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当前位置:首页  »  实验视频  »  Neuroscience  »  Primary Neuronal Cultures from the Brains of Late Stage Drosophila Pupae

  • Primary Neuronal Cultures from the Brains of Late Stage Drosophila Pupae


    时长:未知

    类型:Neuroscience

    时间:2007

    国家:美国

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    播放次数:加载中

    包包:20免费充值包包

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    主讲人:请看下面内容介绍  

播放地址1: FLV文件(说明: 无需安装任何插件,即可快速播放 )

内容介绍

    Beatriz Sicaeros, Jorge M. Campusano, Diane K. O'DowdDepartment of Development and Cell Biology, Department of Anatomy and Neurobiology, University of California, Irvine In this video, we demonstrate the preparation of primary neuronal cultures from the brains of late stage Drosophila pupae. The procedure begins with the removal of brains from animals at 70-78 hrs after puparium formation. The isolated brains are shown after brief incubation in papain followed by several washes in serum-free growth medium. The process of mechanical dissociation of each brain in a 5 ul drop of media on a coverslip is illustrated. The axons and dendrites of the post-mitotic neurons are sheered off near the soma during dissociation but the neurons begin to regenerate processes within a few hours of plating. Images show live cultures at 2 days. Neurons continue to elaborate processes during the first week in culture. Specific neuronal populations can be identified in culture using GAL4 lines to drive tissue specific expression of fluorescent markers such as GFP or RFP. Whole cell recordings have demonstrated the cultured neurons form functional, spontaneously active cholinergic and GABAergic synapses. A short video segment illustrates calcium dynamics in the cultured neurons using Fura-2 as a calcium indicator dye to monitor spontaneous calcium transients and nicotine evoked calcium responses in a dish of cultured neurons. These pupal brain cultures are a useful model system in which genetic and pharmacological tools can be used to identify intrinsic and extrinsic factors that influence formation and function of central synapses.

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