title: Systematic screening of Drosophila deficiency mutations for embryonic phenotypes and orphan receptor ligands.
abstract: 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 approximately 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 approximately 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.
authors.affiliation: Division of Biology, California Institute of Technology, Pasadena, California, United States of America.
authors.forename: Ashley P
authors.initials: AP
authors.lastname: Wright
authors.forename: A Nicole
authors.initials: AN
authors.lastname: Fox
authors.forename: Karl G
authors.initials: KG
authors.lastname: Johnson
authors.forename: Kai
authors.initials: K
authors.lastname: Zinn
mesh_terms: D000818:Animals; D000941:Antigens / Q000378:metabolism; D001369:Axons / Q000378:metabolism; D002490:Central Nervous System / Q000166:cytology / Q000254:growth & development / Q000378:metabolism; D004331:Drosophila melanogaster / Q000196:embryology* / Q000235:genetics*; D005786:Gene Expression Regulation; D017344:Genes, Insect / Q000235:genetics*; D006801:Humans; D008024:Ligands; D009046:Motor Neurons / Q000166:cytology / Q000378:metabolism; D009457:Neuroglia / Q000378:metabolism; D010641:Phenotype; D011956:Receptors, Cell Surface / Q000378:metabolism*; D017384:Sequence Deletion; D013997:Time Factors
chemical_list: D000941:Antigens; D008024:Ligands; D011956:Receptors, Cell Surface
references.citation: Science. 1994 Apr 29;264(5159):677-83
references.pmid: 8171319
references.citation: Cell. 1995 Sep 22;82(6):1013-23
references.pmid: 7553843
references.citation: Curr Biol. 2004 Apr 20;14(8):659-67
references.pmid: 15084280
references.citation: Genome Biol. 2007;8(10):R216
references.pmid: 17927810
references.citation: Nature. 1980 Oct 30;287(5785):795-801
references.pmid: 6776413
references.citation: Cell. 1995 Sep 22;82(6):1025-36
references.pmid: 7553844
references.citation: EMBO J. 2001 Sep 3;20(17):4664-73
references.pmid: 11532931
references.citation: Development. 1996 Jan;122(1):131-9
references.pmid: 8565824
references.citation: Cell. 1998 Jan 23;92(2):205-15
references.pmid: 9458045
references.citation: J Biol Chem. 2001 Mar 23;276(12):9158-65
references.pmid: 11124260
references.citation: Development. 1994 Oct;120(10):2957-66
references.pmid: 7607085
references.citation: Cell. 1996 Feb 23;84(4):599-609
references.pmid: 8598046
references.citation: Development. 1997 May;124(10):1941-52
references.pmid: 9169841
references.citation: Mech Dev. 2003 Apr;120(4):401-13
references.pmid: 12676319
references.citation: Dev Biol. 2001 Oct 1;238(1):47-63
references.pmid: 11783993
references.citation: Dev Biol. 2002 Aug 15;248(2):369-83
references.pmid: 12167411
references.citation: Genes Dev. 1994 Apr 15;8(8):981-94
references.pmid: 7926782
references.citation: Neuron. 2000 Dec;28(3):753-66
references.pmid: 11163264
references.citation: Cell. 1988 Jan 15;52(1):143-51
references.pmid: 3345560
references.citation: Development. 2000 Jan;127(2):393-402
references.pmid: 10603355
references.citation: Genes Dev. 2004 Mar 15;18(6):687-99
references.pmid: 15075295
references.citation: J Vis Exp. 2009 Dec 29;(34):
references.pmid: 20040910
references.citation: Cell. 1993 Jun 18;73(6):1137-53
references.pmid: 8513498
references.citation: Nat Genet. 2004 Mar;36(3):288-92
references.pmid: 14981519
references.citation: Genes Dev. 1994 Nov 1;8(21):2588-601
references.pmid: 7958918
references.citation: Cell. 2001 Apr 6;105(1):57-67
references.pmid: 11301002
references.citation: Cell. 1999 Mar 19;96(6):785-94
references.pmid: 10102267
references.citation: Neuron. 1993 Mar;10(3):409-26
references.pmid: 8461134
references.citation: Science. 2007 Apr 20;316(5823):417-21
references.pmid: 17412918
references.citation: Development. 1995 Feb;121(2):317-32
references.pmid: 7768175
references.citation: Curr Biol. 2005 Oct 11;15(19):1701-11
references.pmid: 16213816
references.citation: Cell. 1988 Jan 15;52(1):133-41
references.pmid: 3345559
references.citation: Annu Rev Neurosci. 1996;19:545-75
references.pmid: 8833454
references.citation: Cell. 1987 Mar 27;48(6):975-88
references.pmid: 3548998
references.citation: J Cell Biol. 2008 Dec 1;183(5):805-18
references.pmid: 19047461
references.citation: Development. 2001 Nov;128(21):4371-82
references.pmid: 11684671