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In summary these data show that agonist
In summary, these data show that agonist exposure induces internalization and reduction in cell surface HA-mAPJ expression that occurs relatively rapidly during agonist exposure and recovers more slowly after agonist removal. Moreover the ppERK response of mAPJ-HEK293 cells to [Pyr1]apelin-13 desensitizes rapidly during sustained stimulation and this desensitization is due to upstream APJ-specific, rather than down-stream ERK-specific, adaptive changes. We show that GRK2-mediated phosphorylation targets mAPJ to CCVs that are internalized from the cell surface in a β−arrestin-independent, EPS15- and dynamin-dependent manner. Our data indicates that receptor internalization is not required for mAPJ desensitization of ppERK responses in the mAPJ-HEK293 cell line. This multifaceted system may be indicative of a complex mechanism in controlling the physiological functions of endogenous apelin and may be important in conditions where there are elevated circulating or tissue levels of APJ. Further study of the signalling and regulation of APJ will help develop ligands for use in potential therapeutic intervention for dysfunctions of physiological homeostasis such as hypertensive disease.
Conflict of interest
Author contributions
Acknowledgements
This work was supported by the Biotechnology and Biological Sciences Research Council (grant No. BB/D00196X/1).
Introduction
Hypertension is one of the main risk factors for the development and progression of heart failure, myocardial infarction (MI) and Stroke [1]. As apelin/APJ system involves in the regulation of blood pressure/myocardial contractility, and apelin receptors are expressed on cardiomyocytes and cardiac endothelium, it is not surprising that the apelin/APJ system plays direct and indirect roles on the structure development and function of the heart [2].
Opioids are mostly known as analgesics, but their role in modulation of Rizatriptan Benzoate mg activity has been emphasized in the recent years [3]. These drugs are currently used in control of clinical conditions such as MI and Stroke.
APJ and opioid receptors (OPRs) belong to class A subfamily of GPCRs superfamily [4], and both receptors transduce signals through Gi and phosphorylation of extra-cellular regulated kinase (ERK1/2) [5], [6]. APJ form heterodimer with the kappa opioid receptors (KORs) in human embryonic kidney cells (HEK-293) [4].
It has been shown that ERK1/2 and protein kinase C (PKC) are the signaling pathways of cardiac contractility [7]. Apelin has positive cardiac inotropic effects through PKC activation and phosphorylation of ERK1/2 [8], and the heterodimerization of APJ and KORs increases PKC in response to apelin [4]. Therefore the two receptors may influence the physiological effects of each other, such as modulation of cardiac contractility. Our previous study indicated that in rats with acute renovascular hypertension, interaction between the APJ and KORs intensified the impact of apelin on blood pressure and this impact was dose-dependent [9]. As the chronic hypertension is more clinically relevant condition, the present study aimed at evaluating the heterodimerization between KOR and APJ and assessing the impact of low and high doses of apelin on the level of heterodimerization in the myocardium in chronic renovascular hypertension. In addition, as the role of ERK1/2 phosphorylation and PKC in mediating the cardiac impacts of apelin in rats with renovascular hypertension and the role of KORs in this regard is unclear, the expression of KORs, the role of these intracellular mediators and the interaction between KORs and APJ on the cardiac impacts of apelin in a chronic renovascular model of hypertension was also investigated.
Materials and methods
Results
Discussion
Apelin 60μg/kg decreases the contractility index and increases tau at its peak effect (min 1), which indicates that apelin causes negative inotropic and lusitropic effects at this dose and time (Fig. 5). The previous study in our lab also indicated that high dose of apelin exerts negative inotropic and lusitropic effects [19].