Locomotor Activity to Study Circadian Rhythms and Sleep Parameters in Drosophila at JoVE.com

这项工作是支持由美国国立卫生研究院补助NIH34958一，E和NS061952 JC

<ol>
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P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analysis+of+locomotor+activity+rhythms+in+Drosophila.">Analysis of locomotor activity rhythms in Drosophila.</a> Nature Protocols. 1, 559-568 (2006).</li><li>Sehadova, H., Glaser, F. T., Gentile, C., Simoni, A., Giesecke, A., Albert, J. T., Stanewsky, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Temperature+entrainment+of+Drosophila's+circadian+clock+involves+the+gene+nocte+and+signaling+from+peripheral+sensory+tissues+to+the+brain.">Temperature entrainment of Drosophila's circadian clock involves the gene nocte and signaling from peripheral sensory tissues to the brain.</a> Neuron. 64, 251-266 (2009).</li><li>Zimmerman, J. E., Raizen, D. M., Maycock, M. H., Maislin, G., Pack, A. 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在这个协议中，我们描述了测量果蝇自发活动的节奏，一个可靠的输出时钟功能的标准读数蝇生物钟的行为的程序。这已用于检测新的时钟突变体（如Konopka和1971年本泽 ; Dubruille等2009）的大型屏幕，并不断地剖析和了解在体内的时钟功能。它也被用来研究果蝇的睡眠觉醒周期，尽管最近的报告表明，视频数字化分析是更可靠的量化的睡眠比使用自主活动的节奏（ 齐默尔曼等人，2008年） 。当使用自主活动的节奏来分析睡眠，在白天的睡眠比例往往被高估。 为了确保本议定书的成功和可重复性，它是至关重要的测定年龄，遗传背景相似的苍蝇，并在同等条件下饲养，水果中的行为表型的苍蝇，如昼夜节律和睡眠/休息活动是非常敏感所有这些因素。在一次实验中使用多个孵化器，重要的是要确保所有孵化器在预期的温度作为温度的函数，因为有些昼夜参数可能会改变。一个字谨慎考虑购买苍蝇孵化器时，并非所有的平等。虽然我们毫不犹豫地推荐任何特定的单位有很多的选择。在&lt;www.flybase.org&gt;提供一个很好的资源， 寻找果蝇工作的孵化器公司出售。有些公司甚至出售“ 果蝇生物钟”的孵化器，其中附加功能，如已为Trikinetics系统和温度斜坡（例如，Tritech）有线。重要功能包括周日光控的能力和良好的温度控制在对果蝇的生理范围（〜15-30 ℃）。孵化器的价格和大小有很大的差异，但是从Trikinetics新的活动监视器，即使是小规模的孵化器，可以容纳相当多的这些设备的数量。此外，虽然湿度控制的孵化器，可用于，这个新增的功能是没有必要的，只要你放在一个小泛水提供湿度（50-70％的罚款）。最后，虽然我们经常使用的数据分析，在本议定书FaasX和失眠，有替代方案和软件（2006年Rosato和Kyriacou），如ClockLab（ActiMetrics），布兰代斯节奏包（D.惠勒，医学院，休斯敦贝勒大学），MAZ（Zordan 等，2007）。

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		<div>
		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>Drosophila activity monitor (DAM)</td>
<td>&nbsp;</td>
<td>Trikinetics Inc.; Waltham, MA</td>
<td>DAM2 or DAM5</td>
<td>DAM2 monitors are more compact, and more can fit into a single incubator</td>
</tr>
<tr>
<td>Power supply interface unit (for DAM system)</td>
<td>&nbsp;</td>
<td>Trikinetics Inc.; Waltham, MA</td>
<td>PSIU9</td>
<td>Includes PS9-1 AC Power Supply</td>
</tr>
<tr>
<td>Light controller</td>
<td>&nbsp;</td>
<td>Trikinetics Inc.; Waltham, MA</td>
<td>LC6</td>
<td>&nbsp;</td>
<tr>
<td>Pyrex glass tubes</td>
<td>&nbsp;</td>
<td>Trikinetics Inc.; Waltham, MA</td>
<td>PGT5, PGT7, and PGT10</td>
<td>&nbsp;</td>
<tr>
<td>Plastic activity tube caps</td>
<td>&nbsp;</td>
<td>Trikinetics Inc.; Waltham, MA</td>
<td>CAP5</td>
<td>Yarn can be used instead of plastic caps.</td>
</tr>
<tr>
<td>DAM System data collection software</td>
<td>&nbsp;</td>
<td>Trikinetics Inc.; Waltham, MA</td>
<td>&nbsp;</td>
<td>Versions available for both Mac and PC</td>
</tr>
<tr>
<td>FaasX software</td>
<td>&nbsp;</td>
<td>M. Boudinot and F. Rouyer, Centre National de la Recherche Scientifique, Gif-sur-Yvette Cedex, France</td>
<td>&nbsp;</td>
<td>Only for Mac</td>
</tr>
<tr>
<td>Insomniac 2.0 software</td>
<td>&nbsp;</td>
<td>Leslie Ashmore, Department of Pharmacology &amp; Chemical Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA</td>
<td>&nbsp;</td>
<td>Runs on Matlab. Can be used on both PC and Macintosh.</td>
</tr>
<tr>
<td>Environmental incubator with temperature and diurnal control, e.g. Percival incubator</td>
<td>&nbsp;</td>
<td>Percival Scientific, Inc.</td>
<td>I-30BLL</td>
<td>Interior space dimension:
Width: 65cm;
Height: 86cm;
Depth: 55cm</td>
</tr>
<tr>
<td>Environmental incubator with temperature and diurnal control, e.g. DigiTherm Heating/Cooling Incubator with Circadian Timed Lighting and Timed Temperature</td>
<td>&nbsp;</td>
<td>Tritech Research, Inc.</td>
<td>05DT2CIRC001</td>
<td>Interior space dimension:
Width: 36m;
Height: 56m;
Depth: 28cm</td>
</tr>
<tr>
<td>APC Smart-UPS 2200VA 120V (Emergency power backup unit)</td>
<td>&nbsp;</td>
<td>APC</td>
<td>SU2200NET</td>
<td>Output Power Capacity of 1600 Watts</td>
</tr>
<tr>
<td>Sucrose</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>S7903</td>
<td>&nbsp;</td>
<tr>
<td>Bacto Agar</td>
<td>&nbsp;</td>
<td>Becton Dickinson</td>
<td>214010</td>
<td>&nbsp;</td>
<tr>
<td>TissuePrep Paraffin pellets</td>
<td>&nbsp;</td>
<td>Fisher Scientific</td>
<td>T565</td>
<td>Melting point 56&deg;C-57&deg;C</td>
</tr>
<tr>
<td>Block heater </td>
<td>&nbsp;</td>
<td>VWR</td>
<td>12621-014</td>
<td>&nbsp;</td>		</tbody>
		</table>
		</div>

assaying locomotor activity to study circadian rhythms and sleep parameters in drosophila

大多数生命形式表现出的细胞，生理和行为现象，是由内源性昼夜驱动（≡24小时）心脏起搏器或时钟的生活节奏。在人体生理系统的故障与许多疾病或功能紊乱。对我们的基本生理节律的机制认识出现了很大的进展，从遗传筛选，据此作为时钟功能读出一个容易测量的行为节奏。利用果蝇的研究，我们了解细胞和生化的昼夜节律的基地作出了开创性的贡献。在果蝇中读出测量的标准生物钟的行为是自发活动。在一般情况下，监控系统包括专门设计的设备，可以测量对果蝇的运动运动。这些设备被安置在环境控制的孵化器坐落在暗室和使用红外光束中断记录的小管内所载的个人苍蝇的自发活动。当过多少天计算， 果蝇表现出每天的活动和不活动周期，行为的节奏，是由动物的内源性昼夜节律系统管辖。与市售的自发活动的监控设备的出现和发展的数据分析软件方案，整个程序已经简化。我们使用系统的Trikinetics公司，这是这里所描述的过程，是目前全世界使用最流行的系统。最近，相同的监控设备已被用于研究在果蝇的睡眠行为。由于日常唤醒许多果蝇的睡眠周期可以同时测量只有1至2周，连续的自发活动数据通常是足够的，这个系统是理想的大型屏幕，以确定果蝇表现改变生物钟或睡眠属性。

Watch this Scientific Journal Video about Assaying Locomotor Activity to Study Circadian Rhythms and Sleep Parameters in Drosophila at JoVE.com

Assaying Locomotor Activity to Study Circadian Rhythms and Sleep Parameters in Drosophila

我们描述用于记录日常的自发活动节律的过程<em&gt;果蝇</em&gt;和随后的数据分析。自发活动节律是一个可靠的动物生物钟的行为输出，时钟功能的标准读数研究昼夜的突变体或环境研究如何调节昼夜节律系统时使用。

含量自发活动，研究昼夜节律和睡眠参数果蝇</em

Most life forms exhibit daily rhythms in cellular, physiological and behavioral phenomena that are driven by endogenous circadian (&equiv;24 hr) ...

Research

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Neuroscience

Assaying Locomotor Activity to Study Circadian Rhythms and Sleep Parameters in Drosophila

Laser Capture Microdissection of Drosophila Peripheral Neurons

Laser Capture Microdissection of Drosophila Peripheral Neurons

激光捕获显微切割技术的果蝇外周神经元

The dendritic arborization (da) neurons of the Drosophila peripheral nervous system (PNS) provide an excellent model system in which to investigate the ...

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Circadian rhythms are physiological functions that cycle over a period of approximately 24 hours (circadian- circa: approximate and diem: day)1, 2.  They ...

Assaying Locomotor, Learning, and Memory Deficits in Drosophila Models of Neurodegeneration

Assaying Locomotor, Learning, and Memory Deficits in Drosophila Models of Neurodegeneration

含量的运动能力，学习和记忆的赤字果蝇</em神经退行性疾病>模型

Advances in genetic methods have enabled the study of genes involved in human neurodegenerative diseases using Drosophila as a model system1. Most of ...

Non-Laser Capture Microscopy Approach for the Microdissection of Discrete Mouse Brain Regions for Total RNA Isolation and Downstream Next-Generation Sequencing and Gene Expression Profiling

Non-Laser Capture Microscopy Approach for the Microdissection of Discrete Mouse Brain Regions for Total RNA Isolation and Downstream Next-Generation Sequencing and Gene Expression Profiling  |

总RNA的分离和下游下一代测序和基因表达谱的非激光捕获显微切割离散小鼠脑地区显微镜方法

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Simultaneous Electroencephalography, Real-time Measurement of Lactate Concentration and Optogenetic Manipulation of Neuronal Activity in the Rodent Cerebral Cortex

同时脑电图，实时测量的啮齿动物的大脑皮质中的神经元活动的乳酸浓度与Optogenetic的操作

Although the brain represents less than 5% of the body  by mass, it utilizes approximately one quarter of the glucose used by the body  at rest1. The ...

Recording and Analysis of Circadian Rhythms in Running-wheel Activity in Rodents

运行轮活动在啮齿类动物的昼夜节律的记录和分析

When rodents have free access to a running wheel in their home cage, voluntary use of this wheel will depend on the time of day1-5. Nocturnal rodents, ...

Light Preference Assay to Study Innate and Circadian Regulated Photobehavior in Drosophila Larvae

Light Preference Assay to Study Innate and Circadian Regulated Photobehavior in Drosophila Larvae

光偏好的检测，以研究的先天和昼夜调节感光特性<em&gt;果蝇</em&gt;幼虫

Light acts as environmental signal to control animal behavior at various levels. The Drosophila larval nervous system is used as a unique model to answer ...

Determination of the Spontaneous Locomotor Activity in Drosophila melanogaster

Determination of the Spontaneous Locomotor Activity in Drosophila melanogaster

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Drosophila melanogaster has been used as an excellent model organism to study environmental and genetic manipulations that affect behavior. One such ...

High-throughput Analysis of Locomotor Behavior in the Drosophila Island Assay

High-throughput Analysis of Locomotor Behavior in the Drosophila Island Assay

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Advances in next-generation sequencing technologies contribute to the identification of (candidate) disease genes for movement disorders and other ...

Single-cell Resolution Fluorescence Live Imaging of Drosophila Circadian Clocks in Larval Brain Culture

Single-cell Resolution Fluorescence Live Imaging of Drosophila Circadian Clocks in Larval Brain Culture

单细胞分辨荧光活成像在幼虫脑培养中的果蝇昼夜节律时钟

The circadian pacemaker circuit orchestrates rhythmic behavioral and physiological outputs coordinated with environmental cues, such as day/night cycles. ...