这项工作是由美国国立卫生研究院支持的授予F32 NS078958（DTB）和R01 AG033620（BG）。

1。组装测试飞行<ol><li>安全飞行缸环站1用链钳。 （留约3厘米的圆柱体重量菜的下面。） （注:飞行汽缸我们用90厘米长，直径13.5厘米）。 </li><li>插入称量皿上一薄层矿物油的飞行筒的下方。 </li><li>安全漏斗环站2使用环钳爪和钳。调整漏斗的高度，使得漏斗的底部是与飞行圆筒的顶部齐平。 （注:该漏斗的前端的直径必须小于放入滴管，使得小瓶不会掉下通过小瓶的外径）。 </li><li>插入落料管进入漏斗的顶部上，并用一个卡爪夹紧。 （注:我们使用滴管即25厘米长滴飞含小瓶从这个高度可以让所有的苍蝇与受力均匀一致弹射落管的内径应为史赖。TLY比小瓶允许小瓶以自由落下的外径大。） </li><li>切割聚丙烯片材（S）到合适的大小。 （注:为了有助于插入和移除的薄片，其宽度应比飞行筒的内周面稍小）。 </li><li>涂抹薄薄的一层缠结陷阱的表。允许使用前坐了1小时。 （注:保留足够的空间，在表的顶部和底部（大约3公分）无涂层把握片插入/删除）。 </li><li>将飞行气缸内的聚丙烯酰胺片。 </li><li>用松木支架组装相机的轨道。 （注:确保轨道的底部可以支持摄像头没有挡住了镜头参见图1B。） </li><li>添加瓶塞和螺旋到位。 （注:发生在位置的限位器，将让相机以查看全景模式下，整个塑料片。） </li></ol> 2。运行试验<ol> &lt;李&gt;收集苍蝇被测试的小瓶。为了获得最佳效果，请使用不超过20苍蝇/瓶。 </li><li>轻轻拍打苍蝇小瓶的底部，拔出，再插入滴管和释放瓶中。 （注:将小瓶落下的落料管，直到它碰到漏斗窄口当小瓶击中漏斗，苍蝇被喷入气缸的飞行。） </li><li>提起落管，以去除空瓶。 （注:苍蝇相同的测试组的多个小瓶可以在一个单一的聚丙烯酰胺片被检测我们发现，高达200苍蝇（10瓶20苍蝇每个）可以用于测试和成像随手在一张纸上。 </li><li>取出塑料片，将它放在一个平坦的白色表面。 （注:白色的海报板可以使用，如果台面是深色。） </li><li>组装相​​机在轨道的塑料片。相机应足够高的片材具有在视场中的片材的顶部和底部之上。 </li><li>沿滑动个摄像头E导轨抱着"捕获"按钮获取全景图像时。 </li><li>苍蝇降落在油的数量可以手动对每个试验进行计数。 </li><li>用于在给定的实验条件下都重复步骤2.2-2.7。苍蝇可以从每次试验之间的片材被移除。可选地，几个薄片可以被使用，以对每个试验的新表。 </li></ol> 3。数据收集<ol><li>使用ImageJ软件打开图像文件。 </li><li>如果需要的话，只包括着陆表面积作物图像。 （这是涂在缠结陷阱的区域。） </li><li>图像转换成8位灰度。 </li><li>创建一个"门槛"，以筛选出的白色背景。 （图像→调整→阈值）。 </li><li>设置参数来识别每个使用"分析粒子"菜单苍蝇。 （分析→分析颗粒）定义用于识别粒子的参数。与我们建立，我们发现使用5-90像素2和0.4-1.0的圆形区域将准确地识别所有的样品。 </li><li>采用坐标为每个粒子的生成的列表中的每个测量苍蝇的位置。的像素的x坐标可以被转换成厘米来计算降落高度。 </li><li>表导入到电子表格（如Microsoft Excel）。 </li></ol>

<ol>
	<li>Benzer, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+dissection+of+behavior.">Genetic dissection of behavior.</a> Sci. Am. 229, 24-37 (1973).</li><li>Ali, Y. O., Escala, W., Ruan, K., Zhai, R. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assaying+locomotor,+learning,+and+memory+deficits+in+Drosophila+models+of+neurodegeneration.">Assaying locomotor, learning, and memory deficits in Drosophila models of neurodegeneration.</a> J. Vis. Exp. , e2504 (2011).</li><li>de Vries, S. E., Clandinin, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optogenetic+Stimulation+of+Escape+Behavior+in+Drosophila+melanogaster.">Optogenetic Stimulation of Escape Behavior in Drosophila melanogaster.</a> J. Vis. Exp. , e50192 (2013).</li><li>Mundiyanapurath, S., Certel, S., Kravitz, E. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Studying+aggression+in+Drosophila+(fruit+flies).">Studying aggression in Drosophila (fruit flies).</a> J. Vis. Exp. , e155 (2007).</li><li>Nichols, C. D., Becnel, J., Pandey, U. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Methods+to+assay+Drosophila+behavior.">Methods to assay Drosophila behavior.</a> J. Vis. Exp. , e3795 (2012).</li><li>Shiraiwa, T., Carlson, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Proboscis+extension+response+(PER)+assay+in+Drosophila.">Proboscis extension response (PER) assay in Drosophila.</a> J. Vis. Exp. , e193 (2007).</li><li>Lloyd, T. E., Taylor, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Flightless+flies:+Drosophila+models+of+neuromuscular+disease.">Flightless flies: Drosophila models of neuromuscular disease.</a> Ann. N.Y. Acad. Sci. 1184, e1-e20 (2010).</li><li>Atkinson, N. 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=Molecular+separation+of+two+behavioral+phenotypes+by+a+mutation+affecting+the+promoters+of+a+Ca-activated+K+channel.">Molecular separation of two behavioral phenotypes by a mutation affecting the promoters of a Ca-activated K channel.</a> J. Neurosci. 20, 2988-2993 (2000).</li><li>Atkinson, N. S., Robertson, G. A., Ganetzky, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+component+of+calcium-activated+potassium+channels+encoded+by+the+Drosophila+slo+locus.">A component of calcium-activated potassium channels encoded by the Drosophila slo locus.</a> Science. 253, 551-555 (1991).</li><li>Elkins, T., Ganetzky, B., Wu, C. F. <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+mutation+that+eliminates+a+calcium-dependent+potassium+current.">A Drosophila mutation that eliminates a calcium-dependent potassium current.</a> Proc. Natl. Acad. Sci. U.S.A. 83, 8415-8419 (1986).</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, 386-395 (2005).</li><li>Fry, S. N., Rohrseitz, N., Straw, A. D., Dickinson, M. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Visual+control+of+flight+speed+in+Drosophila+melanogaster.">Visual control of flight speed in Drosophila melanogaster.</a> J. Exp. Biol. 212, 1120-1130 (2009).</li><li>Mronz, M., Lehmann, F. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+free-flight+response+of+Drosophila+to+motion+of+the+visual+environment.">The free-flight response of Drosophila to motion of the visual environment.</a> J. Exp. Biol. 211, 2026-2045 (2008).</li></ol>

该作者有没有利益冲突披露。

使用这里介绍的方法，我们已经能够快速评估大量的果蝇突变体的飞行性能，提供比以前更高的效率。对于我们的实验中，我们经常分开的男性和女性，并提高他们在密度低（小于20苍蝇/瓶），以限制攻击可能会破坏翅膀。另一个重要的考虑是控制适当的，由于遗传背景的差异在飞行性能上的差异。我们还发现它有助于让苍蝇至少24小时的麻醉与二氧化碳在飞行试验之前恢复。另外，苍蝇可能暴露于低温（4℃），以便更快的恢复没有可能影响行为进行麻醉。 在这个协议中的限速步骤是从试验之间的塑料片材除去苍蝇。一种方法来提高效率是使用大量Ò同时f床单，撇开，因为它们使用的床单和清洗一次全部以下数据集合。该落下至底部苍蝇将需要手工计数，因为它们将不包括在片材上。尽管如此，与手工计算落地的高度相比，这很容易。重新申请缠结陷阱给每个表需要会因涂层厚也有所不同。根据我们的经验，个人结算将持续一个月了新的镀膜之前需要。 一个附加的优点，以使用缠结陷阱以上的矿物油，是从纸张恢复活苍蝇的能力。由于苍蝇只是坚持缠结陷阱的表面，而不是变得浸泡，个别苍蝇可以很容易地删除。 "飞的"苍蝇落到底部，也可通过与空烧瓶代替矿物油托盘回收。 我们相信，这里所描述的飞行行为的自动测量提供了一个NUMBER优于先前方法的优点，允许对遗传筛选更高层次的吞吐量，重现性和准确性。自动评分也被用于提高吞吐量进行行为分析，如环法11。此外，直接测量着陆高度提供了比简单的合格/不合格测量（％传单等 ）有较大的敏感性，使我们能够检测到更细微的差别在飞行性能。 这里所描述的实验可以由测量的飞行行为更为复杂的方面，包括飞行速度12和自由飞行应对运动13视觉控制后续进一步分析加以补充。虽然这些测试是更费时的并且不适合于大规模的遗传筛选，它们可帮助提供有关在一个运动反应的特定基因的功能的详细信息。

果蝇一直被用来研究行为1的遗传基础，而且研究人员已经设计出多种方法来分析不同类型的行为2-6。苍蝇一直在提供神经肌肉疾病7的有用模型特别有用。用来研究自发行为的普遍分析是飞行性能。原飞航测试仪是用于识别飞行缺陷突变体和对飞行能力1量化考核有用的，但它有几个缺点，限制了它的高通量筛选应用:采用涂油气缸是凌乱和繁琐，某些功能如气缸和介绍的苍蝇入管具有可变的力的长度减少量化精度，以及它是很难从测试仪恢复活苍蝇。为了克服这些限制，我们修改了飞行测试，包括了一些改进。我们增加了一个"降恩被"引入果蝇消除实验和用户之间的可变性，我们使用涂有粘合剂，其允许更容易清洗和个人蝇恢复可拆卸的压克力板，我们已经增加了飞行管的长度，以提高定量精度和可靠性。最后，我们使用数码相机和成像软件计算苍蝇降落的高度，我们相信这些改进将是有益的兴趣在飞行性能缺陷进行大规模遗传筛选任何实验室。

<table><tbody><tr><td>Putty knife</td><td>Home Depot</td><td>630147</td><td>&lt;a href=&quot;http://www.homedepot.com&quot;&gt;www.homedepot.com&lt;/a&gt;</td></tr><tr><td>Pine back band moulding (2x)</td><td>Home Depot</td><td>156469</td><td>&lt;a href=&quot;http://www.homedepot.com&quot;&gt;www.homedepot.com&lt;/a&gt;</td></tr><tr><td>Furring Strip Board</td><td>Home Depot</td><td>164704</td><td>&lt;a href=&quot;http://www.homedepot.com&quot;&gt;www.homedepot.com&lt;/a&gt;</td></tr><tr><td>Tangle-Trap Insect Trap Coating</td><td>BioControl Network</td><td>268941</td><td>&lt;a href=&quot;http://www.biconet.com&quot;&gt;www.biconet.com&lt;/a&gt;</td></tr><tr><td>Laptop Computer&amp;nbsp;</td><td>Apple</td><td></td><td>&lt;a href=&quot;http://www.apple.com/mac/&quot;&gt;www.apple.com/mac/&lt;/a&gt;</td></tr><tr><td>Mineral oil</td><td>Fisher Scientific</td><td>BP26291</td><td>&lt;a href=&quot;http://www.fishersci.com&quot;&gt;www.fishersci.com&lt;/a&gt;</td></tr><tr><td>White poster board</td><td>Staples</td><td>247403</td><td>&lt;a href=&quot;http://www.staples.com&quot;&gt;www.staples.com&lt;/a&gt;</td></tr><tr><td>Polystyrene weighing dish</td><td>Fisher Scientific</td><td>S67091A</td><td>&lt;a href=&quot;http://www.fishersci.com&quot;&gt;www.fishersci.com&lt;/a&gt;</td></tr><tr><td>ImageJ Software</td><td>National Institutes of Health</td><td></td><td>&lt;a href=&quot;http://rsb.info.nih.gov/ij/&quot;&gt;http://rsb.info.nih.gov/ij/&lt;/a&gt;</td></tr><tr><td>Digital camera</td><td>Sony</td><td>DSC-TX7</td><td>&lt;a href=&quot;http://www.store.sony.com&quot;&gt;www.store.sony.com&lt;/a&gt;</td></tr><tr><td>Fine forceps</td><td>Fine Science Tools</td><td></td><td>&lt;a href=&quot;http://www.finescience.com&quot;&gt;www.finescience.com&lt;/a&gt;</td></tr><tr><td>Polycarbonate cylinder (drop tube)</td><td>McMaster-Carr</td><td>8585K62</td><td>&lt;a href=&quot;http://www.mcmaster.com&quot;&gt;www.mcmaster.com&lt;/a&gt;</td></tr><tr><td>Flight cylinder (acrylic)</td><td>McMaster-Carr</td><td>8486K943</td><td>&lt;a href=&quot;http://www.mcmaster.com&quot;&gt;www.mcmaster.com&lt;/a&gt;</td></tr><tr><td>Polycarbonate sheets</td><td>McMaster-Carr</td><td>85585K25</td><td>&lt;a href=&quot;http://www.mcmaster.com&quot;&gt;www.mcmaster.com&lt;/a&gt;</td></tr><tr><td>ring stand (2x)</td><td>Fisher Scientific</td><td>S47808</td><td>&lt;a href=&quot;http://www.fishersci.com&quot;&gt;www.fishersci.com&lt;/a&gt;</td></tr><tr><td>Ring support</td><td>Fisher Scientific</td><td>S47791</td><td>&lt;a href=&quot;http://www.fishersci.com&quot;&gt;www.fishersci.com&lt;/a&gt;</td></tr><tr><td>Three-prong extension clamps (x2)</td><td>Fisher Scientific</td><td>05-769-7Q</td><td>&lt;a href=&quot;http://www.fishersci.com&quot;&gt;www.fishersci.com&lt;/a&gt;</td></tr><tr><td>Funnel</td><td>Fisher Scientific</td><td>10-500-3</td><td>&lt;a href=&quot;http://www.fishersci.com&quot;&gt;www.fishersci.com&lt;/a&gt;</td></tr><tr><td>chain clamps (2x)</td><td>VWR</td><td>21573-275</td><td>&lt;a href=&quot;http://www.vwr.com&quot;&gt;www.vwr.com&lt;/a&gt;</td></tr><tr><td>Glass vials</td><td>VWR</td><td>66020-198</td><td>&lt;a href=&quot;http://www.vwr.com/&quot;&gt;www.vwr.com&lt;/a&gt;</td></tr></tbody></table>

an improved method for accurate and rapid measurement of flight performance in drosophila

果蝇已经被证明是对行为的分析，一个有用的模型系统，包括飞行。在初始飞行测试涉及的滴飞入的油包覆的量筒;降落高度产生足够的推力，使其与圆筒的内壁接触之前提供通过评估多远苍蝇落入飞行性能的量度。在这里，我们描述了飞行测试仪有四个重大改进的更新版本。首先，我们增加了一个"落管"，以确保所有的苍蝇进入飞行圆柱体试验之间类似的速度，用户之间消除了变异。其次，我们与涂在缠结陷阱，旨在捕捉活虫粘合剂可拆卸的塑料片代替油涂层。第三，我们使用一个较长的气缸以使飞行能力更精确鉴别。第四，我们使用数码相机和成像软件，可以自动飞行性能得分。这些改进允许说唱ID，的飞行行为的定量评估，对于大型数据集和大规模遗传筛选有用的。

<html> 图1A示出了更新后的航班仪组件的示意图。 图1B示出的轨道设计允许相机拍摄的全景图像而不阻塞的视野。代表性的结果示于图2，其中slowpoke突变体果 ​​蝇，它具有已知的飞行缺陷8-10的飞行性能，进行比较，野生型广-S苍蝇。控制苍蝇一贯降落靠近圆筒的顶部，与个体之间的非常小的扩散和73±2.0厘米平均降落高度。相比之下，slowpoke苍蝇显示更加多样化登陆蔓延，土地显著降低，平均44±4.1厘米所有的苍蝇都3天，在室温（23℃）升高。 s/ftp_upload/51223/51223fig1.jpg"宽度="600px的"/&gt; 图1。飞行测试仪，摄像头，并跟踪图。（A）插图的设置为更新后的飞行测试。环支架1成立90厘米高的飞行缸;环站2持有漏斗和10厘米长的"滴管"。摄像头（二）图和线路上使用，以产生一个全景图像。该相机应该由轨道不从镜头阻碍观点得到支持。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/51223/51223fig1highres.jpg" target="_blank">点击这里查看大图</a> 。 <img alt="图2" fo:content-width="6in" fo:src="/files/ftp_upload/51223/51223fig2highres.jpg" src="/files/ftp_upload/51223/51223fig2.jpg" width="600px" /> 图2。从样品的飞行试验代表性的结果。的3日龄钙飞行能力的比较广-S控制飞往slowpoke突变体（SLO TS1）。 （A）屏幕捕获的slowpoke和控制苍蝇显示个别苍蝇降落高度。每一个蓝色圆圈代表一个人飞的位置。这些降落高度被用来计算平均降落高度（B），以及对每种基因型总体分布（C）。雄性和雌性果蝇汇集在一起​​每个样品中。误差棒代表平均值的标准误差。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/51223/51223fig2highres.jpg" target="_blank">点击这里查看大图</a> 。

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An Improved Method for Accurate and Rapid Measurement of Flight Performance in Drosophila

这里，我们描述了一种用于飞行性能果蝇快速和准确的测量，从而实现高通量筛选。

在对飞行性能的准确，快速测定方法的改进<em&gt;果蝇</em

Drosophila has proven to be a useful model system for analysis of behavior, including flight. The initial flight tester involved dropping flies into an ...

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An Improved Method for Accurate and Rapid Measurement of Flight Performance in Drosophila

11.8K 次观看.  University of Wisconsin-Madison. 这里，我们描述了一种用于飞行性能果蝇快速和准确的测量，从而实现高通量筛选。 

视频: An Improved Method for Accurate and Rapid Measurement of Flight Performance in Drosophila

The TreadWheel: Interval Training Protocol for Gently Induced Exercise in Drosophila melanogaster

The incidence of complex metabolic diseases has increased as a result of a widespread transition towards lifestyles of increased caloric intake and lowered activity levels. These multifactorial diseases arise from a combination of genetic, environmental, and behavioral factors. One such complex disease is Metabolic Syndrome (MetS), which is a cluster of metabolic disorders, including hypertension, hyperglycemia, and abdominal obesity. Exercise and dietary intervention are the primary treatments recommended by doctors to mitigate obesity and its subsequent metabolic diseases. Exercise intervention, in particular aerobic interval training, stimulates favorable changes in the common risk factors for Type 2 Diabetes Mellitus (T2DM), Cardiovascular Disease (CVD), and other conditions. With the influx of evidence describing the therapeutic effect exercise has on metabolic health, establishing a system that models exercise in a controlled setting provides a valuable tool for assessing the effects of exercise in an experimental context. Drosophila melanogaster is a great tool for investigating the physiological and molecular changes that result from exercise intervention. The flies have short lifespans and similar mechanisms of metabolizing nutrients when compared to humans. To induce exercise in Drosophila, we developed a machine called the TreadWheel, which utilizes the fly&#39;s innate, negative geotaxis tendency to gently induce climbing. This enables researchers to perform experiments on large cohorts of genetically diverse flies to better understand the genotype-by-environment interactions underlying the effects of exercise on metabolic health.

The TreadWheel: Interval Training Protocol for Gently Induced Exercise in Drosophila melanogaster

The TreadWheel uses rotational motion to gently induce exercise in adult Drosophila melanogaster by exploiting flies&#39; innate, negative geotaxis. It allows for analysis of the interactions between exercise and factors, such as genotype, sex, and diet, and their effect on physiological and molecular assays to assess metabolic health.

TreadWheel:果蝇轻度诱导运动的间歇训练协议

The incidence of complex metabolic diseases has increased as a result of a widespread transition towards lifestyles of increased caloric intake and ...

科研

遗传学

An Automated Rapid Iterative Negative Geotaxis Assay for Analyzing Adult Climbing Behavior in a Drosophila Model of Neurodegeneration

Neurodegenerative diseases are frequently associated with a progressive loss of movement ability, reduced life span, and age-dependent neurodegeneration. To understand the mechanism of these cellular events, and their causal relationships with each other, Drosophila melanogaster, with its sophisticated genetic tools and diverse behavioral features, are used as disease models for assessing neurodegenerative phenotypes. Here we describe a high-throughput method to analyze Drosophila adult negative geotaxis behavior, as an indication for possible motor defects associated with neurodegeneration. An automated machine is designed and developed to drive fly synchronization using an initial electric impulse, later allowing the recording of negative geotaxis behavior over a course of secs to mins. Images from the digitally recorded video are then processed with the self-designed RflyDetection software for statistical data manipulation. Different from the manually controlled negative geotaxis assay based on single fly, this precise, fast, and high-throughput protocol allows data acquisition from more than hundreds of flies simultaneously, providing an efficient approach to advance our understanding in the underlying mechanism of locomotor deficits associated with neurodegeneration.

An Automated Rapid Iterative Negative Geotaxis Assay for Analyzing Adult Climbing Behavior in a Drosophila Model of Neurodegeneration

This step-by-step protocol analyzes Drosophila negative geotaxis behavior using an automated multi-cylinder system that hosts hundreds of flies and synchronizes their action by an electric motor. Upon synchronization, fly negative geotaxis behavior is assayed, digitally recorded, and analyzed using the self-designed RflyDetection software.

一种自动快速迭代阴性 Geotaxis 分析方法在神经的果蝇模型中的成人攀爬行为

Neurodegenerative diseases are frequently associated with a progressive loss of movement ability, reduced life span, and age-dependent neurodegeneration. ...

行为学

High-Resolution Video Tracking of Locomotion in Adult Drosophila Melanogaster

Flies provide an important model for studying complex behavior due to the plethora of genetic tools available to researchers in this field. Studying locomotor behavior in Drosophila melanogaster relies on the ability to be able to quantify changes in motion during or in response to a given task. For this reason, a high-resolution video tracking system, such as the one we describe in this paper, is a valuable tool for measuring locomotion in real-time. Our protocol involves the use of an initial air pulse to break the flies momentum, followed by a thirty second filming period in a square chamber. A tracking program is then used to calculate the instantaneous speed of each fly within the chamber in 10 msec increments. Analysis software then compiles this data, and outputs a variety of parameters such as average speed, max speed, time spent in motion, acceleration, etc. This protocol will discuss proper feeding and management of flies for behavioral tasks, handling flies without anesthetization or immobilization, setting up a controlled environment, and running the assay from start to finish.

The study of complex locomotor behavior in Drosophila melanogaster is dependent upon the ability to quantify changes in a given fly's movement. This article demonstrates how to do this using a high-resolution tracking system.

高分辨率视频跟踪移动的，在成年果蝇。

Flies provide an important model for studying complex behavior due to the plethora of genetic tools available to researchers in this field.  Studying ...

生物学

Methods to Assay Drosophila Behavior

Drosophila melanogaster, the fruit fly, has been used to study molecular mechanisms of a wide range of human diseases such as cancer, cardiovascular disease and various neurological diseases1. We have optimized simple and robust behavioral assays for determining larval locomotion, adult climbing ability (RING assay), and courtship behaviors of Drosophila. These behavioral assays are widely applicable for studying the role of genetic and environmental factors on fly behavior. Larval crawling ability can be reliably used for determining early stage changes in the crawling abilities of Drosophila larvae and also for examining effect of drugs or human disease genes (in transgenic flies) on their locomotion. The larval crawling assay becomes more applicable if expression or abolition of a gene causes lethality in pupal or adult stages, as these flies do not survive to adulthood where they otherwise could be assessed. This basic assay can also be used in conjunction with bright light or stress to examine additional behavioral responses in Drosophila larvae. Courtship behavior has been widely used to investigate genetic basis of sexual behavior, and can also be used to examine activity and coordination, as well as learning and memory. Drosophila courtship behavior involves the exchange of various sensory stimuli including visual, auditory, and chemosensory signals between males and females that lead to a complex series of well characterized motor behaviors culminating in successful copulation. Traditional adult climbing assays (negative geotaxis) are tedious, labor intensive, and time consuming, with significant variation between different trials2-4. The rapid iterative negative geotaxis (RING) assay5 has many advantages over more widely employed protocols, providing a reproducible, sensitive, and high throughput approach to quantify adult locomotor and negative geotaxis behaviors. In the RING assay, several genotypes or drug treatments can be tested simultaneously using large number of animals, with the high-throughput approach making it more amenable for screening experiments.

Methods to Assay Drosophila Behavior

Drosophila melanogaster is a genetically and behaviorally tractable model system that has been used to understand the molecular and cellular basis of many important biological processes for over a century 1. Drosophila has been well exploited to gain insights into the genetic basis of fly behavior.

对检测方法果蝇行为

Drosophila melanogaster, the fruit fly, has been used to study molecular mechanisms of a wide range of human diseases such as cancer, cardiovascular ...

神经科学

An Automated Method to Determine the Performance of Drosophila in Response to Temperature Changes in Space and Time

Temperature is a ubiquitous environmental factor that affects how species distribute and behave. Different species of Drosophila fruit flies have specific responses to changing temperatures according to their physiological tolerance and adaptability. Drosophila flies also possess a temperature sensing system that has become fundamental to understanding the neural basis of temperature processing in ectotherms. We present here a temperature-controlled arena that permits fast and precise temperature changes with temporal and spatial control to explore the response of individual flies to changing temperatures. Individual flies are placed in the arena and exposed to pre-programmed temperature challenges, such as uniform gradual increases in temperature to determine reaction norms or spatially distributed temperatures at the same time to determine preferences. Individuals are automatically tracked, allowing the quantification of speed or location preference. This method can be used to rapidly quantify the response over a large range of temperatures to determine temperature performance curves in Drosophila or other insects of similar size. In addition, it can be used for genetic studies to quantify temperature preferences and reactions of mutants or wild-type flies. This method can help uncover the basis of thermal speciation and adaptation, as well as the neural mechanisms behind temperature processing.

An Automated Method to Determine the Performance of Drosophila in Response to Temperature Changes in Space and Time

Here we present a protocol to automatically determine the locomotor performance of Drosophila at changing temperatures using a programmable temperature-controlled arena that produces fast and accurate temperature changes in time and space.

一种自动测定果蝇在空间和时间温度变化中的性能的方法

Temperature is a ubiquitous environmental factor that affects how species distribute and behave. Different species of Drosophila fruit flies have specific ...

Determination of the Spontaneous Locomotor Activity in Drosophila melanogaster

Drosophila melanogaster has been used as an excellent model organism to study environmental and genetic manipulations that affect behavior. One such behavior is spontaneous locomotor activity. Here we describe our protocol that utilizes Drosophila population monitors and a tracking system that allows continuous monitoring of the spontaneous locomotor activity of flies for several days at a time. This method is simple, reliable, and objective and can be used to examine the effects of aging, sex, changes in caloric content of food, addition of drugs, or genetic manipulations that mimic human diseases.

Determination of the Spontaneous Locomotor Activity in Drosophila melanogaster

Drosophila melanogaster are useful in studying genetic or environmental manipulations that affect behaviors such as spontaneous locomotor activity.&#160; Here we describe a protocol that utilizes monitors with infrared beams and data analysis software to quantify spontaneous locomotor activity.&#160;

在自发活动的测定<em&gt;果蝇</em

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

A Low-cost Method for Analyzing Seizure-like Activity and Movement in Drosophila

Video tracking systems have been used widely to analyze Drosophila melanogaster movement and detect various abnormalities in locomotive behavior. While these systems can provide a wealth of behavioral information, the cost and complexity of these systems can be prohibitive for many labs. We have developed a low-cost assay for measuring locomotive behavior and seizure movement in D. melanogaster. The system uses a web-cam to capture images that can be processed using a combination of inexpensive and free software to track the distance moved, the average velocity of movement and the duration of movement during a specified time-span. To demonstrate the utility of this system, we examined a group of D. melanogaster mutants, the Bang-sensitive (BS) paralytics, which are 3-10 times more susceptible to seizure-like activity (SLA) than wild type flies. Using this novel system, we were able to detect that the BS mutant bang senseless (bss) exhibits lower levels of exploratory locomotion in a novel environment than wild type flies. In addition, the system was used to identify that the drug metformin, which is commonly used to treat type II diabetes, reduces the intensity of SLA in the BS mutants.

A Low-cost Method for Analyzing Seizure-like Activity and Movement in Drosophila

Using a webcam and a combination of free and inexpensive software programs, locomotive patterns in Drosophila melanogaster can be analyzed to detect differences in the speed, distance, and time of various motor activities.

在分析癫痫样活动和运动的一种低成本方法<em&gt;果蝇</em

Video tracking systems have been used widely to analyze Drosophila melanogaster movement and detect various abnormalities in locomotive behavior. While ...

A Simple Technique to Assay Locomotor Activity in Drosophila

Drosophila melanogaster is an ideal model organism for studying various diseases due to its abundance of advanced genetic manipulation techniques and diverse behavioral features. Identifying behavioral deficiency in animal models is a crucial measure of disease severity, for example, in neurodegenerative diseases where patients often experience impairments in motor function. However, with the availability of various systems to track and assess motor deficits in fly models, such as drug-treated or transgenic individuals, an economical and user-friendly system for precise evaluation from multiple angles is still lacking. A method based on the AnimalTracker application programming interface (API) is developed here, which is compatible with the Fiji image processing program, to systematically evaluate the movement activities of both adult and larval individuals from recorded video, thus allowing for the analysis of their tracking behavior. This method requires only a high-definition camera and a computer peripheral hardware integration to record and analyze behavior, making it an affordable and effective approach for screening fly models with transgenic or environmental behavioral deficiencies. Examples of behavioral tests using pharmacologically treated flies are given to show how the techniques can detect behavioral changes in both adult flies and larvae in a highly repeatable manner.

A Simple Technique to Assay Locomotor Activity in Drosophila

The present protocol assesses the locomotor activity of Drosophila by tracking and analyzing the movement of flies in a hand-made arena using open-source software Fiji, compatible with plugins to segment pixels of each frame based on high-definition video recording to calculate parameters of speed, distance, etc.

一种测定果蝇运动活性的简单技术

Drosophila melanogaster is an ideal model organism for studying various diseases due to its abundance of advanced genetic manipulation techniques and ...

Measurement of Metabolic Rate in Drosophila using Respirometry

Metabolic disorders are a frequent problem affecting human health. Therefore, understanding the mechanisms that regulate metabolism is a crucial scientific task. Many disease causing genes in humans have a fly homologue, making Drosophila a good model to study signaling pathways involved in the development of different disorders. Additionally, the tractability of Drosophila simplifies genetic screens to aid in identifying novel therapeutic targets that may regulate metabolism. In order to perform such a screen a simple and fast method to identify changes in the metabolic state of flies is necessary. In general, carbon dioxide production is a good indicator of substrate oxidation and energy expenditure providing information about metabolic state. In this protocol we introduce a simple method to measure CO2 output from flies. This technique can potentially aid in the identification of genetic perturbations affecting metabolic rate.

Measurement of Metabolic Rate in Drosophila using Respirometry

Metabolic disorders are among one of the most common diseases in humans. The genetically tractable model organism D. melanogaster can be used to identify novel genes that regulate metabolism. This paper describes a relatively simple method which allows studying the metabolic rate in flies by measuring their CO2 production.

代谢率的测量<em&gt;果蝇</em&gt;使用呼吸运动

Metabolic disorders are a frequent problem affecting human health. Therefore, understanding the mechanisms that regulate metabolism is a crucial ...

Measurement of Larval Activity in the Drosophila Activity Monitor

Drosophila larvae are used in many behavioral studies, yet a simple device for measuring basic parameters of larval activity has not been available. This protocol repurposes an instrument often used to measure adult activity, the TriKinetics Drosophila activity monitor (MB5 Multi-Beam Activity Monitor) to study larval activity. The instrument can monitor the movements of animals in 16 individual 8 cm glass assay tubes, using 17 infrared detection beams per tube. Logging software automatically saves data to a computer, recording parameters such as number of moves, times sensors were triggered, and animals&#8217; positions within the tubes. The data can then be analyzed to represent overall locomotion and/or position preference as well as other measurements. All data are easily accessible and compatible with basic graphing and data manipulation software. This protocol will discuss how to use the apparatus, how to operate the software and how to run a larval activity assay from start to finish.

Measurement of Larval Activity in the Drosophila Activity Monitor

This report describes a method for measuring Drosophila larval activity using the TriKinetics Drosophila Activity Monitor. The device employs infrared beams to detect movements of up to 16 individual animals. Data can be analyzed to represent motion parameters including rates and the positions of the animals within the assay chambers.

幼虫活动的测量<em&gt;果蝇</em&gt;活动监视器

Drosophila larvae are used in many behavioral studies, yet a simple device for measuring basic parameters of larval activity has not been available. This ...

在对飞行性能的准确，快速测定方法的改进

摘要

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此视频中的章节

TreadWheel:果蝇轻度诱导运动的间歇训练协议

一种自动快速迭代阴性 Geotaxis 分析方法在神经的果蝇模型中的成人攀爬行为

高分辨率视频跟踪移动的，在成年果蝇。

对检测方法果蝇行为

一种自动测定果蝇在空间和时间温度变化中的性能的方法

在自发活动的测定

在分析癫痫样活动和运动的一种低成本方法

一种测定果蝇运动活性的简单技术

代谢率的测量

幼虫活动的测量