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Published August 19, 2010 | Supplemental Material + Published
Journal Article Open

Systematic Screening of Drosophila Deficiency Mutations for Embryonic Phenotypes and Orphan Receptor Ligands


This paper defines a collection of Drosophila deletion mutations (deficiencies) that can be systematically screened for embryonic phenotypes, orphan receptor ligands, and genes affecting protein localization. It reports the results of deficiency screens we have conducted that have revealed new axon guidance phenotypes in the central nervous system and neuromuscular system and permitted a quantitative assessment of the number of potential genes involved in regulating guidance of specific motor axon branches. Deficiency "kits" that cover the genome with a minimum number of lines have been established to facilitate gene mapping. These kits cannot be systematically analyzed for phenotypes, however, since embryos homozygous for many deficiencies in these kits fail to develop due to the loss of key gene products encoded within the deficiency. To create new kits that can be screened for phenotype, we have examined the development of the nervous system in embryos homozygous for more than 700 distinct deficiency mutations. A kit of ~400 deficiency lines for which homozygotes have a recognizable nervous system and intact body walls encompasses >80% of the genome. Here we show examples of screens of this kit for orphan receptor ligands and neuronal antigen expression. It can also be used to find genes involved in expression, patterning, and subcellular localization of any protein that can be visualized by antibody staining. A subset kit of 233 deficiency lines, for which homozygotes develop relatively normally to late stage 16, covers ~50% of the genome. We have screened it for axon guidance phenotypes, and we present examples of new phenotypes we have identified. The subset kit can be used to screen for phenotypes affecting all embryonic organs. In the future, these deficiency kits will allow Drosophila researchers to rapidly and efficiently execute genome-wide anatomical screens that require examination of individual embryos at high magnification.

Additional Information

© 2010 Wright et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Received: May 10, 2010; Accepted: July 21, 2010; Published: August 19, 2010. Editor: Brian D. McCabe, Columbia University, United States of America. Funding: NIH RO1 grants NS028182 and NS062821 to Kai Zinn, NSF grant IOS-0841551 to Karl Johnson and NRSA postdoctoral fellowship to A. Nicole Fox. The funders had no role in study design, collection and analysis, decision to publish, or preparation of the manuscript. We thank Kevin Cook (Bloomington) for providing extensive information on deletions, and the Bloomington, Szeged, and Kyoto stock centers for stocks. We also thank Peter Lee for helpful discussions, Kaushiki Menon and Angela Stathopoulos for comments on the manuscript, Elena Armand for stock maintenance, and Violana Nesterova for figure preparation. Confocal imaging was performed at the Caltech Biological Imaging Facility. 1D4, BP102, and 7G10 supernatants were obtained from cell lines grown by the Caltech Monoclonal Antibody Facility. RPTP-AP fusion proteins were obtained from baculovirus infected cell supernatants generated by the Caltech Protein Expression Center. Author Contributions: Conceived and designed the experiments: APW ANF KZ. Performed the experiments: APW ANF KGJ KZ. Analyzed the data: APW ANF KZ. Contributed reagents/materials/analysis tools: APW ANF KGJ KZ. Wrote the paper: APW KZ.

Attached Files

Published - Wright2010p11253PLoS_ONE.pdf

Supplemental Material - journal.pone.0012288.s001.xls

Supplemental Material - journal.pone.0012288.s002.xls

Supplemental Material - journal.pone.0012288.s003.xls


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August 19, 2023
October 20, 2023