Research experience
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Sep 2010–
presentResearch: Universitat de Barcelona
Universitat de Barcelona · Departament de Patologia i Terapèutica Experimental · Neuropharmacology and painSpain · Barcelona
Publications (24) View all
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Article: Dopamine D2 Receptor-Mediated Modulation of Adenosine A2A Receptor Agonist Binding within the A2AR/D2R Oligomer Framework.
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ABSTRACT: The molecular interaction between adenosine A2A and dopamine D2 receptors (A2ARs and D2Rs, respectively) within an oligomeric complex has been postulated to play a pivotal role in the adenosine-dopamine interplay in the central nervous system, in both normal and pathological conditions (e.g. Parkinson's disease). While the effects of A2AR challenge on D2R functioning have been largely studied, the reverse condition is still unexplored, a fact that might have impact in therapeutics. Here, we aimed to examine in a real-time mode the D2R-mediated allosteric modulation of A2AR binding when an A2AR/D2R oligomer is established. Thus, we synthesized fluorescent A2AR agonists and evaluated, by means of a flow cytometry homogeneous no-wash assay and a real-time fluorescence resonance energy transfer (FRET)-based approach, the effects on A2AR binding of distinct antiparkinsonian drugs in current clinical use (i.e. pramipexole, rotigotine and apomorphine). Our results provided evidence for the existence of a differential D2R-mediated negative allosteric modulation on A2AR agonist binding that was oligomer-formation dependent, and with apomorphine being the best antiparkinsonian drug attenuating A2AR agonist binding. Overall, the here-developed methods were found valid to prospect the ability of drugs acting on D2Rs to modulate A2AR binding, thus featuring as possible helpful tools for the preliminary selection of D2R-like candidate drugs in the management of Parkinson's disease.Neurochemistry International 04/2013; · 2.86 Impact Factor -
Article: The Parkinson's Disease-Associated Gpr37 Receptor-Mediated Cytotoxicity Is Controlled By Its Intracellular Cysteine-Rich Domain.
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ABSTRACT: GPR37, also known as Pael-R (parkin associated endothelin-like receptor), is an orphan GPCR that aggregates intracellularly in a juvenile form of Parkinson's disease. However, little is known about the structure or function of this receptor. Here, in order to better understand the functioning of this receptor, we focused on the GPR37 C-terminal tail, in particular on a cystein enriched region. Thus, we aimed to reveal the role of these residues on receptor plasma membrane expression and function, and also whether the presence of this cysteine-rich domain is linked to the previously described receptor-mediated cytotoxicity. Interestingly, while the deletion of six cysteine residues within this region did not affect receptor internalization it promoted GPR37 plasma membrane expression and signalling. Furthermore, the removal of the C-terminal cysteine-rich domain protected against GPR37-mediated apoptosis and cell death. Overall, we identified a GPR37 domain, namely the C-terminal tail cysteine-rich domain, which played a critical role in receptor cell surface expression, function and GPR37-mediated cytotoxicity. These results might contribute to better comprehend the pathophysiology (i.e. in Parkinson's disease) of this rather unknown member of the GPCR family. © 2013 International Society for Neurochemistry, J. Neurochem. (2013) 10.1111/jnc.12196.Journal of Neurochemistry 02/2013; · 4.06 Impact Factor -
Article: Molecular determinants of A(2A) R-D(2) R allosterism: role of the intracellular loop 3 of the D(2) R.
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ABSTRACT: In the CNS, an antagonistic interaction has been shown between adenosine A(2A) and dopamine D(2) receptors (A(2) (A) Rs and D(2) Rs) that may be relevant both in normal and pathological conditions (i.e., Parkinson's disease). Thus, the molecular determinants mediating this receptor-receptor interaction have recently been explored, as the fine tuning of this target (namely the A(2) (A) R/D(2) R oligomer) could possibly improve the treatment of certain CNS diseases. Here, we used a fluorescence resonance energy transfer-based approach to examine the allosteric modulation of the D(2) R within the A(2) (A) R/D(2) R oligomer and the dependence of this receptor-receptor interaction on two regions rich in positive charges on intracellular loop 3 of the D(2) R. Interestingly, we observed a negative allosteric effect of the D(2) R agonist quinpirole on A(2) (A) R ligand binding and activation. However, these allosteric effects were abolished upon mutation of specific arginine residues (217-222 and 267-269) on intracellular loop 3 of the D(2) R, thus demonstrating a major role of these positively charged residues in mediating the observed receptor-receptor interaction. Overall, these results provide structural insights to better understand the functioning of the A(2) (A) R/D(2) R oligomer in living cells.Journal of Neurochemistry 08/2012; 123(3):373-84. · 4.06 Impact Factor -
Article: Transcriptional profiling of striatal neurons in response to single or concurrent activation of dopamine D(2), adenosine A(2A) and metabotropic glutamate type 5 receptors: Focus on beta-synuclein expression.
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ABSTRACT: G protein-coupled receptor oligomerization is a concept which is changing the understanding of classical pharmacology. Both, oligomerization and functional interaction between adenosine A(2A,) dopamine D(2) and metabotropic glutamate type 5 receptors have been demonstrated in the striatum. However, the transcriptional consequences of receptors co-activation are still unexplored. We aim here to determine the changes in gene expression of striatal primary cultured neurons upon isolated or simultaneous receptor activation. Interestingly, we found that 95 genes of the total analyzed (15,866 transcripts and variants) changed their expression in response to simultaneous stimulation of all three receptors. Among these genes, we focused on the β-synuclein (β-Syn) gene (SCNB). Quantitative PCR verified the magnitude and direction of change in expression of SCNB. Since β-Syn belongs to the homologous synuclein family and may be considered a natural regulator of α-synuclein (α-Syn), it has been proposed that β-Syn might act protectively against α-Syn neuropathology.Gene 08/2012; 508(2):199-205. · 2.34 Impact Factor -
Article: Fluorescence resonance energy transfer-based technologies in the study of protein-protein interactions at the cell surface.
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ABSTRACT: Understanding of the molecular mechanisms of protein-protein interactions (PPIs) at the cell surface of living cells is fundamental to comprehend the functional meaning of a large number of cellular processes. Here we discuss how new methodological strategies derived from non-invasive fluorescence-based approaches (i.e. fluorescence resonance energy transfer, FRET) have been successfully developed to characterize plasma membrane PPIs. Importantly, these technologies alone - or in concert with complementary methods (i.e. SNAP-tag/TR-FRET, TIRF/FRET) - can become extremely powerful approaches for visualizing cell surface PPIs, even between more than two proteins and also in native tissues. Interestingly, these methods would also be relevant in drug discovery in order to develop new high-throughput screening approaches or to identify new therapeutic targets. Accordingly, herein we provide a thorough assessment on all biotechnological aspects, including strengths and weaknesses, of these fluorescence-based methodologies when applied in the study of PPIs occurring at the cell surface of living cells.Methods 06/2012; 57(4):467-72. · 4.01 Impact Factor
