Consuelo Guerri

Foetal Alcohol Spectrum Disorders and Alterations in Brain and Behaviour

The term Foetal Alcohol Spectrum Disorders (FASD) refers to the range of disabilities that may result from prenatal alcohol exposure. This article reviews the effects of ethanol on the developing brain and its long-term structural and neurobehavioural consequences. Brain imaging, neurobehavioural and experimental studies demonstrate the devastating consequences of prenatal alcohol exposure on the developing central nervous system (CNS), identifying specific brain regions affected, the range of severity of effects and mechanisms involved. In particular, neuroimaging studies have demonstrated overall and regional volumetric and surface area reductions, abnormalities in the shape of particular brain regions, and reduced and increased densities for white and grey matter, respectively. Neurobehaviourally, FASD consists of a continuum of long-lasting deficits affecting multiple aspects of cognition and behaviour. Experimental studies have also provided evidence of the vulnerability of the CNS to the teratogenic effects of ethanol and have provided new insight on the influence of risk factors in the type and severity of observed brain abnormalities. Finally, the potential molecular mechanisms that underlie the neuroteratological effects of alcohol are discussed, with particular emphasis on the role of glial cells in long-term neurodevelopmental liabilities.

Effects of prolonged ethanol exposure on the glial fibrillary acidic protein-containing intermediate filaments of astrocytes in primary culture: a quantitative immunofluorescence and immunogold electron microscopic study.

We investigated the effects of ethanol exposure on the shape of the cell and the morphology of intermediate filaments (IF) of cortical astrocytes in primary culture. The content and distribution of glial fibrillary acidic protein (GFAP), the major component of glial IF, was assessed using an anti-GFAP monoclonal antibody and fluorescence scanning densitometry together with quantitative pre- and post-embedding immunogold electron microscopy. The astrocytes were from 21-day-old fetuses obtained from both control and chronic alcoholic rats and were cultured for 28 days in the absence or presence of ethanol (25 mM). The main findings were: (a) ethanol-exposed astrocytes failed to develop processes or to acquire a filamentous IF distribution pattern; (b) these cells showed less GFAP than astrocytes without alcohol; (c) ethanol interfered with the reorganization of the anti-GFAP binding sites from clustered to random; and (d) astrocytes from alcohol-exposed fetuses cultured in the absence of ethanol also showed these alterations, suggesting initial damage to astrocyte precursor cells. Since the glial filaments play a crucial role in creating a scaffolding that guides neuronal migration, the effect of ethanol on astrocyte IF may possibly be correlated with the mechanisms underlying mental retardation and motor dysfunction which are characteristics of fetal alcohol syndrome.

Neuroimmune Activation and Myelin Changes in Adolescent Rats Exposed to High-Dose Alcohol and Associated Cognitive Dysfunction: A Review with Reference to Human Adolescent Drinking

AIMSnThe aim of the study was to assess whether intermittent ethanol administration to adolescent rats activates innate immune response and TLRs signalling causing myelin disruption and long-term cognitive and behavioural deficits.nnnMETHODSnWe used a rat model of intermittent binge-like ethanol exposure during adolescence.nnnRESULTSnBinge-like ethanol administration to adolescent rats increased the gene expression of TLR4 and TLR2 in the prefrontal cortex (PFC), as well as inflammatory cytokines TNFα and IL-1β. Up-regulation of TLRs and inflammatory mediators were linked with alterations in the levels of several myelin proteins in the PFC of adolescent rats. These events were associated with previously reported long-term cognitive dysfunctions. Conversely, the same ethanol treatment did not cause significant changes in either inflammatory mediators or myelin changes in the brain of adult rats.nnnCONCLUSIONnActivation of innate immune receptors TLRs in the PFC appears to be involved in the neuroinflammation and demyelination processes induced by ethanol exposure during adolescence. The findings support the vulnerability of the juvenile brain to the effects of ethanol and the long-term cognitive consequences of binge drinking. In addition, ethanol-induced PFC dysfunctions might underlie the propensity of adolescents for impulsivity and to ignore the negative consequences of their behaviour, both of which could increase the risk of substance abuse.

Evolution of several cytoskeletal proteins of astrocytes in primary culture: Effect of prenatal alcohol exposure

In the present work we have analyzed, using immunoblotting and immunofluorescence techniques, the evolution of several cytoskeletal proteins during the development of astrocytes in primary culture. The effect of prenatal exposure to alcohol on these proteins was also evaluated. Microtubular protein α-tubulin decreased approximately 47% from 4 to 7 days after which its content remained practically constant. Immunofluorescence studies showed also that the content of α-tubulin was greater at day 4 of culture. This increase in fluorescence was coincident with the presence of globular particles which were found in interphase astrocytes and stained with both anti α- and anti-β-tubulin. These structures appeared only in proliferating cells. Glial fibrillary acidic protein (GFAP) and vimentin were analyzed as intermediate filament (IF) proteins. GFAP, in cytoskeletal preparations, increased regularly for 14 days followed by a decrease to day 21. In contrast, vimentin showed a progressive increase throughout the entire culture period. Fluorescence studies revealed some differences between the IF distribution patterns of GFAP and vimentin.In astrocytes obtained from rats prenatally exposed to ethanol, decreases in the amounts of all the cytoskeletal proteins studied were found during the entire culture period. In these cells a striking disorganization of cytoskeleton was also observed. The alcohol-induced decrease of GFAP in cultured astrocytes was also found when this protein was studied in preparations from whole brain developed “in vivo”.

Plasma profile of pro-inflammatory cytokines and chemokines in cocaine users under outpatient treatment: influence of cocaine symptom severity and psychiatric co-morbidity.

The treatment for cocaine use constitutes a clinical challenge because of the lack of appropriate therapies and the high rate of relapse. Recent evidence indicates that the immune system might be involved in the pathogenesis of cocaine addiction and its co‐morbid psychiatric disorders. This work examined the plasma pro‐inflammatory cytokine and chemokine profile in abstinent cocaine users (nu2009=u200982) who sought outpatient cocaine treatment and age/sex/body mass‐matched controls (nu2009=u200965). Participants were assessed with the diagnostic interview Psychiatric Research Interview for Substance and Mental Diseases according to the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, Text Revision (DSM‐IV‐TR). Tumor necrosis factor‐alpha, chemokine (C‐C motif) ligand 2/monocyte chemotactic protein‐1 and chemokine (C‐X‐C motif) ligand 12 (CXCL12)/stromal cell‐derived factor‐1 (SDF‐1) were decreased in cocaine users, although all cytokines were identified as predictors of a lifetime pathological use of cocaine. Interleukin‐1 beta (IL‐1β), chemokine (C‐X3‐C motif) ligand 1 (CX3CL1)/fractalkine and CXCL12/SDF‐1 positively correlated with the cocaine symptom severity when using the DSM‐IV‐TR criteria for cocaine abuse/dependence. These cytokines allowed the categorization of the outpatients into subgroups according to severity, identifying a subgroup of severe cocaine users (9–11 criteria) with increased prevalence of co‐morbid psychiatric disorders [mood (54%), anxiety (32%), psychotic (30%) and personality (60%) disorders]. IL‐1β was observed to be increased in users with such psychiatric disorders relative to those users with no diagnosis. In addition to these clinical data, studies in mice demonstrated that plasma IL‐1β, CX3CL1 and CXCL12 were also affected after acute and chronic cocaine administration, providing a preclinical model for further research. In conclusion, cocaine exposure modifies the circulating levels of pro‐inflammatory mediators. Plasma cytokine/chemokine monitoring could improve the stratification of cocaine consumers seeking treatment and thus facilitate the application of appropriate interventions, including management of heightened risk of psychiatric co‐morbidity. Further research is necessary to elucidate the role of the immune system in the etiology of cocaine addiction.

Prenatal exposure to alcohol alters the Golgi apparatus of newborn rat hepatocytes: a cytochemical study.:

The effect of prenatal exposure to ethanol on the Golgi apparatus of newborn rat hepatocytes has been studied cytochemically using several trans-Golgi markers (thiamine pyrophosphatase, uridine diphosphatase, inosine diphosphatase, acid phosphatase, and 5-nucleotidase) as well as a cis-side marker (osmium impregnation). The amount of cerium phosphate formed in the cytochemical reactions was roughly quantitated by stereologic methods. The Golgi apparatus of about 40% of the hepatocytes appeared disorganized after alcohol treatment, and in the other 60%, the electron density of reaction product deposits for all phosphatases investigated was decreased. 5-Nucleotidase was completely absent in cisternae of Golgi apparatus of treated cells. In control cells impregnated with osmium tetroxide, reduced osmium compounds were observed in most Golgi cisternae and in nearby vesicles. In contrast, only small vesicles appeared positive in treated hepatocytes. These results suggest that prenatal alcohol exposure alters some Golgi functions. Thus, the decrease in nucleoside diphosphatase and 5-nucleotidase cytochemical activities after ethanol exposure strongly suggests that this treatment could affect glycosylation in the Golgi apparatus of newborn rat hepatocytes.

Intermittent ethanol exposure increases long-lasting behavioral and neurochemical effects of MDMA in adolescent mice

RationaleHeavy binge drinking is increasingly frequent among adolescents, while ethanol (EtOH) is often used in combination with 3,4-methylenedioxymethamphetamine (MDMA).ObjectivesThe long-lasting effects of intermittent exposure to EtOH and MDMA during adolescence on motor activity, anxiety, and social behavior were evaluated in adult mice. The concentration of brain monoamines in the striatum, cortex, and hippocampus was measured following the behavioral test.MethodsAdolescent OF1 mice were exposed to ethanol (1.25xa0g/kg) on two consecutive days at 48-h intervals over a 14-day period (from PND 29 to 42). A total of eight injections of MDMA (10 or 20xa0mg/kg) were administered twice daily at 4-h intervals over two consecutive days, and this schedule was repeated 6xa0days later (PND 33, 34, 41, and 42). Behavioral tests and analysis of brain monoamines took place on PND 64 to 67.ResultsExposure to MDMA during adolescence increased the anxiogenic response in the elevated plus maze, with adult mice spending less time in the open arms of the maze and exhibiting lower concentrations of DA in the striatum. A pattern of ethanol administration modeling binge drinking during adolescence enhanced these effects and undermined the hyperthermic response induced by MDMA. Passive avoidance was affected only when EtOH was administered alone.ConclusionsJuvenile administration of MDMA and alcohol was found to cause a decrease in monoamine levels in adulthood, as well as changes in social interaction behaviors, locomotor activity, increase measures of anxiety in the elevated plus maze (EPM), and decrease step-through latencies in passive avoidance test.

Fetal alcohol effects : Potential treatments from basic science

This article represents the proceedings of a symposium presented at the 2004 annual meeting of the International Society for Biomedical Research on Alcoholism, held in Mannheim, Germany. The presentations were as follows: 1) “Antioxidants Prevent Ethanol-Induced Cell Death in Developing Brain and in Cultured Neural Cells” by M. Pascual, M. C. Garcia-Minguillan, and Consuelo Guerri; 2) “Rational Development of Ethanol Antagonists” by Michael E. Charness and Michael F. Wilkemeyer; 3) “Choline Supplementation as a Treatment for Fetal Alcohol Effects” by Jennifer D. Thomas and Hector D. Dominguez; 4) “Cerebellar and Cortical Plasticity After Neonatal Alcohol Exposure: Model of Intervention” by Anna Y. Klintsova, Charles R. Goodlett, and William T. Greenough; and 5) “Circadian Rhythms in Prenatally Ethanol-Exposed Rats” by Hiromi Sakata-Haga.

SY13-3IS THE NEUROIMMUNE RESPONSE INVOLVED IN FETAL ALCOHOL SPECTRUM DISORDERS?

Inflammation during brain development is involved in the pathogenesis of early brain injury and cognitive dysfunctions. We reported that ethanol can activate the TLR4 immune receptors causing neuroinflammation, myelin dysfunction, brain damage and cognitive effects in adults and adolescents. However, it is presently unknown the potential role of neuroimmune-TLR4 response in the …

P-74AUTOPHAGY CONSTITUTES A PROTECTIVE MECHANISM AGAINST ETHANOL TOXICITY IN MOUSE GLIAL CELLS

Ethanol induces brain damage and neurodegeneration by triggering inflammatory processes in glial cells through the activation of Toll-like receptor 4 (TLR4) signaling. Recent evidence indicates the role of protein degradation pathways in neurodegeneration and in alcoholic liver disease, but how these processes affect the brain remains elusive. We have recently …