Dual-labeling method for electron microscopy to characterize synaptic connectivity using genetically encoded fluorescent reporters in Drosophila.

Dual-labeling method for electron microscopy to characterize synaptic connectivity using genetically encoded fluorescent reporters in Drosophila.
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电子显微镜的双标记方法,使用果蝇中的遗传编码的荧光报道表征突触连通性。

DOI:
10.1016/j.jneumeth.2010.10.020
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发表时间:
2011-01-15
影响因子:
3
通讯作者:
Stopfer, Mark
Stopfer, Mark
中科院分区:
医学4区
文献类型:
--
作者:
Tanaka, Nobuaki K.;Dye, Louis;Stopfer, Mark

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光学和电子显微镜(LM和EM)都为表征完整大脑中的神经元回路提供了重要的优势:LM可以揭示大脑区域之间神经元追踪的一般模式,EM可以确认小区域内已识别神经元之间的突触连接。在少数物种中,荧光蛋白的遗传标记已被用于LM来可视化多种神经元并分析其形态和投影模式。然而,将这些大规模图案与EM分析中可用的精细细节相结合一直是一项技术挑战。为了用EM分析表达荧光标记物的神经元的突触连接,我们开发了一种用于预嵌入大脑的双标记方法。在表达遗传标记的果蝇中,也注射了标记物,我们观察了AL中两个神经元群体之间的突触连接,其中一个已被证明介导了一种特定的功能,即气味诱发的神经振荡。
Light and electron microscopy (LM and EM) both offer important advantages for characterizing neuronal circuitry in intact brains: LM can reveal the general patterns neurons trace between brain areas, and EM can confirm synaptic connections between identified neurons within a small area. In a few species, genetic labeling with fluorescent proteins has been used with LM to visualize many kinds of neurons and to analyze their morphologies and projection patterns. However, combining these large-scale patterns with the fine detail available in EM analysis has been a technical challenge. To analyze the synaptic connectivity of neurons expressing fluorescent markers with EM, we developed a dual-labeling method for use with pre-embedded brains. In Drosophila expressing genetic labels and also injected with markers we visualized synaptic connections among two populations of neurons in the AL, one of which has been shown to mediate a specific function, odor evoked neural oscillation.
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