Name: a single-fly assay for foraging behavior in drosophila
Uploaded: 2013-11-04T21:30:00
Description: Watch this Scientific Journal Video about A Single-fly Assay for Foraging Behavior in Drosophila at JoVE.com

这项工作是由研究经费JWW从美国国立卫生研究院（R01DK092640）和美国国家科学基金会（0920668）的支持。

1。粉煤灰收集和饥饿<ol><li>后面的实验苍蝇在控制温度和湿度条件下（ 如 21℃，50〜60％相对湿度）在12小时亮/暗周期。 </li><li>收集雌蝇在羽化的日子，把它们摆放，伴随着4-5的男性，成为新的食品小瓶（最多30个每小瓶）。年龄过得2-5天。 </li><li>准备室的苍蝇饥饿。 <ol><li>推单一组织（4.8×8.4英寸）下降到一个空的塑料小瓶的底部。完全浸泡的组织用蒸馏水冲洗。使用对象向下推纸巾?...

<ol>
	<li>Dethier, V. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The hungry fly : a physiological study of the behavior associated with feeding. , (1976).</li><li>Root, C. M., Ko, K. I., Jafari, A., Wang, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Presynaptic+facilitation+by+neuropeptide+signaling+mediates+odor-driven+food+search.">Presynaptic facilitation by neuropeptide signaling mediates odor-driven food search.</a> Cell. 145, 133-144 (2011).</li><li>Root, C. M., 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+presynaptic+gain+control+mechanism+fine-tunes+olfactory+behavior.">A presynaptic gain control mechanism fine-tunes olfactory behavior.</a> Neuron. 59, 311-321 (2008).</li><li>Semmelhack, J. L., Wang, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Select+Drosophila+glomeruli+mediate+innate+olfactory+attraction+and+aversion.">Select Drosophila glomeruli mediate innate olfactory attraction and aversion.</a> Nature. 459, 218-223 (2009).</li><li>Faucher, C., Forstreuter, M., Hilker, M., de Bruyne, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+responses+of+Drosophila+to+biogenic+levels+of+carbon+dioxide+depend+on+life-stage,+sex+and+olfactory+context.">Behavioral responses of Drosophila to biogenic levels of carbon dioxide depend on life-stage, sex and olfactory context.</a> J. Exp. Biol. 209, 2739-2748 (2006).</li><li>Quinn, W. G., Harris, W. A., 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=Conditioned+behavior+in+Drosophila+melanogaster.">Conditioned behavior in Drosophila melanogaster.</a> Proc. Natl. Acad. Sci. U.S.A. 71, 708-712 (1974).</li><li>Fishilevich, E., Domingos, A. I., Asahina, K., Naef, F., Vosshall, L. B., Louis, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chemotaxis+behavior+mediated+by+single+larval+olfactory+neurons+in+Drosophila.">Chemotaxis behavior mediated by single larval olfactory neurons in Drosophila.</a> Curr. Biol. 15, 2086-2096 (2005).</li><li>Venken, K. J., Simpson, J. H., 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=Genetic+manipulation+of+genes+and+cells+in+the+nervous+system+of+the+fruit+fly.">Genetic manipulation of genes and cells in the nervous system of the fruit fly.</a> Neuron. 72, 202-230 (2011).</li><li>Hamada, F. N., 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=An+internal+thermal+sensor+controlling+temperature+preference+in+Drosophila.">An internal thermal sensor controlling temperature preference in Drosophila.</a> Nature. 454, 217-220 (2008).</li><li>Kitamoto, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Conditional+modification+of+behavior+in+Drosophila+by+targeted+expression+of+a+temperature-sensitive+shibire+allele+in+defined+neurons.">Conditional modification of behavior in Drosophila by targeted expression of a temperature-sensitive shibire allele in defined neurons.</a> J. Neurobiol. 47, 81-92 (2001).</li><li>Sweeney, S. T., Broadie, K., Keane, J., Niemann, H., O'Kane, C. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Targeted+expression+of+tetanus+toxin+light+chain+in+Drosophila+specifically+eliminates+synaptic+transmission+and+causes+behavioral+defects.">Targeted expression of tetanus toxin light chain in Drosophila specifically eliminates synaptic transmission and causes behavioral defects.</a> Neuron. 14, 341-351 (1995).</li><li>Brand, A. H., Perrimon, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Targeted+gene+expression+as+a+means+of+altering+cell+fates+and+generating+dominant+phenotypes.">Targeted gene expression as a means of altering cell fates and generating dominant phenotypes.</a> Development. 118, 401-415 (1993).</li></ol>

在这个协议中，我们描述了食物搜索行为分析一步一步的过程。除了食物有关的气味，它也可适于用于苍蝇的定位其他气味物体的能力的研究。例如，它可能会朝雄蝇3在队友的本地化行为的研究应用还有几个额外的考虑这个协议，我们将在这里提到有关此过程： 首先，饲养温度决定实验苍蝇多久应该进行老化测试之前。建议年龄的范围进行检查，以确定最合适的年?...

饥饿的国家促进两种欲求行为：寻找食物和食品消费1。这个简单的行为测定法是与觅食2,3相关的趋化行为的研究非常有用。具体来说，它跟踪飞位置，行走速度和延迟定位食物气味的目标。食品发现的延迟作为度量测量的变化在苍蝇的气味探测系统的变化在其内部欲求状态的下游的灵敏度。此法的手动版是以前用来显示GABA-B受体信号是很重要的气味本地化行为，成蝇3。检测的电流自动版本是有助的在多短神经肽F（sNPF）信令重塑地图嗅觉的果蝇和影响欲求行为2的研究。 测试是在黑暗的，温度和湿度控制的房间进行。数字上面的透明亚克力板的测试设置摄像机跟踪苍蝇由660 nm的LED照明背光。从相机信息是实时由驻旁边的测试区域的计算机进行处理。我们使用的数据采集软件在测试期间录制和保存苍蝇位置的坐标。 在这个范例中，受试者被释放到包含在该中心食品气味的舞台;气味对象创建诱导苍蝇的食物搜索行为竞技场内食品串味梯度。类似的气味搜索协议已对chemosensation在单一果蝇幼虫7的研究应用。而其他行为分析，如四场嗅觉4,5或T型迷宫6评估厌恶的气味或退房的行为，这种模式是最适合评估嗅觉的灵敏度和趋化行为。 几个关键的优势伴随着这个ASSA年。首先，它允许快速获取大数据集，因为数据收集和分析大多是自动化的。第二，该测定分离和测量单个苍蝇的行为，从而消除社会嗅觉可能影响他们的行为。三，协议和简单的实验设计的简单性使检测效率，容易教别人。 另外，此法可用于进一步探讨神经回路基本食品的搜索行为由它提供给果蝇 8的广泛的遗传工具箱结合。转基因的靶向表达，沉默或激发的神经元可以与如GAL4-UAS系统，以及无人机，shibire TS1，UAS-破伤风毒素，和UAS-TRPA1（B）工具来实现转基因9-12。

<table border="1">
	<tbody>
		<tr>
			<td>Name of Reagent/Material</td>
			<td>Company </td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Apple Cider Vinegar</td>
			<td>Spectrum</td>
			<td></td>
			<td>commercially available</td>
		</tr>
		<tr>
			<td>Agarose, Type VII</td>
			<td>Sigma-Aldrich</td>
			<td>A0701</td>
			<td>low gelling temperature agarose</td>
		</tr>
		<tr>
			<td>Acrylic Testing Plate</td>
			<td></td>
			<td>custom</td>
			<td>Plate contains 6 arenas. Each arena is 60 mm in diameter 6 mm in height. See testing plate diagrams for specific measurements.</td>
		</tr>
		<tr>
			<td>LabVIEW V.8.5</td>
			<td>National Instruments</td>
			<td>776670-09</td>
			<td>platform for programs: PositioningTool.vi, FlyTracking--Six Zones.vi NOTE: &#34;elapsed time.vi&#34;, &#34;time into file.vi&#34;, and &#34;two object detect.vi&#34; are included subroutines that must be available in order for the main data acquisition program &#34;FlyTracking--Six zones.vi&#34; to run.</td>
		</tr>
		<tr>
			<td>LabVIEW Vision 8.5</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>LabVIEW Vision Acquisition Software 8.5</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>LabVIEW Vision Builder AI 3.5</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Igor Pro V.6</td>
			<td>Wavemetric, Inc.</td>
			<td></td>
			<td>platform for macro: Data Analysis for Fly Tracking--Six Zones</td>
		</tr>
		<tr>
			<td>Basler scA1390-17fm</td>
			<td>National Instruments</td>
			<td>779980-01</td>
			<td>Digital Camera NOTE: driver for camera available at Baslerweb.com</td>
		</tr>
		<tr>
			<td>8 mm lens</td>
			<td>National Instruments</td>
			<td>780024-01</td>
			<td>Lens for Basler Digital Camera</td>
		</tr>
		<tr>
			<td>Ground Glass Diffuser Plate</td>
			<td>Edmund Optics</td>
			<td>custom</td>
			<td>Diffuses light, 25 cm x 30 cm</td>
		</tr>
		<tr>
			<td>US Std. No. 100</td>
			<td>Fischer Scientific</td>
			<td>04-881X</td>
			<td>Sieve with nominal opening of 150 &#956;m</td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td>Lighting Option 1</td>
		</tr>
		<tr>
			<td>LED backlight 660 nm (20 cm x 20 cm)</td>
			<td>Spectra West</td>
			<td>BL47192</td>
			<td>a simpler but more expensive lighting option.</td>
		</tr>
		<tr>
			<td>Power Supply for LED Backlight</td>
			<td>Spectra West</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td>Lighting Option 2</td>
		</tr>
		<tr>
			<td>660 nm LEDs</td>
			<td>Superbrightleds</td>
			<td>RL5R1330</td>
			<td>Wavelength 660 nm (approximately 7 x 7 LED array for a 14.7 inch x 9.75 inch panel)</td>
		</tr>
		<tr>
			<td>Linear DC Power Supply</td>
			<td>GW Instek</td>
			<td>GPS-1830D</td>
			<td>Power supply for LED Panel</td>
		</tr>
		<tr>
			<td>Solderless Breadboard</td>
			<td>Digikey</td>
			<td>922354-ND</td>
			<td>Breadboard for LEDs</td>
		</tr>
	</tbody>
</table>

a single-fly assay for foraging behavior in drosophila

对于许多动物，饥饿促进了有利于寻找适当的食物来源的方式变化的嗅觉系统。在这个视频文章中，我们描述了一个自动检测测量饥饿或过饱的嗅觉依赖食物搜索行为的影响在成年果蝇果蝇 。在一个不透光的盒子由红灯是看不见的果蝇照明，链接到自定义数据采集软件的摄像机同时监控6苍蝇的位置。每个飞局限于走在包含在该中心食物的气味个别领域。测试领域休息多孔层的功能，以防止气味的积累。延迟来定位气味源，它反映在不同的生理状态嗅觉灵敏度的度量，是通过软件分析确定。在这里，我们讨论运行此行为范式的关键力学和涉及有关飞LOADI具体问题纳克，气味污染，检测温度，数据质量和统计分析。

数据分析软件和布局，其中的一个例子可在​​图1中可以看出，是根据一组数据的标准使用其10分钟试验期间，以评估每个蝇的效能。下列标准用于确定是否从每个苍蝇的数据将被用于数据分析，其目的是消除这些苍蝇是无法执行食品搜索任务因受伤，疾病，压力，或缺乏动力。 苍蝇不活动的时间超过300秒被认为是“无效”，并从数据集将被拒绝，除非它们一）...

Watch this Scientific Journal Video about A Single-fly Assay for Foraging Behavior in Drosophila at JoVE.com

A Single-fly Assay for Foraging Behavior in Drosophila

在这个视频文章中，我们描述了一个自动检测测量饥饿或过饱的嗅觉依赖食物搜索行为的影响在成年果蝇果蝇。

单苍蝇检测方法的觅食行为的<em&gt;果蝇</em

For many animals, hunger promotes changes in the olfactory system in a manner that facilitates the search for appropriate food sources. In this video ...

Research

JoVE Journal

Neuroscience

A Single-fly Assay for Foraging Behavior in Drosophila

Single Sensillum Recordings in the Insects Drosophila melanogaster and Anopheles gambiae

Single Sensillum Recordings in the Insects Drosophila melanogaster and Anopheles gambiae

在昆虫单感器记录果蝇按蚊</em

The sense of smell is essential for insects to find foods, mates, predators, and oviposition sites3. Insect olfactory sensory neurons (OSNs) are enclosed ...

科研

神经科学

Methods to Assay Drosophila Behavior

Methods to Assay Drosophila 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 ...

Optogenetic Stimulation of Escape Behavior in Drosophila melanogaster

Optogenetic Stimulation of Escape Behavior in Drosophila melanogaster

Optogenetic刺激的逃避行为<em&gt;果蝇</em

A  growing number of genetically encoded tools are becoming available that allow non-invasive  manipulation of the neural activity of specific neurons in ...

A Procedure for Implanting a Spinal Chamber for Longitudinal In Vivo Imaging of the Mouse Spinal Cord

A Procedure for Implanting a Spinal Chamber for Longitudinal In Vivo Imaging of the Mouse Spinal Cord

一个程序，植入脊柱商会纵<em&gt;在体内</em&gt;鼠标脊髓成像

Studies in the mammalian neocortex have enabled unprecedented resolution of cortical structure, activity, and response to neurodegenerative insults by ...

Dyeing Insects for Behavioral Assays: the Mating Behavior of Anesthetized Drosophila

Dyeing Insects for Behavioral Assays: the Mating Behavior of Anesthetized Drosophila

染色昆虫行为检测：麻醉的交配行为<em&gt;果蝇</em

Mating experiments using Drosophila have contributed greatly to the understanding of sexual selection and behavior. Experiments often require simple, easy ...

A Behavioral Assay for Mechanosensation of MARCM-based Clones in Drosophila melanogaster

A Behavioral Assay for Mechanosensation of MARCM-based Clones in Drosophila melanogaster

一个行为分析对机械感觉MARCM为基础的克隆<em&gt;果蝇</em

Because of the structural and functional homology to the hair cells of the mammalian inner ear, the neurons that innervate the Drosophila external sense ...

Foraging Path-length Protocol for Drosophila melanogaster Larvae

Foraging Path-length Protocol for Drosophila melanogaster Larvae

觅食路径长度协议<em&gt;果蝇</em&gt;幼虫

The Drosophila melanogaster larval path-length phenotype is an established measure used to study the genetic and environmental contributions to behavioral ...

A Simple Way to Measure Alterations in Reward-seeking Behavior Using Drosophila melanogaster

A Simple Way to Measure Alterations in Reward-seeking Behavior Using Drosophila melanogaster

一个简单的方法来衡量在改建悬赏寻求行为使用<em&gt;果蝇</em

We describe a protocol for measuring ethanol self-administration in fruit flies (Drosophila melanogaster) as a proxy for changes in reward states. We ...

Quantification of Drosophila Grooming Behavior

果蝇美容行为的量化

Drosophila grooming behavior is a complex multi-step locomotor program that requires coordinated movement of both forelegs and hindlegs. Here we present a ...

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

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

果蝇岛试验中运动行为的高通量分析

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