Organization of cyclic AMP-dependent connexin 43 in Swiss 3T3 cells attached to a cellulose substratum

• Published in: Biomaterials 2002, Vol.23 (2), p.413-421
• Main Authors: Faucheux, N, Dufresne, M, Nagel, M.D
• Format: Article
• Language: English
• Subjects: 3T3 Cells; Animals; Brefeldin A - pharmacology; Cell aggregation; Cellulose - metabolism; Connexin; Connexin 43 - metabolism; Cuprophan; Cyclic AMP; Cyclic AMP - metabolism; Gap junction; Gap Junctions - metabolism; Gap Junctions - ultrastructure; Mice; Microscopy, Electron; Protein Binding
• ISSN: 0142-9612; 1878-5905
• DOI: 10.1016/S0142-9612(01)00120-X

We have previously shown that the adenylyl cyclase, which produces cyclic AMP (cAMP) in Swiss 3T3 cells, is activated by their attachment to a cellulose substratum (Cuprophan, CU). This substratum adsorbs vitronectin poorly, prevents cell spreading and causes them to aggregate. By contrast, cells spread out on polystyrene and contain low concentrations of cAMP. We have found that Connexin 43 (Cx 43) gap junction plaques are involved in this cell aggregation. MDL 12330 A, a specific inhibitor of adenylyl cyclase, prevented cell aggregation on CU and abolished Cx 43 channel clustering. But forskolin, a direct activator of adenylyl cyclase, and 8Br cAMP, a cell-permeable analogue of cAMP, caused Cx 43 channel clustering in cells attached to polystyrene. Hence, Cx 43 channel clustering is regulated by cAMP in Swiss 3T3 cells. In addition, neither brefeldin A nor monensin (inhibitors of transit through the endoplasmic reticulum and Golgi apparatus), abolished Cx 43 channel clustering in cells aggregated on CU. Thus, the Cx 43 that form clusters in cells attached to CU are not dependent upon the trafficking of Cx 43 from intracellular storage sites, but are probably reorganised from the plasma membrane.