2010. Vol. 52, N 5, p. 412-419
NUCLEAR MATRIX ORGANIZATION OF THE CHROMOCENTERS IN CULTURED MURINE FIBROBLASTS

E. V. Sheval, V. Y. Polyakov

A. N. Belozersky Institute of Physico-Chemical Biology, Moscow State University
and Institute of Agricultural Biochemistry, Moscow;
e-mail: sheval_e@genebee.msu.su

In the current work, the structural organization of nuclear matrix of pericentromeric heterochromatin blocks (chromocenters) inside cultured murine fibroblasts was investigated. After 2 M NaCl extraction without DNase I treatment, chromocenters were extremely swelled, and it was impossible to detect them using conventional electron microscopy. Using immunogolding with anti-topoisomerase IIα antibody, we demonstrated that residual chromocenters were subdivided into numerous discrete aggregates. After 2 M NaCl extraction with DNase I treatment, the residual chromocenters appeared as a dense meshwork of thin fibers, and using this feature, the residual chromocenters were easily distinguished from the rest of nuclear matrix. After extraction with dextran sulfate and heparin, the chromocenters were decondensed, and chromatin complexes having rosette organization (central core from which numerous DNA fibers radiated) were seen. Probably, the appearance of these rosettes was a consequence of incomplete chromatin extraction. Thus, the nuclear matrix of pericentromeric chromosome regions in cultured murine fibroblasts differs morphologically from the rest of nuclear matrix.

Key words:  nuclear matrix, pericentromeric heterochromatin blocks, murine fibroblasts


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