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Red de Trastornos Adictivos
Granada, Palacio de Congresos
22-23 de Septiembre de 2015
Reunión científica 2015
En asociación con
Reunión científica 2015
PROGRAMA
A. COMUNICACIONES ORALES (I)
Día 22 de Septiembre
9:30 BIENVENIDA Y PRESENTACIÓN DE LA REUNIÓN
9:45-11:30
COMORBILIDAD PSIQUIÁTRICA EN ADICCIÓN A COCAINA:
ESTUDIOS EN HUMANOS
1. ALCOHOL Y COCAINA EN CONDUCTORES DE VEHICULO EN
ESPAÑA: 2008/9-2013. I. Fierro Lorenzo, T. Gómez Talegón, F.J.
Álvarez González.
2. SEVERITY OF COCAINE DEPENDENCE PREDICTS ATTENTION,
COGNITIVE INFLEXIBILITY AND DECISION-MAKING IMPAIRMENTS.
C. Martín Pérez, J.F. Navas, J. Verdejo-Román, C. Rojas-González, I.
Muela, R. Vilar-López.
3. PROTEOMIC PROFILE OF NITRATION AND S-NITROSYLATION
STRESS IN ABSTINENT COCAINE ADDICTS. F.J. Pavón, E. Baixeras,
P. Araos, M. Pedraz, F. Rodríguez de Fonseca, E. Fernández Espejo.
4. SEX DIFFERENCES IN PSYCHIATRIC COMORBIDITY AND PLASMA
BIOMARKERS FOR COCAINE ADDICTION IN ABSTINENT
COCAINE-ADDICTED SUBJECTS IN OUTPATIENT SETTINGS. M.
Pedraz, P. Araos, N. García-Marchena, A. Serrano, P. Romero-Sanchiz,
J. Suárez, E. Castilla-Ortega, F. Mayoral-Cleries, J.J. Ruiz, A. Pastor, V.
Barrios, J.A. Chowen, J. Argente, M. Torrens, R. de la Torre, F.
Rodríguez de Fonseca, F.J. Pavón.
5. PLATELET 5-HT2A RECEPTOR, GRK2 KINASE AND I1-IMIDAZOLINE
RECEPTOR IN FORMER COCAINE ABUSERS WITH MAJOR
DEPRESSION: EFFECTS OF 5-HT DEPLETION. B. Keller, R.
Rodríguez-Minguela, M. Álvaro-Bartolomé, C. Pérez-Mañá, M. Farré,
M.J. García-Fuster, M. Torrens, J.A. García-Sevilla.
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6. TRIER SOCIAL STRESS TEST IN COCAINE DEPRESSED PATIENTS.
M. Torrens Melich, R. Rodríguez Minguela, J.I. Mestre-Pinto, C. PérezMaña, L. Galindo Guarin, E. Papaseit Fontanet, M. Farré Albaladejo
11:30-12:00 CAFÉ
12:00-14:00 ESTUDIOS EN MODELOS ANIMALES
7. POSITIVE
AND
NEGATIVE
SYMPTOMS
IN
A
NEURODEVELOPMENTAL MODEL OF SCHIZOPHRENIA DO NOT
TRANSLATE INTO HIGHER COCAINE CONSUMPTION. R. Capellán,
R. Santos-Toscano, J. Orihuel, E. Borcel, M. Ucha, D. Roura-Martínez,
A. Contreras, A. Higuera-Matas, E. Ambrosio.
8. EFECTOS A LARGO PLAZO DEL ESTRÉS SOCIAL REPETIDO
DURANTE LA ADOLESCENCIA SOBRE EL CONSUMO DE ETANOL
Y COCAÍNA. M.C. Blanco, M. Rodríguez-Arias, S. Montagud-Romero,
M.A. Aguilar, R. Maldonado, M.L. Laorden, M.I. Colado, J. Manzanares,
J. Miñarro.
9. EFECTO DE LAS EXPERIENCIAS TEMPRANAS COMO FACTORES
DE VULNERABILIDAD PARA EL DESARROLLO DE ALTERACIONES
EMOCIONALES Y CONSUMO DE DROGAS. I. Gracia-Rubio, M.
Moscoso-Castro, A. Esteve, J. Miñarro, R. Nadal, M.V. Milanés, M.L.
Laorden, O. Valverde.
10. ROLE OF FRACTALKINE RECEPTOR IN COCAINE-INDUCED
CONDITIONED PLACE PREFERENCE. F. Rodríguez de Fonseca, E.
Castilla-Ortega, L. Santín, A. Serrano, J. Suárez, C. Guerri, M.
Rodríguez-Arias, J. Miñarro, F.J. Pavón.
11. ROLE OF CANNABINOID CB2 RECEPTORS IN BRAIN DAMAGE
FOLLOWING HYPOXIA-ISCHEMIA IN ADULT MICE. E. Kossatz, P.
Robledo, R. Maldonado.
12. DELTA-9-TETRAHYDROCANNABINOL CAUSES SYNAPTIC AND
MOTOR COORDINATION IMPAIRMENTS THROUGH A MECHANISM
UNDERLYING COX-2 ACTIVATION AND MICROGLIAL REACTIVITY.
L. Cutando, V. Salgado-Mendialdúa, R. Maldonado, A. Ozaita.
14:00-15:30 COMIDA
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15:30-18:30 CONSECUENCIAS MÉDICAS DEL ALCOHOLISMO:
ESTUDIOS EN HUMANOS
13. COSTES SANITARIOS ASOCIADOS AL CONSUMO DE ALCOHOL
EN DOS MILLONES DE CATALANES ATENDIDOS EN ATENCIÓN
PRIMARIA. L. Miquel, E. Vela, M. Bustins, L. Segura, J. Colom, A. Gual.
14. MARCADORES DE ACTIVACIÓN LINFOCITARIA EN PACIENTES
CON ALCOHOLISMO CRÓNICO. P. Zuluaga, A. Teniente, A.
Sanvisens, D. Fuster, I. Rivas, F. Bolao, E. Martínez-Cáceres, J. Tor, R.
Muga.
15. GENDER DIFFERENCES IN ETHANOL-INDUCED TLR4-DEPENDENT
INFLAMMATORY
RESPONSE
IN
HUMAN
AND
ANIMAL
ADOLESCENTS WITH BINGE DRINKING. M. Pascual, J. Montesinos,
M. Marcos, J. Laso, C. Guerri.
16. PLASMA CHEMOKINES IN ABSTINENT ALCOHOL USERS:
IDENTIFICATION OF EOTAXIN-1 (CCL11) AS A POTENTIAL
INDICATOR OF PSYCHIATRIC COMORBIDITY. F.J. Pavón, N. GarcíaMarchena, V. Barrios, G. Ponce, P. Araos, M. Pedraz, A. Serrano, F.
Arias, P. Romero-Sanchiz, J. Suárez, J.A. Chowen, J. Argente, M.
Torrens, G. Rubio, F. Rodríguez de Fonseca.
17. OLEOYLETHANOLAMIDE AS A BIOMARKER OF ETHANOL
CONSUMPTION AND ABSTINENCE: COMPARISON WITH OTHER
ACYLETHANOLAMIDES. A. Serrano, N. García-Marchena, F.J. Pavón,
A. Pastor, P. Araos, M. Pedraz, M. Calado, J. Suárez, M. Torrens, G.
Rubio, R. de la Torre, F. Rodríguez de Fonseca.
18. RELACIÓN
DE
LA
LONGITUD
TELOMÉRICA
Y
LOS
POLIMORFISMOS DE LA TELOMERASA CON EL ALCOHOLISMO
H. Llorente, M. Marcos, I. Pastor, M.A. Pérez Nieto, J. Fernández
Mateos, C. Cieza Borrella, R. González-Sarmiento, F. Javier Laso.
18:00-19:00 SESIÓN DE PÓSTERES
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B. COMUNICACIONES ORALES (II)
Día 23 de Septiembre
9:30-11:30 ESTUDIOS EN MODELOS ANIMALES (I)
19. THE CANNABINOID
Manzanares Robles.
CB2
RECEPTOR
AND
ADDICTION.
J.
20. EFECTOS
TRANSGENERACIONALES
DEL
CONSUMO
DE
ALCOHOL EN LA AUTOADMINISTRACIÓN OPERANTE DE
SACARINA Y DE ALCOHOL. S. Fuentes, N. Daviu, A. Armario, R,
Nadal.
21. LONG
TERM
SYNAPTIC
PLASTICITY
DEPENDENT
ON
CANNABINOID CB1 RECEPTORS ACTIVATION IS ALTERED IN THE
DENTATE GYRUS OF ADULT MICE EXPOSED TO ETHANOL
DURING ADOLESCENCE. S. Peñasco, N. Puente, A. Ramos, N. Royo,
A. Gutiérrez, I. Bonilla, L. Reguero, M.J. Canduela, J. MendizabalZubiaga, F. Rodríguez de Fonseca, J. Suárez, I. Elezgarai, P. Grandes.
22. BINGE-LIKE ETHANOL EXPOSURE IN MICE DECREASES D1/5
RECEPTORS IN NUCLEUS ACCUMBENS BUT NOT IN
PREFRONTAL CORTEX. M.D. Gutiérrez-López, R. Vidal, F. Porcu, M.I.
Colado, E. O’Shea.
23. BRAIN-DERIVED NEUROTROPHIC FACTOR (BDNF) EXPRESSION
IS INCREASED IN BASOLATERAL AMYGDALA AFTER REPEATED
SOCIAL DEFEAT STRESS IN ADULTS MICE BUT NOT IN
ADOLESCENTS. J. Navarro-Zaragoza, P. Almela, C. Núñez, M.V.
Milanés, M.L. Laorden, J. Miñarro.
11:30-12:00 CAFÉ
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12:00-14:00 ESTUDIOS EN MODELOS ANIMALES (II)
24. CONSEQUENCES OF ALCOHOL CONSUMTPION THROUGHOUT
LIFESPAN-INVESTIGATION OF FACTORS OF VULNERABILITY. E.M.
Marco, C. Irala, M.D. Hernández, J.A. Ballesta, M.E. Serrano, S.
Peñasco, C. Guerri, M. López-Gallardo, M.P. Viveros.
25. NALMEFENE IS EFFECTIVE IN TREATING ALCOHOL-COCAINE
INTERACTIONS: ASSOCIATION WITH GENE EXPRESSION
PROFILES OF HISTONE DEACETYLASES. J. Calleja-Conde, V.
Echeverry-Alzate, E. Giné, K.M. Bühler, M.P. Viveros, R. Nadal, R.
Maldonado, F. Rodríguez de Fonseca, A. Gual, J.A. López-Moreno.
26. EVALUACIÓN DE LA PARTICIPACIÓN DE LOS RECEPTORES
CANNABINOIDES CB1 EN LAS PROPIEDADES REFORZANTES DEL
ALCOHOL Y LOS FACTORES QUE MODIFICAN EL RIESGO DE
CONSUMO EN RATONES. S. Mendonça Netto, R. Martín, R.
Maldonado.
27. PHARMACOLOGICAL ACTIVATION OF EITHER CANNABINOID CB1
OR CB2 RECEPTORS COUNTERACTS THE ETHANOL-INDUCED
IMPAIRMENT OF HIPPOCAMPAL CELL PROLIFERATION IN ADULT
RAT BRAIN. P. Rivera, L. Bindila, F. Alen, A. Vargas, F.J. Pavón, A.
Serrano, L. Rubio, B. Lutz, F. Rodríguez de Fonseca, J. Suárez.
28. BOTH GENETIC DELETION AND PHARMACOLOGICAL BLOCKADE
OF LYSOPHOSPHATIDIC ACID LPA1 RECEPTOR RESULTS IN
INCREASED ALCOHOL CONSUMPTION. A. Serrano, E. CastillaOrtega, F.J. Pavón, L. Sánchez-Marín, G. Estivill-Torrús, C. Pedraza, E.
Blanco, J. Suárez, L. Santín, F. Rodríguez de Fonseca.
29. LPA1 RECEPTOR KNOCKOUT MICE SHOW STRONG DEFICIT ON
GLUTAMATE BIOSYNTHETIC ENZYME GLUTAMINASE (GLS, KGA)
AND ALTERED MORPHOLOGY OF HIPPOCAMPAL AND CORTICAL
DENDRITIC SPINES. A. Peñalver, J.A. Campos-Sandoval, R. SánchezVaro, C. Cardona, L. Castilla, E. Blanco, M. Martín-Rufián, J.A. Segura,
J.M. Matés, F.J. Alonso, A. Gutiérrez, F. Rodríguez de Fonseca, J.
Márquez.
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RESÚMENES DE LAS COMUNICACIONES ORALES
1. ALCOHOL Y COCAINA EN CONDUCTORES DE VEHICULO EN
ESPAÑA: 2008/9-2013. I. Fierro Lorenzo, T. Gómez Talegón, F.J.
Álvarez González.
Centro de Estudios de Alcohol y Drogas, Farmacología, Facultad de Medicina,
Universidad de Valladolid.
Introducción: Conducir con la presencia de alcohol, drogas y ciertos medicamentos es
frecuente en los países desarrollados y en particular en España. Intervenir en este
campo es una prioridad.
Objetivo: Conocer la evolución en la prevalencia de conductores españoles que
conducen con la presencia de alcohol y cocaína entre 2008-9 y 2013.
Métodos: Tanto en el estudio de 2008-9 (n=3302), como en 2013 (n=2932) la población
diana fueron los conductores de vehículos ligeros de motor, que circulaban por vías
públicas españolas, tanto en un ámbito urbano como interurbano. Se consideraron
casos positivos en alcohol, a aquellos conductores con una cantidad de alcohol
superior a 0.05 mg por litro de aire espirado, y para la cocaína si la concentración de
dicha sustancia en el fluido oral superaba 170 ng/ml; se realizó un segundo análisis de
confirmación y cuantificación mediante cromatografía.
Resultados: Se ha observado un descenso en los casos positivos solo a alcohol (%,
[95% IC], 2008/9: 4.92% [4.18–5.66], 2013: 3.41% [2.27–4.07], p <0.05), pero no de los
casos positivos solo a cocaína (respectivamente, 1.28% [0.90–1.67], 0.87% [0.54–
1.21]).
Conclusiones: A pesar de este descenso, la presencia de alcohol (4.16%) y
cualquierdroga (5.79%) en los conductores españoles es aún elevada. La realización
de controles de drogas en carretera, es la causa disuasoria más probable para explicar
este descenso en la prevalencia de conductores con presencia de sustancias.
Agradecimientos: Este estudio ha sido financiado por la Dirección General de Tráfico
(0100DGT23059) y por el Instituto de Salud Carlos III, Redes Temáticas de
Investigación Cooperativa, Red de Trastornos Adictivos Project No. RD12/0028/00212.
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2.
SEVERITY OF COCAINE DEPENDENCE PREDICTS ATTENTION,
COGNITIVE INFLEXIBILITY AND DECISION-MAKING IMPAIRMENTS.
C. Martín Pérez, J.F. Navas, J. Verdejo-Román, C. Rojas-González, I.
Muela, R. Vilar-López.
Facultad de Psicología y Centro de Investigación Mente, Cerebro y Comportamiento
(CIMCYC), Universidad de Granada, Granada
Objectives: Cocaine dependence has been related to neuropsychological impairments
such as attention, cognitive flexibility and decision-making. The aim of this study was to
analyze the relationship between the severity of cocaine dependence and the
disadvantageous performance on attention, cognitive flexibility and decision-making
tasks. Our hypothesis is that cocaine, due to its neurotoxic effect, will show a dosedependent effect. Thus, higher scores on dependence will relate to worse
neuropsychological functioning.
Material and Methods: Thirty-six cocaine dependent patients participated in the study
[33 males; mean age 41.19 (9.69); mean years of schooling 11.14 (2.69)]. All
participants were following outpatient treatment. Severity of cocaine dependence was
assessed with the Substance Dependence Severity Scale (SDSS). Attention was
measured with the Test d2; cognitive inflexibility was measured with the Probabilistic
Reversal Learning Task; and decision-making was assessed with the Iowa Gambling
Task (IGT) versions A and E (related to sensitivity to reward and punishment,
respectively). Four linear regression analyses were conducted to check if cocaine
dependence predicted neuropsychological deficits.
2
Results: Severity of cocaine dependence predicted scores on the Test d2 (R =.19; F=
2
7.37; p=.009), Probabilistic Reversal Learning Task (R =.12; F= 4.29; p=.046), and
2
decision making as measured with the IGT version E (R =.14; F= 5.15; p=.030) and
2
marginally version A (R =.11; F= 3.61; p=.066).
Conclusions: These findings support previous evidence of alterations in executive
functioning in cocaine dependent individuals and point to a dose-dependent relationship
between cocaine use and neuropsychological problems in attention, cognitive flexibility
and decision making.
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3. PROTEOMIC PROFILE OF NITRATION AND S-NITROSYLATION
STRESS IN ABSTINENT COCAINE ADDICTS. F.J. Pavón, E. Baixeras,
P. Araos, M. Pedraz, F. Rodríguez de Fonseca, E. Fernández Espejo.
Instituto de Investigación Biomédica de Málaga (IBIMA), Hospital Regional Universitario
de Málaga. Universidad de Sevilla, Sevilla.
Objectives. Cocaine abuse is known to alter redox homeostasis inducing excess of
reactive nitrogen species (RNS) leading to RNS-induced stress (1). The main types of
RNS-induced stress are nitration of tyrosine residues (nitrative stress) and Snitrosylation of cysteine residues (S-nitrosylation stress).
Redox-sensitive posttranslational modifications on tyrosine and cysteine residues could impact on the
structure and function of addiction related signaling proteins. The objective was to
analyze in plasma of abstinent cocaine addicts and controls: 1) the presence of
nitration/S-nitrosylation stress; b) redox-altered proteins, and 3) clinical correlations.
Material and methods. Plasma proteins were analyzed with Western-blotting, and
proteomic techniques. Informed consent was obtained from all patients and controls.
Two variables were considered: cocaine symptom severity [DSM-IV criteria for cocaine
abuse (4 criteria) and dependence (7 criteria)] and time of abstinence (days).
Results. No general clinical differences were observed between addicts and controls.
The study revealed the presence of nitrosylation stress in abstinent cocaine addicts
with a high level of cocaine addiction, but not a low one (DSM-IV criterion > 5; p<0.02
versus controls). Nitrative stress was not observed in any case. Abstinent subjects with
a high addiction criterion showed strongly S-nitrosylated proteins bands at ~250,
~230, 68 (albumin) and ~25 kDa. In addition, degree of S-nitrosylation stress correlated
with the degree of addiction (R=0.62, p<0.01, Pearson’s test). Specifically, density
levels of S-nitrosylation of ~250 and ~130-kDa proteins showed positive correlation with
degree of addiction (~250 kDa, R=0.48, p<0.02; ~130 kDa, R=0.41, p<0.05).
Conclusions. Plasma of abstinent cocaine addicts with high number of DMS-IV criteria
for cocaine abuse and dependence (> 5) shows evidence of S-nitrosylation stress and
enhanced S-nitrosylation of several proteins, without nitrative stress. These findings
suggest that cocaine abuse leads to RNS-induced changes of cysteine rather than
tyrosine residues of proteins. This difference is important on the light that nitrosylation
stress, unlike nitrative stress, is reversible. S-nitrosylation during abstinence correlated
with the DMS-IV degree of cocaine addiction. These proteins are under study.
(1)Uys JD, Mulholland PJ, Townsend DM. Glutathione and redox signaling in substance
abuse. Biomed Pharmacother. 2014; 68(6):799-807.
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4. SEX DIFFERENCES IN PSYCHIATRIC COMORBIDITY AND PLASMA
BIOMARKERS FOR COCAINE ADDICTION IN ABSTINENT
COCAINE-ADDICTED SUBJECTS IN OUTPATIENT SETTINGS. M.
Pedraz, P. Araos, N. García-Marchena, A. Serrano, P. Romero-Sanchiz,
J. Suárez, E. Castilla-Ortega, F. Mayoral-Cleries, J.J. Ruiz, A. Pastor, V.
Barrios, J.A. Chowen, J. Argente, M. Torrens, R. de la Torre, F.
Rodríguez de Fonseca, F.J. Pavón.
Unidad Gestión Clínica de Salud Mental, Instituto de Investigación Biomédica de
Málaga (IBIMA), Hospital Regional Universitario de Málaga, Universidad de
Málaga, Málaga, Spain
There are sex differences in the progression of drug addiction, relapse, and response to
therapies. Because biological factors participate in these differences, they should be
con-sidered when using biomarkers for addiction. In the current study, we evaluated the
sex differences in psychiatric comorbidity and the concentrations of plasma mediators
that have been reported to be affected by cocaine. Fifty-five abstinent cocaine-addicted
subjects diag-nosed with lifetime cocaine use disorders (40 men and 15 women) and 73
healthy controls (48 men and 25 women) were clinically assessed with the diagnostic
interview “Psychi-atric Research Interview for Substance and Mental Disorders.”
Plasma concentrations of chemokines, cytokines, N-acyl-ethanolamines, and 2-acylglycerols were analyzed accord-ing to history of cocaine addiction and sex, controlling
for covariates age and body mass index (BMI). Relationships between these
concentrations and variables related to cocaine addiction were also analyzed in
addicted subjects. The results showed that the concentra-tions of chemokine (C-C
motif) ligand 2/monocyte chemotactic protein-1 (CCL2/MCP-1) and chemokine (C-X-C
motif) ligand 12/stromal cell-derived factor-1 (CXCL12/SDF-1) were only affected by
history of cocaine addiction. The plasma concentrations of interleukin 1-beta (IL-1b), IL6, IL-10, and tumor necrosis factor-alpha (TNFa)were affected by history of cocaine
addiction and sex. In fact, whereas cytokine concentrations were higher in control
women relative to men, these concentrations were reduced in cocaine-addicted women
without changes in addicted men. Regarding fatty acid derivatives, history of cocaine
addiction had a main effect on the concentration of each acyl derivative, whereas Nacyl-ethanolamines were increased overall in the cocaine group, 2-acyl-glycerolswere
decreased. Interestingly, N-palmitoleoyl-ethanolamine (POEA) was only increased in
cocaine-addicted women. The covariate BMI had a significant effect on POEA and Narachidonoyl-ethanolamine con-centrations. Regarding psychiatric comorbidity in the
cocaine group, women had lower incidence rates of comorbid substance use disorders
than did men. For example, alco-hol use disorders were found in 80% of men and 40%
of women. In contrast, the addicted women had increased prevalences of comorbid
psychiatric disorders (i.e., mood, anxiety, and psychosis disorders). Additionally,
cocaine-addicted subjects showed a rela-tionship between the concentrations of Nstearoyl-ethanolamine and 2-linoleoyl-glycerol and diagnosis of psychiatric
comorbidity.These results demonstrate the existence of a sex influence on plasma
biomarkers for cocaine addiction and on the presence of comorbid psychopathologies
for clinical purposes.
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5. PLATELET 5-HT2A RECEPTOR, GRK2 KINASE AND I1-IMIDAZOLINE
RECEPTOR IN FORMER COCAINE ABUSERS WITH MAJOR
DEPRESSION: EFFECTS OF 5-HT DEPLETION. B. Keller, R.
Rodríguez-Minguela, M. Álvaro-Bartolomé, C. Pérez-Mañá, M. Farré,
M.J. García-Fuster, M. Torrens, J.A. García-Sevilla.
Laboratory of Neuropharmacology/IUNICS and RETICS-RTA, Universitat de les Illes
Balears, Palma de Mallorca. IMIM-Hospital del Mar Medical Research Institute and
RETICS-RTA, Universitat Autònoma de Barcelona.
The association of cocaine use disorder (CUD) with major depression (MD) is a
frequent diagnosis in which specific biomarkers for independent MD or cocaine-induced
MD are needed. Putative biomarkers for these disorders include dysregulation of brain
and platelet 5-HT2A receptors, G protein-coupled receptor kinase 2 (GRK2) and I1imidazoline receptor (I1-IR) or nischarin. These targets were quantified (Western blot
with validated antibodies) in platelet membranes of CUD/MD patients (n=23, in
remission of cocaine use and depressive symptoms) and matched controls (n=10)
before (basal) and after tryptophan (Tryp)/serotonin (5-HT) depletion (5 and 24 h),
which was induced using the Acute Tryptophan Depletion Test (plasma Tryp depleted
by >85% at 5 h; recovery at 24 h). Data were analyzed using one-sample t-test (control:
100%) or two-tailed t-test. Basal platelet 5-HT2A monomeric, but not dimeric, receptor
form was decreased in CUD/MD patients (n=17) regardless of antidepressant (AD)
medication or cocaine-induced MD (16-57%). In controls (n=10), Tryp/5-HT depletion
increased platelet 5-HT2A dimeric (45%), but not momomeric, receptor form. In AD drugfree (n=7), but not in AD-treated (n=13), CUD/MD patients, Tryp/5-HT depletion further
decreased platelet 5-HT2A monomeric/dimeric receptor forms (13-46%). Platelet GRK2
was not altered after Tryp/5-HT depletion in control (n=10) and CUD/MD subjects
(n=23). No basal differences were found for the I1-IR (nischarin) in control and CUD/MD
subjects. However, Tryp/5-HT depletion reduced its content in AD drug-free (15-45%,
n=8) and treated CUD/MD patients with induced MD (37% at 24h, n=4), whereas ADtreated patients with MD not induced by cocaine had increased (30-35%, n=9) I1-IR
contents (no changes in control subjects). These results indicate a dysregulation of
platelet 5-HT2A receptor (reduced content) in CUD/MD and further suggest that the I 1-IR
could be useful for discrimination between independent (n=15) and cocaine-induced
(n=8) MD. Supported by RTA-RD12/0028/0011 (JAGS) and RTA-RD12/0028/0009
(MT) (MINECO, ISCIII, FEDER).
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6. TRIER SOCIAL STRESS TEST IN COCAINE DEPRESSED PATIENTS.
M. Torrens Melich, R. Rodríguez Minguela, J.I. Mestre-Pinto, C. PérezMaña, L. Galindo Guarin, E. Papaseit Fontanet, M. Farré Albaladejo
Parc de Salut Mar,
Institut de Neuropsiquiatria i Addiccions,
Barcelona,
Spain
Institut Municipal d'Investigació Médica IMIM,
Barcelona,
Spain.
hospital Germans Trías i Pujol, Servei de Farmacología Clínica, Barcelona, Spain
Universitat Autònoma de Barcelona,
Departament de Psiquiatria i Medicina Legal,
Barcelona, Spain
Aim:
To study clinical and neurobiological differences among Primary Major
Depression and Cocaine-Induced Depression in subjects with Cocaine Dependence
Disorder, by the study of stress response mean the Trier Social Stress Test.
Methods Subjects: A total of 15 patients diagnosed of Cocaine Dependence and
Primary Major Depression; 9 patients diagnosed of Cocaine Dependence and
Substance-Induced Depression; 10 patients with Cocaine Dependence without any
other psychiatric diagnoses, and 20 healthy controls. The patients with cocaine
dependence must have an abstinence of consume at least of 2 weeks long. The
patients with major depression must be in a remission episode. Instruments: psychiatric
diagnoses mean Psychiatric Research Interview for Substance and Mental Disorders,
personality traits by Temperament and Character Inventory-R, Hamilton Rating Scale
for Depression, State-Trait Anxiety Inventory and visual analogic scales.
Functional test: Trier social stress test
Experimental session’s variables: Cortisol plasma level, heart rate, systolic and
diastolic blood pressure, breathing frequency, temperature, STAI-E and Visual analog
scales (sadness, irritability, anxiety, happiness, upset stomach, dizziness)
Preliminary Results: The three groups of patients showed a flatter response on
cortisol plasma levels after the TTS than Healthy Controls. The changes in cortisol
levels after TTS , did not differ among patients with Cocaine Dependence and Primary
Major Depression or Cocaine-Induced Depression The performance on the Test
Tryptophan Depletion did not affect the response to the TTS.
Conclusions In cocaine dependent patients they are not differences in the stress
response among concurrent cocaine-induced depression and primary major
depression.
Acknowledgments: Supported in part by grants from the Instituto de Salud Carlos III
and FEDER (ISC-III, FIS 12/01838 and 09/02121), and RETIC RTA RD12/0028/0009.
Liliana Galindo and Esther Papaseit are Rio Hortega fellows (ISC-III, CM14/00111 and
CM13/00016).
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7. POSITIVE
AND
NEGATIVE
SYMPTOMS
IN
A
NEURODEVELOPMENTAL MODEL OF SCHIZOPHRENIA DO NOT
TRANSLATE INTO HIGHER COCAINE CONSUMPTION. R. Capellán,
R. Santos-Toscano, J. Orihuel, E. Borcel, M. Ucha, D. Roura-Martínez,
A. Contreras, A. Higuera-Matas, E. Ambrosio.
Facultad de Psicología. UNED, Madrid. Brain and Mind Institute, École Polytechnique
Fédéral de Lausanne, Suiza.
Introduction: Epidemiological data suggest that infections during pregnancy might
render the developing fetus more susceptible to schizophrenic disorders in adulthood.
In addition, there is a higher incidence of addictive disorders among schizophrenic
patients. The aim of this work was to use a prenatal infection model of schizophrenia
(acute i.p. administration of LPS during gestation) to study in male and female rats the
relationship between cognitive, negative and positive symptoms and the propensity to
consume cocaine later in life.
Hypothesis: LPS-exposed offspring would show deficits related to positive, negative
and cognitive symptoms of schizophrenia, as well as an increased pattern of cocaine
consumption.
Methods: Pregnant female Sprague-Dawley rats were injected with the
lipopolysaccharide –LPS- (GD15,16; 100 µg/kg i.p.) or saline and upon reaching
adulthood, cognitive (working memory in a T maze), negative (deficits in social
interaction) and positive symptoms (deficits in sensory-motor gating) were assessed.
Cocaine self-administration, dose-response patterns and extinction were then
examined.
Results: LPS administration resulted in higher levels of plasma TNFα, decreased
weight gain and hypothermia in the mothers. As adults, LPS-exposed offspring showed
impaired working memory and a deficit in sensory-motor gating (regardless of the sex).
No alterations were observed in the social interaction test. Both LPS- and salineexposed rats self-administered cocaine with no differences between groups. Similar
dose-response curves were observed in the four groups.
Conclusions: These results do not support the association between schizophrenic
symptoms and the higher prevalence of cocaine use disorders.
Supported by grants from the Ministerio de Ciencia e Innovación ( SAF2013-47520-P);
Ministerio de Sanidad, Servicios Sociales e Igualdad (Red de Trastornos AdictivosRTA-RD12/028/0020 -Instituto de Salud Carlos III- and Plan Nacional sobre Drogas,
2012I057); Dirección General de Investigación de la Comunidad de Madrid ( S2011/BMD-2308; Programa de Actividades I+D+I CANNAB-CM); UNED (Plan de
Promoción de la Investigación); and European Union (JUST/2013/DPIP/AG/4823-EU
MADNESS).
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Reunión científica 2015
8. EFECTOS A LARGO PLAZO DEL ESTRÉS SOCIAL REPETIDO
DURANTE LA ADOLESCENCIA SOBRE EL CONSUMO DE ETANOL
Y COCAÍNA. M.C. Blanco, M. Rodríguez-Arias, S. Montagud-Romero,
M.A. Aguilar, R. Maldonado, M.L. Laorden, M.I. Colado, J. Manzanares,
J. Miñarro.
Unidad de Investigación Psicobiología de las Drogodependencias, Departamento de
2
Psicobiología, Facultad de Psicología, Universitat de València. Laboratory of
Neuropharmacology, Departament de Ciencies Experimentals i de la Salut, Universitat
3
Pompeu Fabra, Barcelona. Departamento de Farmacología, Facultad de Medicina,
4
Campus Espinardo, Universidad de Murcia. Departamento de Farmacología, Facultad
de Medicina, Universidad Complutense, Instituto de Investigación Sanitaria Hospital 12
5
de Octubre, Madrid, Instituto de Neurociencias, Universidad Miguel Hernández-CSIC,
Alicante.
El estrés social durante la adolescencia incrementa el riesgo de sufrir trastornos
mentales en la edad adulta. El modelo intruso-residente se considera actualmente
como el mejor para estudiar las consecuencias neurobiológicas del acoso escolar o
bullying. Los ratones adultos que experimentaron repetidas derrotas sociales en la
adolescencia no muestran alteraciones en su nivel de ansiedad ni déficits cognitivos,
pero si presentan conductas depresivas. Estos ratones también presentan un
incremento en la auto-administración de etanol, mostrando mayor número de
respuestas efectivas, una ingesta significativamente mayor y una mayor motivación
por el etanol. También observamos que estos ratones son mas sensibles a los efectos
reforzantes condicionados de la cocaína, ya que desarrollan condicionamiento de la
preferencia de lugar con dosis inefectivas de este psicoestimulante, requieren muchas
más sesiones de extinción tras recibir una dosis efectiva y también son más sensible a
la reinstauración con dosis priming de cocaína. Sin embargo, muestran un retraso en al
adquisición de la auto-administración de una dosis efectiva de cocaína. Los eventos
estresante alteran la reorganización cerebral que ocurre durante la adolescencia,
afectando entre otros al sistema dopaminérgico. Hemos observado que la derrota
social induce un incremento en la tiroxin hidroxilasa y disminución en los factores de
transcripción NUR1 y PIXT3 en el área tegmental ventral. Los ratones derrotados
también muestran un incremento en la permeabilidad de la barrera hematoencefálica,
con una disminución de las proteínas de unión y degradación de la lámina basal en el
núcleo accumbens y el hipocampo. Por lo tanto la exposición a la derrota social durante
la adolescencia produce profundos cambios cerebrales que perduran hasta la edad
adulta y que presumiblemente son los responsables del incremento en los efectos
reforzantes de la cocaína y del etanol.
Agradecimientos: Ministerio de Economía y Competitividad (MINECO), PSI201124762,PSI2014-51847-R, Instituto de Salud Carlos III, Red de Trastornos Adictivos
(RTA) RD12/0028/0005, RD12/0028/0023, RD12/0028/0002, RD12/0028/0003,
RD12/0028/ 0019, y Unión Europea, Fondos FEDER “una manera de hacer Europa”
Generalitat Valenciana, Conselleria de Educación, PROMETEOII/2014/063.
The European Foundation for Alcohol Research (ERAB), EA13 08.
13
Reunión científica 2015
9. EFECTO DE LAS EXPERIENCIAS TEMPRANAS COMO FACTORES
DE VULNERABILIDAD PARA EL DESARROLLO DE ALTERACIONES
EMOCIONALES Y CONSUMO DE DROGAS. I. Gracia-Rubio, M.
Moscoso-Castro, A. Esteve, J. Miñarro, R. Nadal, M.V. Milanés, M.L.
Laorden, O. Valverde.
Grupo de investigación Neurobiología del comportamiento. Departamento de Ciencias
Experimentales y de la Salud. Universitat Pompeu Fabra, Barcelona. Programa de
Neurociencias. Instituto Hospital del Mar de Investigaciones Médicas. IMIM. Barcelona.
Departamento de Psicobiología. Universidad de Valencia. Instituto de Neurociencias.
5
Universidad Autónoma de Barcelona. Departamento de Farmacología. Facultad de
Medina. Universidad de Murcia.
Las experiencias tempranas juegan un papel importante en el desarrollo cerebral,
pudiendo afectar las respuestas emocionales y la motivación. Los eventos adversos
durante la infancia y la exposición temprana a drogas de abuso representan factores de
riesgo para el desarrollo ulterior de enfermedades psiquiátricas, en especial depresión
y trastornos por uso de sustancias. En este estudio, hemos evaluado los efectos de
vulnerabilidad a la presencia de alteraciones emocionales conferidos por dos
situaciones experimentales en el ratón. Una de ellas es el modelo de estrés crónico
postnatal, basado en la separación maternal (SM). El segundo modelo ha sido un
modelo de consumo intermitente y voluntario de una solución alcohólica (20% v/v), el
llamado “drinking in the dark test” (DID test), diseñado para producir un consumo de
alcohol en atracón. Este segundo modelo se ha aplicado a ratones en la época
adolescente. En este contexto, el objetivo de nuestro estudio ha sido evaluar cada una
de las experiencias tempranas (SM y consumo de alcohol en la adolescencia) sobre la
conducta emocional y el consumo de cocaína a lo largo de la vida del animal. En el
caso del estudio de la SM, hemos evaluado el impacto del modelo en machos y
hembras. Nuestros resultados muestran una mayor reactividad a las situaciones de
estrés en los animales expuestos a la SM, que comienza a observarse en los animales
adolescentes y se prolonga hasta la edad adulta. Los efectos observados fueron más
intensos en el caso de las hembras. Además, los animales expuestos a la SM
mostraron respuestas de anhedonia en el test de la sacarina y una disminución de los
efectos de recompensa de la cocaína en el modelo de preferencia de lugar sin que se
afectara la conducta de autoadministración de cocaína a la dosis evaluada (0.75
mg/kg/inf). El segundo paradigma utilizado como factor de vulnerabilidad, la exposición
al consumo de alcohol durante la adolescencia, produjo en los animales adultos un
incremento de los efectos hiperlocomotores agudos de la cocaína, así como una mayor
sensibilización a dichos efectos hiperlocomotores. Por último, los ratones expuestos al
consumo intermitente de alcohol durante la adolescencia, incrementaron sus tasas de
consumo de cocaína en el modelo de auto-administración. En conclusión, nuestros
estudios demuestran que la exposición temprana a estrés crónico, así como la
exposición al modelo de consumo de alcohol voluntario alteran la conducta emocional y
motivacional del animal en la edad adulta. En el caso de los animales expuestos a la
MS predominan las alteraciones emocionales y la anhedonia, mientras que la
exposición temprana al alcohol parece incrementar el consumo de cocaína en el ratón.
Este estudio ha sido financiado por el Ministerio de Economía e Innovación y FEDER
(SAF2013-41761-R), el Ministerio de de Sanidad, Servicios Sociales e Igualdad (PNSD
015-0020) (Red Temática de Investigación Cooperativa en Salud (ISCIIIFEDER)
(RETIC-Trastornos adictivos RTA), y la Generalitat de Catalunya (2014SGR34,
2014SGR14).
14
Reunión científica 2015
10. ROLE OF FRACTALKINE RECEPTOR IN COCAINE-INDUCED
CONDITIONED PLACE PREFERENCE. F. Rodríguez de Fonseca, E.
Castilla-Ortega, L. Santín, A. Serrano, J. Suárez, C. Guerri, M.
Rodríguez-Arias, J. Miñarro, F.J. Pavón.
Unidad de Gestión Clínica de Salud Mental. Laboratorio de Neuropsicofarmacología.
Hospital regional universitario de Málaga
Fractalkine [chemokine C-X3-C motif ligand-1 (CX3CL1)] concentration has been
reported to be associated with the cocaine symptom severity in abstinent cocaine users.
In preclinical studies, plasma CX3CL1 is also affected in cocaine-treated mice but we
have no data about the cocaine effects on the CX 3CL1 concentrations in the CNS.
Because CX3CL1 is expressed by neurons and regulates cell proliferation in the
hippocampus through the CX3C receptor-1 (CX3CR1), we examined the hippocampal
concentrations of CX3CL1 in acute and chronic cocaine-treated mice. Complementary
to CX3CL1, we measured the chemokine C-X-C motif ligand 12 (CXCL12) and the
interleukin-1 beta (IL-1β). Next, we evaluated the role of CX3CL1 in the rewarding
properties of cocaine in the conditioned place preference (CPP) paradigm using Cx3cr1
knockout mice. Prior to CPP, these mice were evaluated in the elevated plus maze, the
open-field test and the Y-maze. We found an increase in CX3CL1, CXCL12 and IL-1β
concentrations after the cocaine injection with maximum concentrations at 60-120 min.
Interestingly, the repeated administration of cocaine produced a marked increase in the
hippocampal concentrations of CX3CL1 compared to the acute treatment (t=0,
p<0.001). During the behavioral characterization of Cx3cr1 knockout mice, we found no
differences in exploratory and anxiety-like behaviors compared to the wild-type mice.
However, the Cx3cr1 knockout mice were unable to discriminate the novel arm from the
familiar ones in the Y-maze. Regarding the cocaine-induced CPP, we observed an
increased locomotor activity and a notably impaired extinction in the Cx3cr1 knockout
mice. Collectively, these results indicate that CX 3CL1 is altered in the hippocampus of
cocaine-treated mice and the CX3CL1/CX3CR1 signaling is required for cocaineinduced CPP.
Acknowledgments
This work was supported by The Fondo de Investigación Sanitaria (Red de Trastornos
Adictivos, FEDER, RD12/0028/001 to F.R. de F,
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11. ROLE OF CANNABINOID CB2 RECEPTORS IN BRAIN DAMAGE
FOLLOWING HYPOXIA-ISCHEMIA IN ADULT MICE. E. Kossatz, P.
Robledo, R. Maldonado.
Laboratory of Neuropharmacology, University Pompeu Fabra, Barcelona IMIM-Hospital
del Mar Research Institute, Barcelona
The endogenous cannabinoid system seems to play an important role in the
neuropathology associated with brain ischemia. The aim of this study was to evaluate
the neuroprotective effects of cannabinoid CB2 receptors in the behavioural and
biochemical alterations induced following hypoxia-ischemia. Mice lacking CB2
receptors (KO) and control littermates (WT) were anesthetized, and the left common
carotid artery was permanently ligated. Following recovery, mice were placed in a
hypoxia chamber with 10% oxygen for 60 min. Behavioural measurements in the
rotarod, beam walking, object recognition, open field, and Irwin tests were carried out
24 h, 72 h and 7 days after this procedure. After testing, brains were prepared for
histological and immunohistochemistry analysis. Hypoxia-ischemia induced brain
damage ipsilateral to the carotid ligation, although significant differences in lesion size
were observed in both genotypes. WT mice showed small damage in the hippocampus
and cortex, while KO mice exhibited larger lesions in hippocampus, striatum, cortex and
amygdala. Behavioural alterations were observed in both genotypes. However, WT
mice progressively recovered motor functionality, while KO mice showed persistent
deficits in motor learning, coordination and balance. A significantly higher expression of
astrocytes and microglia in the hippocampus, striatum and cortex was observed in KO
with respect to WT mice, consistent with the greater lesion size observed in these
animals. Memory deficits in the object recognition test were observed 72 h following
hypoxia-ischemia in KO and WT mice. However, no recovery in this function was
observed in either genotype, suggesting that CB2 receptors may not exert
neuroprotective effects on memory dysfunction. Our results indicate that CB2 receptors
may have a specific neuroprotective role in motor learning, coordination and balance
deficits following hypoxia-ischemia insult, and strongly suggest that they may be
important new targets to accelerate the recovery of these brain functions after brain
damage occurs.
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12. DELTA-9-TETRAHYDROCANNABINOL CAUSES SYNAPTIC AND
MOTOR COORDINATION IMPAIRMENTS THROUGH A MECHANISM
UNDERLYING COX-2 ACTIVATION AND MICROGLIAL REACTIVITY.
L. Cutando, V. Salgado-Mendialdúa, R. Maldonado, A. Ozaita.
Laboratori de Neurofarmacologia. Departament de Ciències Experimentals i de la Salut,
Universitat Pompeu Fabra, 08003-Barcelona
Chronic use of cannabis has been associated to cerebellar dysfunction in humans. We
previously demonstrated in mice that exposure to delta-9-tetrahydrocannabinol (THC),
the main psychoactive component of cannabis, triggers microglial reactivity in the
cerebellar molecular layer, and enhances the expression of neuroinflammatory
molecules, such as interleukin 1β (IL-1β) cytokine or cyclooxygenase-2 (COX-2)
enzyme. In the cerebellum, COX-2 is found in Purkinje cells and its expression is
generally up-regulated following brain insults, via glutamatergic and inflammatory
mechanisms. COX-2 synthesizes prostaglandins, which modulate synaptic plasticity
and microglial activation through their action on EP prostaglandin receptors, such as
EP2 receptors, that are mainly expressed on activated microglia. We evaluated the role
of COX-2 in the cerebellar deficits associated with cannabis consumption and
withdrawal. THC administration in mice (5mg/kg; 5.5 days; twice per day) produced
COX-2 and EP2 enhanced expression in the cerebellum, which were still increased 5
days after THC-treatment. THC withdrawal was characterized by fine motor
coordination deficits in the footprint, beam walking and hanger test performance. We
evaluated the expression of ionotropic glutamate receptor 2 (GluR2) and glutamate
receptor delta 2 (GluRδ2), both involved in cerebellar long-term depression (LTD)
performance. Both receptors were modified in the cerebellum of THC-withdrawn mice.
Interestingly, the sub-chronic administration of the COX-2 inhibitor NS-398 (10mg/kg; 5
days; once per day) after THC exposure, prevented the alterations in fine motor
coordination observed in the THC-withdrawn mice and normalized EP2, GluR2 and
GluRδ2 expression levels. In addition, NS-398 also prevented microglial reactivity in the
cerebellar molecular layer. These results suggest that COX-2 plays an important role in
the control of microglial reactivity in the cerebellum during THC-withdrawal conditions,
revealing COX-2 as crucial in the cerebellar deficits associated to repeated cannabis
exposure.
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13. COSTES SANITARIOS ASOCIADOS AL CONSUMO DE ALCOHOL
EN DOS MILLONES DE CATALANES ATENDIDOS EN ATENCIÓN
PRIMARIA. L. Miquel, E. Vela, M. Bustins, L. Segura, J. Colom, A. Gual.
Grup de Recerca en Addiccions Clínic. IDIBAPS. Red de Trastornos Adictivos. Divisió
d'anàlisis i de gestió de registres de l’activitat, Servei Català de la Salut. Departament
de Drogodependències. Agencia de Salut Pública. Govern de Catalunya.
Introducción: El alcoholismo representa una de las principales causas de
morbimortalidad, pérdida de años de calidad de vida y genera un elevado gasto
sanitario (1% aproximadamente del PIB). Sin embargo, se desconoce la relación entre
los costes y el patrón de consumo de alcohol. El objetivo del estudio es conocer la
relación entre la cantidad de alcohol consumido y los costes sanitarios.
Hipótesis: Se espera encontrar un incremento del gasto sanitario a un mayor
consumo de alcohol.
Métodos: Estudio transversal. Se incluyó a 1.909.358 pacientes mayores de edad
atendidos en los centros de atención primaria de Cataluña de los que se disponía de
datos de consumo de alcohol en el año 2011 o 2012. Los datos sociodemográficos,
clínicos, de uso de recursos y costes se obtuvieron mediante el Sistema de Información
para el Desarrollo de la Investigación en Atención Primaria (SIDIAP) y el registro de
Morbilidad. Se analizó el gasto sanitario del año 2013 (visitas en atención primaria,
visitas en atención especializada, hospitalizaciones, gasto farmacéutico, gasto en
pruebas complementarias).
Resultados: El 54,2% fueron mujeres. La edad media de la muestra de 55,6 (DE 18,6)
años. El 3,4% de la población atendida en primaria eran bebedores de riesgo. El 61,7%
de la muestra presentaban una enfermedad totalmente o parcialmente atribuible al
alcohol. De media los bebedores de riesgo gastaron más que los bebedores de bajo
riesgo (925,1€ (IC95% 910,6-939,6€) vs 859,2€ (IC95% 854,9-863,6€)). Se observó un
ligero incremento de los costes sanitarios al incrementar el consumo de alcohol.
Conclusiones: Según la edad y el sexo, a mayor cantidad de alcohol consumido, se
observa un incremento de los costes sanitarios. Las estrategias dirigidas a la reducción
del consumo en la población general pueden ayudar a la reducción del gasto sanitario.
Agradecimientos: Este trabajo también ha sido financiado por Lundbeck (CP041374)
y el proyecto RD12/0028/0016, integrado en el Plan Nacional de I+D+I y cofinanciado
por el ISCII-Subdirección General de Evaluación y el Fondo Europeo de Desarrollo
Regional (FEDER).
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14. MARCADORES DE ACTIVACIÓN LINFOCITARIA EN PACIENTES
CON ALCOHOLISMO CRÓNICO. P. Zuluaga, A. Teniente, A.
Sanvisens, D. Fuster, I. Rivas, F. Bolao, E. Martínez-Cáceres, J. Tor, R.
Muga.
2
Servicio de Medicina Interna y Servicio de Inmunología, Hospital Universitari Germans
Trias i Pujol, Universitat Autònoma de Barcelona, Badalona. Centro Delta de Atención a
las Drogodependencias, Institut Municipal de Serveis Personals, Badalona Servicio de
Medicina Interna, Hospital Universitari de Bellvitge, Universitat de Barcelona,
L’Hospitalet de Llobregat
INTRODUCCIÓN: Los resultados sobre la activación de linfocitos T (LT) en el
alcoholismo son contradictorios; estudios in vitro han demostrado que el consumo
+
crónico de alcohol es capaz de disminuir la activación de los LTCD4 . Así mismo, la
+
activación de LTCD8 se ve comprometida, demostrándose un defecto en la habilidad
de liberar gránulos citotóxicos y en neutralizar antígenos específicos.
+
+
OBJETIVOS Determinar el estado de activación de LTCD4 y LTCD8 mediante
marcadores de superficie (CD38 y HLD-DR) en pacientes con trastorno por uso de
alcohol.
MATERIAL Y MÉTODO Estudio transversal en pacientes que ingresan para
desintoxicación de alcohol entre 3/2013 y 3/2015 en el Hospital Universitari Germans
Trias i Pujol, Badalona. Al ingreso se realizó una historia clínica para consumo de
alcohol, antecedentes patológicos y grado de comorbilidad; se analizaron muestras de
sangre para hematología, bioquímica e inmunología. Se excluyeron los pacientes con
infección por VIH, cáncer, enfermedades autoinmunes o bajo tratamiento
inmunosupresor. La inmunidad celular se determinó mediante inmunofenotipado de
+
+
sangre periférica. Se obtuvieron valores absolutos y porcentajes de LT (CD3 ), LTCD4
+
+
+
y LTCD8 y de los diferentes estados de activación para LTCD4 y LTCD8 : (I) no
+
activos (HLA-DR CD38 ), (II) activación temprana (HLA-DR CD38 ), (III) activación
+
+
+
tardía (HLA-DR CD38 ) y (IV) doblemente activados (HLA-DR CD38 ). Las
subpoblaciones descritas se compararon con las de 50 individuos sanos.
RESULTADOS Se incluyeron 54 pacientes (88% H). La edad al ingreso fue 48 años
(RIQ:34-54 años), el consumo de alcohol fue de 145 g/día (RIQ:87-202 gramos).
+
+
9
La media (±DE) de linfocitos T, LTCD4 y LTCD8 fue de 1.370 ± 674x10 /L, 845 ±
9
9
+
466x10 /L y 451 ± 240x10 /L, respectivamente. Los LTCD4 se distribuyeron, según
9
su estado de activación, de la siguiente forma: (I):546 ± 291x10 /L, (II):244 ±
9
9
9
213x10 /L, (III):43 ± 30x10 /L y (IV):15 ± 10x10 /L
+
Los LTCD8 se distribuyeron, según su estado de activación, de la siguiente forma:
9
9
9
9
(I):315 ±178 x10 /L, (II):51 ±52 x10 /L, (III):59 ± 61x10 /L y (IV):20 ± 16x10 /L
En comparación con individuos sanos, los LTCD4+ de pacientes con trastorno por uso
de alcohol presentaron menor activación temprana (p=0.092) y mayor activación tardía
+
(p=0.076). En cuanto a LTCD8 los casos con alcoholismo presentaron mayor número
de linfocitos con activación temprana (p=0.037), activación tardía (p=0.000), y
doblemente activados (p=0.082).
+
+
CONCLUSIONES La activación tardía de LTCD4 y LTCD8 es frecuente, así como la
+
activación temprana de LTCD8 . Estos hallazgos sugieren un estado de inflamación
crónica y mayor perfil citotóxico de los pacientes con trastorno por uso de alcohol.
Financiación: Trabajo parcialmente financiado por el Ministerio de Ciencia e
Innovación (RETICS RD12/0028/0006), Ministerio de Sanidad (PNSD 2014|042).
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15. GENDER DIFFERENCES IN ETHANOL-INDUCED TLR4-DEPENDENT
INFLAMMATORY
RESPONSE
IN
HUMAN
AND
ANIMAL
ADOLESCENTS WITH BINGE DRINKING. M. Pascual, J. Montesinos,
M. Marcos, J. Laso, C. Guerri.
Department of Cellular Pathology, Príncipe Felipe Research Center, Valencia
Department of Internal Medicine, University Hospital of Salamanca
Alcohol is one of the first drugs of choice among adolescents, and heavy binge drinking
is the most frequent pattern of drinking among teenagers. One important long-lasting
consequence of alcohol use during adolescence is the higher risk of developing alcohol
abuse and dependence in adulthood, being female adolescents more vulnerable than
male to the impairing effects of alcohol. The aim of this study is to research the possible
gender differences in the plasma levels of cytokines and chemokines in human
adolescents after acute alcohol intoxication and to correlate these results with the TLR4
response, using WT and TLR4-KO male and female adolescent mice with binge ethanol
treatment. Plasma samples from both male and female adolescent with symptoms of
acute alcohol intoxication were collected in a department of emergency medicine.
Plasma from abstainers adolescents of the same age were used as controls. For the
experimental studies, WT and TLR4-KO female adolescent mice (PND 30) treated with
or without one dose of ethanol (3 g/kg) or intermittent ethanol treatment for 2 weeks (8
doses) were used. The levels of cytokines and chemokines were assessed in the
Prefrontal Cortex (PFC) and in plasma. Our results show that whereas in human
females there is an increase of several cytokine and chemokine levels (IL-1β, MIP-1α,
MCP-1, IL-17A, fractalkine, IFN-, IL-2, IL-5, IL-8, IL-10), only the levels of fractalkine
were increased in human male adolescents. Using female adolescent mice, we
observed that while one dose of ethanol increased the levels of IL-1β, MIP-1α, MCP-1,
IL-17A in serum but not in the PFC, intermittent ethanol treatment (8 doses) significantly
increased IL-1β, MIP-1α, MCP-1 and IL-17A levels in the PFC of WT mice. Conversely,
no differences were observed between control and ethanol-treated TLR4-KO mice after
either one dose or intermittent ethanol treatment. Thus, the present findings reveal that
females are more vulnerable than males to the inflammatory effects of binge ethanol
drinking and confirm that TLR4 is an important target of ethanol-induced
neuroinflammation. (Supported by ISCIII/FEDER-RTA, RTA-Network RD12-0028-007,
SAF2012-33747, ERAB, PNSD2014).
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Reunión científica 2015
16. PLASMA CHEMOKINES IN ABSTINENT ALCOHOL USERS:
IDENTIFICATION OF EOTAXIN-1 (CCL11) AS A POTENTIAL
INDICATOR OF PSYCHIATRIC COMORBIDITY. F.J. Pavón, N. GarcíaMarchena, V. Barrios, G. Ponce, P. Araos, M. Pedraz, A. Serrano, F.
Arias, P. Romero-Sanchiz, J. Suárez, J.A. Chowen, J. Argente, M.
Torrens, G. Rubio, F. Rodríguez de Fonseca.
Hospital Regional Universitario de Málaga, Málaga, Spain; Hospital Infantil Universitario
Niño Jesús, Madrid, Spain; Hospital Universitario 12 de Octubre, Madrid, Spain: Institut
Hospital del Mar d’Investigacions Mèdiques (IMIM), Barcelona, Spain.
Alcohol and other drugs alter the peripheral production of inflammatory mediators such
as cytokines and chemokines. Chemokines play an important role in neuroplasticity and
synaptic remodeling, an essential step in behavioral consolidation. Alcohol-induced
changes in chemokine levels might potentiate or promote addictive behaviors in a brain
region specific manner but also contributing to the high prevalence of psychiatric
comorbidity observed in alcohol users. In fact, recent studies have proposed to
chemokines as pathologically relevant biomarkers or therapeutic targets for psychiatric
disorders such as depression, bipolar disorder, schizophrenia and Alzheimer’s disease.
The overall aim of this study was to evaluate peripheral levels of chemokines in alcohol
users under treatment taking into account alcohol-related variables and psychiatric
comorbidity.
Eighty-five chronic alcohol users, who were recruited from outpatient programs for
alcoholism (Hospital Universitario 12 de Octubre, Madrid), and 55 healthy control
subjects were psychiatrically evaluated with the interview PRISM (Psychiatric Research
Interview for Substance and Mental disorders) for substance and mental disorders.
Plasma samples were obtained to quantify chemokine levels using a multiplex
suspension protein array. Analyses of the means, correlation studies and generalized
linear models were performed to analyze the present data. Alcohol users displayed a
high prevalence of comorbid substance (45%; mainly cocaine and cannabis) and
mental disorders (72%; mainly mood disorders, ADHD and personality disorders).
CCL3, CCL2, CXCL12 and CX3CL1 were found altered in alcohol users (p<0.01,
p<0.05, p<0.01 and p<0.05, respectively) compared with controls. CCL2 levels were
associated with time of abstinence, especially in alcoholics without other substance use
disorders (rs=-0.40, p<0.01). Although CCL11 had not altered in alcohol users, the
positive correlation with age in the control group (r=+0.42, p<0.01) was abolished in
alcoholics. Regarding comorbidity, whereas alcoholics with other substance use
disorders had no changes in the chemokine levels, alcohol users with non-substance
psychiatric disorders had decreased CCL11 levels (p<0.01) compared with alcoholics
without disorders. The changes of CCL11 in comorbid users were mainly related to
primary mood disorders (p<0.01). In addition to psychiatric comorbidity, sex had a main
effect on CCL11 levels in alcoholics and women displayed a sustained decrease.The
present data indicate that alcohol use disorders affect the circulating levels of
chemokines. Additionally, the diagnosis of comorbid non-substance psychiatric
disorders was associated with changes in CCL11. Further research is necessary to
elucidate the role of chemokines in the etiology of alcohol addiction and psychiatric
comorbidity.
Acknowledgements: Instituto de Salud Carlos III (ISCIII), Red de Trastornos Adictivos
UE-FEDER 2012 (RD12/0028); Ministerio de Economía y Competitividad (PI13/02261);
Plan Nacional sobre Drogas 049/2009 and 049/2013; Consejería de Salud y Bienestar
Social, Junta Andalucía (PI0228-2013 and PI0823-2012). Contracts “Miguel Servet”
(CP14/00212 and CP14/00173) and “Río Hortega” (CM13/0115) from ISCIII.
21
Reunión científica 2015
17. OLEOYLETHANOLAMIDE AS A BIOMARKER OF ETHANOL
CONSUMPTION AND ABSTINENCE: COMPARISON WITH OTHER
ACYLETHANOLAMIDES. A. Serrano, N. García-Marchena, F.J. Pavón,
A. Pastor, P. Araos, M. Pedraz, M. Calado, J. Suárez, M. Torrens, G.
Rubio, R. de la Torre, F. Rodríguez de Fonseca.
UGC de Salud Mental, Instituto IBIMA, Hospital Regional Universitario de Málaga,
Spain; IMIM, Barcelona, Spain; Universitat Autonoma de Barcelona, Spain; INAD,
Barcelona, Spain; Universidad Complutense, Madrid, Spain; Instituto de Investigación
Hospital 12 de Octubre, Madrid, Spain; Universidat Pompeu Fabra, Barcelona, Spain.
Oleoylethanolamide (OEA) is a bioactive lipid mediator that belongs to the fatty acid
ethanolamide (FAE) family. Recently, we have demonstrated that alcohol administration
causes the release of OEA in rodents and it contributes to the regulation of alcohol
intake, the acute motivational response to alcohol and the onset of withdrawal
symptoms after cessation of chronic alcohol consumption. The present work
characterizes plasma-free FAEs in abstinent alcohol users from a specific alcohol
program in Hospital Universitario 12 de Octubre (Madrid, Spain) meeting criteria for
alcohol use disorders (AUDs, n=79), and age-/gender-/body mass-matched healthy
control volunteers (n=79). Substance and mental disorders that commonly occur with
substance abuse were assessed by the semi-structured interview “Psychiatric Research
Interview for Substance and Mental Diseases” according to the “Diagnostic and
th
Statistical Manual of Mental Disorders, 4 Edition, Text Revision” (DSM-IV-TR) and
plasma-free FAEs were quantified by a liquid chromatography-tandem mass
spectrometry system. The results indicated that plasma OEA as well as other FAEs
were increased in alcohol users. Multivariate predictive model based on these FAEs as
explanatory variables were developed to distinguish AUD subjects from controls
providing high discriminatory power. However, these alterations were not influenced by
the severity of alcohol symptoms, duration of problematic alcohol use or by the
diagnosis of some comorbid psychiatric disorders. In contrast, we observed the plasma
concentration of FAEs was affected by the time of abstinence. Thus, we found that
plasma
levels
of
OEA,
arachidonoylethanolamide
(AEA)
and
docosatetraenoylethanolamide (DEA) were higher in subjects were short-term
abstinence (3 or less months) than in subjects with long-term abstinence (12 or more
months). In summary, alcohol consumption modifies the circulating levels of OEA and
other FAEs, and these changes are associated with the length of abstinence. These
results identified OEA as a reliable biomarker of alcohol consumption and abstinence.
Acknowledgements: the present study has been supported by the Ministerio de
Economía y Competitividad (MEC) and Instituto de Salud Carlos III (ISC-III) (project
PI13/02261); MEC, ISCIII and Red de Trastornos Adictivos UE-FEDER/EU-ERDF 2012
(RD12/0028); Ministerio de Sanidad, Servicios Sociales e Igualdad and Plan Nacional
Sobre Drogas (projects 049/2009 and 049/2013); Junta de Andalucía and Plan Andaluz
de Investigación, Desarrollo e Innovación UE-FEDER/EU-ERDF (CTS-433); Junta de
Andalucía and Consejería de Economía, Innovación y Ciencia (Project PI45403); Junta
de Andalucía and Consejería de Igualdad, Salud y Políticas Sociales (projects PI08232012 and PI0228-2013); contracts ’Miguel Servet’ from ISC-III (CP14/00173,
CP14/00212 and CP12/03109).
22
Reunión científica 2015
18. RELACIÓN
DE
LA
LONGITUD
TELOMÉRICA
Y
LOS
POLIMORFISMOS DE LA TELOMERASA CON EL ALCOHOLISMO
H. Llorente, M. Marcos, I. Pastor, M.A. Pérez Nieto, J. Fernández
Mateos, C. Cieza Borrella, R. González-Sarmiento, F. Javier Laso.
Unidad de Alcoholismo. Departamento de Medicina Interna. Hospital Clínico
Universitario de Salamanca. Unidad de Medicina Molecular. Universidad de
Salamanca.
Introducción: Los telómeros son unos complejos ribonucleoproteicos que protegen al
cromosoma durante replicación del ADN. Variaciones en la longitud telomérica así
como la presencia de determinados polimorfismos del complejo telomerasa se han
asociado con diversas enfermedades inflamatorias. El alcoholismo crónico produce
una activación de la respuesta inflamatoria.
Hipótesis: existe una asociación entre la longitud telomérica y el alcoholismo, con
una posible modulación por polimorfismos del complejo telomerasa.
Material y métodos: estudiamos la longitud telomérica en 98 pacientes alcohólicos
(con abuso o dependencia según el DSM-IV) de sexo masculino con edades entre 40
y 70 años, remitidos a la Unidad de Alcoholismo del Hospital Universitario de
Salamanca o ingresados en dicho Hospital, frente a 99 controles sanos del mismo
rango de edad, con igual proporción de fumadores. Mediante la técnica de PCR
comparativa cuantitativa obtuvimos la longitud telomérica de cada individuo frente a
una muestra de referencia. El estudio de los polimorfismos de los genes TERT y
TERC, que codifican proteínas implicadas en el complejo telomerasa se realizó
mediante sondas TaqMan. El análisis estadístico se realizó con el programa GenEx v6.
Resultados: encontramos una menor longitud telomérica en alcohólicos que en
controles (P < 0.01), diferencia con los controles que se mantuvo al analizar los
subgrupos de pacientes con abuso o dependencia de alcohol o los pacientes con
cirrosis o sin hepatopatía alcohólica. La presencia del alelo G del polimorfismo
rs2293607 del gen TERC se asoció con una menor longitud telomérica y con un mayor
riesgo de alcoholismo.
Conclusión: nuestros resultados indican una posible asociación entre el alcoholismo
crónico y el acortamiento de los telómeros, independientemente del patrón de
consumo enólico y de la presencia de cirrosis hepática. La presencia del polimorfismo
rs2293607 del gen TERC favorece el desarrollo de alcoholismo y está asociado con
una menor longitud telomérica.
23
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19. THE CANNABINOID
Manzanares Robles.
CB2
RECEPTOR
AND
ADDICTION.
J.
Instituto de Neurociencias y Facultad de Farmacia de la Universidad Miguel Hernández
de Alicante.
In the last years, a number of studies have changed the concept that the CB2r were
exclusively located in the spleen and lymphocytes. Nowadays, the results of different
studies have revealed the presence of CB2r in a wide variety of brain regions
suggesting their potential implications in the regulation of different neuropsychiatric
disorders, including drug addiction.
The use of genetically modified mice or pharmacological manipulations of this receptor
by agonists and antagonists further evidences the important role that CB2r play in the
vulnerability and treatment of anxiety, depression, schizophrenia, impulsivity likebehaviors, cognitive–related disorders and in the development or regulation of addictive
behaviors induced by drugs such as alcohol, cocaine and nicotine.
The behavioral alterations depending upon deletion or overexpression of CB2r predicts
distinct vulnerability to drugs of abuse. In this respect, we have found that mice
overexpressing CB2r (CB2xP) presented reduced cocaine-induced motor sensitization,
cocaine-induced conditioned place aversion and self-administered less cocaine that WT
mice. On the other hand, CB2KO mice presented increased ethanol-conditioned place
preference, voluntary ethanol consumption and preference, acquisition of ethanol self–
administration and increased motivation to drink alcohol compared to WT mice. In
contrast, CB2KO mice did not show nicotine-induced place conditioning and self–
administered significantly less nicotine. Somatic signs of nicotine withdrawal increased
significantly in WT but were absent in CB2KO mice. Interestingly, the administration of
AM630 blocked the nicotine withdrawal and failed to alter basal behavior in salinetreated WT mice.Taken together, these results provide sufficient evidence to consider
that CB2r function is closely involved in the vulnerability and development of behavioral
alterations and drug abuse. Furthermore, it seems that pharmacological manipulation of
this receptor may be useful as a potential new valuable target for the treatment of
psychiatric disorders and drug dependence.
24
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20. EFECTOS
TRANSGENERACIONALES
DEL
CONSUMO
DE
ALCOHOL EN LA AUTOADMINISTRACIÓN OPERANTE DE
SACARINA Y DE ALCOHOL. S. Fuentes, N. Daviu, A. Armario, R,
Nadal.
Institut de Neurociències; Unidad de Psicobiología (Facultad de Psicología); Unidad de
Fisiología Animal (Facultad de Biociencias), Universitat Autònoma de Barcelona, 08192
Cerdanyola del Vallès
La exposición a drogas de abuso puede inducir efectos transgeneracionales,
probablemente asociados a cambios epigenéticos, que perduren sin exposiciones
adicionales a la droga. En el presente estudio nuestro objetivo fue investigar el efecto
en la descendencia de la exposición paterna repetida a etanol (consumo forzado). Se
utilizaron ratas macho Long-Evans que fueron expuestas o no durante 14 días a etanol
en la dieta, y tras un período de washout se cruzaron con hembras siempre controles.
Se estudió como se modificaba en las madres la conducta materna hacia las crías
potencialmente percibidas como distintas, y el efecto en la conducta de
autadministración operante de etanol y de sacarina en la descendencia masculina y
femenina una vez adulta. Como procedimiento de autoadministración operante se
utilizó el de substitución progresiva por sacarina, y se estudió en grupos
independientes de animales la autoadministración de alcohol y la de sacarina en
programas de razón fija 3, razón progresiva, extinción y recaída inducida por estímulos
condicionados. Las madres manifestaron una menor conducta materna hacia las crías
engendradas de padres que habían consumido alcohol. Los resultados indicaron
además que aunque la autoadministración de alcohol no se modificó de una forma
importante, se indujo un claro efecto anhedónico en la descendencia masculina (no en
la femenina) manifestado por una menor autoadministración de soluciones dulces.
Estos resultados pueden tener implicaciones en el estudio de los factores de
vulnerabilidad a inducir fenotipos depresivos en hijos de padres alcohólicos.
Agradecimientos: ISCIII, FEDER, Plan Nacional sobre Drogas, MICINN, SGR e
ICREA-Acadèmia (Generalitat de Catalunya)
25
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21. LONG
TERM
SYNAPTIC
PLASTICITY
DEPENDENT
ON
CANNABINOID CB1 RECEPTORS ACTIVATION IS ALTERED IN THE
DENTATE GYRUS OF ADULT MICE EXPOSED TO ETHANOL
DURING ADOLESCENCE. S. Peñasco, N. Puente, A. Ramos, N. Royo,
A. Gutiérrez, I. Bonilla, L. Reguero, M.J. Canduela, J. MendizabalZubiaga, F. Rodríguez de Fonseca, J. Suárez, I. Elezgarai, P. Grandes.
Faculty of Medicine and Dentistry, University of the Basque Country UPV/EHU, E48940 Leioa Achucarro Basque Center for Neuroscience, Bizkaia Science and
Technology Park, Zamudio Hospital Regional Universitario de Málaga, Instituto de
Investigación Biomédica de Málaga (IBIMA), Málaga
Alcohol drinking, especially among adolescents and young adults, is a serious public
health concern. Ethanol interacts with the endocannabinoid system (ECS) whose
function may be altered in ethanol dependence. Here, we investigated the effect of
ethanol consumption on excitatory synaptic transmission and plasticity mediated by the
cannabinoid CB1 receptor in dentate gyrus (DG).
Male C57BL6 mice were exposed to intermittent ethanol intake (20% (v/v) in tap water)
using a 4 days drinking-in-the-dark procedure during adolescence (PD 30±2 to 54±2).
Animals were given access to ethanol (or water) for 2h sessions during 3 days, and 4h
session on the 4th day. At 18-21 days withdrawal from ethanol, adult mice were
sacrificed. Electrophysiological, immunohistochemical, and molecular techniques were
applied.
Excitatory postsynaptic potentials (fEPSPs) were evoked after stimulation of the medial
perforant path and recorded in the supragranular zone of the dentate molecular layer
(ML) in the presence of the GABAA antagonist picrotoxin. CB1 activation by CP55,940
(10μM) inhibited fEPSPs in controls (26.43±2.77% of baseline) as already shown.
However, this effect was not observed in ethanol-exposed mice (4.9±7.47% of
baseline). Furthermore, ML synaptic stimulation (10min, 10Hz) triggered a long term
depression (LTD) of the excitatory transmission that was absent in adult mice after
***
ethanol consumption during adolescence (2.7±3.12% of inhibition; p<0.0001 ). This
plasticity was CB1 dependent as the AM251 antagonist (4µM) abolished LTD (8±6.6% of
inhibition).
CB1 immunoreactivity decreased in ML of ethanol-exposed (87.47±0.58%) vs control
(100±0.77%) mice. Also, the relative mRNA and CB 1 protein significantly decreased,
while a significant increase in MAGL (mRNA and protein) was detected.
Altogether, repetitive exposure to ethanol during adolescence leads to a deficit of
endocannabinoid-dependent LTD in adult DG excitatory synapses, probably due to a
down-regulation of CB1 receptors and a reduction of the endocannabinoid tone by an
increase of MAGL.
Funded by RETICS, ISCIII (RD12/0028/0001; RD12/0028/0004); MINECO (BFU201233334); Basque Government (IT764-13); UPV/EHU (UFI11/41); ISCIII (MINECO)
(CP12/03109, PI13/02261); JA (SAS111224); JA UE/ERDF (PI45403, CTS-8221).
26
Reunión científica 2015
22. BINGE-LIKE ETHANOL EXPOSURE IN MICE DECREASES D1/5
RECEPTORS IN NUCLEUS ACCUMBENS BUT NOT IN
PREFRONTAL CORTEX. M.D. Gutiérrez-López, R. Vidal, F. Porcu, M.I.
Colado, E. O’Shea.
Facultad de Medicina, Universidad Complutense de Madrid, Madrid
Introduction: Binge drinking is a common practice among younger sectors of the
population. The intensive consumption of alcohol during this period produces both
behavioural and neurobiological changes in the maturing brain which could lead to
increased susceptibility to drug abuse in adulthood.
Objectives: The aim of this study was to evaluate the effect of intensive ethanol (EtOH)
consumption in mice on the density of D1/D5 and D2/D3 receptors in nucleus
accumbens and prefrontal cortex and on D5 receptor mRNA levels in peripheral blood.
Methods: Mice (male C57BL/6J, 4 weeks old) were exposed to 4 repeated cycles of
the Drinking in the Dark paradigm (cycle = substitution of drinking water with EtOH 20%
during 2h for 3 days, followed by 1 day of 4h of EtOH followed by 3 days of rest) and
were killed 24h after the last EtOH exposure. D1/5 and D2/3 receptor density and D2/3
receptor signalling were quantified in the nucleus accumbens and prefrontal cortex by
receptor and functional autoradiography respectively. In addition, D5 mRNA levels were
determined in peripheral blood by RT-PCR.
Results: Intensive EtOH decreased D1/5 receptor density in the nucleus accumbens
(41%) but not in the prefrontal cortex compared to controls 24h after last EtOH
exposure. No changes in D2/3 receptor density or signalling were observed. Intensive
EtOH also decreased D5 receptor mRNA in peripheral blood (66%).
Conclusions: These data suggest that intensive and repetitive binge-like EtOH
consumption may change the capacity of neurons to respond to dopamine by
modulating expression of D1/5 receptors in nucleus accumbens but not in prefrontal
cortex. The fact that D5 receptor mRNA expression is concomitantly decreased in
peripheral blood suggests that this parameter could serve as a peripheral biomarker of
EtOH-induced changes in D1/5 receptor density.
Acknowledgements: Ministerio de Sanidad, Servicios Sociales e Igualdad (PNSD
2013I077), ISCIII RETICS RTA (RD12/0028/0002) and Fondo Europeo de Desarrollo
Regional (FEDER).
27
Reunión científica 2015
23. BRAIN-DERIVED NEUROTROPHIC FACTOR (BDNF) EXPRESSION
IS INCREASED IN BASOLATERAL AMYGDALA AFTER REPEATED
SOCIAL DEFEAT STRESS IN ADULTS MICE BUT NOT IN
ADOLESCENTS. J. Navarro-Zaragoza, P. Almela, C. Núñez, M.V.
Milanés, M.L. Laorden, J. Miñarro.
Facultad de Medicina. Universidad de Murcia, Murcia, Spain
Unidad de Investigación Psicobiología de las Drogodependencias, Departamento de
Psicobiologia, Facultad de Psicología, Universidad de Valencia, Spain
It has been shown previously that stress has a prominent role in drug addiction
influencing in an increased drug-taking and the resulting addiction. Regarding this, one
of the major adaptations that occur to develop addiction refers to learning and memory
processes which results in long-term memory of drug reward. It has been shown that
social defeat stress increases the rewarding effects of psychostimulant drugs such as
cocaine. The aim of this study was to evaluate the effects of repeated social defeat
(RSD) on the conditioned rewarding effects of cocaine using conditioned place
preference (CPP) paradigm. Adolescent (PND 27-36) and adult (PND 47-56) mice were
exposed to four episodes of social defeat and were conditioned three weeks later with
cocaine (1 mg/kg, i.p.). The effects of social defeat on the brain-derived neurotrophic
factor (BDNF), molecular mediator of memory consolidation processes, pERK and
pCREB in the basolateral amygdala (BLA) and in the hippocampal CA 2 and CA3 were
also evaluated. Adolescent and adult mice exposed to RSD showed an increase in the
conditioned reinforcing effects of cocaine. Adults exposed to social defeat displayed
significantly higher levels of BDNF in the BLA but no changes were seen in adolescents
when compared to controls. However, pCREB levels were decreased in adolescent
mice compared to adults in hippocampal CA2 and CA3. These results support the idea
that RSD stress increases the rewarding effects of cocaine in parallel with increased
BDNF in the BLA and decreased pCREB in the hippocampal region both nuclei which
are known to contribute to the formation of memories. Together with previous results
from our laboratory we suggest that BDNF could be implicated in the increase of the
reinforcing effects observed in mice treated with cocaine and exposed to RSD.
Acknowledgements: Instituto de Salud Carlos III RETICS RD/12/0028/0003, SAF
2013/49076.
28
Reunión científica 2015
24. CONSEQUENCES OF ALCOHOL CONSUMTPION THROUGHOUT
LIFESPAN-INVESTIGATION OF FACTORS OF VULNERABILITY. E.M.
Marco, C. Irala, M.D. Hernández, J.A. Ballesta, M.E. Serrano, S.
Peñasco, C. Guerri, M. López-Gallardo, M.P. Viveros.
Dpto. Fisiología (Fisiología Animal II), Facultad de Biología. Universidad Complutense,
Madrid. Dpto. Fisiología, Facultad de Medicina, Universidad Complutense, Madrid..
Centro de Investigación Príncipe Felipe, Valencia.
Alcohol drinking is a serious public health concern. Remarkably, gender differences
have been described for alcohol consumption and dependence. Since stress during
childhood can be a factor of vulnerability in relation to alcohol consumption, we first
investigated the consequences of early life stress on adolescent alcohol consumption in
both male and female rats. By using a model of early life stress (maternal deprivation,
MD) we found no changes in baseline voluntary alcohol intake during adolescence as a
consequence of MD; however, an increased voluntary alcohol consumption was
reported when animals were exposed to stressful events later in life.
Adolescent alcohol consumption might exhibit detrimental consequences. Considering
current changes in alcohol consumption patterns, we evaluated the effects of
intermittent access to ethanol during adolescence by using a modified drinking-in-thedark procedure. A remarkable deficit in cognition, i.e. decreased recognition memory,
was observed in both male and female animals in the long term despite the animals
self-administered moderate amounts of alcohol. Intermittent alcohol consumption during
adolescence also induced significant sex-dependent changes in hippocampal and
frontal cortex neurotransmitter systems, presynaptic proteins and epigenetic markers.
In addition, depression has been frequently associated with alcohol consumption,
although causality between conditions has not yet been elucidated. In the next study,
male and female Wistar rats were exposed to 6 weeks of chronic unpredictable mild
stress (CUMS) to induce a depressive-like phenotype. Indeed, depressive-like
responses were observed in the Forced Swimming Test, i.e. increased immobility time,
in animals of both sexes, in the absence of changes in anhedonia and general motor
activity. Notably, higher rates of alcohol consumption were exclusively observed in
CUMS females when compared to control females; no differences were observed
among males. CUMS appears to differently affect male and female animals’ behaviour
and sex- and region-dependent differences were also observed when analysing
possible neural correlates.
Acknowledgements: Instituto de Salud “Carlos III” (FIS), RETICS, Red de Trastornos
Adictivos –FEDER- (RD2012/0028/0021). GRUPOS UCM-BSCH: Ref UCM 951579
29
Reunión científica 2015
25.
NALMEFENE IS EFFECTIVE IN TREATING ALCOHOL-COCAINE
INTERACTIONS: ASSOCIATION WITH GENE EXPRESSION PROFILES
OF HISTONE DEACETYLASES. J. Calleja-Conde, V. Echeverry-Alzate, E.
Giné, K.M. Bühler, M.P. Viveros, R. Nadal, R. Maldonado, F. Rodríguez de
Fonseca, A. Gual, J.A. López-Moreno.
Department of Psychobiology, School of Psychology, Campus de Somosaguas,
Complutense University of Madrid, 28223, Madrid, Spain. Department of Cellular Biology,
School of Medicine, Complutense University of Madrid, 28040, Madrid, Spain. Department of
Physiology (Animal Physiology II), School of Biology, Complutense University of Madrid,
28040, Madrid, Spain.Psychobiology Unit, School of Psychology, Institut de Neurociències,
Universitat Autònoma de Barcelona, 08193, Bellaterra, Barcelona, Spain Laboratori de
Neurofarmacologia, Departament de Ciències Experimentals i de la Salut, Universitat
Pompeu Fabra, Barcelona, 08003, SpainFundación IMABIS, Laboratorio de Medicina
Regenerativa, Hospital Regional Universitario Carlos Haya, 29010, Málaga, Spain.
Addictions Unit, Department of Psychiatry, Clinical Institute of Neuroscience, Hospital Clínic,
Barcelona, 08036, Spain
The opioid antagonist nalmefene (selincro®), was approved for alcohol-related disorders by
the European Medicines Agency in 2013. There was no study of effectiveness of nalmefene
in the comorbid use of cocaine and alcohol.Using operant alcohol self-administration in
Wistar rats and qRT-PCR, we (a) evaluated the dose-response curve for subcutaneous and
oral nalmefene throughout four-day periods; (b) the effects on nalmefene on increasing
doses of alcohol; (c) on responses to alcohol with the co-administration of cocaine; and (d)
the association between the gene expression profiles of histone deacetylases (Hdac1-11) in
blood, prefrontal cortex, heart, liver and kidney and the behavioral results. Nalmefene dosedependently reduced the alcohol responses, up to 50.3%, at the subcutaneous (0.01, 0.05,
0.1 mg/kg) and oral (10, 20, 40 mg/kg) doses tested within the treatment period. Nalmefene
reduced systematically the levels of alcohol responses independently of the concentration of
alcohol (10, 15, 20%). Cocaine caused an increase of about 40% in alcohol responses but it
was fully reversed by nalmefene and prevented it within the following four days of nalmefene
withdrawal. When nalmefene was administered 18h before the alcohol session, it was
ineffective for reducing alcohol responses. Except for Hdac2 and 4, nalmefene prevented the
alcohol-induced increase in Hdac gene expression in blood. In kidney, the nalmefene group
showed significant differences compared with the cocaine group in Hdac1, 2, 5 and 8.
Conclusions and implications: Nalmefene was effective for reducing responses to alcohol
even with the co-administration of cocaine.
Acknowledgments This work was supported by The European Foundation for Alcohol
Research (to J.A.L.M., F.R. de F., R.M, R.N., and M.P.V.), the Fondo de Investigación
Sanitaria (Red de Trastornos Adictivos, FEDER, RD12/0028/0015 to J.A.L.M.,
RD12/0028/001 to F.R. de F., RD12/0028/023 to R.M., RD12/0028/0014 to R.N.,
RD12/0028/1021 to M.P.V., RD12/0028/0008 to F.J.L.; and PI10/01692 to M.M.), and
Ministerio de Ciencia e Innovación (SAF2011-26818 to J.A.L.M.).
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26. EVALUACIÓN DE LA PARTICIPACIÓN DE LOS RECEPTORES
CANNABINOIDES CB1 EN LAS PROPIEDADES REFORZANTES DEL
ALCOHOL Y LOS FACTORES QUE MODIFICAN EL RIESGO DE
CONSUMO EN RATONES. S. Mendonça Netto, R. Martín, R.
Maldonado.
Laboratorio de Neurofarmacología, Universitat Pompeu Fabra, Barcelona
El consumo de alcohol por parte de los adolescentes representa un problema de gran
magnitud en España que preocupa de manera importante a los profesionales de la
salud. Los mecanismos neurobiológicos determinantes de este consumo no han sido
aún esclarecidos y su conocimiento podría favorecer el desarrollo de nuevos abordajes
terapéuticos y preventivos. Diversos estudios han demostrado la participación de los
receptores cannabinoides CB1 en las propiedades adictivas del alcohol y sugieren que
dichos receptores pueden estar implicados en la vulnerabilidad de los sujetos jóvenes
al consumo de esta droga. En este trabajo hemos utilizado diferentes líneas de ratones
knockout deficientes de receptores CB1 para esclarecer la implicación de dichos
receptores en los factores que determinan la vulnerabilidad al consumo de alcohol en
sujetos jóvenes mediante el empleo de sofisticadas técnicas de auto-administración
operante. Así, se ha estudiado el papel de los receptores cannabinoides CB1 (CB1R)
en un comportamiento de búsqueda de alcohol. El receptor cannabinoide CB1 es muy
abundante en el sistema nerviosos central y está involucrado en analgesia, ansiedad y
apetito. En una primera serie experimental, hemos estudiado las respuesta operantes
realizadas por ratones CB1R knockout (KO) constitutivos y ratones controles wild-type
(WT) de la misma camada que han sido entrenados en el comportamiento de
búsqueda de alcohol. Hemos evaluado tanto el comportamiento operante de búsqueda
de esta droga como la ingesta activa de la misma en programas de refuerzo de razón
fija 1 (FR1), 3 (FR3) y 5 (FR5) y con concentraciones de sacarina que gradualmente se
substituyeron por concentraciones crecientes de alcohol, observándose cambios
significativos entre los ratones WT y los CB1R KO en las diferentes etapas
experimentales, siendo los animales knockout los que más respondieron. En una
segunda serie experimental, se estudiarán los mismos parámetros en ratones KO
condicionales. Por otra parte, hemos realizado estudios de proteómica para evaluar las
implicaciones de posibles proteínas, hasta ahora desconocidas, que pudieran estar
relacionadas con la implicación de los receptores CB1R en los procesos adictivos.
Este trabajo ha sido financiado por el Plan Nacional Sobre Drogas (#PNSD-2013I068)
31
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27. PHARMACOLOGICAL ACTIVATION OF EITHER CANNABINOID CB1
OR CB2 RECEPTORS COUNTERACTS THE ETHANOL-INDUCED
IMPAIRMENT OF HIPPOCAMPAL CELL PROLIFERATION IN ADULT
RAT BRAIN. P. Rivera, L. Bindila, F. Alen, A. Vargas, F.J. Pavón, A.
Serrano, L. Rubio, B. Lutz, F. Rodríguez de Fonseca, J. Suárez.
UGC Salud Mental, Instituto de Investigación Biomédica de Málaga (IBIMA),
Universidad de Málaga-Hospital Universitario Regional de Málaga, Avda. Carlos Haya
82, Pabellón de Gobierno, 29010, Málaga, Spain. Institute of Physiological Chemistry,
University Medical Center of the Johannes Gutenberg-University of Mainz,
Duesbergweg 6, 55128 Mainz, Germany. Departamento de Anatomía y Medicina Legal,
Universidad de Málaga, 29071, Málaga, Spain.
Chronic alcohol exposure reduces endocannabinoid activity and disrupts adult
neurogenesis in rodents, resulting in structural and functional alterations. As
cannabinoid receptor agonist promotes adult neural progenitor cell (NPC) proliferation,
we evaluated the protective effects of the fatty-acid amide-hydrolase (FAAH) inhibitor
URB597 that enhances the endocannabinoid receptor tone, the selective CB1 receptor
agonist ACEA and the selective CB2 receptor agonist JWH133 on NPC proliferation in
alcoholic rats. We performed immunohistochemical and stereological analyses of cells
expressing the mitotic phosphorylation of histone-3 (phospho-H3+) and the replicating
cell DNA marker 5-bromo-2’-deoxyuridine (BrdU+) in main neurogenic zones of adult
brain: subgranular zone of dentate gyrus (SGZ), subventricular zone of lateral ventricles
(SVZ) and hypothalamus, in URB597, ACEA and JWH133-treated rats with forced
consumption of alcohol (11%) and sucrose liquid diets for two weeks. After a controlled
isocaloric pair-feeding period of both sucrose and alcoholic diets, animals were allowed
ad libitum alcohol intake (7.3±1.1 g/kg/day). Alcohol intake resulted in a reduction in
BrdU+ cells in SGZ, SVZ and hypothalamus. Although all treatments (URB597, ACEA,
JWH133) increased alcohol consumption, the alcohol-induced impairment of NPC
proliferation was reversed by ACEA and JWH133 in SGZ, and was specifically
counteracted by JWH133 in SVZ. URB597 neither affected NPC proliferation nor
reversed alcohol-induced impairment of NPC proliferation. Specific activation of CB1
and CB2 receptors rescued impaired hippocampal NPC proliferation caused by alcohol,
which could be associated with increased alcohol tolerance. This neuroprotective role of
selective CB1 and CB2 receptor agonists may results in a potential clinical interest for
alcoholics in risk of neural damage.
Supported by Instituto de Salud Carlos III, Ministerio de Economía y Competitividad,
UE-ERDF. Grant number: CP12/03109
32
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28. BOTH GENETIC DELETION AND PHARMACOLOGICAL BLOCKADE
OF LYSOPHOSPHATIDIC ACID LPA1 RECEPTOR RESULTS IN
INCREASED ALCOHOL CONSUMPTION. A. Serrano, E. CastillaOrtega, F.J. Pavón, L. Sánchez-Marín, G. Estivill-Torrús, C. Pedraza, E.
Blanco, J. Suárez, L. Santín, F. Rodríguez de Fonseca.
UGC Salud Mental, Instituto de Investigación Biomédica de Málaga (IBIMA),
Universidad de Málaga-Hospital Universitario Regional de Málaga, Avda. Carlos Haya
82, Pabellón de Gobierno, 29010, Málaga, Spain.
Background: The lysophosphatidic acid (LPA) is a new, intercellular signaling molecule
in the brain involves in multiple functions. This study investigates the consequences of
either genetic deletion or pharmacological blockade of lysophosphatidic acid receptor-1
(LPA1) in alcohol consumption.
Methods: The experiments were performed by the administration of LPA1 receptor
antagonist Ki16425 in both mice and rats or by using LPA1-null mice.
Results: In the two-bottle free choice paradigm, the LPA1-null mice of both sexes
preferred the alcohol more than their wild-type counterparts. The male LPA1-null mice
were then further characterized, showing a notably increased ethanol drinking after a
deprivation period and a reduced sleep time after acute ethanol administration. In
addition, LPA1- null mice were more anxious than the wild-type mice in the elevated
plus maze test. For the pharmacological experiments, the acute administration of the
antagonist Ki16425 consistently increased ethanol consumption in both wild-type mice
and rats; while it did not modulate alcohol drinking in the LPA1-null mice and lacked
intrinsic rewarding properties and locomotor effects in a conditioned place preference
paradigm. In addition, LPA1-null mice exhibited a marked reduction on the expression
of glutamate-transmission-related genes similar to those observed in alcohol-exposed
rodents.
Conclusions: Results suggest a relevant role for the LPA/LPA1 signaling
in alcoholism so the LPA1-null mice emerge as a new model for genetic vulnerability to
excessive alcohol drinking (coupled with reduced ethanol sensitivity and altered
emotionality). The pharmacological manipulation of LPA1 receptor arises as a new
target for the study and treatment of alcoholism.
Acknowledgements: the present study has been supported by the Ministerio de
Economía y Competitividad (MEC) and Instituto de Salud Carlos III (ISC-III) (project
PI13/02261); MEC, ISCIII and Red de Trastornos Adictivos UE-FEDER/EU-ERDF 2012
(RD12/0028); Ministerio de Sanidad, Servicios Sociales e Igualdad and Plan Nacional
Sobre Drogas (projects 049/2009 and 049/2013); Junta de Andalucía and Plan Andaluz
de Investigación, Desarrollo e Innovación UE-FEDER/EU-ERDF (CTS-433); Junta de
Andalucía and Consejería de Economía, Innovación y Ciencia (Project PI45403); Junta
de Andalucía and Consejería de Igualdad, Salud y Políticas Sociales (projects PI08232012 and PI0228-2013); contracts ’Miguel Servet’ from ISC-III (CP14/00173,
CP14/00212 and CP12/03109).
33
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29. LPA1 RECEPTOR KNOCKOUT MICE SHOW STRONG DEFICIT ON
GLUTAMATE BIOSYNTHETIC ENZYME GLUTAMINASE (GLS, KGA)
AND ALTERED MORPHOLOGY OF HIPPOCAMPAL AND CORTICAL
DENDRITIC SPINES. A. Peñalver, J.A. Campos-Sandoval, R. SánchezVaro, C. Cardona, L. Castilla, E. Blanco, M. Martín-Rufián, J.A. Segura,
J.M. Matés, F.J. Alonso, A. Gutiérrez, F. Rodríguez de Fonseca, J.
Márquez.
Dpto. Biología Molecular y Bioquímica Facultad de Ciencias, Universidad de Málaga,
Málaga. Red RETICS de trastornos adictivos (RTA). Instituto de Investigación
Biomédica (IBIMA) Departamento de Biología Celular, Genética y Fisiología. Facultad
de Ciencias, Universidad de Málaga. IBIMA. Centro de Investigación Biomédica en Red
sobre Enfermedades Neurodegenerativas (CIBERNED). Laboratorio de Medicina
Regenerativa, Instituto de Investigación Biomédica (IBIMA), Complejo Hospitalario de
Málaga (Hospital Carlos Haya), Pabellón de Gobierno, Málaga.
Objectives: The objective of the present study was to utilize mice with knocked-down
lysophosphatidic acid 1 (LPA1) receptor to ascertain changes in glutamatergic
transmission that may help to explain part of the cognitive and memory deficits shown
by these KO-LPA1 mice.
Material & methods: A well characterized KO-LPA1 mouse strain was used as animal
model and compared with wild-type (WT) and heterozygous animals. Expression
studies were implemented by immunohistochemistry and Western analysis of mouse
brain regions, real-time quantitative RT-PCR of GA isoforms, enzymatic analysis of
regional GA activity and Golgi staining to assess dendritic spine morphology and
density.
Results: A strong reduction of KGA immunoreactivity was mostly revealed in cerebral
cortex and hippocampus of KO-LPA1 mice versus WT and heterozygous animals. In
contrast, neither mRNA levels nor enzyme activity were significantly altered in KO mice
suggesting compensatory mechanisms for neurotransmitter Glu synthesis. Interestingly,
Golgi staining of hippocampal and cortical neurons revealed a clear morphology change
toward a less-mature undifferentiated spine phenotype, without changes in the total
number of spines.
Conclusions: The molecular mechanisms underlying KGA downregulation in null LPA1
mutant mice are unknown. However, LPA increases neuronal differentiation,
arborization and neurite outgrowth of developing neurons, while Gln-derived Glu,
through GA reaction, has been also involved in neuronal growth and differentiation. It is
tempting to speculate that downregulation of KGA protein in KO-LPA1 mice induce
morphological changes in dendritic spines of cortical and hippocampal neurons which,
in turn, may account for memory and cognitive deficits shown by KO-LPA1 mice.
Acknowledgements: Red de Trastornos Adictivos, RTA, (RD12/0028/0013/) RETICS,
ISCIII-FEDER y Consejería Innovación, Ciencia y Empresa, Junta de Andalucía
(Proyecto de Excelencia CVI-6656).
34
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RESÚMENES DE LOS PÓSTERES
1. DIFFERENTIAL MODULATION OF PLASMA CORTICOSTERONE
AND BRAIN NORADRENALINE RECEPTORS DURING THE
INCUBATION OF COCAINE, HEROIN AND SUCROSE CRAVING. A. P
D. Roura-Martínez, M. Ucha, R. Santos-Toscano, A. Contreras, R.
Capellán, J. Orihuel, E. Ambrosio, A Higuera-Matas.
Department of Psychobiology. School of Psychology. UNED. Madrid, Spain.
Objectives. The progressive increase (incubation) of craving is a phenomenon
commonly suffered by addicts during abstinence periods and it has been reproduced in
animal models. The aim of this study was to describe the changes in plasmatic
corticosterone, and alpha2A and beta1A noradrenaline receptor mRNA expression
levels in ventromedial prefrontal cortex (vmPFC) and basolateral amygdala (BLA)
during the incubation of cocaine, heroin and sucrose craving.
Materials and methods. Male Lewis rats underwent self-administration protocols that
are known to induce incubation of craving: 6 h per day sessions (cocaine, heroin,
saline), or 2 h per day (sucrose, water), for 10 consecutive days. Then, rats from each
group were assigned to two withdrawal conditions: a half of the animals were sacrificed
after one day of withdrawal and the other half after one month of forced abstinence,
collecting blood for corticosterone measuring and adrenal cortex. The brains were also
collected and vmPFC and BLA were dissected for qPCR analysis.
Results. Cocaine and heroin treated rats showed adrenomegalia at early withdrawal as
well as increased corticosterone levels. Conversely, this increase was observed only
after 30 days of withdrawal in the sucrose group. A downregulation of BLA alpha2A
gene expression after cocaine and upregulation of beta1A with heroin were observed,
while in vmPFC sucrose craving induced a downregulation of alpha2A receptor gene
expression.
Conclusions. Although apparently stress is not determinant in cue induced-relapse into
drug-taking with short access protocols, it could be playing an important role during the
incubation of craving.
Supported by grants from the Ministerio de Ciencia e Innovación ( SAF2013-47520-P);
Ministerio de Sanidad, Servicios Sociales e Igualdad (Red de Trastornos AdictivosRTA-RD12/028/0020 -Instituto de Salud Carlos III- and Plan Nacional sobre Drogas,
2012I057); Dirección General de Investigación de la Comunidad de Madrid ( S2011/BMD-2308; Programa de Actividades I+D+I CANNAB-CM); UNED (Plan de
Promoción de la Investigación); and European Union (JUST/2013/DPIP/AG/4823-EU
MADNESS).
35
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2. DIFFERENTIAL CHANGES IN THE PI3K/AKT/MTOR SIGNALING
PATHWAY IN THE BASOLATERAL COMPLEX OF THE AMYGDALA
AFTER COCAINE AND HEROIN SELF-ADMINISTRATION: EFFECTS
OF COMPULSIVE CONSUMPTION AND INCUBATION OF CRAVING.
M. Ucha, D. Roura-Martínez, R. Santos-Toscano, A. Contreras, R.
Capellán; J. Orihuel, A. Higuera-Matas, E. Ambrosio.
Department of Psychobiology, School of Psychology. UNED. 28040, Madrid, Spain.
Objectives: One of the defining features of drug addiction is the high risk of relapse,
even after prolonged abstinence. Such relapse is often triggered by drug-associated
cues that provoke drug craving. Responsiveness to those cues progressively increases
during absence, a phenomenon termed incubation of craving. In this study, we used a
rat model of this incubated craving to study the potential implication of the
PI3K/Akt/mTOR signaling pathway which plays a role in learning and memory and that
has been linked to drug abuse.
Materials and methods: Male Lewis rats self-administered either cocaine, heroin or
saline during 10 days under extended access. We then tested their responsiveness to
drug-associated cues after 1 or 30 days of forced abstinence and we confirmed the
presence of drug craving in our conditions. A different batch of rats followed the same
self-administration protocol and were sacrificed after 1 or 30 days of abstinence. We
analyzed the expression of several genes of the PI3K/Akt/mTOR pathway in the
basolateral complex of the amygdala, which plays a key role in cue-induced relapse.
Results: We found a significant increase in the expression of the p70s6k and 4ebp2
genes (final effectors in this pathway) after heroin self-administration, even after 30
days of abstinence. The gene expression of both Akt2 and igf2r also tended to increase
after heroin, although this trend was not significant. The only significant effect in the
cocaine groups was a decrease in Rictor gene expression, but this effect was only seen
after 30 days of abstinence, suggesting its involvement in the incubation of cocaine
craving.
Conclusions: These results suggest that the PI3K/Akt/mTOR pathway is affected by
heroin self-administration under extended access conditions and that the mTORC
complex 2 (as revealed by the alterations in Rictor gene expression) could regulate the
incubation of cocaine craving.
Supported by grants from the Ministerio de Ciencia e Innovación ( SAF2013-47520-P);
Ministerio de Sanidad, Servicios Sociales e Igualdad (Red de Trastornos AdictivosRTA-RD12/028/0020 -Instituto de Salud Carlos III- and Plan Nacional sobre Drogas,
2012I057); Dirección General de Investigación de la Comunidad de Madrid ( S2011/BMD-2308; Programa de Actividades I+D+I CANNAB-CM); UNED (Plan de
Promoción de la Investigación); and European Union (JUST/2013/DPIP/AG/4823-EU
MADNESS).
36
Reunión científica 2015
3. SEX -DEPENDENT EFFECTS OF UNPREDICTABLE POSTNATAL
MATERNAL STRESS PLUS MATERNAL DEPRIVATION ON GLIAL
CELLS,
AND
CB1
CANNABINOID
RECEPTOR
IN
THE
HIPPOCAMPAL FORMATION OF ADULT RATS. L. Arnaldo, R. Nadal,
S. Fuentes, A. Armario, MP. Viveros, M. López-Gallardo.
Department of Physiology (Animal Physiology II), Faculty of Biology, Universidad
Complutense, Madrid, Spain; Institut de Neurociències and Psychobiology Unit (School
of Psychology), Universitat Autònoma de Barcelona, Spain; Institut de Neurociències.
Animal Physiology (School of Biosciences), Universitat Autònoma de Barcelona, Spain;
Department of Physiology, Faculty of Medicine, Universidad Complutense, Madrid,
Spain.
Traumatic experiences during childhood can increase the risk of suffering
neuropsychiatric diseases in the adulthood. In animals, neonatal stress models have
been developed to mimic early life stress in humans and to investigate the neural basis
of its long term effects. The hippocampal formation appears to be one important target
of the deleterious effects caused by these stress protocols.
In the present study, we have investigated the effects of a combination of unpredictable
postnatal maternal stress (electric shock, restraint, forced swim) 20 minutes/day; and
unpredictable maternal separation, 3 hours/day, from postnatal day (PND) 1 to 14, on
the hippocampal formation of adult Long-Evans rats (PND 90) of both sexes. We
analyzed by immunohistochemistry, neuronal and glial markers, CB1 receptor
expression and several synaptic plasticity players. Notably, the present neonatal stress
model increased active maternal behavior.
In treated animals of both sexes, a decreased number of GFAP+ cells was found in the
polymorphic layer of the Dentate Gyrus. In all areas analyzed, the number of Iba1+ cells
tended to decrease in males and to increase in females. In relation with the stage of
activation of microglia (I corresponding to ramified resting microglia and V to ameboid
microglia, the most active one), an increase of morphotype II versus morphotype I cells
was found in all areas and in both sexes. A decreased expression of CB1 cannabinoid
receptor was found in treated males, mainly in CA3 area. No alterations were found in
the expression levels of NeuN or synaptophysin. In conclusion, the present combined
model of neonatal stress induced long term sexually dimorphic effects in glial cells and
CB1 receptors in the hippocampal formation of adult rats.
Agradecimientos: ISCIII, FEDER, Plan Nacional sobre Drogas, MICINN, MINECO.
37
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4. MMP-3 AND MMP-9 METALOPROTEINASE ACTIVITIES ARE
INCREASED IN PLASMA FROM CHRONIC ALCOHOLIC PATIENTS.
M.D. Gutiérrez, M. Velasco, M. Marcos, E. O’Shea, F.J. Laso, M.I.
Colado.
Departamento de Farmacología, Facultad de Medicina, Universidad Complutense,
Madrid; Departamento de Medicina Interna, Hospital Universitario-Instituto de
Investigación Biomédica, Salamanca.
Introduction: Chronic alcohol consumption impairs executive and cognitive functions.
Alcohol induces a neuroinflammatory process and there is growing evidence of an effect
of alcohol on blood-brain barrier (BBB) integrity. Several metaloproteinases (MMPs) are
involved in the disruption of BBB structure in models of brain damage. In addition, in
certain types of brain damage S100B and neuron specific enolase (NSE) are secreted
and increased plasmatic levels of these molecules have been proposed as potential
biomarkers.
Objectives: The aim of this study was to evaluate the enzymatic activity and expression
of MMP-2, MMP-9 and MMP-3 and to quantify the levels of S100B and NSE in plasma
from actively chronic alcohol-consuming patients.
Methods: Twenty-four chronic alcohol-consuming patients with heavy daily alcohol
consumption and 27 healthy, age-matched subjects were studied. Patients had no
severe medical conditions, liver disease or psychiatric symptoms. Plasma was isolated
from peripheral blood. MMP-9, MMP-2 and MMP-3 enzymatic activities were
determined by zymography. Western blot was used to study MMP-9, MMP-2 and MMP3 expression as well as S100B presence in plasma. S100B and NSE levels were
quantified by ELISA.
Results: Alcohol-consuming patients showed chronic (12-528 months) and heavy daily
alcohol consumption (mean: 176 g/day). Plasma MMP-9 activity was slightly (32%) and
MMP-3 markedly increased (84%). No differences between groups were found for
MMP-2 activity or for protein expression in all cases. S100B and NSE concentration
were similar between groups.
Conclusions: Data indicate that chronic active alcohol intake may be associated with
an increase in MMP-9 and MMP-3 activities in plasma and suggest that these
parameters could be used as potential biomarkers. In addition, and in contrast to
previous data in literature, our results indicate that S100B and NSE cannot be consider
biomarkers in chronic alcohol-consuming patients since no changes were observed
between alcoholic and control groups.
Acknowledgements: ISCIII RETICS RTA (RD12/0028/0002) and Fondo Europeo de
Desarrollo Regional (FEDER).
38
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5. A SINGLE NEUROTOXIC DOSE OF MDMAINCREASESKYNURENINE
PATHWAY METABOLISM IN PLASMA AND HIPPOCAMPUS OF RAT.
R. Vidal, C. Abuin, E. O’Shea, M.I. Colado.
Departamento de Farmacología, Facultad de Medicina, Universidad Complutense,
Madrid
Introduction: The kynurenine pathway is tightly controlled by the peripheral and central
immune systems and dysregulations of the pathway are associated with
neurodegenerative and neurological disorders. MDMA induces neuroinflammation in the
rat brain reflected as an increase in interleukin-1βproduction and microglial activation.
Objective: To evaluate the time-course of MDMA-induced changes in tryptophan
metabolites in plasma and hippocampus of rat. As brain kynurenines are influenced by
the peripheral kynurenine pathway, it seems reasonable to propose that fluctuations in
the blood levels of these metabolites might directly affect central kynurenine
metabolism.
Materials and methods: Male Dark Agouti rats received a single neurotoxic dose of
MDMA (12.5 mg/kg, i.p.) and were sacrificed at 1h, 3h, 6h, 24h and 7 days. Plasma and
hippocampal levels of serotonin, tryptophan, kynurenine and kynurenic acid were
measured by high pressure liquid chromatography (HPLC). The ratios of
serotonin/tryptophan and kynurenine/tryptophan, as anindex of activity of the enzymes
involved in the metabolism of tryptophan, were also calculated.
Results: Plasma kynurenine concentration increased 3h and 6h after MDMA while
tryptophan concentration was reduced at 6h. Consequently, MDMA produced an
increase in the kynurenine/tryptophan ratio 3h and 6h after dosing. The
kynurenine/tryptophan ratio also increased in hippocampus although the effect was only
evident 6h after MDMA. Plasma serotonin concentration decreased 3h and 6h after
MDMA thereby reducing the serotonin/tryptophan ratio at the same time-points. In
hippocampus, serotonin concentration was reduced at all time-points resulting in a
decreased serotonin/tryptophan ratio up to 7 days after MDMA. Kynurenic acid
concentration only increased in plasma 3h after MDMA.
Conclusions: Administration of a neurotoxic dose of MDMA in Dark Agouti rats leads to
an imbalance of the kynurenine pathway resulting in an increase of the
kynurenine/tryptophan ratio in plasma and hippocampus reflecting increased
indoleamine 2,3 dioxygenase activity (enzyme converting tryptophan in kynurenine).
Acknowledgments: MINECO (SAF2013-40592-R), MSSSI (PNSD 2014I015), ISCIII
RETICS RTA (RD12/0028/0002) and Fondo Europeo de Desarrollo Regional (FEDER).
39
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6. LONG-TERM
EFFECTS
OF
ADOLESCENT
INTERMITTENT
ALCOHOL EXPOSURE ON THE SPLENIC ENDOCANNABINOID
SYSTEM AND PLASMA INFLAMMATORY MEDIATORS IN MALE
AND FEMALE RATS. J. Decara, F.J. Pavón, E.M. Marco, M. Vázquez,
V. Mela, F. Alén, J. Suárez, E. Giné, J.A. López Moreno, J. Chowen, F.
Rodríguez de Fonseca, A. Serrano, M.P. Viveros.
UGC Salud Mental, IBIMA, Hospital Universitario Regional de Málaga; Departamento
de Fisiología, Facultad de Biología, Universidad Complutense, Madrid; Departamento
de Biología Celular, Facultad de Psicología, Universidad Complutense, Madrid; Servicio
de Pediatría y Endocrinología Pediátrica, Hospital Infantil Universitario Niño Jesús,
Instituto de Investigación Sanitaria del Hospital Universitario de La Princesa, Madrid.
Intermittent alcohol exposure is a common pattern of alcohol consumption among
adolescents. Alcohol is known to modulate immune function and other signaling
systems associated with inflammation such as the endocannabinoid system (ECS).
Because there are sex differences in immune responses, we evaluated the expression
of the splenic ECS and the plasma concentration of inflammatory mediators in female
and male rats exposed to alcohol during adolescence. A 4-day drinking in the dark
(DID) procedure for 4 weeks was used as a model of intermittent forced-alcohol
administration (20%, v/v) in Wistar rats of both sexes. Plasma, liver and spleen were
collected 2 weeks after the last DID session. The mRNA expression of components of
the ECS in the spleen was analyzed by qRT-PCR analysis. The plasma concentrations
of chemokines (CX3CL1 and CCL2) and pro-inflammatory cytokines (TNF-α, IL-1β, IL-6
and IL-10) were analyzed by multiplex immunoassay. This DID procedure produced a
moderate alcohol consumption during adolescence and it did not cause liver damage.
Analysis of mRNA expression of endocannabinoid components in the spleen indicated
that the cannabinoid receptor CB1 and the nuclear receptor PPARα were affected by
alcohol exposure with a decrease in females and males, respectively. The mRNA levels
of the endocannabinoid-synthesizing enzymes NAPE-PLD and DGLβ in the spleen
were increased by alcohol exposure, but the NAPE-PLD increase was only observed in
female rats. Regarding the plasma concentrations of inflammatory mediators, whereas
CX3CL1 was only affected by sex with lower concentration in males, TNF-α and IL-6
concentrations were significantly affected by alcohol and sex. TNF-α and IL-6 were
higher in females relative to control males, but alcohol abolished this difference. In
conclusion, moderate and intermittent alcohol consumption during adolescence is
sufficient to induce long-term changes in the expression of the ECS in the spleen and
changes in the concentration of pro-inflammatory cytokines in plasma, but this was not
associated with liver damage. Furthermore, we demonstrate sex-differences in the
effects of alcohol on the expression of endocannabinoid-related enzymes and proinflammatory cytokines.
.
Supported by: Instituto de Salud Carlos III (ISC-III), RETICS, Red de Trastornos
Adictivos UE-FEDER 2012 (RD12/0028/0021); GRUPOS UCM-BSCH (UCM 951579),
Ministerio de Economía y Competitividad (PI13/02261); Plan Nacional sobre Drogas
049/2013; Consejería de Economía, Innovación y Ciencia, Junta de Andalucía UEFEDER (CTS-433); Consejería de Salud y Bienestar Social, Junta Andalucía (PI02282013 and PI0823-2012). ISC-III UE-FEDER Servet I (CP14/00212 and CP14/00173).
40
Reunión científica 2015
7. EFFECTS OF ADOLESCENT BINGE DRINKING ON THE
EXPRESSION OF THE ENDOCANNABINOID SYSTEM IN THE BRAIN.
L. Sánchez, J. Decara, F.J. Pavón, F. Rodríguez de Fonseca, A.
Serrano.
UGC de Salud Mental, Instituto IBIMA, Hospital Regional Universitario de Málaga,
Spain.
A common abuse pattern among adolescents is called binge drinking and it contributes
to a main portion of alcohol-related problems. This pattern of alcohol consumption in
adolescent years leads to important structural and functional changes in the developing
brain, increasing the propensity to later abuse. Additionally, the endocannabinoid
system (ECS) is involved in the motivational and addictive properties of ethanol. In the
present study, we evaluated the long-term consequence of adolescent binge drinking on
the expression of genes related to ECS within addiction-related brain circuitries in
Wistar rats.
Animals were divided into 3 groups of ethanol-exposure during adolescence: 1) Free
access to ethanol, using the 2-bottle choice paradigm; 2) Binge-like alcohol, using
ethanol injections; 3) Binge-like alcohol and free access to ethanol. During adulthood,
all groups had free access to ethanol. Animals were sacrificed and brains were removed
to determinate the expression of the ECS using qRT-PCR. Five areas were evaluated:
Prefrontal cortex, Striatum, Amygdala, Hippocampus and Nucleus Accumbens.
We found that adolescent binge drinking was associated with significant changes in the
mRNA expression of enzymes and receptors of the ECS in a cerebral area-dependent
manner. In the Prefrontal Cortex, free access to ethanol increased the mRNA levels of
the cannabinoid CB2 receptor and the enzymes involved in the synthesis (DAGLβ) and
degradation (FAAH and MAGL) of endocannabinoids. In the Striatum, changes in the
mRNA were observed in the group that received ethanol injections and these rats
showed an increase in the mRNA levels of CB2 receptors and all of the enzymes
involved in the monoacylglycerol pathway. In the Amygdala, the free access to ethanol
induced a reduction of CB2, FAAH and DAGLβ mRNA levels. Regarding the
Hippocampus, there was a significant effect on the mRNA expression of CB1 and CB2
receptors, resulting in an increase in animals with free access to alcohol. Finally, we
observed no remarkable changes in the Nucleus Accumbens.
In summary, these results show that adolescent binge drinking produces alterations on
the ECS that are maintained into adulthood. Future studies will elucidate the role of CB2
receptors as primary target in the alcohol-induced changes in the ECS of cerebral areas
related to addiction.
Acknowledgements: the present study has been supported by the Ministerio de
Economía y Competitividad (MEC) and Instituto de Salud Carlos III (ISC-III) (project
PI13/02261); MEC, ISC-III and Red de Trastornos Adictivos UE-FEDER/EU-ERDF
2012 (RD12/0028); Junta de Andalucía and Consejería de Igualdad, Salud y Políticas
Sociales (PI0823-2012 and PI0228-2013); ’Miguel Servet I’ research contracts from
ISC-III and UE-FEDER/EU-ERDF (CP14/00212 and CP14/00173).
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8. PLASMA CONCENTRATIONS OF BDNF AND IGF-1 IN ABSTINENT
COCAINE USERS WITH HIGH PREVALENCE OF SUBSTANCE USE
DISORDERS: RELATIONSHIP TO PSYCHIATRIC COMORBIDITY. P.
Araos, M. Pedraz, A.I. Martín-Velasco, N. García-Marchena, A. Serrano,
P. Romero-Sanchiz, J. Suárez, E. Castilla-Ortega, V. Barrios, R.
Campos-Cloute, J.J. Ruiz, M. Torrens, J.A. Chowen, J. Argente, R. de la
Torre, L.J. Santín, M.Á. Villanúa, F. Rodríguez de Fonseca, F.J. Pavón.
UGC Salud Mental, Instituto IBIMA, Hospital de Málaga; Hospital Infantil Niño Jesús,
Madrid; IMIM, Barcelona; Universidad de Málaga; Universidad Complutense, Madrid.
Objectives and methods. Recent studies have identified biomarkers related to the
severity and pathogenesis of cocaine addiction and common comorbid psychiatric
disorders. Monitoring these plasma mediators may improve the stratification of cocaine
users seeking treatment. Because the neurotrophic factors are involved in neural
plasticity, neurogenesis and neuronal survival, we have determined plasma
concentrations of brain-derived neurotrophic factor (BDNF), insulin- like growth factor 1
(IGF-1) and IGF-1 binding protein 3 (IGFBP-3) in a cross-sectional study with abstinent
cocaine users who sought outpatient treatment for cocaine (n = 100) and age/body
mass matched controls (n = 85). Participants were assessed with the diagnostic
interview ‘Psychiatric Research Interview for Substance and Mental Disorders’.
Results and conclusions. Plasma concentrations of these peptides were not different
in cocaine users and controls. They were not associated with length of abstinence,
duration of cocaine use or cocaine symptom severity. The pathological use of cocaine
did not influence the association of IGF-1 with age observed in healthy subjects, but the
correlation between IGF-1 and IGFBP-3 was not significantly detected. Correlation
analyses were performed between these peptides and other molecules sensitive to
addiction: BDNF concentrations were not associated with inflammatory mediators, lipid
derivatives or IGF-1 in cocaine users, but correlated with chemokines
(fractalkine/CX3CL1 and SDF-1/CXCL12) and N-acyl-ethanolamines (N-palmitoyl-,Noleoyl-, N-arachidonoyl-, N-linoleoyl- and N-dihomo-γ-linolenoyl-ethanolamine) in
controls; IGF-1 concentrations only showed association with IGFBP-3 concentrations in
controls; and IGFBP-3 was only correlated with N-stearoyl-ethanolamine concentrations
in cocaine users. Multiple substance use disorders and life-time comorbid
psychopathologies were common in abstinent cocaine users. Interestingly, plasma
BDNF concentrations were exclusively found to be decreased in users diagnosed with
both primary and cocaine induced disorders for mood and anxiety disorders. In
summary, BDNF, IGF-1 and IGFBP-3 were not affected by a history of pathological use
of cocaine supported by the absence of associations with other molecules sensitive to
cocaine addiction. However, BDNF was affected by comorbid mood disorders. Further
research is necessary to elucidate the role of BDNF and IGF-1 in the transition to
cocaine addiction and associated psychiatric comorbidity.
Supported by: Instituto de Salud Carlos III (ISC-III), Red de Trastornos Adictivos UEFEDER 2012 (RD12/0028); Ministerio de Economía y Competitividad (PI13/02261);
Plan Nacional sobre Drogas 049/2009 and 049/2013; Consejería de Economía,
Innovación y Ciencia, Junta de Andalucía UE-FEDER (CTS-433); Consejería de Salud
y Bienestar Social, Junta Andalucía UE-FEDER (PI0228-2013 and PI0823-2012);
Departament d’Innovació, Universitats i Empresa, Generalitat de Catalunya (2014-SGR680); Servet I, ISC-III/UE-FEDER (CP014/00212).
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9. PSYCHIATRIC COMORBIDITY IN ALCOHOL USERS IN A HOSPITAL
PROGRAM FOR OUTPATIENT TREATMENT.ANALYSIS OF GENDER
DIFFERENCES. N. García-Marchena, P. Araos, F.J. Pavón, G. Ponce,
M. Pedraz, A. Serrano, F. Arias, P. Romero, J. Suárez, M. Torrens, G.
Rubio, F. Rodríguez de Fonseca.
Hospital R. U. de Málaga, Málaga, Spain; Hospital Universitario 12 de Octubre, Madrid,
Spain; Institut Hospital del Mar d’Investigacions Mèdiques (IMIM), Barcelona, Spain.
In order to optimize care and improve prognosis of alcoholism, the psychiatric
characterization of patients is necessary since alcohol addiction is associated with high
psychiatric comorbidity. However, gender differences have received little attention to
improve the stratification of these patients. The overall objective of this study was to
estimate the prevalence of psychiatric comorbidity in a sample outpatient treated for
alcohol use considering gender.
We recruited 162 alcohol users from outpatient programs for alcoholism treatment at
Hospital Universitario 12 de Octubre in Madrid (Spain) based on inclusion and exclusion
criteria for participation in the study.After obtaining informed consent, all participants
were evaluated regarding the prevalence of substance use disorders and psychiatric
comorbidity, according to Diagnostic and Statistical Manual of Mental Disorders DSMIV-TR(Diagnostic and Statistical Manual of Mental Disorders, 4th text revised) with the
semi structured interview PRISM (Psychiatric Research Interview for Substance and
Mental disorders) and the WURS (Wender-Utah Rating Scale).
The sample was composed of 75.3% men and 24.7% women with an average age at
49.3 years old and 15 years of pathological use of alcohol who were diagnosed with
alcohol use disorders (abuse or dependence). The prevalence of lifetime psychiatric
comorbidity was 86.5%, highlighting mood disorders (37%) followed by attention deficit
hyperactivity disorder (24.7%) and anxiety disorders (17.9%).The alcohol-induced
disorders were more frequent in mood and psychotic disorders, whereas the primary
disorders were more prevalent in anxiety disorders. We found significant gender
differences in substance use disorders and psychiatric comorbidity. Regarding the
pattern of alcohol consumption, men started drinking at a younger age and they
progressed faster from the onset to alcohol addiction with a longer and more severe
alcohol use history compared with women. Women showed higher prevalence of
psychiatric comorbidity compared with men (82.5% and 63.9%, respectively) mainly due
to mood disorders. In addition, there was a lower prevalence of other substance-use
disorders (e.g. cocaine and cannabis)in women (25.0%) than men (43.4%). Gender is a
critical factor to be considered for an adequate stratification of alcohol users who are
seeking treatment. The different prevalences of psychiatric comorbidity and substance
use disorders suggest differential therapeutic approaches for alcoholism to treating men
and women.
Instituto de Salud Carlos III (ISC-III), Red de Trastornos Adictivos UE-FEDER 2012
(RD12/0028); Ministerio de Economía y Competitividad (PI13/02261); Plan Nacional
sobre Drogas 049/2009 and 049/2013; Consejería de Economía, Innovación y Ciencia,
Junta de Andalucía UE-FEDER (CTS-433); Consejería de Salud y Bienestar Social,
Junta Andalucía (PI0228-2013 and PI0823-2012). Contracts “Miguel Servet”
(CP14/00212 and CP14/00173) and “Río Hortega” (CM13/0115) from ISC-III-UEFEDER.
43
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10. I1-IMIDAZOLINE RECEPTOR PROTEIN IS REDUCED IN THE
PREFRONTAL CORTEX OF COCAINE ADDICTS. Keller, M. ÁlvaroBartolomé, M.J. García-Fuster, R. La Harpe, L.F. Callado, J.J. Meana,
J.A. García-Sevilla.
Laboratory of Neuropharmacology/IUNICS and RETICS-RTA, Universitat de les Illes
Balears (UIB), Palma de Mallorca; Centre Universitaire Romand de Médecine Légale–
Site Genève, Université de Genève, Switzerland; Departament of Pharmacology and
CIBERSAM, Universidad del País Vasco (UPV/EHU), Leioa.
Cocaine addiction involves a dysregulation of dopamine neurotransmission in the
human brain. Recent studies suggested that I1-imidazoline receptors (IR) participate in
the prevention of cue-induced cocaine relapse in rats.
To test the hypothesis that human cocaine addiction is also associated with malfunction
of I1-IR, the content of two I1-related proteins were quantified in the prefrontal cortex
(PFC) of well characterized cohorts (all addicts: 10 male/9 female; 38±3 yr; 38±6 h PMI,
postmortem interval) of pure cocaine addicts (n=8), mixed cocaine/opiate addicts (n=11)
and matched controls (13 male/6 female; 41±3 yr; 39±6 h PMI) with specific and
validated antibodies (anti-IRBP antibody for 85 kDa I1-IR, and anti-NISCH antibody for
167 kDa I1-IR).
Blood samples of these cocaine abusers revealed high concentrations of cocaine (3-13
microg/ml) and benzoylecgonine (0.6-10 microg/ml), the main active metabolite.
Cocaine and benzoylecgonine were also detected in hair samples (8-310 ng/mg),
indicating chronic exposure to cocaine over the last 6-12 months. As a marker of
cocaine addiction, the density of dopamine D2-like receptors (anti-D2/D3/D4 receptor
antibody) in the PFC of pure cocaine addicts was reduced (29%, n=8, P=0.05) when
compared with that quantified in age-, gender, and PMI-matched controls. The content
of 85 kDa I1-IR was also significantly reduced (one-sample t-test) in the PFC of cocaine
addicts (all cocaine abusers: 27±5%, n=17, P=0.0002; pure cocaine abusers: 19±6%,
n=8, P=0.02; mixed cocaine/opiate abusers: 33±8%, n=9, P=0.005). In contrast, the
content of 167 kDa related to I1-IR (nischarin/IRAS) was not altered (all cocaine
abusers: 3±8%, n=17; pure cocaine abusers: +13±15%, n=8; mixed cocaine/opiate
abusers: 13±9%, n=9).
These results suggest impaired I1-IR-mediated functions (e.g.
neuroplasticity or apoptosis inhibition) in the brain of cocaine addicts.
regulation
of
Supported by RETICS-RTA RD12/0028/011, Plan Nacional sobre Drogas Grant
2012/001 and CIBERSAM (MINECO, ISCIII, FEDER). MJGF is a ‘Ramón y Cajal’
Researcher (MINECO).
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11. ADOLESCENT COCAINE EXPOSURE ALTERS THE PROPENSITY
TO SIGN- AND GOAL-TRACKIN SELECTIVELY BRED RATS. M.J.
Garcia-Fuster, A. Parsegian, S.B. Flagel, H. Akil.
IUNICS, Universidad de las Islas Baleares, Palma de Mallorca, Spain; Molecular &
Behavioral Neuroscience Institute, University of Michigan, Ann Arbor, MI, USA.
Adolescence is a critical period for neurodevelopmental changes and increased
propensity to use drugs. Bred high-responder (bHR) and low-responder (bLR) rats are
selectively bred based on locomotor response to a novel environment, such that bHRs
exhibit increased novelty-induced activity relative to bLRs. These phenotypes also differ
in their propensity to attribute incentive salience to reward cues. bHRs approach, or
sign-track, to cues associated with rewards; whereas bLR rats go to the location of
reward delivery, or goal-track, upon cue presentation. Sign-tracking behavior is
reflective of an increased propensity to attribute incentive salience to reward cues and
has been associated with addiction liability. It is predicted with 90-100% accuracy that a
bHR rat will be a sign-tracker and bLR a goal-tracker.
Thus, this genetic animal model was used to examine the impact of adolescent cocaine
th
exposure. On postnatal days 33-39bHR and bLR rats from the 38 generation were
exposed to a sensitizing regimen of cocaine (15 mg/kg, i.p, 7 days) or vehicle. The
tendency to sign- or goal-track was then examined in adulthood using standard Med
Associates chambers. Each Pavlovian training session consisted of 25 trials, wherein
the brief presentation of an illuminated retractable lever was followed by the responseindependent delivery of a food pellet. Behavior directed towards the lever and the food
cup was recorded as sign- and goal-tracking behavior, respectively.
Interestingly, adolescent cocaine exposure had no effect on sign-tracking behavior, but
altered goal-tracking behavior in both phenotypes. Cocaine exposure attenuated goaltracking behavior for bHRs, but enhanced it for bLRs. Thus, if born with a predisposition
to goal-track, drug exposure in adolescence enhances that tendency. In contrast, if born
with a predisposition to sign-track, adolescent drug exposure has little, if any, effect on
that tendency.
Supported by RETICS-RTA RD12/0028/0011 (ISCIII, MINECO-FEDER), Plan Nacional
sobre Drogas Grant 2012/0011 (MSSSI-FEDER) and by Fundación Alicia Koplowitz to
MJGF, and by NIDA (5P01DA021633) and Office of Naval Research (ONR: N00014-091-0598 and N00014-12-1-0366) to HA. MJGF is a ‘Ramón y Cajal’ Researcher
(MINECO).
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12. ENVIRONMENTAL
ENRICHMENT
REVERSES
COGNITIVE
IMPAIRMENTASSOCIATED
WITH
ALTERATION
OF
CB1DEPENDENT LTD AFTER ETHANOL CONSUMPTION DURING
ADOLESCENCE. I. Rico-Barrio, S. Peñasco, N. Puente, A. Ramos, N.
Royo, A. Gutiérrez, I. Bonilla, L. Reguero, M.J. Canduela, J. MendizabalZubiaga, F. Rodríguez de Fonseca, J. Suárez, I. Elezgarai, P. Grandes.
Department of Neurosciences, Faculty of Medicine and Dentistry, University of the
Basque Country UPV/EHU, E-48940 Leioa, Spain; Achucarro Basque Center for
Neuroscience, Bizkaia Science and Technology Park, Zamudio, Spain; Hospital
Regional Universitario de Málaga, Instituto IBIMA, Málaga, Spain.
Alcohol consumption, especially during adolescence, is one of the main problems that
concern our society.Studies in the last decade indicate that the endocannabinoid
system (ECS) function may be altered in ethanol dependence.An enriched environment
(EE) has been shown to significantly facilitate recovery from brain injury due to
important anatomical, molecular and functional changes that occur along brain
development.However, it is not known whether an EE has any effect on cognitive
impairment associated with ethanol consumption. Our aim is to investigate this during
the adolescence period and the role of an enriched environment to counteract ethanol
administration effects on plasticity mediated by cannabinoid CB 1 receptors in the
hippocampal dentate gyrus.
Male C57BL6 mice were exposed to a 4 days drinking-in-the-dark (DID) procedure
during adolescence (PD 30 ± 2 to 54 ± 2). The animals were given free access to
ethanol (20% (v/v) in tap water) or water for 2-h sessions during three consecutive days,
and a 4-h session on the 4thday. Then, from postnatal 56 to 74withdrawal days, the
animals were reared under two different conditions: standard laboratory condition (SC)
and enriched environment. Recognition memory was assessed by the novel object
recognition test (NORT) in the last three days of the withdrawal period(from p70 to p73).
Specifically, total exploration time, discrimination index % and recognition index were
measured in each group (SC-H2O, SC-OH, EE-H2O and EE-OH). The next day(p74)
adult mice were sacrificed.
Synaptic stimulation (10min, 10Hz) of the medial perforant path triggered a CB 1dependent long term depression (LTD) of the excitatory transmission that was absent in
adult mice after ethanol consumption during adolescence (2.7±3.12% of inhibition;
p<0.0001***). Furthermore, a significant lower recognition memory in the alcohol treated
group (SC-OH) compared to the untreated control group (SC-H2O) was observed.
Interestingly, both enriched groups (EE-H2O and EE-OH) showed higher values for
each of the measured parameters in the novel object recognition test compared to the
SC reared ethanol treated group (SC-OH). These results suggest that this experimental
paradigm can reverse the effects of ethanol consumption on recognition memory that
appears associated with an impairment of CB1-long term synaptic plasticity.
Key words: alcohol consumption, endocannabinoid system, enriched environment,
memory.
Funded by RETICS, ISCIII (RD12/0028/0001; RD12/0028/0004); MINECO (BFU201233334); Basque Government (IT764-13); UPV/EHU (UFI11/41); ISCIII (MINECO)
(CP12/03109, PI13/02261); JA (SAS111224); JA UE/ERDF (PI45403, CTS-8221).
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13. LONG-TERM ANXIETY EFFECTS ARE ASSOCIATED WITH
EPIGENETIC DYSFUNCTIONS INDUCED BY INTERMITTENT
ETHANOL TREATMENT DURING ADOLESCENCE: ROLE OF TLR4
RECEPTORS. M. Pascual, J. Montesinos, M. Rodríguez-Arias, J.
Miñarro, C. Guerri.
Department of CellularPathology, Centro de Investigación Príncipe Felipe, Valencia;
Department of Psychobiology, Facultad de Psicología, Universitat de Valencia,
Valencia.
The adolescence is a vulnerable brain maturation stage to the neurotoxic and
behavioral effects of alcohol. Although the mechanisms of these effects are largely
unknown, we have shown that ethanol by activating innate immune receptors toll-like
receptor 4 (TLR4), induces neuroinflammatory damage and causes cognitive behavioral
dysfunction, events which are maintained in adult mice exposed to ethanol during
adolescence. Therefore, the aim of this study is to evaluate whether the long-term
effects induced by ethanol abuse in adolescent animals are associated with epigenetic
chromatin changes and behavioral dysfunctions and whether these events correlate
with ethanol-induced inflammatory environment.
To answer these questions, wild-type (WT) and TLR4-deficient (TLR4-KO) adolescent
mice (PND 30) were treated intermittently with ethanol (3 g/kg) for 2 weeks, followed by
21 days of abstinence during maturation to young adulthood (65 day-old).
We show that ethanol treatment activates TLR4 signaling and triggers inflammatory
mediators in the adolescent prefrontal cortex. These events were associated with
changes in the histone acetylation (upregulation K9 H3, K5 H4 and K12 H4) and
methylation (downregulation of 3me-K4 H3). Young adult mice, treated with ethanol
during adolescence, showed long-term behavioral effects, as increased both the
anxiety-like behavior (open field behavior and elevated plus maze), the rewarding
effects (as demonstrated with conditioned place preference) and they also increased
the amount of voluntary alcohol intake, using two-bottle choice paradigm. These longterm effects in young adult mice were associated with changes in the histone H4
acetylation within the bdnf and fosb gene promoters, using chromatin
immunoprecipitation studies. Interestingly, the elimination of the TLR4 receptors
restores both the long-term behavior impairments and histone changes induced by
ethanol treatment during the adolescence.
These results support the role of the neuroimmune response and TLR4 signaling in the
epigenetic and behavioral effects of ethanol in the adolescence.
(Supported by ISCIII/FEDER-RTA, RTA-Network RD12-0028-007, SAF2012-33747).
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14. INFLUENCE OF MATERNAL SEPARATION WITH EARLY WEANING
ON THE EXPRESSION OF NEURONAL PLASTICITY IN THE
BASOLATERAL AMYGDALA (BLA). E. Valero, I. García-Rubio, M.L.
Laorden, O. Valverde, M.V. Milanés.
Grupo de Farmacología Celular y Molecular. Facultad de Medicina. Universidad de
Murcia, Murcia; Instituto Murciano de Investigación Biosanitaria (IMIB), Murcia; Grupo
de Neurobiología del Comportamiento (GReNeC), Universitat Pompeu Fabra,
Barcelona; Programa de Investigación de Neurociencia. IMIM (Instituto Hospital del Mar
de Investigaciones Médicas), Barcelona.
Early life experiences induced changes in behavior and brain function. Adverse events
during childhood are associated with increased risk of mood disorders in the adulhood,
such as depression, anxiety and drug addiction. Many studies have shown that
maternal separation, an animal model of early life neglect, can regulate the
hypothalamic-pituitary-adrenal axis (HPA axis) and alter the expression of synaptic
plasticity genes that are critically involved in the establishment of limbic brain circuits
and subsequent brain function and behavior during adulthood. However, the molecular
basis of the long-lasting effects of early life stress on brain function has not been fully
elucidated.
The aim of the present study was to investigate in the basolateral amygdala (BLA), the
influence of the early experiences, such as maternal separation with early weaning or
social interaction, on parameters involved in brain development and plasticity using
conditioned place preference (CPP) paradigm.
Mice were reared in different types of nests: standard nest (SN), maternal separation
with early weaning (MSEW) and communal nest (CN), and were conditioned with 3
mg/Kg or 15 mg/Kg of cocaine when they were adolescent. Present study indicated that
the expression of Arc, norepinephrine (NA) and glucocorticoids receptor (GR) after the
CPP test was modified depending on the type of nest.
These changes along the results of CPP, suggest that glucocorticoids, NA and Arc
might play an important role in processes of memory. In addition, the expression of
brain derived neurotrophic factor (BDNF) decreased in CN, probably because of the
stress for social competencies.
Supported by: MINECO (SAF/FEDER2014-49076); Red de Trastornos Adictivos-ISCIII
(RD12/0028/0003); Fundación Séneca, Agencia Regional de Ciencia y Tecnología
Región de Murcia (15405/PI/10); Instituto Murciano de Investigación Biosanitaria (IMIBHCUVA; GI/IMIB/C040/2011).
48
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15. EXPRESSION OF HEAT SHOCK PROTEIN 27 IN CARDIAC TISSUE
DURING COCAINE WITHDRAWAL. P. Almela, T. Gómez-Morte, B.
Ribeiro, M.V. Milanés, M.L. Laorden.
Department of Pharmacology, Faculty of Medicine, University of Murcia; Department of
Human Anatomy and Psychobiology, Faculty of Psychology, University of Murcia.
Cocaine has been reported to cause similar effects to physiological stressors in the
brain, including heat shock protein (Hsp) expression, although these effects have not
been elucidated in detail.
We have previously demonstrated that morphine withdrawal induces Hsp27 expression
in heart tissue and now hypothesize that spontaneous withdrawal from cocaine could
upregulate this protein expression.
Ascending doses of cocaine hydrochloride (5, 15 and 25 mg/kg) were administered to
mice during 10 days. On day 10 and 1 h after the last cocaine administration, a group of
mice was sacrificed. Other group of animals was killed on day 11, 24 h after the last
cocaine injection. The last group of mice received an acute cocaine injection and was
sacrificed on day 10. Control animals received saline instead of cocaine. Hsp27
expression was evaluated by immunoblotting in right and left ventricle.
Before performing molecular assays, mice weight was checked. Mice treated with
cocaine showed a significantly lower body weight gain than animals receiving saline
injections. Our western blot results show an enhancement in Hsp27 expression in the
left and right ventricle 24 h after the last cocaine injection. However, we did not observe
any change in Hsp27 expression at the 1 h time point or in mice receiving acute
cocaine.
Our findings have significant implications for understanding the cardiac adaptive effects
induced by cocaine treatment and its withdrawal and provide new information into the
possible role of Hsp27 in cardioprotection.
Supported by Red de Trastornos Adictivos-ISCIII (RD12/0028/003), MINECO (Grant
SAF/FEDER 2013-49076-P) and Fundación Séneca (Grant 15405/PI 10).
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16. A TRANSLATIONAL STUDY OF ALCOHOL BINGE IN RATS AND
HUMANS: GENE EXPRESSION PROFILING OF HISTONE
DEACETYLASES IN BLOOD. V. Echeverry-Alzate1, M. Marcos2, J.
Calleja-Conde1, K. M. Bühler1, P. Costa-Alba3, E. Bernardo4, F.J. Laso2,
F. Rodríguez de Fonseca5, R. Nadal6, M. P. Viveros7, R. Maldonado8, E.
Giné9, and J. A. López-Moreno.
Department of Psychobiology, School of Psychology, Campus de Somosaguas,
Complutense University of Madrid, 28223, Madrid, Spain; Alcoholism Unit. Department
of Internal Medicine. University Hospital of Salamanca. Paseo San Vicente 58-186
37007. Salamanca, Spain; Emergency Department. University Hospital of Salamanca.
Paseo San Vicente 58-186 37007. Salamanca, Spain; Department of Vascular Biology
and Inflammation, Fundación Centro Nacional de Investigaciones Cardiovasculares
Carlos III, 28029 Madrid, Spain; Fundación IMABIS, Laboratorio de Medicina
Regenerativa, Hospital Regional Universitario Carlos Haya, 29010, Málaga, Spain;
Psychobiology Unit, School of Psychology, Universitat Autònoma de Barcelona, 08193,
Bellaterra, Barcelona, Spain; Department of Physiology (Animal Physiology II), School
of Biology, Complutense University of Madrid, 28040, Madrid, Spain; Laboratori de
Neurofarmacologia, Departament de Ciències Experimentals i de la Salut, Universitat
Pompeu Fabra, Barcelona, Spain; Department of Cellular Biology, School of Medicine,
Complutense University of Madrid, 28040, Madrid, Spain.
This study aimed to characterize Hdac1-11 gene expression in the rat peripheral blood,
liver, heart, prefrontal cortex and amygdala after repeated alcohol binges, and to
determine the parallelism of Hdac gene expression between rats and humans in
peripheral blood. For this purpose, we examined Hdac gene expression following 1-, 4or 8-alcohol binges (3g/kg) in the rat, in patients who were admitted in the hospital
emergency department for acute alcohol intoxication, and in rats in daily operant alcohol
self-administration. We found that: (a) acute alcohol binge reduced gene expression in
the rat peripheral blood (Hdac1-10), being attenuated this effect after repeated alcohol
binges. In liver, there was also a reduction (Hdac2,3,4) but an increase in heart
(Hdac1,7,8) and amygdala (Hdac1,2,5); (b) there were some correlations in gene
expression among tissues; (c) there was an increase of alcohol concentrations in the
blood 1-4h after repeated alcohol binges, (d) only the 8-alcohol-binges group developed
hepatic steatosis (fatty liver); (e) in humans, alcohol binge increased HDAC gene
expression in peripheral blood (except for HDAC9, 10); and (f) daily operant alcohol
self-administration in rats increased Hdac gene expression in blood similarly to that
observed in human. Our results suggest that an increase in HDAC gene expression in
peripheral blood is associated to an experienced drinker whereas a reduction would be
linked to the first exposures to alcohol.
Acknowledgments
This work was supported by The European Foundation for Alcohol Research (to
J.A.L.M., F.R. de F., R.M, R.N., and M.P.V.), the Fondo de Investigación Sanitaria (Red
de Trastornos Adictivos, FEDER, RD12/0028/0015 to J.A.L.M., RD12/0028/001 to F.R.
de F., RD12/0028/023 to R.M., RD12/0028/0014 to R.N., RD12/0028/1021 to M.P.V.,
RD12/0028/0008 to F.J.L.; and PI10/01692 to M.M.), and Ministerio de Ciencia e
Innovación (SAF2011-26818 to J.A.L.M.).
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17. REPEATED THC ADMINISTRATION AFFECTS STRUCTURAL
PLASTICITY IN THE HIPPOCAMPUS. V. Salgado-Mendialdúa, M.
Gomis-González, R. Maldonado, A. Ozaita.
Facultat de Ciències de la Salut i de la Vida, Universitat Pompeu Fabra, Parc de
Recerca Biomèdica de Barcelona, Barcelona.
Δ9-tetrahydrocannabinol (THC), the main psychoactive component of Cannabis sativa
plant, causes memory impairment through the activation of CB1 cannabinoid receptors.
These amnesic-like effects do not show tolerance after repeated exposure to THC, and
are persistent for a few days after THC withdrawal. Memory formation and consolidation
trigger alterations at excitatory synapses leading to dendritic spine restructuration in
terms of shape and density, a phenomenon called structural plasticity. Structural
plasticity depends on local synaptic regulation of protein synthesis and cytoskeleton rearrangements where actin and its related proteins play an important role. The aim of this
study is to analyze whether repeated exposure to THC has any effect on structural
plasticity in the hippocampus, a brain area involved in memory formation, by using the
Thy1-EGFP transgenic mouse model that expresses enhanced green fluorescent
protein (EGFP) in principal neurons of the brain.
THC sub-chronic administration under conditions that affect object recognition memory
decreased the number of mushroom dendritic spine shape (mature form) and increased
the thin and stubby shapes (immature forms) in the hippocampus. In this brain area
proteins involved in actin cytoskeleton modulations such as myristoylated alanine-rich
C-kinase substrate (MARCKS), neurogranin, cofilin and profilin changed their
expression and/or phosphorylation state after THC treatment.
Our results show that memory impairment produced by chronic THC treatment is
paralleled with hippocampal alterations in structural plasticity and point to the key role
played by actin cytoskeleton and its related proteins in this unwanted side-effect of
THC.
Supported by BFU2012-33500 (MINECO, Spain). V S-MAN-R was supported by a FPI
fellowship (MINECO, Spain).
51
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18. EXPRESSION OF GLS AND GLS2 GLUTAMINASE ISOFORMS IN
ASTROCYTES. C. Cardona, E. Sánchez-Mejías, J.C. Dávila, M. MartínRufián, J.A. Campos-Sandoval, J. Vitorica, F.J. Alonso, J.M. Matés, J.A.
Segura, A. Gutiérrez, J. Márquez.
Departamento de Biología Molecular y Bioquímica, Canceromics Lab. Facultad de
Ciencias, Universidad de Málaga. Instituto de Investigación Biomédica de Málaga
(IBIMA). Dpto. de Biología Celular, Genética y Fisiología. Facultad de Ciencias,
Universidad de Málaga. IBIMA. Centro de Investigación Biomédica en Red sobre
Enfermedades Neurodegenerativas (CIBERNED); Proteomics Lab, Central Facility
Core, Universidad de Málaga; Depto. Bioquímica y Biología Molecular, Facultad de
Farmacia, Universidad de Sevilla. CIBERNED.
Objectives: The expression of glutaminase (GA) in glial cells has been a controversial
issue and matter of debate for many years. Actually, GA is essentially considered as a
neuronal marker in brain. In this work, a comprehensive study was devised to elucidate
expression of GA in neuroglia and, more concretely, in astrocytes.
Material & methods: We employed a combination of complementary approaches,
including immunocytochemistry/immunofluorescence/electron microscopy (EM) with
isoform-specific antibodies, real-time quantitative RT-PCR and in situ activity staining to
check in vivo GA activity.
Results: We demonstrate expression of GLS and GLS2 isoforms in glutamatergic and
GABAergic neurons and astroglial cells. Both GLS (KGA) andGLS2 (GAB/LGA)
isozymes were found to be expressed in cerebral cortex, cerebellar, and hippocampal
astrocytes. Furthermore, a clear segregation was found for each GA isoform, with KGA
appearing only in astrocytic mitochondria while GAB/LGA was preferentially detected in
both nucleus and mitochondria.
Conclusions: GA is no longer a reliable neurochemical marker of glutamatergic
neurons and synapses. Mitochondrial expression of GA isoforms is relevant for both
astrocytic energy metabolism and Glu/Gln cycle while nuclear expression might be
associated with transcriptional regulation. Astrocytes play an integral role in modulating
synaptic transmission and plasticity, both key mechanisms underlying addiction. It
remains to be determined whether inhibition of GA activity/function in mitochondria or
nucleus may contribute to glial cell dysfunction with pathological consequences.
Acknowledgements: Red de Trastornos Adictivos, RTA, (RD12/0028/0013/) RETICS,
ISCIII, y Consejería Innovación, Ciencia y Empresa, Junta de Andalucía (Proyecto de
Excelencia CVI-6656).
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19. DESIGN AND GENERATION OF A GLUTAMINASE Gls2
CONDITIONAL KNOCKOUT MICE. A. Peñalver, M. Tosina, M. MartínRufián, J.A Campos-Sandoval, J.A. Segura, J.M. Matés, M.A. Ramírez,
F.J. Alonso, A. Gutiérrez-Adán, J. Márquez.
Dpto. de Biología Molecular y Bioquímica. Facultad de Ciencias. Universidad de
Málaga. Instituto de Investigación Biomédica (IBIMA); Dpto. de Reproducción Animal.
INIA. Madrid.
Objectives: The objective of the present study is to generate a conditional knockout
(KO) mouse model to elucidate the cerebral function of glutaminase (GA)Gls2 gene.
Material & methods: We achieved modification of the Gls2 mouse gene by gene
targeting since two loxP sites were introduced flanking critical exons. After crossing
these floxed mice with Cre recombinase-expressing mice, we will be able to delete the
gene only in brain cells, as Cre expression is under control of the neuron-specific
synapsin promoter.
Results: First, we obtained murine embryonic stem (ES) mutant cells after
electroporation with the transgenic vector. Then, we obtained chimeric mice containing
the transgenic cassette in the correct mouse gene. After that, we removed the gene-trap
cassette by crossing with Flp-recombinase mice, which reverts the mutation to wild type
(WT) leaving loxP sites on either side of critical exons. Now, we are mating these mice
with Cre-recombinase mice to obtain the brain-specific KO-Gls2 animals.
-/-
Conclusions: Obtaining homozygous Gls2 mice with null expression of Gls2 in brain
will allow us to elucidate the cerebral function of this gene and differentiate it from the
other GA gene (Gls) expressed in brain. We presume this KO mouse would also be a
very useful model for validation of the glutamatergic theory of cocaine addiction.
Acknowledgements: Red de Trastornos Adictivos, RTA, (RD12/0028/0013/) RETICS,
ISCIII, y Consejería Innovación, Ciencia y Empresa, Junta de Andalucía (Proyecto de
Excelencia CVI-6656).
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20. CONSUMO DE ETANOL DURANTE LA ADOLESCENCIA: PAPEL DE
LOS RECEPTORES TLR4 EN LOS EFECTOS REFORZANTES DE LA
COCAÍNA. M. Rodríguez-Arias, C. Guerri, M. Pascual, J. Miñarro.
Unidad de Investigación Psicobiología de las Drogodependencias, Departamento de
Psicobiología, Facultad de Psicología, Universitat de València; Departamento de
Patología Molecular y Celular, Centro de Investigación Príncipe Felipe. Valencia; Red
Temática de Investigación Cooperativa en Salud (RETICS-Trastornos Adictivos),
Instituto de Salud Carlos III, MINECO and FEDER.
Estudios previos han demostrado que el etanol, activando el receptor inmunitario innato
TLR4 en las células gliales induce la liberación de mediadores inflamatorios con el
consecuente daño cerebral. Los receptores TLR ejercen un papel clave en la
regulación del sistema inmune cerebral iniciando una respuesta inmunitaria innata y
adaptativa ante una infección. Utilizando ratones knockout carentes de este receptor,
ya habíamos observado que los efectos perjudiciales sobre la función cognitiva que
induce el consumo de etanol durante la adolescencia están mediados por los
receptores TLR4.
Utilizando igualmente estos ratones modificados genéticamente, en el presente estudio
tratamos de demostrar que los receptores TLR4 también median el incremento de los
efectos reforzantes condicionados de la cocaína tras el consumo de etanol (3 g/kg) en
forma de atracón durante la adolescencia. Tres semanas después de finalizar la
administración de etanol, se realizó un condicionamiento de preferencia de lugar (CPL)
en ratones WT y KO con una dosis no efectiva de cocaína (3 mg/kg).
Confirmando estudios previos, los ratones WT expuestos a etanol durante la
adolescencia, desarrollan CPL y una vez extinguida la preferencia reinstauran dicha
preferencia con dosis de 1,5, 0,75 y 0,375 mg/kg de cocaína. Sin embargo los ratones
KO TLR4 tratados con etanol no desarrollaron preferencia, resultado similar al
observado en los ratones que no recibieron etanol ya fueran WT o KO. Además, los
ratones WT tratados con etanol muestran un perfil anxiogénico en el laberinto elevado
en cruz, perfil que tampoco se observa en los KO TLR4 tratados con etanol ni en los
dos grupos controles.
Nuestros resultados confirman el papel de los receptores TLR4 y sugieren nuevos
tratamientos farmacológicos que puedan normalizar la respuesta inmune responsable
de los efectos a largo plazo del consumo de etanol durante la adolescencia.
Agradecimientos
Ministerio de Economía y Competitividad (MINECO), PSI2011-24762,PSI2014-51847R, Instituto de Salud Carlos III, Red de Trastornos Adictivos (RTA)
RD12/0028/0005,RD12/0028/0007and Unión Europea, Fondos FEDER “una manera de
hacer
Europa”
Generalitat
Valenciana,
Conselleria
de
Educación,
PROMETEOII/2014/063. The European Foundation for Alcohol Research (ERAB),
EA13 08.
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21. LA DERROTA SOCIAL REVIERTE LOS EFECTOS AVERSIVOS
CONDICIONADOSDEL ALCOHOL Y AUMENTA SU CONSUMO EN
RATONES MACHO. M.P. García-Pardo, M. Rodríguez-Arias, J. Miñarro,
M.A. Aguilar.
Unidad de investigación Psicobiología de las Drogodependencias, Departamento de
Psicobiología, Facultad de Psicología, Universitat de Valencia.Red Temática de
Investigación Cooperativa en Salud (RETICS-Trastornos Adictivos), Instituto de Salud
Carlos III, MINECO and FEDER.
Aunque las experiencias estresantes son un factor de riesgo para el abuso de alcohol
en humanos, los estudios en modelos animales han demostrado que la exposición a
estrés social modifica los efectos del alcohol de forma diferencial en función de los
parámetros temporales y los paradigmas empleados para evaluar sus efectos. El
objetivo del presente trabajo fue estudiar los efectos de la exposición a dos
procedimientos diferentes de derrota social (aguda o repetida) en dos paradigmas
usados para evaluar los efectos motivacionales del alcohol (el condicionamiento de
lugar y el “two-bottle choice”) en ratones macho adultos.
En la derrota aguda los animales eran derrotados en un encuentro agonístico en un
área neutral inmediatamente antes de la exposición al alcohol. En la derrota repetida
los animales eran derrotados en un modelo de intruso-residente tres semanas antes de
la exposición al alcohol. Para evaluar los efectos de cada derrota social se usaron seis
grupos de ratones: dos grupos derrotados y dos controles fueron evaluados en el
condicionamiento de lugar (1.25 o 2.5 g/kg de alcohol); un grupo derrotado y otro
control fueron usados para evaluar el consumo voluntario de alcohol.
La dosis alta de alcohol produce aversión condicionada sólo en animales controles.
Además, los animales expuestos a ambos patrones de derrota muestran un incremento
en el consumo de alcohol.
Por tanto, nuestros resultados indican que los efectos del estrés social sobre las
propiedades motivacionales del alcohol en ratones son consistentes utilizando
diferentes paradigmas. El hecho de que la derrota social reduzca los efectos aversivos
del alcohol puede contribuir a que los animales incrementen su consumo. Asimismo
estos resultados sugieren que la exposición al estrés social puede inducir efectos a
corto y largo plazo incrementando la vulnerabilidad de los sujetos a consumir alcohol.
Agradecimientos
Ministerio de Economía y Competitividad (MINECO), Dirección General de
Investigación, PSI2011-24762, PSI2014-51847-R, Instituto de Salud Carlos III, Red de
Trastornos Adictivos (RTA) RD12/0028/0005 and Unión Europea, Fondos FEDER “una
manera de hacer Europa”. Generalitat Valenciana, Conselleria de Educación,
PROMETEOII/2014/063 y VAL+id (MPGP). The European Foundation for Alcohol
Research (ERAB), EA13 08.
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22. HIPOFOLATEMIA
ERITROCITARIA
EN
PACIENTES
CON
TRASTORNO POR USO DE ALCOHOL. P. Zuluaga, A. Sanvisens, F.
Bolao, D. Fuster, I. Rivas, J. Tor, R. Muga.
Servicio de Medicina Interna, Hospital Universitari Germans Trias i Pujol, Universidad
Autónoma de Barcelona, Badalona; Servicio de Medicina Interna, Hospital Universitari
de Bellvitge, Universitat de Barcelona, L’Hospitalet de Llobregat; Centro Delta de
Atención a las Drogodependencias, Institut Municipal de Serveis Personals, Badalona.
Introducción: El déficit de folato es una de las carencias nutricionales características
en pacientes con trastorno por uso de alcohol y se atribuye a dieta pobre, malabsorción
intestinal, alteraciones en la captación y almacenamiento hepático, así como a cambios
en su metabolismo. El objetivo de este estudio es caracterizar la hipofolatemia
eritrocitaria en pacientes con trastorno por uso de alcohol.
Métodos: Estudio transversal en una serie de pacientes que ingresan para tratamiento
del trastorno entre 2007 y 2015. Al ingreso se realizó anamnesis del consumo de
alcohol, antecedentes patológicos, exploración física, datos antropométricos y
extracción de sangre para hemograma, bioquímica y serologías virales. El déficit de
folato eritrocitario se estableció en valores <220 ng/mL. La hepatopatía alcohólica (HA)
se definió mediante criterios clínico-biológicos. Se utilizaron modelos de regresión
logística para establecer factores asociados a hipofolatemia eritrocitaria.
Resultados: 211 pacientes consecutivamente admitidos, (79% H), con una edad de 46
años [RIQ: 40-51 años]; el 38% solicitaban tratamiento por primera vez, 76% iniciaron
el consumo de alcohol <18 años y la cantidad de alcohol ingerida era de 160 g/día
[RIQ: 120-200 g/día]. Globalmente, el 34% de los pacientes presentaba macrocitosis
(VCM >100fl.), 13% anemia (Hb<13 g/dL en H y 12 g/dL en M) y 2,8% anemia
megaloblástica por déficit de folatos. Además, el 17% mostraban hipoalbuminemia
(<35 g/L), 17% infección por VHC y 14% criterios de HA. Hipofolatemia eritrocitaria se
observó en el 18% de los casos y se asoció en el análisis multivariado a macrocitosis
(VCM>100 fl.) (OR=2.2, IC95%: 0.97-5.1) y HA (OR=2.9, IC95%:1.04-8.2). Sin
embargo, tener infección por VHC se asoció a menor probabilidad (OR=0.10
CI95%:0.1-0.81) de presentar hipofolatemia eritrocitaria.
Conclusiones:
Hipofolatemia eritrocitaria es frecuente en el alcoholismo y se asocia a macrocitosis. El
papel de la HA y de la infección por VHC en los depósitos de folato requiere mayor
investigación.
Financiación:
Trabajo parcialmente financiado por el Ministerio de Ciencia e Innovación (RETICS
RD12/0028/0006), Ministerio de Sanidad (PNSD 2014|042).
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23. ALTERACIONES
COMPORTAMENTALES
Y
NEUROINFLAMATORIAS PRODUCIDAS POR EL BLOQUEO
SELECTIVO DE LOS RECEPTORES A2A DE ADENOSINA:
RELEVANCIA EN EL ESTUDIO DE LA ESQUIZOFRENIA. M.
Moscoso-Castro, I. Gracia-Rubio, F. Ciruela, O. Valverde.
Grup de Recerca en Neurobiologia del Comportament (GReNeC), Departament de
Ciències Experimentals i de la Salut, Universitat Pompeu Fabra, Barcelona, España;
Unitat de Farmacologia, Departament Patologia i Terapèutica Experimental, Facultat de
Medicina, IDIBELL, Universitat de Barcelona, L'Hospitalet de Llobregat, España;
Programa de Investigación en Neurociencias. IMIM (Instituto Hospital Mar de
Investigaciones Médicas), Barcelona, España.
La esquizofrenia es un trastorno psiquiátrico crónico, grave y debilitante con un origen
en el desarrollo neurológico y que carece de un tratamiento eficaz. Por otra parte, la
esquizofrenia se asocia con frecuencia con el consumo de drogas de abuso, en
particular nicotina y alcohol. Por lo tanto, la identificación de nuevas dianas
farmacológicas resulta de un gran interés clínico. El nucleósido adenosina modula los
sistemas de dopamina y glutamato, ambos implicados en la fisopatología de la
esquizofrenia y resulta una diana de interés en este contexto. En particular, los
receptores A2A de adenosina, se expresan en regiones cerebrales responsables de
controlar las respuestas emocionales y los procesos cognitivos, incluyendo el estriado,
hipocampo, la corteza cerebral y la glía. Así, el objetivo de nuestro estudio ha sido
evaluaren animales knockout (KO) con inactivación completa y específica del receptor
A2Ael desarrollo de un endofenotipo de esquizofrenia.
Para llevar a cabo nuestro estudio, hemos desarrollado diferentes paradigmas
comportamentales para identificarla presencia de síntomas de tipo esquizofrénico en
ratones macho adulto tanto KO como wild-type (WT).
Nuestros resultados muestran un deterioro general en la filtración de estímulos
sensoriales en los animales KO que fue evaluada por la prueba de inhibición de
prepulso y la inhibición latente. La hiperlocomoción inducida por la administración de
los psicomiméticos d-amfetamina y MK-801 se vio reducida en los animales KO.
Además, los animales A2A mostraron alteraciones motoras y de tipo depresivo,
evaluadas con el rotarod y en la prueba de suspensión de cola respectivamente. La
evaluación cognitiva de los animales demostró la existencia de déficits cognitivos
severos en el laberinto radial y en la prueba de reconocimiento de objetos. Por último,
las alteraciones comportamentales se asociaron con alteraciones en la microglía en
regiones hipocampales de los animales KO.
En resumen, nuestros datos apoyan el papel de la adenosina en la esquizofrenia y
postulan los animales A2A KO como un nuevo modelo de ratón de la enfermedad.
Este estudio ha sido financiado por el Ministerio de Economía e Innovación y FEDER
(SAF2013-41761-R), el Ministerio de de Sanidad, Servicios Sociales e Igualdad (PNSD
015-0020) (Red Temática de Investigación Cooperativa en Salud (ISCIIIFEDER)
(RETIC-Trastornos adictivos RTA, RD12/0028/0024), y la Generalitat de Catalunya
(2014SGR34, 2014SGR14). MM-C ha sido financiada por Retic-ISCIIIRD/12/0028/0024.
57
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24. DYSFUNCTIONAL SOCIAL DECISION-MAKING IN EXCESS WEIGHT
ADULTS. J. Verdejo-Román, J.F. Navas, C. Martín-Pérez, C. RojasGonzález, I. Muela, R. Vilar-López.
Facultad de Psicología y Centro de Investigación Mente, Cerebro y Comportamiento (
Objectives:
According to the World Health Organization (WHO), the prevalence of overweight and
obesity has reached epidemic proportion. Recent neuroscience models posit that
obesity should be conceptualised as a mental disorder involving alterations in
motivation, emotion and decision-making. In this study, we use functional magnetic
resonance imaging (fMRI) to characterize alterations of functional brain activation during
social decision-making in excess weight versus normal weight adults.
Material & methods:
Thirty-nine adults with excess weight (19 with overweight and 20 with obesity) and 39
adults with normal weight were scanned while performing a social decision-making task
(Ultimatum Game). During this task, participants had to accept or reject bids made by
other peer to split monetary stakes. Offers varied in fairness (fair offers: the proposer
shares around 50% of the money; unfair offers: the proposer shares <30% of the
money).In all cases accepting mean both players will be paid, whereas rejecting mean
none of them would get the money. Task related Blood-Oxygen-Level-Dependent
(BOLD) signal during unfair offers compared with fair offers was tested between groups
using Statistical Parametrical Mapping software (SPM8). Correlation analyses were
performed between brain areas activations and body mass index (BMI).
Results:
Excess weight participants accept significantly more unfair offers than normal weight
peers (p=0.011).Excess weight group relative to normal weight, exhibit increased
activation of anterior insula, dorsolateral prefrontal (dlPFC), and anterior cingulate
cortices (ACC)during decision about unfair offers compared to fair offers. Anterior
insula, and dlPFC activations showed significant and positive correlations with BMI
(r=0.356, p=0.001; r=0.303, p=0.007, respectively).
Conclusions:
Excess weight is associated with higher acceptance of unfair offers and increased
activation in emotional perception (anterior insula), cognitive evaluation (dlPFC) and
conflict between emotion and cognition (ACC). These findings suggest dysfunctional
social decision-making circuitry in excess weight group, which increase with severity of
obesity.
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25. SEX-DEPENDENT EFFECTS OF CHRONIC UNPREDIACTABLE
STRESS ON ALCOHOL CONSUMPTION; ANALYSIS OF NEURAL
CORRELATES. E.M. Marco, J.A. Ballesta, M.E. Serrano, C. Irala, M.D.
Hernández, M. López-Gallardo, M.P. Viveros.
Facultad de Biología. Universidad Complutense, Madrid; Facultad de Medicina,
Universidad Complutense, Madrid.
Depression has been frequently associated with alcohol consumption. However, the
temporal relationship between these conditions has not yet been elucidated.
In the present study we aimed to investigate alcohol consumption in an animal model of
depression, analyzing possible neural correlates by western blotting in the frontal cortex
(FCx) and the hippocampal formation (HF). In male and female Wistar rats, exposure to
6 weeks of chronic unpredictable mild stress (CUMS) was employed to induce a
depressive-like phenotype.
As expected, CUMS induced depressive-like responses in the Forced Swimming Test,
i.e. increased immobility time, in animals of both sexes, but did not affect behaviour in
the Sucrose Preference Test or in the Open Field. Decreases in body weight gain and
food intake were only observed among stressed male animals. Higher rates of alcohol
consumption were exclusively observed in CUMS females when compared to control
females, in the absence of differences among males. At least 5 days after the end of the
experimental protocol animals were sacrificed. Male and female animals exposed to
CUMS showed, in the FCx, an increment in GFAP and CB1R expression, together with
a decrease in NCAM expression. In the HF, these animals showed an increase in
PSD95 expression. In addition to these region-dependent effects, sex dependent effects
were also observed. Thus, a decrease in FCx synaptophysin expression and HF CB1R
expression were found in male animals exposed to CUMS, whereas the opposite effects
were observed in females. In turn, hippocampal NCAM expression was increased in
males but decreased in females. A decrease in CB2R expression was only observed in
the HF of stressed females.
In conclusion, CUMS appears to differently affect male and female animals’ behaviour
and the neural parameters analyzed show region and sex dependent effects on
synaptic plasticity markers.
Acknowledgements: Instituto de Salud “Carlos III” (FIS), RETICS, Red de Trastornos
Adictivos (RD2012/0028/0021). GRUPOS UCM-BSCH: Ref UCM 951579.
59