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3 Janelia Publications

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    Simpson Lab
    10/20/11 | Genetic manipulation of genes and cells in the nervous system of the fruit fly.
    Venken KJ, Simpson JH, Bellen HJ
    Neuron. 2011 Oct 20;72(2):202-30. doi: 10.1016/j.neuron.2011.09.021

    Research in the fruit fly Drosophila melanogaster has led to insights in neural development, axon guidance, ion channel function, synaptic transmission, learning and memory, diurnal rhythmicity, and neural disease that have had broad implications for neuroscience. Drosophila is currently the eukaryotic model organism that permits the most sophisticated in vivo manipulations to address the function of neurons and neuronally expressed genes. Here, we summarize many of the techniques that help assess the role of specific neurons by labeling, removing, or altering their activity. We also survey genetic manipulations to identify and characterize neural genes by mutation, overexpression, and protein labeling. Here, we attempt to acquaint the reader with available options and contexts to apply these methods.

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    Simpson LabRubin Lab
    06/01/11 | BrainAligner: 3D registration atlases of Drosophila brains.
    Peng H, Chung P, Long F, Qu L, Jenett A, Seeds AM, Myers EW, Simpson JH
    Nature Methods. 2011 Jun;8:493-500. doi: 10.1038/nmeth.1602

    Analyzing Drosophila melanogaster neural expression patterns in thousands of three-dimensional image stacks of individual brains requires registering them into a canonical framework based on a fiducial reference of neuropil morphology. Given a target brain labeled with predefined landmarks, the BrainAligner program automatically finds the corresponding landmarks in a subject brain and maps it to the coordinate system of the target brain via a deformable warp. Using a neuropil marker (the antibody nc82) as a reference of the brain morphology and a target brain that is itself a statistical average of data for 295 brains, we achieved a registration accuracy of 2 μm on average, permitting assessment of stereotypy, potential connectivity and functional mapping of the adult fruit fly brain. We used BrainAligner to generate an image pattern atlas of 2954 registered brains containing 470 different expression patterns that cover all the major compartments of the fly brain.

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    Looger LabSimpson Lab
    03/01/11 | Drosophila brainbow: a recombinase-based fluorescence labeling technique to subdivide neural expression patterns.
    Hampel S, Chung P, McKellar CE, Hall D, Looger LL, Simpson JH
    Nature Methods. 2011 Mar;8:253-9. doi: 10.1038/nmeth.1566

    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.

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