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Non-classical regulation of β1- and β2-adrenoceptor-mediated inotropic responses in rat heart ventricle by the G protein Gi

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Abstract

Studies suggest that increased activity of Gi contributes to the reduced β-adrenoceptor-mediated inotropic response (βAR-IR) in failing cardiomyocytes and that β2AR-IR but not β1AR-IR is blunted by dual coupling to Gs and Gi. We aimed to clarify the role of Gi upon the β1AR-IR and β2AR-IR in Sham and failing myocardium by directly measuring contractile force and cAMP accumulation. Contractility was measured ex vivo in left ventricular strips and cAMP accumulation in cardiomyocytes from rats with post-infarction heart failure (HF) or sham operates (Sham). The β2AR-IR in Sham and HF was small and was amplified by simultaneously inhibiting phosphodiesterases 3 and 4 (PDE3&4). In HF, the inotropic response and cAMP accumulation evoked by β1AR- or β2AR-stimulation were reduced. Inactivation of Gi with pertussis toxin (PTX) did not restore the β1AR-IR or β2AR-IR in HF to Sham levels but did enhance the maximal β2AR-IR. PTX increased both β1AR- and β2AR-evoked cAMP accumulation more in Sham than that in HF, and HF levels approached those in untreated Sham. The potency of agonists at β1 and at β2ARs (only under PDE3&4 inhibition) was increased in HF and by PTX in both HF and Sham. Without PDE3&4 inhibition, PTX increased only the maximal β2AR-IR, not potency. We conclude that Gi regulates both β1AR- and β2AR-IR independent of receptor coupling with Gi. Gi together with PDE3&4 tonically restrict the β2AR-IR. Gi inhibition did not restore the βAR-IR in HF despite increasing cAMP levels, suggesting that the mechanism of impairment resides downstream to cAMP signalling.

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Abbreviations

AC:

Adenylyl cyclase

ADP:

Adenosine diphosphate

βAR:

β-Adrenoceptor

βAR-IR:

β-Adrenoceptor-mediated inotropic response

Cil:

Cilostamide

(dF/dt) max :

maximal development of force

HF:

Heart failure

IBMX:

3-Isobutyl-1-methylxanthine

LVEDP:

Left ventricular end-diastolic pressure

LVSP:

Left ventricular systolic pressure

PDE:

Phosphodiesterase

PTX:

Pertussis toxin

Rol:

Rolipram

Sham:

Sham-operated

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Acknowledgments

We thank Iwona Gutowska Schiander for excellent technical assistance. This work was supported by the Norwegian Council on Cardiovascular Diseases, the Research Council of Norway, the Stiftelsen Kristian Gerhard Jebsen foundation, the Anders Jahre foundation for the promotion of science, the Family Blix foundation, the Simon-Fougner-Hartmann family foundation and grants from the University of Oslo.

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Correspondence to Finn Olav Levy.

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Caroline Bull Melsom and Rizwan Iqbal Hussain contributed equally to the work.

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Melsom, C.B., Hussain, R.I., Ørstavik, Ø. et al. Non-classical regulation of β1- and β2-adrenoceptor-mediated inotropic responses in rat heart ventricle by the G protein Gi . Naunyn-Schmiedeberg's Arch Pharmacol 387, 1177–1186 (2014). https://doi.org/10.1007/s00210-014-1036-7

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