Indications of Centromere Movement during Interphase and Differentiationa

Abstract

Mouse and human DNA sequences from centromeric and ribosomal domains were labeled with biotinylated deoxynucleotides and hybridized in situ to paraformaldehyde-fixed tissue culture cells. Centromeres were widely dispersed in most of these interphase nuclei. At late G2 phases of the cell cycle, centromeres appeared to coalesce and then to align in an orderly pattern, with discrete positional assignments for individuals chromosomes in metaphase and anaphase. Ribosomal cistrons were also organized in an orderly and defined fashion during mitosis. As soon as the nuclear membrane forms in early G1, centromeres rapidly disperse throughout the nucleus. Centromere patterns during G1 and S were indistinguishable in cultured cells, as determined by double-labeling experiments. Antibodies that bind to centric chromosomal proteins revealed the same patterns in cultured cells as those obtained with DNA sequence-specific probes. Large differentiated neurons display reproducible collections of centromeres in interphase that are very different from those seen in cultured cells. Neurons in widely divergent mammalian species, despite large differences in centromeric DNA sequences, maintain similar nuclear positions for these chromosomal segments. Similarly, ribosomal cistrons are positioned in comparable nuclear locales in neurons of divergent species. It is suggested that such arrangements reflect, or are necessary for, the function of a given cell type. Studies of large cerebellar neurons at critical times in development indicated a relative "movement" of centromeric domains, away from the nuclear membrane and toward the central nucleolar region. It is possible that the orderly and temporal positioning of centromeric, as well as of other chromosomal regions, is based on protein-nucleic acid interactions. Implications for trisomy 21 and other disorders involving chromosomal rearrangements, such as transposition, are considered from this perspective.