Activation of the CREB/c-Fos pathway during long-term synaptic plasticity in the cerebellum granular layer

The induction of long-term potentiation and depression (LTP and LTD) is thought to trigger gene expression and protein synthesis, leading to consolidation of synaptic and neuronal changes. However, while LTP and LTD have been proposed to play important roles for sensori-motor learning in the cerebellum granular layer, their association with these mechanisms remained unclear. Here, we have investigated phosphorylation of the cAMP-responsive element binding protein (CREB) and activation of the immediate early gene c-Fos pathway following the induction of synaptic plasticity by thetaburst stimulation (TBS) in acute cerebellar slices. LTP and LTD were localized using voltage-sensitive dye imaging (VSDi). At two time points following TBS (15 min and 120 min), corresponding to the early and late phases of plasticity, slices were fixed and processed to evaluate CREB phosphorylation (P-CREB) and c-FOS protein levels, as well as Creb and c-Fos mRNA expression. High levels of P-CREB and Creb/c-Fos were detected before those of c-FOS, as expected if CREB phosphorylation triggered gene expression followed by protein synthesis. No differences between control slices and slices stimulated with TBS were observed in the presence of an N-methyl-Daspartate receptor (NMDAR) antagonist. Interestingly, activation of the CREB/c-Fos system showed a relevant degree of colocalization with long-term synaptic plasticity. These results show that NMDAR-dependent plasticity at the cerebellum input stage bears about transcriptional and post-transcriptional processes potentially contributing to cerebellar learning and memory consolidation

Essays in macroeconomics

Esta tesis consta de tres capítulos, cada uno de los cuales analiza temas relevantes en el campo de la Macroeconomía. La investigación realizada en los tres capítulos se basa en la metodología de los modelos dinámicos de equilibrio general. Los fundamentos microeconómicos se encuentran en el núcleo de este tipo de modelos, en los que los agentes forman sus expectativas sobre el futuro de manera racional. Esta metodología ha demostrado ser una herramienta poderosa para abordar un gran conjunto de problemas económicos. El primer capítulo, ``Risk Aversion, Entrepreneurship and Wealth Distribution'', explora el efecto de la heterogeneidad en la aversión al riesgo sobre el emprendimiento, el comportamiento del ahorro y la distribución de la riqueza. La distribución de la riqueza es uno de los temas más debatidos no solo en la política contemporánea, sino también en los medios de comunicación y en ámbitos académicos. En particular, la gran concentración de riqueza en la parte superior de la distribución, que se observa en muchas economías avanzadas, se ha situado en el centro del debate. La relevancia de este problema no se ha visto socavada a pesar de la dificultad que supone estimar los niveles de riqueza acumulada en manos de los hogares más ricos. Apoyado por razones empíricas, el marco teórico para analizar la distribución de la riqueza debe incluir tanto el emprendimiento como la heterogeneidad de las preferencias. Con este objetivo, en el primer capítulo se desarrolla un modelo de elección ocupacional con agentes heterogéneos en el que los hogares varían en su aversión al riesgo, se enfrentan a perturbaciones idiosincrásicas no asegurables, tienen restricciones financieras y deben decidir si convertirse en trabajadores o en emprendedores. Esta decisión se basa en una combinación de la aversión al riesgo individual, las habilidades específicas en cada ocupación y los niveles de riqueza acumulados. Como resultado se obtiene que la aversión al riesgo tiene un efecto directo e indirecto en la decisión de ahorro y en la ocupación de los agentes. Por un lado, una mayor aversión al riesgo disuade a los agentes de convertirse en emprendedores y, por lo tanto, los ahorros destinados a superar las restricciones financieras son menores. Por otro lado, una mayor aversión al riesgo también conduce a un mayor ahorro por motivo de precaución que, indirectamente, atenúa las restricciones financieras y hace que la opción de convertirse en emprendedor sea más atractiva. En el modelo desarrollado, el último efecto domina, y los agentes con mayor aversión al riesgo acumulan mayores niveles de riqueza y también se convierten en emprendedores. La contribución más importante de este modelo es que permite desentrañar los efectos que la aversión al riesgo y las fricciones financieras tienen sobre el emprendimiento, que es un factor clave para explicar la desigualdad en la acumulación de la riqueza. El segundo capítulo se titula ``Long-term business relationships, bargaining and monetary policy'' y es un trabajo conjunto con Mirko Abbritti y Tommaso Trani. Su motivación se deriva de la creciente literatura empírica que documenta la importancia de las relaciones comerciales a largo plazo y los procesos de negociación sobre la rigidez de los precios y las dinámicas empresariales. Este artículo introduce relaciones comerciales entre empresas (B2B) a largo plazo y procesos de negociación de precios en un modelo monetario de equilibrio general dinámico estocástico (DSGE) estándar. El modelo se basa en dos suposiciones. Primero, tanto los productores mayoristas como los minoristas necesitan gastar recursos para formar nuevas relaciones comerciales. Segundo, una vez que se forma una relación comercial, el precio se establece en una negociación bilateral entre las empresas. El modelo proporciona un marco riguroso para estudiar el efecto de las relaciones comerciales a largo plazo y los procesos de negociación sobre la política monetaria y la dinámica del ciclo económico. Además, se demuestra que estas relaciones comerciales reducen tanto el papel de asignación de los precios intermedios como los efectos reales de las perturbaciones de la política monetaria. También encontramos que el modelo hace un gran trabajo al replicar los segundos momentos y las correlaciones cruzadas de los datos, y que mejora con respecto al modelo de referencia Neo Keynesiano en explicar dichos estadísticos. El tercer y último capítulo de esta tesis, ``On Staggered Prices and Optimal Inflation'', es coautoreado con uno de mis directores, Miguel Casares. En este artículo se calcula la tasa de inflación óptima en estado estacionario bajo dos especificaciones de rigidez de precios diferentes, Calvo (1983) y Taylor (1980), en un modelo con competencia monopolística. Encontramos que la tasa óptima de inflación en estado estacionario es siempre positiva. Este resultado es robusto a los cambios en el grado de rigidez de los precios. En ambos casos con rigidez de precios, la tasa de inflación óptima es aproximadamente igual al cociente entre la tasa de descuento y la elasticidad de Dixit-Stiglitz. Para calibraciones estándar, el coste de bienestar de la inflación es cuantitativamente pequeño, pero significativamente más alto si los precios son rígidos a la Calvo que si son rígidos a la Taylor.Fundación Banco Sabadell, Fundación Bancaria Caja Navarra y Universidad Pública de NavarraPrograma de Doctorado en Economía, Empresa y Derecho (RD 99/2011)Ekonomiako, Enpresako eta Zuzenbideko Doktoretza Programa (ED 99/2011

The Neurotoxicity of DOPAL: Behavioral and Stereological Evidence for Its Role in Parkinson Disease Pathogenesis

BACKGROUND: The etiology of Parkinson disease (PD) has yet to be fully elucidated. We examined the consequences of injections of 3,4-dihydroxyphenylacetaldehyde (DOPAL), a toxic metabolite of dopamine, into the substantia nigra of rats on motor behavior and neuronal survival. METHODS/PRINCIPAL FINDINGS: A total of 800 nl/rat of DOPAL (1 µg/200 nl) was injected stereotaxically into the substantia nigra over three sites while control animals received similar injections of phosphate buffered saline. Rotational behavior of these rats was analyzed, optical density of striatal tyrosine hydroxylase was calculated, and unbiased stereological counts of the substantia nigra were made. The rats showed significant rotational asymmetry ipsilateral to the lesion, supporting disruption of dopaminergic nigrostriatal projections. Such disruption was verified since the density of striatal tyrosine hydroxylase decreased significantly (p<0.001) on the side ipsilateral to the DOPAL injections when compared to the non-injected side. Stereological counts of neurons stained for Nissl in pars compacta of the substantia nigra significantly decreased (p<0.001) from control values, while counts of those in pars reticulata were unchanged after DOPAL injections. Counts of neurons immunostained for tyrosine hydroxylase also showed a significant (p=0.032) loss of dopaminergic neurons. In spite of significant loss of dopaminergic neurons, DOPAL injections did not induce significant glial reaction in the substantia nigra. CONCLUSIONS: The present study provides the first in vivo quantification of substantia nigra pars compacta neuronal loss after injection of the endogenous toxin DOPAL. The results demonstrate that injections of DOPAL selectively kills SN DA neurons, suggests loss of striatal DA terminals, spares non-dopaminergic neurons of the pars reticulata, and triggers a behavioral phenotype (rotational asymmetry) consistent with other PD animal models. This study supports the "catecholaldehyde hypothesis" as an important link for the etiology of sporadic PD

Plasma exosome profile in st-elevation myocardial infarction patients with and without out-of-hospital cardiac arrest

The identification of new biomarkers allowing an early and more accurate characterization of patients with ST-segment elevation myocardial infarction (STEMI) is still needed, and exosomes represent an attractive diagnostic tool in this context. However, the characterization of their protein cargo in relation to cardiovascular clinical manifestation is still lacking. To this end, 35 STEMI patients (17 experiencing resuscitated out-of-hospital cardiac arrest (OHCA-STEMI) and 18 uncomplicated) and 32 patients with chronic coronary syndrome (CCS) were enrolled. Plasma exosomes were characterized by the nanoparticle tracking analysis and Western blotting. Exosomes from STEMI patients displayed a higher concentration and size and a greater expression of platelet (GPIIb) and vascular endothelial (VE-cadherin) markers, but a similar amount of cardiac troponin compared to CCS. In addition, a difference in exosome expression of acute-phase proteins (ceruloplasmin, transthyretin and fibronectin) between STEMI and CCS patients was found. GPIIb and brain-associated marker PLP1 accurately discriminated between OHCA and uncomplicated STEMI. In conclusion, the exosome profile of STEMI patients has peculiar features that differentiate it from that of CCS patients, reflecting the pathophysiological mechanisms involved in STEMI. Additionally, the exosome expression of brain-and platelet-specific markers might allow the identification of patients experiencing ischemic brain injury in STEMI

Modulatory Communication Signal Performance Is Associated with a Distinct Neurogenomic State in Honey Bees

Studies of animal communication systems have revealed that the perception of a salient signal can cause large-scale changes in brain gene expression, but little is known about how communication affects the neurogenomic state of the sender. We explored this issue by studying honey bees that produce a vibratory modulatory signal. We chose this system because it represents an extreme case of animal communication; some bees perform this behavior intensively, effectively acting as communication specialists. We show large differences in patterns of brain gene expression between individuals producing vibratory signal as compared with carefully matched non-senders. Some of the differentially regulated genes have previously been implicated in the performance of other motor activities, including courtship behavior in Drosophila melanogaster and Parkinson's Disease in humans. Our results demonstrate for the first time a neurogenomic brain state associated with sending a communication signal and provide suggestive glimpses of molecular roots for motor control

The mechanism of functional up-regulation of P2X3 receptors of trigeminal sensory neurons in a genetic mouse model of Familial Hemiplegic Migraine type 1 (FHM-1)

A knock-in (KI) mouse model of FHM-1 expressing the R192Q missense mutation of the Cacna1a gene coding for the \u3b11 subunit of CaV2.1 channels shows, at the level of the trigeminal ganglion, selective functional up-regulation of ATP -gated P2X3 receptors of sensory neurons that convey nociceptive signals to the brainstem. Why P2X3 receptors are constitutively more responsive, however, remains unclear as their membrane expression and TRPV1 nociceptor activity are the same as in wildtype (WT) neurons. Using primary cultures of WT or KI trigeminal ganglia, we investigated whether soluble compounds that may contribute to initiating (or maintaining) migraine attacks, such as TNF\u3b1, CGRP, and BDNF, might be responsible for increasing P2X3 receptor responses. Exogenous application of TNF\u3b1 potentiated P2X3 receptor-mediated currents of WT but not of KI neurons, most of which expressed both the P2X3 receptor and the TNF\u3b1 receptor TNFR2. However, sustained TNF\u3b1 neutralization failed to change WT or KI P2X3 receptor currents. This suggests that endogenous TNF\u3b1 does not regulate P2X3 receptor responses. Nonetheless, on cultures made from both genotypes, exogenous TNF\u3b1 enhanced TRPV1 receptor-mediated currents expressed by a few neurons, suggesting transient amplification of TRPV1 nociceptor responses. CGRP increased P2X3 receptor currents only in WT cultures, although prolonged CGRP receptor antagonism or BDNF neutralization reduced KI currents to WT levels. Our data suggest that, in KI trigeminal ganglion cultures, constitutive up-regulation of P2X3 receptors probably is already maximal and is apparently contributed by basal CGRP and BDNF levels, thereby rendering these neurons more responsive to extracellular ATP. \ua9 2013 Hullugundi et al

Transplantation of Neuronal-Primed Human Bone Marrow Mesenchymal Stem Cells in Hemiparkinsonian Rodents

Bone marrow-derived human mesenchymal stem cells (hMSCs) have shown promise in in vitro neuronal differentiation and in cellular therapy for neurodegenerative disorders, including Parkinson' disease. However, the effects of intracerebral transplantation are not well defined, and studies do not agreed on the optimal neuronal differentiation method. Here, we investigated three growth factor-based neuronal differentiation procedures (using FGF-2/EGF/PDGF/SHH/FGF-8/GDNF), and found all to be capable of eliciting an immature neural phenotype, in terms of cell morphology and gene/protein expression. The neuronal-priming (FGF-2/EGF) method induced neurosphere-like formation and the highest NES and NR4A2 expression by hMSCs. Transplantation of undifferentiated and neuronal-primed hMSCs into the striatum and substantia nigra of 6-OHDA-lesioned hemiparkinsonian rats revealed transient graft survival of 7 days, despite the reported immunosuppressive properties of MSCs and cyclosporine-immunosuppression of rats. Neither differentiation of hMSCs nor induction of host neurogenesis was observed at injection sites, and hMSCs continued producing mesodermal fibronectin. Strategies for improving engraftment and differentiation post-transplantation, such as prior in vitro neuronal-priming, nigral and striatal grafting, and co-transplantation of olfactory ensheathing cells that promote neural regeneration, were unable to provide advantages. Innate inflammatory responses (Iba-1-positive microglia/macrophage and GFAP-positive astrocyte activation and accumulation) were detected around grafts within 7 days. Our findings indicate that growth factor-based methods allow hMSC differentiation toward immature neuronal-like cells, and contrary to previous reports, only transient survival and engraftment of hMSCs occurs following transplantation in immunosuppressed hemiparkinsonian rats. In addition, suppression of host innate inflammatory responses may be a key factor for improving hMSC survival and engraftment

Genome-Wide Gene-Environment Study Identifies Glutamate Receptor Gene GRIN2A as a Parkinson's Disease Modifier Gene via Interaction with Coffee

Our aim was to identify genes that influence the inverse association of coffee with the risk of developing Parkinson's disease (PD). We used genome-wide genotype data and lifetime caffeinated-coffee-consumption data on 1,458 persons with PD and 931 without PD from the NeuroGenetics Research Consortium (NGRC), and we performed a genome-wide association and interaction study (GWAIS), testing each SNP's main-effect plus its interaction with coffee, adjusting for sex, age, and two principal components. We then stratified subjects as heavy or light coffee-drinkers and performed genome-wide association study (GWAS) in each group. We replicated the most significant SNP. Finally, we imputed the NGRC dataset, increasing genomic coverage to examine the region of interest in detail. The primary analyses (GWAIS, GWAS, Replication) were performed using genotyped data. In GWAIS, the most significant signal came from rs4998386 and the neighboring SNPs in GRIN2A. GRIN2A encodes an NMDA-glutamate-receptor subunit and regulates excitatory neurotransmission in the brain. Achieving P2df = 10−6, GRIN2A surpassed all known PD susceptibility genes in significance in the GWAIS. In stratified GWAS, the GRIN2A signal was present in heavy coffee-drinkers (OR = 0.43; P = 6×10−7) but not in light coffee-drinkers. The a priori Replication hypothesis that “Among heavy coffee-drinkers, rs4998386_T carriers have lower PD risk than rs4998386_CC carriers” was confirmed: ORReplication = 0.59, PReplication = 10−3; ORPooled = 0.51, PPooled = 7×10−8. Compared to light coffee-drinkers with rs4998386_CC genotype, heavy coffee-drinkers with rs4998386_CC genotype had 18% lower risk (P = 3×10−3), whereas heavy coffee-drinkers with rs4998386_TC genotype had 59% lower risk (P = 6×10−13). Imputation revealed a block of SNPs that achieved P2df<5×10−8 in GWAIS, and OR = 0.41, P = 3×10−8 in heavy coffee-drinkers. This study is proof of concept that inclusion of environmental factors can help identify genes that are missed in GWAS. Both adenosine antagonists (caffeine-like) and glutamate antagonists (GRIN2A-related) are being tested in clinical trials for treatment of PD. GRIN2A may be a useful pharmacogenetic marker for subdividing individuals in clinical trials to determine which medications might work best for which patients