Gap junction communication between cells aggregated on a cellulose-coated polystyrene: influence of connexin 43 phosphorylation.
The appropriate functioning of tissues and organ systems depends on intercellular communication such as gap junctions formed by connexin (Cx) protein channels between adjacent cells. We have previously shown that Swiss 3T3 cells aggregated on hydrophilic cellulose substratum Cuprophan (CU) establish short linear gap junctions composed of Cx 43 in cell surface plaques. This phenomenon seems to depend on the high intracellular cyclic AMP (cAMP) concentration triggered by attachment of the cells to CU. We have now used a cellulose-coated polystyrene inducing the same cell behaviour to analyse the gap junction communication between aggregated cells. The transfer of the dye Lucifer Yellow (LY) between cells showed that cells aggregated on cellulose substratum rapidly (within 90 min) establish functional gap junctions. Inhibitors of cAMP protein kinase (PKI) or protein kinase C (GF109203X) both inhibited the diffusion of LY between neighbouring cells. Western blot analysis showed that this change in permeability was correlated with a decrease in Cx 43 phosphorylation. Thus, cellulose substrata seem to induce cell-cell communication through Cx 43 phosphorylation modulated by PKA and PKC. To understand the mechanisms by which a substratum regulates gap junctional communication is critically important for the emerging fields of tissue engineering and biohybrid devices.
Faucheux
N
N
Domaine Biomatériaux-Biocompatibilité, Université de Technologie de Compiègne, UMR CNRS 6600, BP20529, Compiègne Cédex 60205, France.
Zahm
J M
JM
Bonnet
N
N
Legeay
G
G
Nagel
M D
MD
D000818:Animals; D015153:Blotting, Western; D002450:Cell Communication; D002482:Cellulose / Q000737:chemistry*; D018031:Connexin 43 / Q000378:metabolism*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D017629:Gap Junctions; D051379:Mice; D010766:Phosphorylation; D011137:Polystyrenes / Q000737:chemistry*; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism; D041701:Swiss 3T3 Cells
D018031:Connexin 43; D004791:Enzyme Inhibitors; D011137:Polystyrenes; D002482:Cellulose; D017868:Cyclic AMP-Dependent Protein Kinases; D011493:Protein Kinase C