Evaluation of chromosome painting to assess the induction and persistence of chromosome aberrations in mouse bone marrow cells by chemical agents

Fluorescence in situ hybridization with chromosome-specific painting probes (FISH painting) has been successfully applied to detect radiation-induced stable aberrations in humans and mice, whereas a few mouse studies with chemicals mostly failed to show any increase in chromosome-painting-detectable changes, especially in bone marrow cells. To further explore the feasibility of the painting approach to detect chemically induced stable aberrations, we treated mice with a single high dose of benzene, a potent bone-marrow-targeting clastogenic chemical and sacrificed them 24, 36h or 15 days later to collect bone marrow cells and analyze chromatid- and chromosome-type aberrations by FISH painting. In addition, we treated another group of mice with 18 daily low doses to show the potential for aberration induction and accumulation under chronic exposure. Chromatid-type aberrations were significantly increased 24 and 36h after acute treatment while chromosome-type ones were elevated above control values 36h and 15 days after exposure, showing that at least part of benzene-induced chromatid exchanges were converted into potentially stable chromosome aberrations. The most common aberration was an extra copy of one painted chromosome in a metaphase with the euploid number of centromeres which was interpreted as the consequence of a symmetric recombination between pericentromeric regions of one painted and one unpainted chromatid. Under chronic exposure, neither chromosome- nor chromatid-type aberrations were significantly elevated over control values, suggesting that the probability of enough primary lesions and secondary DNA double strand breaks occurring close enough together in time to allow chromosome exchanges to form is a critical limiting factor especially in a cycling cell population.