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First published online July 31, 2003
doi: 10.1242/10.1242/jcs.00656
Research Article |
1 Department of Human Morphology, Faculty of Medicine
2 Department of Biology, Faculty of Arts and Sciences, American University of
Beirut, PO Box 11-0236, Beirut, Lebanon
* Authors for correspondence (e-mail: me00{at}aub.edu.lb; rtalhouk{at}aub.edu.lb)
Accepted 7 May 2003
The relationship between gap junctional intercellular communication (GJIC)
and mammary cell (CID-9) differentiation in vitro was explored. CID-9 cells
differentiate and express ß-casein in an extracellular matrix (ECM)- and
hormone-dependent manner. In response to interaction with the ECM, cells in
culture modulated the expression of their gap junction proteins at the
transcriptional and post-translational levels. In the presence of EHS-matrix,
connexins (Cx)26, 32 and 43 localized predominantly to the plasma membrane,
and enhanced GJIC [as measured by Lucifer Yellow (LY) dye transfer assays] was
noted. Inhibition of GJIC of cells on EHS-matrix with 18
glycyrrhetinic
acid (GA) resulted in reversible downregulation of ß-casein expression.
In the presence of cAMP, cells cultured on plastic expressed ß-casein,
upregulated Cx43 and Cx26 protein levels and enhanced GJIC. This was reversed
in the presence of 18 GA. cAMP-treated cells plated either on a
non-adhesive PolyHEMA substratum or on plastic supplemented with
function-blocking anti-ß1 integrin antibodies, maintained
ß-casein expression. These studies suggest that cell-ECM interaction
alone may induce differentiation through changes in cAMP levels and formation
of functional gap junctions. That these events are downstream of ECM
signalling was underscored by the fact that enhanced GJIC induced partial
differentiation in mammary epithelial cells in the absence of an exogenously
provided basement membrane and in a ß1-integrin- and
adhesion-independent manner.
Key words: Connexins, Differentiation, ECM, GJIC, Mammary
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