The X chromosome frequently lags behind in female lymphocyte anaphase

Abstract

Pancentromeric FISH and X-chromosome painting were used to characterize anaphase aberrations in 2,048 cultured lymphocytes from a healthy 62-year-old woman. Of 163 aberrant anaphases, 66.9% contained either chromosomes or their fragments that lagged behind. Characterization of 200 laggards showed that 49% were autosomes, 33. 5% were autosomal fragments, and 17.5% were X chromosomes. The X chromosome represented one-fourth of all lagging chromosomes and was involved much more often than would be expected by chance (1/23). Labeling of the late-replicating inactive X chromosome with 5-bromo-2'-deoxyuridine revealed that both X homologues contributed equally to the laggards. Among 200 micronuclei examined from interphase cells, the proportion of the X chromosome (31%) and autosomal fragments (50%) was higher than among anaphase laggards, whereas autosomes were involved less often (19%). These findings may reflect either selection or the fact that lagging autosomes, which were more proximal to the poles than were lagging X chromosomes, were more frequently included within the main nucleus. Our results suggest that the well-known high micronucleation and loss of the X chromosome in women's lymphocytes is the result of frequent distal lagging behind in anaphase and effective micronucleation of this chromosome. This lagging appears to affect the inactive and active X chromosomes equally.

Figure 1

Characterization of anaphase/telophase laggards in female lymphocytes. A, Metaphase in which the X chromosome and centromeres are identified, by means of FISH (red), and (B) the late-replicating inactive X chromosome (uniform green label) is distinguished from the active X chromosome (discontinuous green label), by means of BrdU labeling. C, Anaphase with a lagging X chromatid (red) that represents the inactive X chromosome (D) (green). E, Anaphase with a lagging X chromatid (red) that represents the active X chromosome (F).