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Fluorescence cytometry of microtubules and nuclear DNA during cell‐cycle and reverse‐transformation
Author(s) -
Vergani L.,
Gavazzo P.,
Facci P.,
Diaspro A.,
Mascetti G.,
Nicolini C.,
Are.,
Gaspa L.
Publication year - 1992
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.240500210
Subject(s) - flow cytometry , biology , microtubule , cell cycle , dapi , population , chromatin , cell , microbiology and biotechnology , cytoplasm , cytometry , staining , chemistry , dna , biochemistry , genetics , medicine , environmental health
Synchronized CHO‐K1 cells and their dibutyryl c‐AMP treated counterparts have been characterized by means of static and flow fluorescence cytometry at the level of nuclear DNA and cytoplasmic microtubules. In order to confirm earlier findings on synchronized population, Carnoy fixed and hydrolyzed, several new findings are here reported at the level of single intact cell. The fluorescence intensity of DAPI‐stained glutaraldehyde fixed 2C cells correlates well with the average absorbance of the corresponding Feulgen‐stained cells, thereby appearing also to be a measure of chromatin condensation during the G1 phase. In the early part of G1, the drastic alteration in anti‐β tubulin immunostaining is shown to parallel microtubule depolymerization induced by calcium or colcemide. The known 1–2 h lengthening of the G1 period after reverse‐transformation appears to correlate with a similar delay in the abrupt chromatin decondensation. The above results are discussed in terms of the role of microtubules and nuclear morphometry (and their coupling) in the control of cell cycle progression of transformed vs. fibroblast‐like cells. © 1992 Wiley‐Liss, Inc.