ASSAYS AND GENES FOR CELL DEATH

摘要

Quick and simple assays for apoptotic cell deaths and apoptotic and necrotic cell deaths are described, which are based on selective staining by the dyes toluidine blue, acridine orange, and Nile blue. Various tissue preparations, including live embryos and fixed tissue sections, can be assayed. Use of this assay to study cell deaths during Drosophila embryogenesis is described. Other uses of the assays, including environmental toxicity testing, identification of cell death-defective mutants, mapping of patterns of cell death in tissues, drug screening, and potential diagnostic and therapeutic uses, are also described. The present invention also relates to cell death genes, which are genes required for programmed cell deaths; mutant organisms, in which embryonic programmed cell death occurs to a less than normal extent; proteins encoded by the cell death genes; antibodies which bind the cell death gene products; and agents which alter the ability of cell death genes to cause programmed death of cells. As described herein, applicants have identified two genes which function in the initiation of apoptosis or programmed cell death. These two genes, referred to respectively as the reaper (rpr) gene and the head involution defective (hid) gene, map to position 75C1,2 on the third chromosome in Drosophila (D.) melanogaster and exibit expression patterns related to the pattern of cell death during Drosophila embryogenesis; mutations in each gene reduce levels of cell deaths or abolish cell death.