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Journal of Bioenergetics and Biomembranes

, Volume 45, Issue 4, pp 409–419 | Cite as

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

  • Daniela Begandt
  • Almke Bader
  • Linda Gerhard
  • Julia Lindner
  • Lutz Dreyer
  • Barbara Schlingmann
  • Anaclet NgezahayoEmail author
Article

Abstract

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.

Keywords

Dipyridamole Gap junction cAMP Endothelial cells Connexin 43 

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Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Daniela Begandt
    • 1
  • Almke Bader
    • 1
  • Linda Gerhard
    • 1
  • Julia Lindner
    • 1
  • Lutz Dreyer
    • 1
    • 3
  • Barbara Schlingmann
    • 1
    • 2
  • Anaclet Ngezahayo
    • 1
    • 2
    Email author
  1. 1.Institute of BiophysicsLeibniz University HannoverHannoverGermany
  2. 2.Center for Systems Neuroscience HannoverUniversity of Veterinary Medicine Hannover FoundationHannoverGermany
  3. 3.Institute for Multiphase ProcessesLeibniz University HannoverHannoverGermany

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