Dipyridamole-related enhancement of gap junction coupling in the GM-7373 aortic endothelial cells correlates with an increase in the amount of connexin 43 mRNA and protein as well as gap junction plaques

作者：Begandt Daniela; Bader Almke; Gerhard Linda; Lindner Julia; Dreyer Lutz; Schlingmann Barbara; Ngezahayo Anaclet^*

来源：Journal of Bioenergetics and Biomembranes, 2013, 45(4): 409-419.

DOI：10.1007/s10863-013-9518-8

• Dipyridamole
• Gap junction
• cAMP
• Endothelial cells
• Connexin 43

Previous data showed that dipyridamole enhanced gap junction coupling in vascular endothelial and smooth muscle cell lines by a cAMP-dependent mechanism. The present study investigates the level at which dipyridamole affects gap junction coupling. In the GM-7373 endothelial cell line, scrape loading/dye transfer experiments revealed a rapid increase in gap junction coupling induced during the first 6 h of dipyridamole treatment, followed by a slow increase induced by further incubation. Immunostaining analyses showed that the rapid enhancement of gap junction coupling correlated with an increased amount of Cx43 gap junction plaques and a reduced amount of Cx43 containing vesicles, while the amount of Cx43 mRNA or protein was not changed during this period, as found by semiquantitative RT-PCR and Western blot. Additionally, brefeldin A did not block this short-term-induced enhancement of gap junction coupling. Along with the dipyridamole-induced long-term enhancement of gap junction coupling, the amount of Cx43 mRNA and protein additionally to the amount of Cx43 gap junction plaques were increased. Furthermore, the anti-Cx43 antibody detected only two bands at 42 kDa and 44 kDa in control cells and cells treated with dipyridamole for 6 h, while long-term dipyridamole-treated cells showed a third band at 46 kDa. We propose that a dipyridamole-induced cAMP synthesis increased gap junction coupling in the GM-7373 endothelial cell line at different levels: the short-term effect is related to already oligomerised connexins beyond the Golgi apparatus and the long-term effect involves new expression and synthesis as well as posttranslational modification of Cx43.

• 出版日期2013-8