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doi:10.1534/genetics.104.035691
A more recent version of this article appeared on June 1, 2005.
REGULAR RESEARCH PAPERS |
A Genetic Screen for Suppressors of Drosophila NSF2 Neuromuscular Junction Overgrowth
Matthew J. Laviolette 1, Paula Nunes 1, Jean-Baptiste Peyre 2, Toshiro Aigaki 2 and Bryan A. Stewart 1*
1 University of Toronto at Scarborough
2 Tokyo Metropolitan University
* To whom correspondence should be addressed. E-mail: stewart{at}utsc.utoronto.ca.
Submitted on September 8, 2004
Revised on September 27, 2004
Accepted on 15 February 2005
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 understand 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 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 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. Reassuringly, we recovered a few genes that had been previously isolated in similar gain of function screens for genes affecting neural development. 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 and it potentially points to new roles for NSF.
Key Words: hypersprouting, neuromuscular junction, neuron, synapse
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  P. Nunes, N. Haines, V. Kuppuswamy, D. J. Fleet, and B. A. Stewart Synaptic Vesicle Mobility and Presynaptic F-Actin Are Disrupted in a N-ethylmaleimide-sensitive Factor Allele of Drosophila Mol. Biol. Cell, November 1, 2006; 17(11): 4709 - 4719. [Abstract] [Full Text] [PDF]
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