Increased Gi protein signaling potentiates the negative chronotropic effect of adenosine in the SHR right atrium

Original Article
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Abstract

Hypertension is a risk factor for cardiovascular diseases, which have been associated with dysfunction of sympathetic and purinergic neurotransmission. Therefore, herein, we evaluated whether modifications of adenosine receptor signaling may contribute to the cardiac dysfunction observed in hypertension. Isolated right atria from spontaneously hypertensive (SHR) or normotensive Wistar rats (NWR) were used to investigate the influence of adenosine receptor signaling cascade in the cardiac chronotropism. Our results showed that adenosine, the endogenous agonist of adenosine receptors, and CPA, a selective agonist of A1 receptor, decreased the atrial chronotropism of NWR and SHR in a concentration- and time-dependent manner, culminating in cardiac arrest (0 bpm). Interestingly, a 3-fold lower concentration of adenosine was required to induce the negative chronotropic effect in SHR atria. Pre-incubation of tissues from both strains with DPCPX, a selective A1 receptor antagonist, inhibited the negative chronotropic effect of CPA, while simultaneous inhibition of A2 and A3 receptors, with ZM241385 and MRS1523, did not change the adenosine chronotropic effects. Moreover, 1 μg/ml pertussis toxin, which inactivates the Gαi protein subunit, reduced by 80% the negative chronotropic effects of adenosine in the NWR atrium, with minor effects in SHR tissue. These data indicate that the negative chronotropic effect of adenosine in right atrium depends exclusively on the activation of A1 receptors. Moreover, the distinct responsiveness of NWR and SHR atria to pertussis toxin reveals that the enhanced negative chronotropic response of SHR right atrium is probably due to an increased activity of Gαi protein-mediated.

Keywords

Adenosine receptor G protein Heart rate SHR Right atria 

Notes

Acknowledgements

We thank Enio S. A. Pacini and Edilson D. S. Junior for helpful suggestions on the experimental design.

Compliance with ethical standards

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted.

Supplementary material

210_2018_1482_Fig8_ESM.gif (73 kb)
Supplementary Fig. 1

Chronotropic effect of adenosine and CPA is unchanged after consecutive agonist curves. Adenosine (0.1 – 1000 μM) (a and b) or the adenosine A1 receptor agonist CPA (0.1 nM – 30 μM) (c and d) effect on chronotropism was evaluated in right atria from normotensive (NWR, a and c) and spontaneously hypertensive rats (SHR, b and d) after two consecutive agonist curves. After the first curve (open circle), preparation was washed out with nutritive solution and the second curve (black circle) was repeated after 1 hour of equilibration time. Potency (pEC50) of the agonists was compared using Student’s t-test. Chronotropism is expressed as mean ± S.E.M. of basal value (100%) obtained before addition of adenosine (n=6) or CPA (n=3) (GIF 73 kb)

210_2018_1482_MOESM1_ESM.tif (99 kb)
High Resolution Image (TIFF 98 kb)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Pharmacology, Escola Paulista de MedicinaUniversidade Federal de São Paulo (UNIFESP)São PauloBrazil

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