PREPARATION AND CHARACTERIZATION OF NUCLEAR MATRIX/CHROMOSOME SCAFFOLD
IN SITU
E. V. Sheval,1, 2 S. Yu. Kurchashova,1 E. R. Timirbulatova,1
V. Yu. Polyakov 1, 2
1 A. N. Belozcrsky Institute of Physico-Chcmical Biology, Moscow State University,
and 2 All-Russian Institute of Agricultural Biotechnology, Moscow, Russia;
e-mail: esheval@mail.ru
A method of nuclear matrix and chromosomal scaffold preparation from cultured animal cells was developed. After the
high-salt extraction, interphase and mitotic cells were not detached from the coverslips that enabled us to analyse the nuclear
matrix and chromosomal scaffold in cells at all mitotic phases. Morphological methods (phase contrast microscopy and electron
microscopy of ultrathin sections) did not reveal any structures that could be identified as a chromosomal scaffold. However, after
staining with antibodies to XCAP-E and topoisomerase Ha some structures were revealed in metaphase cells having both localization
and morphology of a chromosomal scaffold. The cell residuals were not stained with antibodies to XCAP-E and topoisomerase IIα,
if the nuclear matrix and chromosomal scaffold were destabilized by addition of β-mercaptoethanol.
Key words: chromosome, scaffold, nuclear matrix
|
