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

摘要

pLoss of the iY/i chromosome in iDrosophila/i 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 iin vivo/i chromosome loss in cells of the developing iDrosophila/i wing primordia, we first engineered a iY/i chromosome with an iattP/i docking site. By making use of the ?|C31 integrase system, we site-specifically integrated a genomic transgene encompassing the imultiple wing hair/i (imwh/i) locus into this iattP/i site, leading to a imwh/isup+/supiY/i chromosome. This chromosome fully rescues the imwh/i mutant phenotype, an excellent recessive wing cell marker mutation. Loss of this imwh/isup+/supiY/i chromosome in wing primordial cells then leads to manifestation of the imwh/i mutant phenotype in imwh/i-homozygous cells. The forming imwh/i clones permit us to quantify the effect of agents and genetic alterations by assaying frequency and size of the imwh/i mosaic spots. To illustrate the use of the imwh/isup+/supiY/i loss system, the effects of four known mutagens (X-rays, colchicine, ethyl methanesulfonate, and formaldehyde) and two genetic conditions (loss- and gain-of-function ilodestar/i mutant alleles) are documented. The procedure is simple, sensitive, and inexpensive./p