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Biphasic increase of gap junction coupling induced by dipyridamole in the rat aortic A-10 vascular smooth muscle cell line

  • Daniela Begandt
  • Almke Bader
  • Lutz Dreyer
  • Natalie Eisert
  • Thilo Reeck
  • Anaclet NgezahayoEmail author
Research Article

Abstract

The rat aortic smooth muscle cell line A-10 was used to investigate the effect of dipyridamole on the gap junction coupling of smooth muscle cells. The scrape loading/dye transfer (SL/DT) technique revealed that dipyridamole concentrations between 5 μM and 100 μM significantly increased gap junction coupling. The adenosine receptor antagonist MRS 1754, as well as the PKA inhibitors Rp-cAMPS and H-89 were able to inhibit the dipyridamole-related increase in coupling, while forskolin and Br-cAMP also induced an enhancement of the gap junction coupling. Regarding the time-dependent behaviour of dipyridamole, a short-term effect characterised by an oscillatory reaction was observed for application times of less than 5 h, while applications times of at least 6 h resulted in a long-term effect, characterised by a constant increase of gap junction coupling to its maximum levels. This increase was not altered by prolonged presence of dipyridamole. In parallel, a short application of dipyridamole for at least 15 min was found to be sufficient to evoke the long-term effect measured 6 h after drug washout. We propose that in both the short-term and long-term effect, cAMP-related pathways are activated. The short-term phase could be related to an oscillatory cAMP effect, which might directly affect connexin trafficking, assembly and/or gap junction gating. The long-term effect is most likely related to the new expression and synthesis of connexins. With previous data from a bovine aortic endothelial cell line, the present results show that gap junction coupling of vascular cells is a target for dipyridamole.

Keywords

cAMP Dipyridamole Gap junction PKA Vascular smooth muscle cells 

Abbreviations

Br-cAMP

8-Bromoadenosine-3′ 5′-cyclic monophosphate

cAMP

Cyclic adenosine-3′ 5′-cyclic monophosphate

Cx

Connexin

Dip.

Dipyridamole

ENT

Equilibrative nucleoside transporter

Eth.

0.5% Ethanol

For.

Forskolin

H-89

N-[2-(p-Bromocinnamylamino)ethyl]-5-isoquinolinesulfonamide

LY

Lucifer Yellow

PKA

Protein kinase A

Rp-cAMPS

Rp-adenosine-3′,5′-cyclic mono-phosphorothioate triethylamine salt

SL/DT

Scrape loading/dye transfer

Notes

Acknowledgments

This work was supported by Boehringer Ingelheim International GmbH. The authors thank Prof. Dr. Wolfgang Eisert for discussion and comments on the manuscript.

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Copyright information

© The International CCN Society 2013

Authors and Affiliations

  • Daniela Begandt
    • 1
  • Almke Bader
    • 1
  • Lutz Dreyer
    • 1
    • 2
  • Natalie Eisert
    • 1
  • Thilo Reeck
    • 1
  • Anaclet Ngezahayo
    • 1
    • 3
    Email author
  1. 1.Institute of BiophysicsLeibniz University HannoverHannoverGermany
  2. 2.Institute for Multiphase ProcessesLeibniz University HannoverHannoverGermany
  3. 3.Center for Systems Neuroscience HannoverHannoverGermany

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