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首页> 外文会议>Photons and neurons >Fluorescence lifetime images of different green fluorescent proteins in fly brain
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Fluorescence lifetime images of different green fluorescent proteins in fly brain

机译：苍蝇脑中不同绿色荧光蛋白的荧光寿命图像

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摘要

The mechanisms of learning and memory are the most important functions in an animal brain. Investigating neuron circuits and network maps in a brain is the first step toward understanding memory and learning behavior. Since Drosophila brain is the major model for understanding brain functions, we measure the florescence lifetimes of different GFP-based reporters expressed in a fly brain. In this work, two Gal4 drivers, OK 107 and MZ 19 were used. Intracellular calcium ([Ca~(2+)]) concentration is an importation indicator of neuronal activity. Therefore, several groups have developed GFP-based calcium sensors, among which G-CaMP is the most popular and reliable. The fluorescence intensity of G-CaMP will increase when it binds to calcium ion; however, individual variation from different animals prevents quantitative research. In this work, we found that the florescence lifetime of G-CaMP will shrink from 1.8 ns to 1.0 ns when binding to Ca~(2+). This finding can potentially help us to understand the neuron circuits by fluorescence lifetime imaging microscopy (FLIM). Channelrhodopsin-2 (ChR2) is a light-activated ion-channel protein on a neuron cell membrane. In this work, we express ChR2 and G-CaMP in a fly brain. Using a pulsed 470-nm laser to activate the neurons, we can also record the fluorescence lifetime changes in the structure. Hence, we can trace and manipulate a specific circuit in this animal. This method provides more flexibility in brain research.
机译：学习和记忆的机制是动物大脑中最重要的功能。研究大脑中的神经元电路和网络图是了解记忆和学习行为的第一步。由于果蝇大脑是了解大脑功能的主要模型，因此我们测量了在苍蝇大脑中表达的不同基于GFP的报告基因的荧光寿命。在这项工作中，使用了两个Gal4驱动程序OK 107和MZ 19。细胞内钙（[Ca〜（2+）]）浓度是神经元活性的重要指标。因此，几个小组开发了基于GFP的钙传感器，其中G-CaMP是最受欢迎和最可靠的。当G-CaMP与钙离子结合时，其荧光强度将增加；但是，来自不同动物的个体变异会阻止定量研究。在这项工作中，我们发现当与Ca〜（2+）结合时，G-CaMP的荧光寿命将从1.8 ns缩短至1.0 ns。这一发现可能会帮助我们通过荧光寿命成像显微镜（FLIM）了解神经元回路。 Channelrhodopsin-2（ChR2）是神经元细胞膜上的光激活离子通道蛋白。在这项工作中，我们在苍蝇大脑中表达ChR2和G-CaMP。使用470 nm脉冲激光激活神经元，我们还可以记录结构中荧光寿命的变化。因此，我们可以追踪和操纵该动物中的特定电路。这种方法为大脑研究提供了更大的灵活性。

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  • 来源
    《Photons and neurons》|2009年|71800D.1-71800D.8|共8页
  • 会议地点 San Jose CA(US)
  • 作者

    Sih-Yu Lai; Y. Y. Lin; A. S. Chiang; Y. C. Huang;

  • 作者单位

    Institute of Biotechnology and Department of Life Science, National Tsing Hua University, Hsinchu 30013, Taiwan Brain Research Center, National Tsing Hua University, University System of Taiwan, Hsinchu 30013, Taiwan;

    Institute of Photonics Technologies, Department of Electrical engineering, National Tsing-hua University, Hsinchu 30013, Taiwan Brain Research Center, National Tsing Hua University, University System of Taiwan, Hsinchu 30013, Taiwan;

    Institute of Biotechnology and Department of Life Science, National Tsing Hua University, Hsinchu 30013, Taiwan Brain Research Center, National Tsing Hua University, University System of Taiwan, Hsinchu 30013, Taiwan;

    Institute of Photonics Technologies, Department of Electrical engineering, National Tsing-hua University, Hsinchu 30013, Taiwan;

  • 会议组织
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    fluorescence lifetime; green florescent protein;

    机译：荧光寿命绿色荧光蛋白;

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