简单的检测方法的社交回避的量化<em>果蝇</em
Summary
Here, we present a protocol to quantify the avoidance of stressed individuals. This paradigm is powerful yet user-friendly and can be used to assess the influence of genes and environment on one kind of social interaction in Drosophila melanogaster.
Abstract
Drosophila melanogaster is an emerging model to study different aspects of social interactions. For example, flies avoid areas previously occupied by stressed conspecifics due to an odorant released during stress known as the Drosophila stress odorant (dSO). Through the use of the T-maze apparatus, one can quantify the avoidance of the dSO by responder flies in a very affordable and robust assay. Conditions necessary to obtain a strong performance are presented here. A stressful experience is necessary for the flies to emit dSO, as well as enough emitter flies to cause a robust avoidance response to the presence of dSO. Genetic background, but not their group size, strongly altered the avoidance of the dSO by the responder flies. Canton-S and Elwood display a higher performance in avoiding the dSO than Oregon and Samarkand strains. This behavioral assay will allow identification of mechanisms underlying this social behavior, and the assessment of the influence of genes and environmental conditions on both emission and avoidance of the dSO. Such an assay can be included in batteries of simple diagnostic tests used to identify social deficiencies of mutants or environmental conditions of interest.
Introduction
该方法的目标是很容易量化的果蝇简单的社会行为，独立于求偶和侵略一个新的方面。
社会互动是至关重要的内一个社会的正常发育和健康个体，以及一个社会群体作为一个整体的功能。这些相互作用的高复杂性就必须大的样本大小和一个系统，它允许为行为简化为社会行为的遗传和神经基础仍然知之甚少。 果蝇是可用于识别基因的有力的遗传模型，并社会互动的神经基础。事实上，D.果蝇有复杂的社会行为剧目和社会化的一些直接测量已经完成1-7。不过，这些努力都集中在相对复杂的社会行为，如aggres西伯相互作用3,6，求爱3,8-12的各个方面，以及如何社会经验影响其他行为，如学习，或昼夜节律13-17。此外，许多这些测定依赖于分析蝇基的复杂的交互模式，利用视频跟踪和计算机软件来分析得到的数据的丰度。这样的分析是非常宝贵的，并导致重要的新见解，如在组7苍蝇飞相互作用的动态。一个限制，但是，这些检测到社会大众的交通不便，和基本认同他人的机制的了解有限。换句话说，信号由一个单独的和其识别由另一个发射的基础仍然知之甚少18。
相比之下，苍蝇也表现出一个简单的行为，社交回避，使个人从强调苍蝇发出的信号搬开：该D.黑素ogaster应力加臭剂或DSO 19。在高通量测定法中，此行为可以量化为避免由其他蝇，或社交回避19射出的应力的信号。苍蝇被放置在一个T型迷宫装置和给予的选择，以避免含有DSO的小瓶。使用此测定，CO 2被证明是在DSO的组分必要响应为CO 2解剖19所述的神经电路，和一部分。
这里介绍的社交回避法类似概念在西摩·本泽的实验室开发的简单行为检测的启用几代科研人员解剖复杂的行为20。社交回避的分析，可以进行低成本地使用T型迷宫测定中，允许社会行为的更广泛的研究。例如，使用这种测定法，我们最近证明了孤独症不同的遗传风险在社会B对比效果ehavior检测。突变的候选基因自闭症- neurobeachin 21,22 -目前存在的缺陷无论是在社会空间（23别处描述）和社交回避24。异常的多巴胺信号也提出了自闭症在人类25,26的病因中发挥作用。与此相反，以与neurobeachin所获得的结果，我们发现，社交回避表现是不受增加或多巴胺能细胞减少果蝇囊泡单胺转运（VMAT）的水平，尽管社会空间直接相关，这些水平VMAT 27。与neurobeachin和VMAT得到的对比结果强调了识别各种形式的反社会行为，并且因此不同的底层神经电路系统调节响应于他人的可能性。
Protocol
Representative Results
Discussion
本协议描述为社会避免检测的详细过程。广-S只会避免在飞行以前一直强调机械小瓶和应答者的性别和数量不影响社交回避的性能。然而，反应的遗传背景有重要的影响。
以下是几个关键的步骤成功完成这个实验：1）实验前总是转移苍蝇2小时，并确保不影响他们的环境; 2）在一天的同一时间进行的实验总是理想地之间Zeitgeber时间光的发病后ZT5（小时）和ZT9减少的变化表现挂?…
Declarações
The authors have nothing to disclose.
Acknowledgements
We thank Rachelle Kanippayoor for her help in identifying the new wild-type strain as being of the melanogaster species. R.W.F, O.F. and A.F.S were responsible for research design; R.W.F, M.N. and O.F. performed the experiments. R.W.F, M.N., O.F. and A.F.S. analyzed the data; R.W.F., I.S.M. and A.F.S. wrote the manuscript.
This work was supported by PSC-CUNY research awards, jointly funded by The Professional Staff Congress and The City University of New York to A.F.S.; by internal funding from Western University to A.F.S. and I.S.M.; by a training support from the National Alliance for Hispanic Health’s Alliance/Merck Ciencia (Science) Hispanic Scholars Program and a University Fellowship from the Yale Graduate School of Arts and Sciences to R.W.F.
Materials
| Stereo Zoom Microscope | Nikon | SMZ-645 | Any other standard scope for fly handling would work |
| Small paint brushes | for pushing flies | ||
| Porous Polyethylene, 12" x 12" Sheet | Flystuff – Genesee | 46-100 | http://www.flystuff.com/ProductInfo.php?productID=46-100 Porous Plastic sheet for the cold anesthesia box |
| Mini-Alarm Timer/Stopwatch | |||
| Sharpie pens | |||
| Adhesive Tape | |||
| Mini vortex | Fisher | 14-955-151 | http://www.fishersci.com/ecomm/servlet/itemdetail For mechanical agitation of the flies – any vortex would work. |
| Corning Life Sciences DL No.:352017, Falcon test tube; round bottom; disposable; no closure, 14mL; 17 x 100mm | Fisher | 14-959-8 | http://www.fishersci.com/ecomm/servlet/itemdetail?storeId=10652&langId=-1&catalogId=29104&productId=2771811&distype=0&highlightProductsItemsFlag=Y&fromSearch=1&searchType=PROD&hasPromo=0 These snap in place in the in-house made T-maze and counter-current apparatus (see text) |
| cotton balls | to close the vials after the experiment. | ||
| trifold board and white bench cover | to provide a white background, and a homogeneous light. | ||
| white bench cover | |||
| pounding pad | any mouse pad works. | ||
| large black cloth | to cover the counter-current apparatus in phototaxis response. | ||
| cool-white light | Home Depot | 1000516563 | http://www.homedepot.ca/product/illume-26-fluorescent-plug-in-linear/911423 any similar linear light with fluorescent light bulb cool-white at 13-15W would work |
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