Name: high-resolution quantification of odor-guided behavior in drosophila melanogaster using the flywalk paradigm
Uploaded: 2015-12-11T15:00:00
Description: Watch this Scientific Journal Video about High-resolution Quantification of Odor-guided Behavior in Drosophila melanogaster Using the Flywalk Paradigm at JoVE.com

我们感谢丹尼尔·维特技术援助和佩德罗戈维亚在Electricidade EM宝（electricidadeempo.net）定制的跟踪软件为我们的要求。我们也感谢汤姆Retzke支持在拍摄过程。这项研究是由马克斯 - 普朗克协会的支持。

Flywalk的结构和技术细节已描述在别处26（如果是建立这一设置，进一步的信息可以从MK获得的任何问题）。在这里，我们重点的模式，这将有助于获得可靠的结果的处理的详细说明。 1.飞处理<ol><li>后飞低到下一个12小时的食品中27中密度培养：12小时暗条件在23-25℃，相对湿度70％。为此，让20-30新出现的成蝇在一个大的食品小瓶重现1周，然后丢弃成蝇?...

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虽然Flywalk系统似乎相当复杂乍看之下，一旦建立并运行它易于使用，并产生非常强大的结果。强调与生物测定所产生的结果的一致性，可以说的是，这里示出的代表性的结果差不多2年后一些与使用新的跟踪软件和光源的变形设置示出在先前的研究29的结果获得的。尽管如此，引诱反应是 - 尽管略高反应振幅 - 非常类似于以前发表的关于他们的动态。 有哪些应被视为?...

任何神经行为学研究的总体目标是建立单个神经元或神经元电路的活动状态和生物体的行为之间的因果关系。为了实现这一目标的神经元活动和行为应该在相同的激励条件被监控并且这些刺激条件下理想地应当类似于受到审查神经系统发展到使感。特别是当它涉及到行为的生物测定，这些需求在历史上被证明相当苛刻果蝇嗅觉神经行为学。 一旦从源释放，气味羽状迅速分解成薄细丝造成空气运动是气味分布1的主要决定因素湍流扩散。其结果是，昆虫导航朝向气味源经历间歇刺激与穿插的洁净空气可变间隔气味包。都步行和飞行昆虫-包括果蝇 -已被证实通过在羽遭遇逆风飙升，主要是移动的侧风在没有气味2利用这一间歇刺激制度导航- 5。而在生理实验刺激程序基本上模仿那些昆虫可以通过提供单个抽吸气味穿插的洁净空气或动态刺激序列6长时间在其天然环境体验- 11，许多行为的生物测定在果蝇神经行为使用，诸如阱测定，旷场阿里纳斯或T型迷宫依靠气味梯度的12 - 15。但是，由于气味梯度的定义是在浓度可变根据来自气味源的距离，特定行为不能归因于使用这些范例精确臭气浓度。另外，该斜率气味梯度关键取决于臭气物质的物理化学性质。高挥发性化合物的梯度将比由低挥发性化合物创建并因此也更难跟踪生物体依靠测量浓度差在空间导航16的唯一手段较浅- 20，这可能导致一个曲解嗅觉喜好特别是在选择检测。这个效果也非常有害调查朝气味的混合物时的行为，因为它导致不同混合组分比率在空间中的每个点，因此，再次排除生理和行为之间有明显的相关性。 虽然醋蝇倾向于聚集在发酵的水果，它们是独居在他们对食品来源和产卵场所导航。然而，用在果蝇 neuroetholo而不是测试个别动物的许多行为范式gy的检查蝇同伙的气味引导行为和吸引力的得分为苍蝇选择气味过控制刺激的分数。这些人群实验极大地促进了飞神经行为学的理解，许多使用他们提出的意见可以在单飞行实验来证实。然而，已经观察到，苍蝇可以影响每个other&#39;s 第 21和在极端情况下的气味的评价，可以从漠不关心切换撤销取决于人口密度22。此外，从这类实验结果往往提供的行为决定，而不是观察什么苍蝇做，而它是这样做，试图与相关神经元活动的行为时，这将是可取的序列的只有终点。这些相当低解析度人群实验由高分辨率的单飞的方法，例如允许系留飞行竞技场和跑步机对比在当时直接观察的行为反应刺激呈现20,23,24。然而，队列实验仍然很受欢迎，因为他们是非常有效的，并提供强大的结果，即使在相对较低的样本大小，因为个体间和审间的变化被部分平均出由于观察人口过长时间。而系留飞行和跑步机可能提供关于刺激呈现和时间分辨率的金标准，所使用的领域被设计用于单一动物并且因此费时，得到样品大小所需的统计分析。若干其他方法最近已开发出允许一个高效采集高分辨率行为数据的结合定义良好的刺激机制。这些包括无监督3D跟踪多个醋蝇在风洞中组合与气味羽5的精确的3D模型</SUP&gt;，多个单独的苍蝇在气流从两侧25和 Flywalk范例26供应商会选择跟踪。 在Flywalk，15个人的苍蝇都坐落在小玻璃管，并通过高架摄像头下的红色光条件下连续监测。气味被添加到20厘米/秒的连续气流，并通过玻璃管以恒定的速度行进。气流是通过250毫升瓶进入气味输送系统之前，含蒸馏水（加湿器）传递给它加湿。所述flies&#39;位置的感兴趣区（ROI）包围大部分的气味管的长度（但不包括管子（大约5毫米处每侧的外边缘的正方形区域），其中所述蝇不能移动进一步上调或内记录顺风）周围气味介绍（图1A，B）的时间。飞身份保持恒定由跟踪系统吨hroughout与其Y位（即他们的玻璃管限制）的基础上进行实验。刺激气味是使用一种多组分的刺激装置，该装置允许多达8个单气味和所有可能的混合物26,29（图1B）的介绍实现。一个实验的过程是由计算机调节气味递送系统以及收集的温度和湿度信息（计算机1， 图1C）来控制。这台计算机还控制数据记录器（开始/停止录音）它连续不断地跟踪每秒20帧（电脑2）飞行位置的第二台计算机上。飞位置，气味阀门状态（ 即时间点阀门开度），气味ID，温度和湿度周围的气味刺激周期登录计算机2这样的信息对气味和飞位置是同步的，并出口为.csv文件，文件，这可进一步加工，并使用自定义编写分析程序进行分析。因为整个系统是计算机控制的，没有人的干预的实验会话期间是必要的。

<table border="0" cellpadding="0" cellspacing="0" width="866">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Flywalk setup</td>
			<td style="width:325px;">Custom</td>
			<td style="width:119px;"></td>
			<td style="width:207px;">details available upon request</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">stimulus device</td>
			<td style="width:325px;">Custom</td>
			<td style="width:119px;"></td>
			<td>details available upon request</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">LED cluster</td>
			<td style="width:325px;">Custom</td>
			<td style="width:119px;"></td>
			<td>details available upon request</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">HD Pro Webcam C920</td>
			<td style="width:325px;">Logitech, Lausanne, Switzerland</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">2 Computers</td>
			<td style="width:325px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="18">
			<td height="18" style="height:18px;width:216px;">Flywalk Reloaded v1.0 software</td>
			<td style="width:325px;">Electricidade Em P&#243; (electricidadeempo.net)</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Labview 11.0 software</td>
			<td style="width:325px;">National Instruments, Austin, TX</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Standard fly food</td>
			<td style="width:325px;">Custom</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="18">
			<td height="18" style="height:18px;width:216px;">Standard fly vials</td>
			<td style="width:325px;">Greiner bio-one GmbH, Frickenhausen, Germany</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Standard fly vials</td>
			<td style="width:325px;">Greiner bio-one GmbH, Frickenhausen, Germany</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">aspirator</td>
			<td style="width:325px;">Custom</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">mineral oil</td>
			<td style="width:325px;">Sigma-Aldrich (www.sigmaaldrich.com)</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">odors</td>
			<td style="width:325px;">Sigma-Aldrich (www.sigmaaldrich.com)</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="18">
			<td height="18" style="height:18px;width:216px;">200 &#181;l PCR reaction tubes</td>
			<td style="width:325px;">Biozym Scientific GmbH, Oldendorf, Germany</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
	</tbody>
</table>

high-resolution quantification of odor-guided behavior in drosophila melanogaster using the flywalk paradigm

在自然环境中，昆虫等的飞醋果蝇轰炸了大量的不同化学气味物质。为了进一步使问题复杂化，由昆虫的神经系统检测到的气味通常不是单一化合物而是混合物，其成分和浓度比例有所不同。这导致具有由神经系统进行评估不同嗅觉刺激的几乎无限量。 为了理解其中的刺激气味的各方面通过飞确定其评价，因此，希望能够有效地检查气味引向许多增味剂和气味的混合物的行为。要直接关联的行为神经元的活动，行为应该量化可比较的时间框架，并在相同的刺激条件下在神经生理学实验。然而，在果蝇神经行为学许多目前使用的嗅觉生物测定相当斯佩恰lized无论是对效率或实现的分辨率。 Flywalk，一个自动气味递送和跟踪系统，桥梁效率和分辨率之间的差距。它允许什么时候气味包刺激了行走自如飞行，并确定宠动态行为反应的决心。

因为果蝇被允许通过玻璃管气味脉冲和气味脉冲行进之间自由分发他们的玻璃试管内以恒定的速度苍蝇遇到气味在不同的时间取决于它们的x位置处的刺激的时间。其结果是，通过一个有吸引力的10 -3稀释的乙酸乙酯的500毫秒脉冲诱发逆风轨迹的起始点是由约1秒，在他们的玻璃管的顺风端相比，这些苍蝇坐在接近的延迟为蝇逆风端以20厘米/秒和20厘米的玻璃管长度（图2A）的风?...

Watch this Scientific Journal Video about High-resolution Quantification of Odor-guided Behavior in Drosophila melanogaster Using the Flywalk Paradigm at JoVE.com

High-resolution Quantification of Odor-guided Behavior in Drosophila melanogaster Using the Flywalk Paradigm

自动跟踪系统Flywalk是用于在果蝇气味引导行为高分辨率定量。

在气味引导行为的高分辨率定量<em&gt;果蝇</em&gt;使用<em&gt; Flywalk</em&gt;范式

In their natural environment, insects such as the vinegar fly Drosophila melanogaster are bombarded with a huge amount of chemically distinct odorants. To ...

Research

JoVE Journal

Neuroscience

High-resolution Quantification of Odor-guided Behavior in Drosophila melanogaster Using the Flywalk Paradigm

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

Simultaneous Recording of Calcium Signals from Identified Neurons and Feeding Behavior of Drosophila melanogaster

Simultaneous Recording of Calcium Signals from Identified Neurons and Feeding Behavior of Drosophila melanogaster

从查明的神经元和取食行为的钙信号的同时记录果蝇</em

To study neuronal networks in terms of their function in behavior, we must analyze how neurons operate when each behavioral pattern is generated.  Thus, ...

High-resolution Live Imaging of Cell Behavior in the Developing Neuroepithelium

高分辨率实时成像发展中的神经上皮细胞的行为

The embryonic spinal cord consists of cycling neural progenitor cells that give rise to a large percentage of the neuronal and glial cells of the central ...

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 ...

Determination of the Spontaneous Locomotor Activity in Drosophila melanogaster

Determination of the Spontaneous Locomotor Activity in Drosophila melanogaster

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 ...

Straightforward Assay for Quantification of Social Avoidance in Drosophila melanogaster

Straightforward Assay for Quantification of Social Avoidance in Drosophila melanogaster

简单的检测方法的社交回避的量化<em&gt;果蝇</em

Drosophila melanogaster is an emerging model to study different aspects of social interactions. For example, flies avoid areas previously occupied by ...

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 ...

The CApillary FEeder Assay Measures Food Intake in Drosophila melanogaster

The CApillary FEeder Assay Measures Food Intake in Drosophila melanogaster

毛细管馈线分析测量的摄食<em&gt;果蝇</em

For most animals, feeding is an essential behavior for securing survival, and it influences development, locomotion, health and reproduction. Ingestion of ...

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-resolution Confocal Imaging of the Blood-brain Barrier: Imaging, 3D Reconstruction, and Quantification of Transcytosis

血脑屏障的高分辨率共聚焦成像: 成像、3D 重建和 Transcytosis 的量化

The blood-brain barrier (BBB) is a dynamic multicellular interface that regulates the transport of molecules between the circulation and the brain. ...