Flybow: genetic multicolor cell labeling for neural circuit analysis in Drosophila melanogaster

被引:153
作者
Hadjieconomou, Dafni [1 ]
Rotkopf, Shay [2 ]
Alexandre, Cyrille [3 ]
Bell, Donald M. [4 ]
Dickson, Barry J. [2 ]
Salecker, Iris [1 ]
机构
[1] Natl Inst Med Res, MRC, Div Mol Neurobiol, London NW7 1AA, England
[2] Res Inst Mol Pathol, A-1030 Vienna, Austria
[3] Natl Inst Med Res, MRC, Div Dev Neurobiol, London NW7 1AA, England
[4] Natl Inst Med Res, MRC, Confocal Image Anal Lab, London NW7 1AA, England
关键词
SITE-SPECIFIC RECOMBINATION; FLUORESCENT PROTEIN; TRANSGENE EXPRESSION; FLP RECOMBINASE; MOSAIC ANALYSIS; NERVOUS-SYSTEM; VISUAL-SYSTEM; NEURONS; MANIPULATION; GUIDANCE;
D O I
10.1038/NMETH.1567
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
To facilitate studies of neural network architecture and formation, we generated three Drosophila melanogaster variants of the mouse Brainbow-2 system, called Flybow. Sequences encoding different membrane-tethered fluorescent proteins were arranged in pairs within cassettes flanked by recombination sites. Flybow combines the Gal4-upstream activating sequence binary system to regulate transgene expression and an inducible modified Flp-FRT system to drive inversions and excisions of cassettes. This provides spatial and temporal control over the stochastic expression of one of two or four reporters within one sample. Using the visual system, the embryonic nervous system and the wing imaginal disc, we show that Flybow in conjunction with specific Gal4 drivers can be used to visualize cell morphology with high resolution. Finally, we demonstrate that this labeling approach is compatible with available Flp-FRT-based techniques, such as mosaic analysis with a repressible cell marker; this could further support the genetic analysis of neural circuit assembly and function.
引用
收藏
页码:260 / U111
页数:9
相关论文
共 42 条
[1]   An optimized transgenesis system for Drosophila using germ-line-specific φC31 integrases [J].
Bischof, Johannes ;
Maeda, Robert K. ;
Hediger, Monika ;
Karch, Francois ;
Basler, Konrad .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2007, 104 (09) :3312-3317
[2]  
BRAND AH, 1993, DEVELOPMENT, V118, P401
[3]   Talking about a revolution: The impact of site-specific recombinases on genetic analyses in mice [J].
Branda, CS ;
Dymecki, SM .
DEVELOPMENTAL CELL, 2004, 6 (01) :7-28
[4]  
BREWSTER R, 1995, DEVELOPMENT, V121, P2923
[5]   A genome-wide transgenic RNAi library for conditional gene inactivation in Drosophila [J].
Dietzl, Georg ;
Chen, Doris ;
Schnorrer, Frank ;
Su, Kuan-Chung ;
Barinova, Yulia ;
Fellner, Michaela ;
Gasser, Beate ;
Kinsey, Kaolin ;
Oppel, Silvia ;
Scheiblauer, Susanne ;
Couto, Africa ;
Marra, Vincent ;
Keleman, Krystyna ;
Dickson, Barry J. .
NATURE, 2007, 448 (7150) :151-U1
[6]   Eye evolution at high resolution: The neuron as a unit of homology [J].
Erclik, Ted ;
Hartenstein, Volker ;
McInnes, Roderick R. ;
Lipshitz, Howard D. .
DEVELOPMENTAL BIOLOGY, 2009, 332 (01) :70-79
[7]  
FISCHBACH KF, 1989, CELL TISSUE RES, V258, P441, DOI 10.1007/BF00218858
[8]  
Goedhart J, 2010, NAT METHODS, V7, P137, DOI [10.1038/NMETH.1415, 10.1038/nmeth.1415]
[9]   THE FLP RECOMBINASE OF YEAST CATALYZES SITE-SPECIFIC RECOMBINATION IN THE DROSOPHILA GENOME [J].
GOLIC, KG ;
LINDQUIST, S .
CELL, 1989, 59 (03) :499-509
[10]   Reducing the environmental sensitivity of yellow fluorescent protein - Mechanism and applications [J].
Griesbeck, O ;
Baird, GS ;
Campbell, RE ;
Zacharias, DA ;
Tsien, RY .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2001, 276 (31) :29188-29194