Name: measuring exercise levels in drosophila melanogaster using the rotating exercise quantification system (reqs)
Uploaded: 2018-05-27T15:00:00
Description: Watch this Scientific Journal Video about Measuring Exercise Levels in Drosophila melanogaster Using the Rotating Exercise Quantification System REQS at JoVE.com

这项工作得到了来自美国国立糖尿病和消化和肾脏疾病研究所的奖项编号 P30DK056336 的支持, 该研究中心是在伯明翰的阿拉巴马大学营养和肥胖研究所提供的试验补助金。

满足要求由果蝇活动监控单元 (用于源信息, 请参阅材料表) 安装在由马达单元控制的旋转臂上 (图1)。活动监视器确定在给定时间范围内, 在检测管中间的激光束阵列被打乱的频率。有关详细的绘图和深入的描述, 请参见我们以前的出版物9。虽然我们的系统使用 LAM25H 单元, 满足要求可以修改, 以适应其他果蝇活动监测单位以及。
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<ol>
	<li>Rieder, L. E., Larschan, E. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Wisdom+from+the+fly.">Wisdom from the fly.</a> Trends Genet. 30 (11), 479-481 (2014).</li><li>Ugur, B., Chen, K., Bellen, H. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Drosophila+tools+and+assays+for+the+study+of+human+diseases.">Drosophila tools and assays for the study of human diseases.</a> Dis Model Mech. 9 (3), 235-244 (2016).</li><li>Hales, K. G., Korey, C. A., Larracuente, A. M., Roberts, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetics+on+the+fly:+A+primer+on+the+Drosophila+Model+System.">Genetics on the fly: A primer on the Drosophila Model System.</a> Genetics. 201 (3), 815-842 (2015).</li><li>Piazza, N., Gosangi, B., Devilla, S., Arking, R., Wessells, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Exercise-training+in+young+Drosophila+melanogaster+reduces+age-related+decline+in+mobility+and+cardiac+performance.">Exercise-training in young Drosophila melanogaster reduces age-related decline in mobility and cardiac performance.</a> PLoS One. 4 (6), e5886 (2009).</li><li>Gargano, J. W., Martin, I., Bhandari, P., Grotewiel, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rapid+iterative+negative+geotaxis+(RING):+a+new+method+for+assessing+age-related+locomotor+decline+in+Drosophila.">Rapid iterative negative geotaxis (RING): a new method for assessing age-related locomotor decline in Drosophila.</a> Exp Gerontol. 40 (5), 386-395 (2005).</li><li>Tinkerhess, M. J., Ginzberg, S., Piazza, N., Wessells, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endurance+training+protocol+and+longitudinal+performance+assays+for+Drosophila+melanogaster.">Endurance training protocol and longitudinal performance assays for Drosophila melanogaster.</a> J Vis Exp. (61), (2012).</li><li>Mendez, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+TreadWheel:+A+novel+apparatus+to+measure+genetic+variation+in+response+to+gently+induced+exercise+for+Drosophila.">The TreadWheel: A novel apparatus to measure genetic variation in response to gently induced exercise for Drosophila.</a> PLoS One. 11 (10), e0164706 (2016).</li><li>Katzenberger, R. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Drosophila+model+of+closed+head+traumatic+brain+injury.">A Drosophila model of closed head traumatic brain injury.</a> Proc Natl Acad Sci U S A. 110 (44), E4152-E4159 (2013).</li><li>Watanabe, L. P., Riddle, N. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+the+Rotating+Exercise+Quantification+System+(REQS),+a+novel+Drosophila+exercise+quantification+apparatus.">Characterization of the Rotating Exercise Quantification System (REQS), a novel Drosophila exercise quantification apparatus.</a> PLoS One. 12 (10), e0185090 (2017).</li><li>Reed, L. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genotype-by-diet+interactions+drive+metabolic+phenotype+variation+in+Drosophila+melanogaster.">Genotype-by-diet interactions drive metabolic phenotype variation in Drosophila melanogaster.</a> Genetics. 185 (3), 1009-1019 (2010).</li><li>Bartholomew, N. R., Burdett, J. M., VandenBrooks, J. M., Quinlan, M. C., Call, G. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impaired+climbing+and+flight+behaviour+in+Drosophila+melanogaster+following+carbon+dioxide+anaesthesia.">Impaired climbing and flight behaviour in Drosophila melanogaster following carbon dioxide anaesthesia.</a> Sci Rep. 5, 15298 (2015).</li><li>Huang, W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Natural+variation+in+genome+architecture+among+205+Drosophila+melanogaster+Genetic+Reference+Panel+lines.">Natural variation in genome architecture among 205 Drosophila melanogaster Genetic Reference Panel lines.</a> Genome Res. 24 (7), 1193-1208 (2014).</li><li>Mackay, T. F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Drosophila+melanogaster+Genetic+Reference+Panel.">The Drosophila melanogaster Genetic Reference Panel.</a> Nature. 482 (7384), 173-178 (2012).</li><li>Berlandi, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Swing+Boat:+Inducing+and+recording+locomotor+activity+in+a+Drosophila+melanogaster+model+of+Alzheimer's+disease.">Swing Boat: Inducing and recording locomotor activity in a Drosophila melanogaster model of Alzheimer's disease.</a> Front Behav Neurosci. 11, 159 (2017).</li><li>Faville, R., Kottler, B., Goodhill, G. J., Shaw, P. J., van Swinderen, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=How+deeply+does+your+mutant+sleep?+Probing+arousal+to+better+understand+sleep+defects+in+Drosophila.">How deeply does your mutant sleep? Probing arousal to better understand sleep defects in Drosophila.</a> Sci Rep. 5, 8454 (2015).</li></ol>

如有代表性的结果表明, 满足要求能够准确测量果蝇的运动活动。满足要求是灵活的, 并允许研究人员解决与运动生物学或运动干预有关的各种研究问题。要突出显示的协议中有两个关键步骤。首先, 必须测试满足要求的设置, 以确保从满足要求到 DAMSystem308 的数据传输正常工作。如果没有正确设置, 数据线可能会在旋转过程中发生纠缠, 有时满足要求和 DAMSystem308 之间的连接由于磨损而在旋转连接处...

最近, 研究人员已经开始使用果蝇果蝇来研究运动生物学。"黑腹"是100年来1、2的遗传模型系统。然而, 果蝇的研究不仅为遗传学做出了贡献, 也为其他学科包括神经生物学、行为生物学和生理学提供了帮助3。2009年, 第一台用于果蝇的运动机器被描述为4。电力塔利用动物的消极 geotaxis 反应。当扰时 , 果蝇倾向于移动到其外壳的顶端。这种反应是建立良好的, 是流行的 "环" (快速迭代阴性 Geotaxis5) 的基础, 用于估计果蝇的攀爬能力和/或体能。该电源塔使用与电机单元相连的机械臂, 在其外壳内反复举起一组动物, 并将它们放回地面以诱导负 geotaxis 响应 (Tinkerhess et 20126提供一个视频说明使用的电源塔)。在电力塔上进行长时间的治疗, 从而增加了动物的体力活动量 (跑步或飞行), 与未经处理的控制动物相比, 随着时间的推移, 提高了身体健康的环检测的性能4。因此, 这项工作证明了利用果蝇作为运动生物学模型的可行性。
为了扩大果蝇运动研究的可用工具, 在 2016年, 门德斯和同事们描述了第二个果蝇运动机器, Treadwheel7。与动力塔类似, Treadwheel 利用果蝇的负 geotaxis 反应。然而, 这种反应是由动物外壳的持续旋转引起的, 而不是像在动力塔中那样把它们举起来。这种感应方法是温和的, 并允许一个更耐用的运动机制, 以避免任何物理创伤可能发生在电力塔运动 (见 Katzenberger, r.j. et 等20138的影响反复物理果蝇健康的创伤)。与动力塔4类似, 在 Treadwheel 上对动物的运动治疗会导致多种生理反应, 包括身体健康、甘油三酯水平和体重7的变化。因此, 果蝇生物学家研究运动有两种互补的方法。
动力塔和 Treadwheel 的一个限制是无法测量运动治疗引起的活动量。从 Treadwheel 的录像分析显示, 不同果蝇菌株对运动治疗的反应有显著性差异7。具体而言, 研究的菌株在受刺激的7时所执行的额外活动有多大不同。这一观察促使我们开发了第三个运动系统, 即旋转运动定量系统 (满足要求), 它允许我们在旋转诱导运动中测量动物活动水平 9.满足要求利用一个商业上可用的活动监测单元, 安装在旋转臂上, 通过旋转来刺激运动, 如在 Treadwheel 中。与满足要求的初步工作证实, 基因上不同的果蝇菌株-和性别-对旋转刺激有明显不同的反应, 因此, 不同基因型的运动所引起的运动量不完全相同 9.因此, 满足要求现在使果蝇生物学家能够测量治疗引起的运动量, 在运动领域开辟了各种新的研究途径。
这里我们详细描述了如何使用满足要求来量化旋转运动。满足要求诱导旋转运动, 同时测量被治疗动物的活动水平。满足要求可以容纳各种各样的运动项目, 从简单的2小时连续锻炼机制这里展示到更加复杂的间隔训练方法如门德斯和同事7所描述, 并且刺激可以通过调整通过转速 (约1-13 旋转每分钟)。根据用于生产满足要求的活动监测单元, 该方法适用于对单飞蝇或大型动物种群的分析。由于这种多功能性, 满足要求为果蝇研究人员提供了一系列的机会来研究, 例如, 不同的锻炼制度, 饮食干预, 或人口密度的影响。

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measuring exercise levels in drosophila melanogaster using the rotating exercise quantification system (reqs)

果蝇是运动生物学研究的一个新的模型有机体。迄今为止, 已经介绍了两个主要的运动系统、电力塔和 Treadwheel。然而, 还缺乏一种方法来测量通过运动治疗引起的额外动物活动量。旋转运动量化系统 (满足要求) 填补了这种需求, 为经历旋转运动的动物提供了动物活动的量度。本协议详细介绍了如何使用满足要求来评估旋转运动过程中的动物活动, 并说明了可以生成的数据类型。在这里, 我们展示了如何使用满足要求来衡量在运动诱发活动中的性别和应变特定的差异。满足要求还可以用来评估不同的实验参数, 如年龄, 饮食, 或人口大小对运动诱发活动的影响。此外, 它还可用于比较不同运动训练方案的功效。重要的是, 它提供了一个机会, 以规范的运动治疗之间的菌株, 使研究员能够达到同等数量的活动之间的群体, 如果需要的话。因此, 满足要求是一个值得注意的新资源为运动生物学家与果蝇模型系统和补充现有的运动系统。

使用满足要求的单个运行的输出是 DAMSystem308 软件生成的数据表, 默认情况下将标记为 "Monitor1.txt" (例如, 请参见补充文件1)。此类表的摘录显示在表 1中。每一列都包含来自单个检测管的数据, 而行包含从实验开始 (顶端) 到末尾 (底部) 的每个时间间隔中测量的活动。如果有数据切口 (如表 2中所示), 则应监视前三个数据点, 在这种情况下...

Watch this Scientific Journal Video about Measuring Exercise Levels in Drosophila melanogaster Using the Rotating Exercise Quantification System REQS at JoVE.com

Measuring Exercise Levels in Drosophila melanogaster Using the Rotating Exercise Quantification System REQS

旋转运动定量系统 (满足要求) 可以通过旋转来诱导果蝇运动, 同时测量动物的活动量. 在这里, 我们提出一个点对点协议, 详细说明如何测量动物的活动水平, 使用满足要求的旋转运动治疗。

使用旋转运动量化系统测量果蝇中的运动水平 (满足要求)

Drosophila melanogaster is a new model organism for studies in exercise biology. To date, two main exercise systems, the Power Tower and the Treadwheel ...

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Measuring Exercise Levels in Drosophila melanogaster Using the Rotating Exercise Quantification System (REQS)

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