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Ventura M.,University of Barcelona | Mateo F.,University of Barcelona | Serratosa J.,Institute Investigaciones Biomedicas Of Barcelona | Salaet I.,University of Barcelona | And 3 more authors.
International Journal of Biochemistry and Cell Biology | Year: 2010

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is considered a housekeeping glycolitic enzyme that recently has been implicated in cell signaling. Under apoptotic stresses, cells activate nitric oxide formation leading to S-nitrosylation of GAPDH that binds to Siah and translocates to the nucleus. The GAPDH-Siah interaction depends on the integrity of lysine 227 in human GAPDH, being the mutant K227A unable to associate with Siah. As lysine residues are susceptible to be modified by acetylation, we aimed to analyze whether acetylation could mediate transport of GAPDH from cytoplasm to the nucleus. We observed that the acetyltransferase P300/CBP-associated factor (PCAF) interacts with and acetylates GAPDH. We also found that over-expression of PCAF induces the nuclear translocation of GAPDH and that for this translocation its intact acetylase activity is needed. Finally, the knocking down of PCAF reduces nuclear translocation of GAPDH induced by apoptotic stimuli. By spot mapping analysis we first identified Lys 117 and 251 as the putative GAPDH residues that could be acetylated by PCAF. We further demonstrated that both Lys were necessary but not sufficient for nuclear translocation of GAPDH after apoptotic stimulation. Finally, we identified Lys 227 as a third GAPDH residue whose acetylation is needed for its transport from cytoplasm to the nucleus. Thus, results reported here indicate that nuclear translocation of GAPDH is mediated by acetylation of three specific Lys residues (117, 227 and 251 in human cells). Our results also revealed that PCAF participates in the GAPDH acetylation that leads to its translocation to the nucleus. © 2010 Elsevier Ltd.

Mlguez D.G.,Institute Biologla Molecular Of Barcelona | Mlguez D.G.,Autonomous University of Madrid | Gil-Guinon E.,Institute Biologla Molecular Of Barcelona | Pons S.,Institute Biologla Molecular Of Barcelona | And 2 more authors.
Journal of Cell Science | Year: 2013

The transforming growth factor beta (TGF-b) pathway plays key roles in development and cancer. TGF-b signaling converges on the Smad2 and Smad3 effectors, which can either cooperate or antagonize to regulate their transcriptional targets. Here we performed in vivo and in silico experiments to study how such cooperativity and antagonism might function during neurogenesis. In vivo electroporation experiments in the chick embryo neural tube show that Smad2 and Smad3 cooperate to promote neurogenesis, as well as the transcription of Smad3-specific targets. Knockdown of Smad2 enhances neurogenesis and the transcription of Smad3-specific targets. A mathematical model of the TGF-b pathway fits the experimental results and predicts that the proportions of the three different trimeric complexes formed dictates the transcriptional responses of the R-Smad proteins. As such, Smad2 targets are activated solely by the Smad2-Smad2-Smad4 complex, whereas Smad3 targets are activated both by Smad2-Smad3-Smad4 and Smad3-Smad3-Smad4 trimers. We have modeled the Smad responses onto arbitrary genes and propose that this mechanism might be extended to additional activities of TGF-b in development and disease. © 2013. Published by The Company of Biologists Ltd.

Moreno M.,University of Almeria | Gutierrez-Ferre V.E.,University of Almeria | Ruedas L.,University of Almeria | Campa L.,Institute Investigaciones Biomedicas Of Barcelona | And 2 more authors.
Psychopharmacology | Year: 2012

Rationale: Schedule-induced polydipsia (SIP), characterized by the development of excessive drinking under intermittent food reinforcement schedules, has been proposed as a model for obsessive-compulsive disorder, schizophrenia and drug abuse. Objectives: The purpose of this study is to investigate if individual differences in SIP reflect psychopathological behavioural traits related to lack of inhibitory control and reactivity to novelty, and if these differences have neurochemical correlates. Methods: Outbred Wistar rats were selected for being either high (HD) or low (LD) drinkers according to their SIP behaviour. We tested locomotor reactivity to a novel environment and inhibitory control on the five-choice serial reaction time task (5-CSRTT), under baseline vs. extinction conditions and following challenge with d-amphetamine (saline, 0.5 or 1 mg/kg). Post-mortem analyses of the monoaminergic levels in different brain regions were also analysed. Results: Compared to LD animals, HD rats exhibiting SIP acquisition showed no differences in spontaneous locomotor reactivity to novelty. On the 5-CSRTT, HD rats showed a greater increase in perseverative responses under extinction, a trend towards elevated premature responses on baseline, and a significantly greater elevation of premature responses to d-amphetamine 0.5 mg/kg. The HD animals also exhibited increased serotonin activity in the amygdala, and correlational analyses between the rate of drinking on SIP and monoamine levels also revealed altered dopaminergic mesolimbic function. Conclusions: These findings show that HD rats selected by SIP exhibit compulsive and impulsive behaviour based on measures of performance on the five-choice serial reaction time task and associated with changes in monoaminergic systems in limbic-striatal circuitry. © 2011 Springer-Verlag.

Veny M.,Institute Investigaciones Biomedicas Of Barcelona | Esteller M.,CIBER ISCIII | Ricart E.,CIBER ISCIII | PiquE J.M.,CIBER ISCIII | And 2 more authors.
Alimentary Pharmacology and Therapeutics | Year: 2010

Aliment Pharmacol Ther 31, 561-572 SummaryBackground Th1 and Th17 cells have been implicated in Crohn's disease (CD) pathophysiology and may play a role in disease persistence. Aim To determine Th1 and Th17 responses in intestine and peripheral blood of early (<32 weeks since initial symptoms) and late (>2 years) CD patients. Methods Cytokine mRNA in intestinal biopsies was determined by RT-PCR. Cytokine concentration in culture was measured by ELISA and cytokine-producing cells were identified by intracellular staining. Results The inflamed mucosa showed significantly increased IL-17 mRNA levels compared with non-inflamed areas, both in early and late CD patients. However, only patients with late (n = 12), but not early (n = 9), active disease showed increased IL-17 production, as well as a significantly higher percentage of IL-17+CD4+ cells in blood, compared with controls (n = 12) or patients in remission (n = 13). Moreover, cultured peripheral CD4 + cells from late active CD patients presented significantly higher percentages of IL-17+, IL-22+ and IFN-γ+ and a significantly increased production of IL-17 and IL-22, but not IFN-γ+. Conclusions Increased IL-17 gene transcription is common to early and late CD mucosa. However, exacerbated Th17 responses in the peripheral blood appear only in late disease. We propose that this population may constitute a mechanism of perpetuating the disease. © 2010 Blackwell Publishing Ltd.

Jimenez-Sanchez L.,Institute Investigaciones Biomedicas Of Barcelona | Jimenez-Sanchez L.,Research Center Biomedica En Red Of Salud Mental Cibersam | Jimenez-Sanchez L.,Institute dinvestigacions Biomediques August Pi i Sunyer IDIBAPS | Campa L.,Institute Investigaciones Biomedicas Of Barcelona | And 6 more authors.
Neuropsychopharmacology | Year: 2014

Paradoxically, N-methyl-D-aspartate (NMDA) receptor antagonists are used to model certain aspects of schizophrenia as well as to treat refractory depression. However, the role of different subunits of the NMDA receptor in both conditions is poorly understood. Here we used biochemical and behavioral readouts to examine the in vivo prefrontal efflux of serotonin and glutamate as well as the stereotypical behavior and the antidepressant-like activity in the forced swim test elicited by antagonists selective for the GluN2A (NVP-AAM077) and GluN2B (Ro 25-6981) subunits. The effects of the non-subunit selective antagonist, MK-801; were also studied for comparison. The administration of MK-801 dose dependently increased the prefrontal efflux of serotonin and glutamate and markedly increased the stereotypy scores. NVP-AAM077 also increased the efflux of serotonin and glutamate, but without the induction of stereotypies. In contrast, Ro 25-6981 did not change any of the biochemical and behavioral parameters tested. Interestingly, the administration of NVP-AAM077 and Ro 25-6981 alone elicited antidepressant-like activity in the forced swim test, in contrast to the combination of both compounds that evoked marked stereotypies. Our interpretation of the results is that both GluN2A and GluN2B subunits are needed to induce stereotypies, which might be suggestive of potential psychotomimetic effects in humans, but the antagonism of only one of these subunits is sufficient to evoke an antidepressant response. We also propose that GluN2A receptor antagonists could have potential antidepressant activity in the absence of potential psychotomimetic effects.

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