Janelia Farm Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, USA.
Nat Methods. 2011 Mar;8(3):253-9. doi: 10.1038/nmeth.1566. Epub 2011 Feb 6.
We developed a multicolor neuron labeling technique in Drosophila melanogaster that combines the power to specifically target different neural populations with the label diversity provided by stochastic color choice. This adaptation of vertebrate Brainbow uses recombination to select one of three epitope-tagged proteins detectable by immunofluorescence. Two copies of this construct yield six bright, separable colors. We used Drosophila Brainbow to study the innervation patterns of multiple antennal lobe projection neuron lineages in the same preparation and to observe the relative trajectories of individual aminergic neurons. Nerve bundles, and even individual neurites hundreds of micrometers long, can be followed with definitive color labeling. We traced motor neurons in the subesophageal ganglion and correlated them to neuromuscular junctions to identify their specific proboscis muscle targets. The ability to independently visualize multiple lineage or neuron projections in the same preparation greatly advances the goal of mapping how neurons connect into circuits.
我们开发了一种在黑腹果蝇中的多色神经元标记技术，它结合了特异性靶向不同神经元群体的能力和随机颜色选择提供的标记多样性。这种对脊椎动物 Brainbow 的改编利用重组来选择三种可通过免疫荧光检测的表位标记蛋白之一。该构建体的两个拷贝产生六种明亮、可分离的颜色。我们使用 Drosophila Brainbow 来研究同一制备物中多个触角叶投射神经元谱系的神经支配模式，并观察单个单胺能神经元的相对轨迹。可以使用明确的颜色标记来跟踪神经束，甚至是数百微米长的单个神经突。我们追踪了食管下神经节中的运动神经元，并将它们与肌神经连接起来，以确定它们特定的喙肌靶标。在同一制备物中独立可视化多个谱系或神经元投射的能力极大地推进了将神经元连接成回路的映射目标。
