高通量检测，以检查个体产蛋首<em>果蝇</em
Summary
This protocol describes a high throughput assay for testing egg-laying preferences of Drosophila melanogaster at single-animal resolution. This assay provides a simple, efficient, and scalable platform to identify genes and circuit components that control a simple decision-making process.
Abstract
近日， 果蝇产卵偏好已经成为一个听话的基因模型，研究简单的决策过程的神经基础。当选择的网站存入他们的蛋，雌蝇能居其选择的相对吸引力和选择的“较大的两种商品。”然而，如果一个人想采取系统的遗传筛选方法的搜索电路基础底层这个简单的决策过程，因为它们是基于人群的和费力的建立最产蛋偏好测定法是不实际的。为了增加单个雌性产卵偏好学习的吞吐量，我们开发了长达三十个人的每个可以同时测定产蛋偏好蝇以及确保每个女性具有高产蛋率的协议自定义腔室（使他们的偏好是容易辨别和更有说服力）。我们的方法是简单的执行并产生非常一致的结果。此外，这些室可以配备不同的附件，以允许视频记录产蛋动物，并为光遗传学研究提供光。本文提供了制造这些商会和程序准备在这些商会进行检测苍蝇的蓝图。
Introduction
Drosophila melanogaster is a powerful genetic model organism to study the neural basis of behaviors. The rapid developments of genetic tools to manipulate neurons in a targeted manner and the emergence of sophisticated behavioral analysis tools have significantly improved our ability to dissect the circuit mechanisms that underlie the sensory-motor transformation processes of several innate and learned behaviors1-3.
Drosophila egg-laying is a suitable model to study the neural basis of simple decision making processes. In particular, Drosophila females have been shown to possess the ability to compare and rank their options before “committing” to depositing an egg onto a given option4-8. For example, when given only a plain (sucrose-free) substrate or only a sucrose-containing substrate, females readily accept either option for egg-laying. However, when presented with both options, females robust reject the sucrose substrate in some contexts7,9,10. Relatively little is known about the neural mechanisms that allow females to “choose the greater of two goods”, however. A major obstacle has been the lack of an efficient method to assay egg-laying preferences such that one can use a systematic genetic screening approach to study this problem.
In this report, we describe the protocol we developed that allows egg-laying preferences of females to be assayed at single-animal resolution and with substantially improved throughput and consistency over previous methods. Specifically, we provide the blueprints for constructing the chambers we designed, the protocol for preparing the females so that each is primed to lay many eggs, and the protocol for using the chambers.
Protocol
Representative Results
Discussion
该商会在这里描述的协议具有比以前产蛋测定了一些改进。首先，它们提高显著测定单个动物的偏好的吞吐量。每个房间都可以分析30单身女性，它需要不到一个小时来设置。第二，它们增加超过以前的方法产蛋偏好的一致性。竞技场的尺寸的标准化，产蛋基板尺寸，和基片之间的距离可以更容易地不同实验天之间，并从不同的研究小组比较结果。第三，腔室可以配备附件以容纳更多的分析?…
Declarações
The authors have nothing to disclose.
Acknowledgements
We thank the Duke Physics Shop, especially Phil Lewis, for helping us build the behavioral apparatus and attachments and creating the drawing. This work is funded by the National Institutes of Health under award number R01GM100027.
Materials
| UltraPure Agarose | Invitrogen | 16500-500 | |
| Sucrose | Sigma | S0389 | |
| Water bath | Fisher | 15-462-6Q | |
| LifeCam Cinema webcam | Microsoft | H5D-00013 | |
| Red LEDs | Cree | C503B-RAN-CA0B0AA1 | |
| Egg-laying chambers | Custom Built | ||
| Camera holders | Custom Built | ||
| LED holders | Custom Built | ||
| Fly vials (narrow) | Genesee | 32-116BC |
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