Chromosome Organization in Meiosis

Abstract

Our objective is to understand the mechanics of homologous chromosome pairing during meiosis. Aberrant pairing can result in nondisjunction and birth defects in humans. This study used yeast, Saccharomyces cerevisiae, with chromosomally‐integrated arrays of tetO operators that bind TetR repressor proteins fused to GFP to produce a fluorescent signal. In diploid cells, the tetO/TetR‐GFP system allows homologous chromosomes to be identified as two foci (unpaired) or one focus (paired) as they progress through meiosis. We conducted three replicate timecourse experiments, analysing three different stages of meiosis, t=0 hours: pre‐meiotic, t=3 hours: pairing transition, and t=5 hours: synapsis. At each stage, the cells were imaged for 25 minutes, with z‐stacks taken at 30 second intervals. To analyse individual cells, we developed a 4D image analysis pipeline in MATLAB that allowed us to calculate the mean squared change in distance (MSCD), a metric describing the distance between two foci, and analyse deviations from normal diffusive motion.