A door opener for future research: agonist-induced β3-adrenoceptor desensitization in HEK cells but not CHO cells

Seifert, Roland

doi:10.1007/s00210-013-0884-x

Cited by 3 publications

(2 citation statements)

References 14 publications

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“…(expression and density) but the signaling cascade instead (Seifert, 2013). While long-term mechanisms might have a more profound and generalized effect over signaling of GPCRs coupled to Gas, RH driven desensitization by GRK2 might be more specific since soluble GRK2 translocates to plasmatic membrane after receptor activation, affecting only the signaling of the stimulated GPCR.…”

Section: Discussionmentioning

confidence: 99%

The Regulator of G Protein Signaling Homologous Domain of G Protein-Coupled Receptor Kinase 2 Mediates Short-Term Desensitization of β3-Adrenergic Receptor

et al. 2020

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G protein coupled receptor (GPCR) kinases (GRKs) are key regulators of GPCR signaling. Canonical mechanism of GPCR desensitization involves receptor phosphorylation by GRKs followed by arrestin recruitment and uncoupling from heterotrimeric G protein. Although b3-adrenergic receptor (b3AR) lacks phosphorylation sites by GRKs, agonist treatment proved to induce b3AR desensitization in many cell types. Here we show that GRK2 mediates short-term desensitization of b3AR by a phosphorylation independent mechanism but mediated by its domain homologous to the regulator of G protein signaling (RGS). HEK293T cells overexpressing human b3AR presented a short-term desensitization of cAMP response stimulated by the b3AR agonist, BRL37344, and not by forskolin. We found that b3AR desensitization was higher in cells co-transfected with GRK2. Similarly, overexpression of the RGS homology domain but not kinase domain of GRK2 increased b3AR desensitization. Consistently, stimulation of b3AR increased interaction between GRK2 and Gas subunit. Furthermore, in rat cardiomyocytes endogenously expressing b3AR, transfection with dominant negative mutant of RH domain of GRK2 (GRK2/D110A) increased cAMP response to BRL37344 and inhibited receptor desensitization. We expect our study to be a starting point for more sophisticated characterization of the consequences of GRK2 mediated desensitization of the b3AR in heart function and disease.

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Section: Discussionmentioning

confidence: 99%

The Regulator of G Protein Signaling Homologous Domain of G Protein-Coupled Receptor Kinase 2 Mediates Short-Term Desensitization of β3-Adrenergic Receptor

et al. 2020

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“…Overall, these findings suggest that agonist-induced desensitization of β 3 -adrenoceptors occurs in a tissue-or cell type-dependent manner [27].…”

mentioning

confidence: 75%

Human β3-Adrenoreceptor is Resistant to Agonist-Induced Desensitization in Renal Epithelial Cells

Milano

Gerbino

Schena

et al. 2018

Cell Physiol Biochem

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Background/Aims: We recently showed that the β₃-adrenoreceptor (β₃AR) is expressed in mouse kidney collecting ducts (CD) cells along with the type-2 vasopressin receptor (AVPR2). Interestingly, a single injection of a β₃AR selective agonist promotes a potent antidiuretic effect in mice. Before considering the feasibility of chronic β₃AR agonism to induce antidiuresis in vivo, we aimed to evaluate in vitro the signaling and desensitization profiles of human β₃AR. Methods: Human β₃AR desensitization was compared with that of human AVPR2 in cultured renal cells. Video imaging and FRET experiments were performed to dissect β₃AR signaling under acute and chronic stimulation. Plasma membrane localization of β₃AR, AVPR2 and AQP2 after agonist stimulation was studied by confocal microscopy. Receptors degradation was evaluated by Western blotting. Results: In renal cells acute stimulation with the selective β₃AR agonist mirabegron, induced a dose-dependent increase in cAMP. Interestingly, chronic exposure to mirabegron promoted a significant increase of intracellular cAMP up to 12 hours. In addition, a slow and slight agonist-induced internalization and a delayed downregulation of β₃AR was observed under chronic stimulation. Furthermore, chronic exposure to mirabegron promoted apical expression of AQP2 also up to 12 hours. Conversely, long-term stimulation of AVPR2 with dDAVP showed short-lasting receptor signaling, rapid internalization and downregulation and apical AQP2 expression for no longer than 3 h. Conclusions: Overall, we conclude that β₃AR is less prone than AVPR2 to agonist-induced desensitization in renal collecting duct epithelial cells, showing sustained cAMP production, preserved membrane localization and delayed degradation after 12 hours agonist exposure. These results may be important for the potential use of chronic pharmacological stimulation of β₃AR to promote antidiuresis overcoming in vivo renal concentrating defects caused by inactivating mutations of the AVPR2.

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Do β-adrenoceptor agonists induce homologous or heterologous desensitization in rat urinary bladder?

Michel

2013

Naunyn-Schmiedeberg's Arch Pharmacol

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β3-Adrenoceptor agonists have recently been introduced for the symptomatic treatment of the overactive bladder syndrome. As such treatment is not curative, long-term treatment is anticipated to be required. As the susceptibility of β3-adrenoceptors to undergo agonist-induced desensitization is cell type- and tissue-dependent, we have explored whether pre-treatment with a β-adrenoceptor agonist will attenuate subsequent relaxation responses to freshly added agonist using rat urinary bladder as a model. We have used the prototypical β-adrenoceptor agonist isoprenaline, the β2-selective fenoterol and the β3-selective CL 316,243 and mirabegron as well as the receptor-independent bladder relaxant forskolin. We show that a 6-h pre-treatment with agonist can significantly reduce subsequent relaxation against KCl-induced smooth muscle tone, but agonist-induced desensitization was also observed with longer pre-treatments or against passive tension. The agonist-induced desensitization was prominent for the β2 component of rat bladder relaxation but much weaker or even absent for the β3 component. Moreover, β-adrenoceptor agonist pre-treatment reduced contractile responses to the muscarinic agonist carbachol and the receptor-independent stimulus KCl. Taken together these data do not support the hypothesis that the long-term clinical efficacy of β3-adrenoceptor agonists in the treatment of the overactive bladder syndrome will be limited by receptor desensitization. Rather they raise the possibility that such treatment may not only cause smooth muscle relaxation but also may attenuate hyper-contractility of the bladder.

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A door opener for future research: agonist-induced β3-adrenoceptor desensitization in HEK cells but not CHO cells

Cited by 3 publications

References 14 publications

The Regulator of G Protein Signaling Homologous Domain of G Protein-Coupled Receptor Kinase 2 Mediates Short-Term Desensitization of β3-Adrenergic Receptor

The Regulator of G Protein Signaling Homologous Domain of G Protein-Coupled Receptor Kinase 2 Mediates Short-Term Desensitization of β3-Adrenergic Receptor

Human β3-Adrenoreceptor is Resistant to Agonist-Induced Desensitization in Renal Epithelial Cells

Do β-adrenoceptor agonists induce homologous or heterologous desensitization in rat urinary bladder?

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