An Assay to Detect In Vivo Y Chromosome Loss in Drosophila Wing Disc Cells

G3: Genes, Genomes, Genetics

Janos Szabad¹  Koen J. T. Venken²  Hugo J. Bellen³ 

[1] Department of Biology, University of Szeged, H-6720 Szeged, HungaryDepartment of Biology, University of Szeged, H-6720 Szeged, HungaryDepartment of Biology, University of Szeged, H-6720 Szeged, Hungary;Department of Molecular and Human GeneticsDepartment of Molecular and Human GeneticsDepartment of Molecular and Human Genetics;Department of Molecular and Human GeneticsHoward Hughes Medical InstituteProgram in Developmental Biology, Baylor College of Medicine and Neurological Research Institute, Houston, Texas 77030Department of Molecular and Human GeneticsDepartment of Molecular and Human GeneticsHoward Hughes Medical InstituteProgram in Developmental Biology, Baylor College of Medicine and Neurological Research Institute, Houston, Texas 77030Howard Hughes Medical InstituteDepartment of Molecular and Human GeneticsHoward Hughes Medical InstituteProgram in Developmental Biology, Baylor College of Medicine and Neurological Research Institute, Houston, Texas 77030Program in Developmental Biology, Baylor College of Medicine and Neurological Research Institute, Houston, Texas 77030Department of Molecular and Human GeneticsHoward Hughes Medical InstituteProgram in Developmental Biology, Baylor College of Medicine and Neurological Research Institute, Houston, Texas 77030

关键词: chromosome loss;    Y chromosome;    multiple wing hair;    wing mosaic spots;    Drosophila;   

DOI  :  10.1534/g3.112.002899

学科分类：生物科学（综合）

来源: Genetics Society of America

【 摘 要 】

Loss of the Y chromosome in Drosophila has no impact on cell viability and therefore allows us to assay the impact of environmental agents and genetic alterations on chromosomal loss. To detect in vivo chromosome loss in cells of the developing Drosophila wing primordia, we first engineered a Y chromosome with an attP docking site. By making use of the ΦC31 integrase system, we site-specifically integrated a genomic transgene encompassing the multiple wing hair (mwh) locus into this attP site, leading to a mwh⁺Y chromosome. This chromosome fully rescues the mwh mutant phenotype, an excellent recessive wing cell marker mutation. Loss of this mwh⁺Y chromosome in wing primordial cells then leads to manifestation of the mwh mutant phenotype in mwh-homozygous cells. The forming mwh clones permit us to quantify the effect of agents and genetic alterations by assaying frequency and size of the mwh mosaic spots. To illustrate the use of the mwh⁺Y loss system, the effects of four known mutagens (X-rays, colchicine, ethyl methanesulfonate, and formaldehyde) and two genetic conditions (loss- and gain-of-function lodestar mutant alleles) are documented. The procedure is simple, sensitive, and inexpensive.

【 授权许可】

Unknown