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

  • Research Article
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Journal of Cell Communication and Signaling Aims and scope

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.

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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

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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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Authors and Affiliations

  1. Institute of Biophysics, Leibniz University Hannover, Herrenhäuserstr. 2, 30419, Hannover, Germany

    Daniela Begandt, Almke Bader, Lutz Dreyer, Natalie Eisert, Thilo Reeck & Anaclet Ngezahayo

  2. Institute for Multiphase Processes, Leibniz University Hannover, Callinstr. 36, 30167, Hannover, Germany

    Lutz Dreyer

  3. Center for Systems Neuroscience Hannover, 30559, Hannover, Germany

    Anaclet Ngezahayo

Authors
  1. Daniela Begandt
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  2. Almke Bader
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  3. Lutz Dreyer
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  4. Natalie Eisert
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  5. Thilo Reeck
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  6. Anaclet Ngezahayo
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Correspondence to Anaclet Ngezahayo.

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Short summary

Dipyridamole is known as an antithrombotic drug, which acts by inhibiting adenosine transport leading to synthesis of cAMP in platelets. The present study shows that dipyridamole also affects vascular smooth muscle cells by enhancing their gap junction coupling through stimulation of the cAMP/PKA pathway. The increase of gap junction coupling was found to be biphasic with a short-term oscillatory behaviour, probably related to oscillatory synthesis of cAMP, which then affected already synthesized connexins and gap junction channels, and a long-term effect, which seemed to depend on a cAMP/PKA-induced expression and synthesis of connexins.

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Begandt, D., Bader, A., Dreyer, L. et al. Biphasic increase of gap junction coupling induced by dipyridamole in the rat aortic A-10 vascular smooth muscle cell line. J. Cell Commun. Signal. 7, 151–160 (2013). https://doi.org/10.1007/s12079-013-0196-4

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  • DOI: https://doi.org/10.1007/s12079-013-0196-4

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