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Originally published as Genetics Published Articles Ahead of Print on April 16, 2005.
Genetics, Vol. 170, 779-792, June 2005, Copyright © 2005
doi:10.1534/genetics.104.035691
A Genetic Screen for Suppressors of Drosophila NSF2 Neuromuscular Junction Overgrowth
Matthew J. Laviolette*,
Paula Nunes*,
Jean-Baptiste Peyre
,
Toshiro Aigaki and
Bryan A. Stewart*,1
Department of Biology, Tokyo Metropolitan University, Tokyo 192-0397, Japan
* Department of Life Sciences and Zoology, University of Toronto at Scarborough, Toronto, Ontario M1C 1A4, Canada
1 Corresponding author: Department of Life Sciences, University of Toronto at Scarborough, 1265 Military Trail, Toronto, ON M1C 1A4, Canada.
E-mail: stewart{at}utsc.utoronto.ca
The Drosophila larval neuromuscular system serves as a valuable model for studying the genes required for synaptic development and function. N-Ethylmaleimide sensitive factor (NSF) is a molecule known to be important in vesicular trafficking but neural expression of a dominant negative form of NSF2 induces an unexpected overgrowth of the Drosophila larval neuromuscular synapse. We have taken a genetic approach to understanding this novel phenotype by conducting a gain-of-function modifier screen to isolate genes that interact with the overgrowth phenotype. Our approach was to directly visualize the neuromuscular junction (NMJ) using a GFP transgene and screen for suppressors of NMJ overgrowth using the Gene Search collection of P-element insertions. Of the 3000 lines screened, we identified 99 lines that can partially restore the normal phenotype. Analysis of the GS element insertion sites by inverse PCR and comparison of the flanking DNA sequence to the Drosophila genome sequence revealed nearby genes for all but 10 of the 99 lines. The recovered genes, both known and predicted, include transcription factors, cytoskeletal elements, components of the ubiquitin pathway, and several signaling molecules. This collection of genes that suppress the NSF2 neuromuscular junction overgrowth phenotype is a valuable resource in our efforts to further understand the role of NSF at the synapse.
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