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4 Publications

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    10/01/10 | Refinement of tools for targeted gene expression in Drosophila.
    Pfeiffer BD, Ngo TB, Hibbard KL, Murphy C, Jenett A, Truman JW, Rubin GM
    Genetics. 2010 Oct;186(2):735-55. doi: 10.1534/genetics.110.119917

    A wide variety of biological experiments rely on the ability to express an exogenous gene in a transgenic animal at a defined level and in a spatially and temporally controlled pattern. We describe major improvements of the methods available for achieving this objective in Drosophila melanogaster. We have systematically varied core promoters, UTRs, operator sequences, and transcriptional activating domains used to direct gene expression with the GAL4, LexA, and Split GAL4 transcription factors and the GAL80 transcriptional repressor. The use of site-specific integration allowed us to make quantitative comparisons between different constructs inserted at the same genomic location. We also characterized a set of PhiC31 integration sites for their ability to support transgene expression of both drivers and responders in the nervous system. The increased strength and reliability of these optimized reagents overcome many of the previous limitations of these methods and will facilitate genetic manipulations of greater complexity and sophistication.

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    10/01/10 | Semi-automated reconstruction of neural circuits using electron microscopy.
    Chklovskii DB, Vitaladevuni S, Scheffer LK
    Current Opinion in Neurobiology. 2010 Oct;20:667-75. doi: 10.1371/journal.pcbi.1001066

    Reconstructing neuronal circuits at the level of synapses is a central problem in neuroscience, and the focus of the nascent field of connectomics. Previously used to reconstruct the C. elegans wiring diagram, serial-section transmission electron microscopy (ssTEM) is a proven technique for the task. However, to reconstruct more complex circuits, ssTEM will require the automation of image processing. We review progress in the processing of electron microscopy images and, in particular, a semi-automated reconstruction pipeline deployed at Janelia. Drosophila circuits underlying identified behaviors are being reconstructed in the pipeline with the goal of generating a complete Drosophila connectome.

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    01/01/10 | Anatomic analysis of Gal4 expression patterns of the Rubin line collection: the central complex.
    Jenett A, Wolff T, Nern A, Pfeiffer BD, Ngo T, Murphy C, Long F, Peng H, Rubin GM
    Journal of Neurogenetics. 2010;24:71-2
    01/01/10 | Increasing depth resolution of electron microscopy of neural circuits using sparse tomographic reconstruction.
    Veeraraghavan A, Genkin AV, Vitaladevuni S, Scheffer L, Xu C, Hess H, Fetter R, Cantoni M, Knott G, Chklovskii DB
    Computer Vision and Pattern Recognition (CVPR). 2010:1767-74. doi: 10.1109/CVPR.2010.5539846