A genetic mosaic approach for neural circuit mapping in Drosophila
Edited by Jeffrey C. Hall, University of Maine, Orono, ME, and approved July 29, 2010 (received for review April 7, 2010)
Abstract
Transgenic manipulation of subsets of brain cells is increasingly used for studying behaviors and their underlying neural circuits. In Drosophila, the GAL4–upstream activating sequence (UAS) binary system is powerful for gene manipulation, but GAL4 expression is often too broad for fine mapping of neural circuits. Here, we describe the development of unique molecular genetic tools to restrict GAL4 expression patterns. Building on the GAL4-UAS system, our method adds two components: a collection of enhancer-trap recombinase, Flippase (ET-FLP), transgenic lines that provide inheritable, reproducible, and tissue-specific FLP and an FRT-dependent GAL80 “flip-in” construct that converts FLP expression into tissue-specific repression of GAL4 by GAL80. By including a UAS-encoded fluorescent protein, circuit morphology can be simultaneously marked while the circuit function is assessed using another UAS transgene. In a proof-of-principle analysis, we applied this ET-FLP-induced intersectional GAL80/GAL4 repression (FINGR) method to map the neural circuitry underlying fly wing inflation. The FINGR system is versatile and powerful in combination with the vast collection of GAL4 lines for neural circuit mapping as well as for clonal analysis based on the infusion of the yeast-derived FRT/FLP system of mitotic recombination into Drosophila. The strategies and tactics underlying our FINGR system are also applicable to other genetically amenable organisms in which transgenes including the GAL4, UAS, GAL80, and FLP factors can be applied.
Acknowledgments
We thank Doug Allan (University of British Columbia, Vancouver), Rich Binari (Harvard Medical School, Boston), Marc Halfon (University of Buffalo, Buffalo, NY), Norbert Perrimon (Harvard Medical School), Francesca Pignoni (Upstate Medical University, Syracuse, NY), David Sinclair (Harvard Medical School), Kristin Scott (University of California, Berkeley, CA), Gary Struhl (Columbia University, New York), and Ben White (National Institute of Mental Health, Bethesda) for flies and reagents; Greg Books, Kyomi Cho, Lawrence Chu, Ysabel Milton, Molly Sichio, and Ian Tran (Brandeis University) for initial contributions; Luke Carter, Dennis Chang, Randy Hewes, Mac Hooks, John Tauber, Phillip Vanlandingham, and Ben White for discussions or comments on the manuscript; and Luke Carter for helping with brain imaging. We especially thank Jeffrey Hall for his guidance and insightful suggestions made on the manuscript. An internal fund (to B.Z.) from the University of Oklahoma and a grant from the National Science Foundation (IOS-1025556, to B.Z. and R.A.B.) supported R.A.B. and this research. National Institutes of Health Grant R01NS060878 (to B.Z.) partially supported R.A.B.; and R.A.B. acknowledges a National Institutes of Health National Research Service Award (T32 NS07292) training grant awarded to Brandeis University and a separate National Institutes of Health Grant R01 GM21473 awarded to Jeffrey Hall at Brandeis, which provided support for the early stages of this study.
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Freely available online through the PNAS open access option.
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Published online: September 1, 2010
Published in issue: September 14, 2010
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Acknowledgments
We thank Doug Allan (University of British Columbia, Vancouver), Rich Binari (Harvard Medical School, Boston), Marc Halfon (University of Buffalo, Buffalo, NY), Norbert Perrimon (Harvard Medical School), Francesca Pignoni (Upstate Medical University, Syracuse, NY), David Sinclair (Harvard Medical School), Kristin Scott (University of California, Berkeley, CA), Gary Struhl (Columbia University, New York), and Ben White (National Institute of Mental Health, Bethesda) for flies and reagents; Greg Books, Kyomi Cho, Lawrence Chu, Ysabel Milton, Molly Sichio, and Ian Tran (Brandeis University) for initial contributions; Luke Carter, Dennis Chang, Randy Hewes, Mac Hooks, John Tauber, Phillip Vanlandingham, and Ben White for discussions or comments on the manuscript; and Luke Carter for helping with brain imaging. We especially thank Jeffrey Hall for his guidance and insightful suggestions made on the manuscript. An internal fund (to B.Z.) from the University of Oklahoma and a grant from the National Science Foundation (IOS-1025556, to B.Z. and R.A.B.) supported R.A.B. and this research. National Institutes of Health Grant R01NS060878 (to B.Z.) partially supported R.A.B.; and R.A.B. acknowledges a National Institutes of Health National Research Service Award (T32 NS07292) training grant awarded to Brandeis University and a separate National Institutes of Health Grant R01 GM21473 awarded to Jeffrey Hall at Brandeis, which provided support for the early stages of this study.
Notes
*This Direct Submission article had a prearranged editor.
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The authors declare no conflict of interest.
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