Adenosine A2A receptor ligand recognition and signaling is blocked by A2B receptors

Hinz, Sonja; Navarro, Gemma; Borroto-Escuela, Dasiel O.; Seibt, Benjamin F.; Ammon, York-Christoph; Filippo, Elisabetta De; Danish, Azeem; Lacher, Svenja; Červinková, Barbora; Rafehi, Muhammad; Schiedel, Anke C.; Franco, Rafael; Müller, Christa E.

doi:10.18632/oncotarget.24423

Cited by 81 publications

(80 citation statements)

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“…This phenomenon is due to allosteric inter-protomer interactions in the heteromer, i.e. the presence of one receptor blocks the signaling of the partner receptor in the complex (Hinz et al, 2018). Interestingly, the presence of an antagonist could make, as in the case of Ang receptor heteromers in striatal neurons, appear AT 2 R functionality back.…”

Section: Discussionmentioning

confidence: 99%

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Angiotensin AT1 and AT2 receptor heteromer expression in the hemilesioned rat model of Parkinson’s disease that increases with levodopa-induced dyskinesia

Rivas‐Santisteban

Rodríguez‐Pérez

Muñoz

et al. 2020

Preprint

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Background/Aims: The renin-angiotensin system (RAS) is altered in Parkinson’s disease (PD), a disease due to substantia nigra neurodegeneration and whose dopamine-replacement therapy, using the precursor levodopa, leads to dyskinesias as the main side effect. Angiotensin AT1 and AT2 receptors, mainly known for their role in regulating water homeostasis and blood pressure and able to form heterodimers (AT1/2Hets), are present in the central nervous system. We assessed the functionality and expression of AT1/2Hets in Parkinson Disease (PD).Methods: Immunocytochemistry was used to analyze the colocalization between angiotensin receptors, bioluminescence resonance energy transfer was used to detect AT1/2Hets. Calcium and cAMP determination, MAPK activation and label-free assays were performed to characterize signaling in homologous and heterologous systems. Proximity ligation assays was used to quantify receptor expression in mouse primary cultures and in rat striatal sections.Results: We confirmed that AT1 and AT2 receptors form AT1/2Hets that are expressed in cells of the central nervous system. AT1/2Hets are novel functional units with particular signaling properties. Importantly, the coactivation of the two receptors in the heteromer reduces the signaling output of angiotensin. Remarkably, AT1/2Hets that are expressed in both striatal neurons and microglia show a cross-potentiation, i.e. candesartan, the antagonist of AT1 increases the effect of AT2 receptor agonists. In addition, the level of striatal expression increased in the unilateral 6-OH-dopamine lesioned rat PD model and was markedly higher in parkinsonian-like animals that did not become dyskinetic upon levodopa chronic administration if compared with expression in those that became dyskinetic.Conclusion: The results indicate that boosting the action of neuroprotective AT2 receptors using an AT1 receptor antagonist constitutes a promising therapeutic strategy in PD.

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Section: Discussionmentioning

confidence: 99%

“…HEK-293T cells were transiently transfected with the corresponding cDNA by the PEI (PolyEthylenImine, SigmaAldrich, St. Louis, MO) method (Franco et al, 2019;Hinz et al, 2018). Brie y, the corresponding cDNA diluted in 150 mM NaCl was mixed with PEI (5.5 mM in nitrogen residues) also prepared in 150 mM NaCl for 10 min.…”

Section: Cell Transfectionmentioning

confidence: 99%

Angiotensin AT1 and AT2 receptor heteromer expression in the hemilesioned rat model of Parkinson’s disease that increases with levodopa-induced dyskinesia

Rivas‐Santisteban

Rodríguez‐Pérez

Muñoz

et al. 2020

Preprint

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“…Cells were grown in Dulbecco's modified Eagle's medium (DMEM) supplemented with 2 mM L-glutamine, 100 U/ml penicillin/streptomycin, and 5% (v/v) heat inactivated fetal bovine serum (FBS) (Invitrogen, Paisley, Scotland, United Kingdom). Cells were maintained in a humid atmosphere of 5% CO 2 at 37 • C. Cells were transiently transfected with the polyethylenimine (PEI, Sigma, St. Louis, MO, United States) method (38)(39)(40). Briefly, cells were incubated (4h) in a serum-starved medium with the corresponding cDNA and with PEI (5.47 mM in nitrogen residues) and 150 mM NaCl.…”

Section: Cell Culture and Transient Transfectionmentioning

confidence: 99%

Specificity and nanomolar potency of melatonin on G-protein coupled melatonin MT1 and MT2 receptors expressed in HEK-293T human embryo kidney cells

Rivas‐Santisteban¹,

Reyes‐Resina²,

Raïch³

et al. 2019

Melatonin Res.

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This is a pre-registered study, i.e. a study whose hypotheses and experiments designed to address these hypotheses have been deposited in a database before starting the experiments. The study aims at assessing the Gs versus Gi coupling and the potency of melatonin in the human version of melatonin MT1 and MT2 G-protein-coupled receptors expressed in HEK-293T cells. The results show that these receptors are Gi but not Gs coupled. By using a standard procedure of modulation of 0.5 µM forskolin-induced cAMP levels, it was found that the potency on MT2 receptor-mediated actions is in the low nanomolar range, but the potency on MT1 receptor is in the high nanomolar range. The potency of melatonin to stimulate the MT2 receptor is similar to that of a selective agonist, N-[2-(2-methoxy-6H-isoindolo[2,1-a]indol-11-yl)ethyl]butanamide (IIK7). Overall, the data on the potency of melatonin on its receptors will provide a new look for melatonin research. It is important to consider this finding for appropriately addressing physiological or therapeutic effects based on melatonin potency. Thus, the low doses of melatonin used in the existing prolonged release preparations or in other supplements should be revisited.

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“…β‐arrestin recruitment was determined as previously described (Hinz et al, ; Navarro et al, ). Briefly, BRET experiments were performed in transfected HEK‐293T cells 48 hr after transfection with the cDNA corresponding to the indicated receptors fused to the YFP and 1‐μg cDNA corresponding to β‐arrestin 2Rluc.…”

Section: Methodsmentioning

confidence: 99%

Adreno–melatonin receptor complexes control ion homeostasis and intraocular pressure ‐ their disruption contributes to hypertensive glaucoma

Alkozi

Navarro

Aguinaga

et al. 2020

British J Pharmacology

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Background and Purpose: Often, glaucoma presents with elevated eye hydrostatic pressure, which is regulated by endogenous melatonin. Phenylephrine increases cytoplasmic [Ca 2+ ], via α 1 -adrenoceptor activation, that is detrimental in glaucoma. The aims of this study were (a) to elucidate the role of melatonin receptors in humour production and intraocular pressure (IOP) maintenance and (b) to identify glaucomarelevant melatonin-adrenoceptor interactions.Experimental Approach: Biophysical and proximity ligation assays were performed to identify interactions in heterologous expression systems, in cell lines and in human eyes. G s /G i /G q signalling was investigated in these systems and in cells producing aqueous humour. IOP was determined in a mice model of glaucoma. Retinography and topically pharmacological treatment were performed in control and in glaucomatous mice.Key Results: α 1 -adreno-and melatonin receptors form functional complexes in which the C-terminal tail of the adrenoceptor plays a role. Remarkably, activation of α 1adrenoceptors in these complexes did not lead to cytosolic Ca 2+ increases, suggesting G s instead of G q coupling is involved. The number of these complexes significantly decreased in models of glaucoma and, importantly, in human samples from glaucoma patients. This has led to hypothesize that melatonin, a hypotensive agent, plus blockade of α 1 -adrenoceptors could normalize pressure in glaucoma. Remarkably, coinstillation of melatonin and prazosin, an α 1 -adrenoceptor antagonist, resulted in long-term decreases in IOP in a well-established animal model of glaucoma.Conclusions and Implications: The findings are instrumental to understand the physiological function of melatonin in the eye and its potential to address eye pathologies by targeting melatonin receptors and their complexes.Abbreviations: 4PPDOT, cis-4-phenyl-2-propionamidotetralin (antagonist of MT2R); AR, adrenoceptor; IIK7, N-[2-(2-methoxy-6H-isoindolo[2,1-a]indol-11-yl)ethyl]butanamide (MT2R selective agonist); IOP, intraocular pressure; MT 1 R, melatonin MT 1 receptor; MT 2 R, melatonin MT 2 receptor; Rluc, Renilla luciferase; YFP, yellow fluorescence protein; α 1A -MT 1 Hets, α 1A -MT 1 receptor heteromers.A first preprint of this paper was deposited in BioRxiv (https://doi.

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Adenosine A2A receptor ligand recognition and signaling is blocked by A2B receptors

Cited by 81 publications

References 58 publications

Angiotensin AT1 and AT2 receptor heteromer expression in the hemilesioned rat model of Parkinson’s disease that increases with levodopa-induced dyskinesia

Angiotensin AT1 and AT2 receptor heteromer expression in the hemilesioned rat model of Parkinson’s disease that increases with levodopa-induced dyskinesia

Specificity and nanomolar potency of melatonin on G-protein coupled melatonin MT1 and MT2 receptors expressed in HEK-293T human embryo kidney cells

Adreno–melatonin receptor complexes control ion homeostasis and intraocular pressure ‐ their disruption contributes to hypertensive glaucoma

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