Scanning electron microscope analysis of structural changes and aberrations in human chromosomes associated with the inhibition and reversal of inhibition of ultraviolet light induced DNA repair

Metaphase chromosomes appear decondensed in preparations from mitotic cells that have been irradiated with ultraviolet light (UV) and incubated with inhibitors of DNA synthesis; under these conditions DNA repair is inhibited and both single and double strand DNA breaks accumulate. After reversal of the inhibition chromosomes are condensed, but are often damaged. In this paper we show by scanning electron microscopy (SEM) that decondensed HeLa chromosomes are composed of fibre clusters similar to those previously described for the large chromosomes of the Indian muntjac. This suggests that the clusters may be a universal higher order packing unit in mammalian metaphase chromosomes. We also examine by SEM the nature of the aberrations that appear after the reversal of periods of inhibited repair; these include gaps, breaks, deletions and telomeric packing abnormalities. SEM analysis allows an extension and reconsideration of conclusions about chromatid continuity based on the study of conventional light microscope (LM) preparations.