Effects of maternal binge alcohol consumption on emotional, cognitive and addictive behaviour in mice

Maternal alcohol binge drinking during pregnancy can be deleterious for the developing foetus, leading to a wide range of long-lasting morphological and neurobehavioural disabilities known as foetal alcohol spectrum disorders, associated with a higher risk of developing substance use disorders later in life. We sought to assess the effects of prenatal and postnatal alcohol exposure on cognitive, emotional, motor and addictive behaviour in mice and its underlying molecular mechanisms. Pregnant C57BL/6 female mice underwent a procedure to model alcohol binge drinking either during gestation or throughout both the gestation and lactation periods. Then, male offspring were assessed for their behaviour at adulthood. Binge alcohol exposure during early brain development induces cognitive deficits, increased anxiety-like behaviour, motor coordination impairments, and age-dependent locomotor activity alterations. Behavioural effects are associated with an upregulation of pro-inflammatory signalling, gliosis, neuronal death, myelin impairments and epigenetic modifications in the prefrontal cortex and hippocampus. Furthermore, early alcohol exposed mice show alterations in brain network connectivity. Curcumin treatment ameliorates anxiety and cognitive dysfunctions, and rescues alcohol-induced neuroinflammation. In addition, mice exposed to alcohol in utero and postnatally show increased susceptibility to later alcohol and cocaine intake compared with their counterparts. Molecular analyses of the prefrontal cortex and striatum of these animals suggest alterations in the glutamatergic excitability within the mesocorticolimbic reward system following cocaine-induced reinstatement. Altogether, our results reveal that maternal binge-like alcohol consumption induces molecular alterations in offspring’s brain that may underlie the long-lasting impairments in offspring’s behaviour.El consum maternal d’alcohol en afartament durant l’embaràs pot resultar perjudicial per al fetus en desenvolupament, donant lloc a una àmplia gamma de discapacitats físiques i mentals conegudes com a trastorns de l’espectre alcohòlic fetal que persisteixen al llarg de la vida i estan associades a un major risc de desenvolupar trastorns d’ús de substàncies en el futur. En aquesta tesi hem tractat d’avaluar els efectes de l’exposició prenatal i postnatal a l’alcohol en la conducta cognitiva, emocional, motora i addictiva en ratolins i els mecanismes moleculars subjacents a aquests. Les femelles de ratolins C57BL/6 embarassades van ser sotmeses a un procediment per modelar el consum d’alcohol en afartament durant la gestació o bé, al llarg dels períodes de gestació i lactància. A continuació es va avaluar el comportament de la descendència masculina a l’edat adulta. S’ha observat que l’exposició d’alcohol en afartament durant el desenvolupament cerebral indueix dèficits cognitius, augment de l’ansietat, alteracions de coordinació motora i de l’activitat locomotora en funció de l’edat. Els efectes del comportament estan associats a un increment de la senyalització proinflamatòria, gliosi, mort neuronal, deteriorament de la mielina i modificacions epigenètiques en el còrtex prefrontal i l’hipocamp, així com també alteracions en la connectivitat de la xarxa neuronal. El tractament de curcumina alleuja l’ansietat i les disfuncions cognitives, i restableix la neuroinflamació induïda per l’alcohol. A més, els ratolins exposats a l’alcohol durant la gestació i la lactància mostren una major susceptibilitat a la ingesta posterior d’alcohol i cocaïna en comparació amb els seus homòlegs. Els anàlisis moleculars de l’escorça prefrontal i de l’estriat d’aquests animals suggereixen la presència d’alteracions en l’excitabilitat glutamatèrgica en el sistema de recompensa mesocorticolimbic després de la recaiguda induïda per cocaïna. En conjunt, els nostres resultats indiquen que el consum maternal d’alcohol en afartament provoca alteracions moleculars en el cervell de la descendència com a mecanisme subjacent a les alteracions relatives al comportament persistents

Neuroinflammatory and behavioral susceptibility profile of mice exposed to social stress towards cocaine effects

Using the social defeat (SD) model, numerous studies have shown that stressed mice display an enhanced response to the motivational effects of cocaine in the self-administration (SA) and conditioned-place preference (CPP) paradigms. However, not all subjects exposed to stress express its harmful effects. Some are particularly susceptible to the deleterious effects of repeated SD, while resilient mice successfully cope with stressful experiences and display adjusted psychological functioning after stress. Vulnerability to develop stress-related disorders, such as depression, has been linked to coping strategies and more recently to individual differences in the immune system. However, no studies have evaluated if coping strategies and immune system reactivity to social stress experiences can also predict susceptibility to stress-induced enhancement of the cocaine response. We evaluated cocaine response in socially defeated mice in the CPP and SA paradigms. To evaluate neuroimmune reactivity to stress the pro-inflammatory cytokine IL-6 and the chemokine CX3CL1 were measured in the striatum and hippocampus. Behavioral phenotype during and after SD episodes was also evaluated. Our results showed that susceptible mice to the depressive-like behaviors effects of stress showed increased conditioned rewarding effects of cocaine in the CPP. In addition, susceptible mice displayed passive-reactive coping behavior during social stress episodes and more pronounced changes in neuroinflammatory markers after the last SD episode, which lasted for one month. Although the complex mechanisms underlying susceptibility or resilience to social stress are still unclear, our results point to multiple adaptive stress responses expressed at different phenotypic levels

Effects of High-Fat Diet and Maternal Binge-Like Alcohol Consumption and Their Influence on Cocaine Response in Female Mice Offspring

Backgroud: Prenatal alcohol exposure is a leading cause of neurobehavioral and neurocognitive deficits collectively known as fetal alcohol spectrum disorders (FASD), including eating disorders and increased risk for substance abuse as very common issues. In this context, the present study aimed to assess the interaction between alcohol exposure during gestation and lactation periods (PLAE) and a high fat diet (HFD) during childhood and adolescence. Methods: Pregnant C57BL/6 mice underwent a procedure for alcohol binge drinking during gestation and lactation periods. Subsequently, PLAE female offspring were fed with a HFD for 8 weeks and thereafter, nutrition-related parameters as well as their response to cocaine were assessed. Results: In our model, feeding young females with a HFD increased their triglyceride blood levels but did not induce an overweight compared to those fed with a standard diet. Moreover, PLAE affected how females responded to the fatty diet as they consumed less amount of food than water-exposed offspring, consistent with a lower gain of body weight. HFD increased the psychostimulant effects of cocaine. Surprisingly, PLAE reduced the locomotor responses to cocaine without modifying cocaine-induced reward. Moreover, PLAE prevented the striatal overexpression of cannabinoid 1 receptors induced by a HFD and induced an alteration of myelin damage biomarker in the prefrontal cortex, an effect that was mitigated by a HFD-based feeding. Conclusion: Therefore, in female offspring, some effects triggered by one of these factors, PLAE or a HFD, were blunted by the other, suggesting a close interaction between the involved mechanisms

Effects of maternal binge alcohol consumption on emotional, cognitive and addictive behaviour in mice

Maternal alcohol binge drinking during pregnancy can be deleterious for the developing foetus, leading to a wide range of long-lasting morphological and neurobehavioural disabilities known as foetal alcohol spectrum disorders, associated with a higher risk of developing substance use disorders later in life. We sought to assess the effects of prenatal and postnatal alcohol exposure on cognitive, emotional, motor and addictive behaviour in mice and its underlying molecular mechanisms. Pregnant C57BL/6 female mice underwent a procedure to model alcohol binge drinking either during gestation or throughout both the gestation and lactation periods. Then, male offspring were assessed for their behaviour at adulthood. Binge alcohol exposure during early brain development induces cognitive deficits, increased anxiety-like behaviour, motor coordination impairments, and age-dependent locomotor activity alterations. Behavioural effects are associated with an upregulation of pro-inflammatory signalling, gliosis, neuronal death, myelin impairments and epigenetic modifications in the prefrontal cortex and hippocampus. Furthermore, early alcohol exposed mice show alterations in brain network connectivity. Curcumin treatment ameliorates anxiety and cognitive dysfunctions, and rescues alcohol-induced neuroinflammation. In addition, mice exposed to alcohol in utero and postnatally show increased susceptibility to later alcohol and cocaine intake compared with their counterparts. Molecular analyses of the prefrontal cortex and striatum of these animals suggest alterations in the glutamatergic excitability within the mesocorticolimbic reward system following cocaine-induced reinstatement. Altogether, our results reveal that maternal binge-like alcohol consumption induces molecular alterations in offspring’s brain that may underlie the long-lasting impairments in offspring’s behaviour.El consum maternal d’alcohol en afartament durant l’embaràs pot resultar perjudicial per al fetus en desenvolupament, donant lloc a una àmplia gamma de discapacitats físiques i mentals conegudes com a trastorns de l’espectre alcohòlic fetal que persisteixen al llarg de la vida i estan associades a un major risc de desenvolupar trastorns d’ús de substàncies en el futur. En aquesta tesi hem tractat d’avaluar els efectes de l’exposició prenatal i postnatal a l’alcohol en la conducta cognitiva, emocional, motora i addictiva en ratolins i els mecanismes moleculars subjacents a aquests. Les femelles de ratolins C57BL/6 embarassades van ser sotmeses a un procediment per modelar el consum d’alcohol en afartament durant la gestació o bé, al llarg dels períodes de gestació i lactància. A continuació es va avaluar el comportament de la descendència masculina a l’edat adulta. S’ha observat que l’exposició d’alcohol en afartament durant el desenvolupament cerebral indueix dèficits cognitius, augment de l’ansietat, alteracions de coordinació motora i de l’activitat locomotora en funció de l’edat. Els efectes del comportament estan associats a un increment de la senyalització proinflamatòria, gliosi, mort neuronal, deteriorament de la mielina i modificacions epigenètiques en el còrtex prefrontal i l’hipocamp, així com també alteracions en la connectivitat de la xarxa neuronal. El tractament de curcumina alleuja l’ansietat i les disfuncions cognitives, i restableix la neuroinflamació induïda per l’alcohol. A més, els ratolins exposats a l’alcohol durant la gestació i la lactància mostren una major susceptibilitat a la ingesta posterior d’alcohol i cocaïna en comparació amb els seus homòlegs. Els anàlisis moleculars de l’escorça prefrontal i de l’estriat d’aquests animals suggereixen la presència d’alteracions en l’excitabilitat glutamatèrgica en el sistema de recompensa mesocorticolimbic després de la recaiguda induïda per cocaïna. En conjunt, els nostres resultats indiquen que el consum maternal d’alcohol en afartament provoca alteracions moleculars en el cervell de la descendència com a mecanisme subjacent a les alteracions relatives al comportament persistents

The pharmacological reduction of hippocampal neurogenesis attenuates the protective effects of cannabidiol on cocaine voluntary intake

The administration of cannabidiol has shown promising evidence in the treatment of some neuropsychiatric disorders, including cocaine addiction. However, little information is available as to the mechanisms by which cannabidiol reduces drug use and compulsive seeking. We investigated the role of adult hippocampal neurogenesis in reducing cocaine voluntary intake produced by repeated cannabidiol treatment in mice. Cocaine intake was modelled using the intravenous cocaine self-administration procedure in CD1 male mice. Cannabidiol (20 mg/kg) reduced cocaine self-administration behaviour acquisition and total cocaine intake and enhanced adult hippocampal neurogenesis. Our results show that a 6-day repeated temozolomide treatment (25 mg/kg/day), a chemotherapy drug that blocks hippocampal neurogenesis, prevented cannabidiol-induced increment in the early stages of neuronal maturation and differentiation, without altering the basal levels of BrdU/NeuN and doublecortin immunostaining. The reduction of total cocaine intake and operant behaviour acquisition observed following cannabidiol exposure was attenuated by temozolomide treatment. Our results also show a similar effect of temozolamide on a cannabidiol-induced improvement of novel object recognition memory, a task influenced by the proneurogenic effects of cannabidiol (10 and 20 mg/kg). The anxiolytic effects of cannabidiol (10 and 20 mg/kg), however, remained unaffected after its proneurogenic effects decreased. The present study confirms that adult hippocampal neurogenesis is one of the mechanisms by which cannabidiol lowers cocaine reinforcement and demonstrates the functional implication of adult hippocampal neurogenesis in cocaine voluntary consumption in mice. Such findings highlight the possible use of cannabidiol for developing new pharmacotherapies to manage cocaine use disorders.This work was supported by Ministerio de Economía y Competitividad (grant number SAF2016-75966-R-FEDER), by the European Union's Horizon 2020 research andinnovation programme 2014–2020 under grant agreement no 634143. M.A.L. received FPU grant (15/02492) from the Ministeriode Educacion, Cultura y Deporte. L.C.received FPI grant(BES-2014-070657) from the Ministerio de Economía y Competitividad.The Department of Experimental and Health Sciences (UPF) is an“Unidad de Excelencia María de Maeztu” funded by the MINECO (Ref. MDM-2014-0370).The authors thank Gerald-Patrick Fannon for his English proofreading and editing of the manuscript, and Phytoplant Research S.L. for providing CBD compound.The authors declare no conflicts of interest. AUTHORS CONTRIBUTION M.A.L. and O.V. were responsible for the study concept and design.M.A.L .and L.C carried out the experimental studies. M.A.L. and O.V.drafted the manuscript and editing of the manuscript, and Phytoplant Research S.L. for providing CBD compound.The authors declare no conflicts of interest

Histone deacetylases inhibitor trichostatin A reverses anxiety-like symptoms and memory impairments induced by maternal binge alcohol drinking in mice

Background: Alcohol exposure during development has detrimental effects, including a wide range of physical, cognitive and neurobehavioural anomalies known as foetal alcohol spectrum disorders. However, alcohol consumption among pregnant woman is an ongoing latent health problem. Aim: In the present study, the effects of trichostatin A (TSA) on emotional and cognitive impairments caused by prenatal and lactational alcohol exposure were assessed. TSA is an inhibitor of class I and II histone deacetylases enzymes (HDAC), and for that, HDAC4 activity was determined. We also evaluated mechanisms underlying the behavioural effects observed, including the expression of brain-derived neurotrophic factor (BDNF) in discrete brain regions and newly differentiated neurons in the dentate gyrus (DG). Methods: C57BL/6 female pregnant mice were used, with limited access to a 20% v/v alcohol solution as a procedure to model binge alcohol drinking during gestation and lactation. Male offspring were treated with TSA during the postnatal days (PD28-35) and behaviourally evaluated (PD36-55). Results: Early alcohol exposure mice presented increased anxiogenic-like responses and memory deterioration - effects that were partially reversed with TSA. Early alcohol exposure produces a decrease in BDNF levels in the hippocampus (HPC) and prefrontal cortex, a reduction of neurogenesis in the DG and increased activity levels of the HDAC4 in the HPC. Conclusions: Such findings support the participation of HDAC enzymes in cognitive and emotional alterations induced by binge alcohol consumption during gestation and lactation and would indicate potential benefits of HDAC inhibitors for some aspects of foetal alcohol spectrum disorders.The author(s) disclosed receipt of the following financial support for the research, authorship and/or publication of this article: This work was supported by the Ministerio de Economía y Competitividad (Spanish Ministry of Economy and Competitiveness – grant number SAF2016-75966-R-FEDER), and the Spanish Ministry of Health (Retic-ISCIII-RD16/0017/0010 and PNSD 2018/007). SM-R received a postdoctoral fellowship from the Conselleria d’Educació, Investigació, Cultura i Esport (APOSTD/2017/102), Generalitat Valenciana, Spain. LC received an FPI grant (BES-2014-070657) from the Ministerio de Economía y Competitividad, The Department of Experimental and Health Sciences (UPF), ‘Unidad de Excelencia María de Maeztu’ funded by the MINECO (ref. MDM-2014-0370)

Gut-derived peptide hormone receptor expression in the developing mouse hypothalamus

OBJECTIVE: In adult organisms, a number of receptors have been identified which modulate metabolic processes related to peptides derived from the intestinal tract. These receptors play significant roles in glucose homeostasis, food intake and energy balance. Here we assess these classical metabolic receptors and their expression as well as their potential role in early development of hypothalamic neuronal circuits. METHODS: Chow-fed C57BL6/N female mice were mated and hypothalamic tissue was collected from offspring across postnatal development (postnatal day 7–21). Subsequent qPCR and Western Blot analyses were used to determine mRNA and protein changes in gut-derived peptide hormone receptors. Correlations to body weight, blood glucose and circulating leptin levels were analyzed. RESULTS: We describe the gene expression and dynamic protein regulation of key gut-derived peptide hormone receptors in the early postnatal period of the mouse brain. Specifically, we show changes to Gastric inhibitory polypeptide receptor (GIPR), glucagon-like peptide 1 receptor (GLP1R), and cholecystokinin receptor 2 (CCK2R) in the developing hypothalamus. The changes to GIPR and InsR seem to be strongly negatively correlated with body weight. CONCLUSIONS: This comprehensive analysis underscores the need to understand the roles of maternal-derived circulating gut hormones and their direct effect on offspring brain development

Altered brain functional connectivity and behaviour in a mouse model of maternal alcohol binge-drinking

Prenatal and perinatal alcohol exposure caused by maternal alcohol intake during gestation and lactation periods can have long-lasting detrimental effects on the brain development and behaviour of offspring. Children diagnosed with Foetal Alcohol Spectrum Disorders (FASD) display a wide range of cognitive, emotional and motor deficits, together with characteristic morphological abnormalities. Maternal alcohol binge drinking is particularly harmful for foetal and early postnatal brain development, as it involves exposure to high levels of alcohol over short periods of time. However, little is known about the long-term effects of maternal alcohol binge drinking on brain function and behaviour. To address this issue, we used pregnant C57BL/6 female mice with time-limited access to a 20% v/v alcohol solution as a procedure to model alcohol binge drinking during gestation and lactational periods. Male offspring were behaviourally tested during adolescence (30 days) and adulthood (60 days), and baseline neural metabolic capacity of brain regions sensitive to alcohol effects were also evaluated in adult animals from both groups. Our results show that prenatal and postnatal alcohol exposure caused age-dependent changes in spontaneous locomotor activity, increased anxiety-like behaviour and attenuated alcohol-induced conditioned place preference in adults. Also, significant changes in neural metabolic capacity using cytochrome c oxidase (CCO) quantitative histochemistry were found in the hippocampal dentate gyrus, the mammillary bodies, the ventral tegmental area, the lateral habenula and the central lobules of the cerebellum in adult mice with prenatal and postnatal alcohol exposure. In addition, the analysis of interregional CCO activity correlations in alcohol-exposed adult mice showed disrupted functional brain connectivity involving the limbic, brainstem, and cerebellar regions. Finally, increased neurogenesis was found in the dentate gyrus of the hippocampus of alcohol-exposed offspring, suggesting neuroadaptive effects due to early alcohol exposure. Our results demonstrate that maternal binge-like alcohol drinking causes long-lasting effects on motor and emotional-related behaviours associated with impaired neuronal metabolic capacity and altered functional brain connectivity.This study was supported by grants from the EuropeanUnion's Horizon 2020 research and innovation programme 2014-2020under Grant Agreement No 634143, the Spanish Ministry of Economy,Innovation and Competitiveness (SAF2016-75966-R), the SpanishMinistryof Education and Science and Innovation and EuropeanRegional Development Fund (PSI2013-45924-P, PSI2015-73111-EXP, PSI2017-83038-P, PSI2017-83893-R, the Plan Nacional sobre Drogas(#2014/020), and the Plan de Ciencia, Tecnología e Innovación delPrincipado de Asturias (FC-15-GRUPIN14-088). Department ofExperimental and Health Sciences is a“Unidad de Excelencia María deMaeztu”funded by the MINECO (MDM-2014-0370). The authors de-clare no potential conflicts of interest

Altered brain functional connectivity and behaviour in a mouse model of maternal alcohol binge-drinking

Prenatal and perinatal alcohol exposure caused by maternal alcohol intake during gestation and lactation periods can have long-lasting detrimental effects on the brain development and behaviour of offspring. Children diagnosed with Foetal Alcohol Spectrum Disorders (FASD) display a wide range of cognitive, emotional and motor deficits, together with characteristic morphological abnormalities. Maternal alcohol binge drinking is particularly harmful for foetal and early postnatal brain development, as it involves exposure to high levels of alcohol over short periods of time. However, little is known about the long-term effects of maternal alcohol binge drinking on brain function and behaviour. To address this issue, we used pregnant C57BL/6 female mice with time-limited access to a 20% v/v alcohol solution as a procedure to model alcohol binge drinking during gestation and lactational periods. Male offspring were behaviourally tested during adolescence (30 days) and adulthood (60 days), and baseline neural metabolic capacity of brain regions sensitive to alcohol effects were also evaluated in adult animals from both groups. Our results show that prenatal and postnatal alcohol exposure caused age-dependent changes in spontaneous locomotor activity, increased anxiety-like behaviour and attenuated alcohol-induced conditioned place preference in adults. Also, significant changes in neural metabolic capacity using cytochrome c oxidase (CCO) quantitative histochemistry were found in the hippocampal dentate gyrus, the mammillary bodies, the ventral tegmental area, the lateral habenula and the central lobules of the cerebellum in adult mice with prenatal and postnatal alcohol exposure. In addition, the analysis of interregional CCO activity correlations in alcohol-exposed adult mice showed disrupted functional brain connectivity involving the limbic, brainstem, and cerebellar regions. Finally, increased neurogenesis was found in the dentate gyrus of the hippocampus of alcohol-exposed offspring, suggesting neuroadaptive effects due to early alcohol exposure. Our results demonstrate that maternal binge-like alcohol drinking causes long-lasting effects on motor and emotional-related behaviours associated with impaired neuronal metabolic capacity and altered functional brain connectivity.This study was supported by grants from the EuropeanUnion's Horizon 2020 research and innovation programme 2014-2020under Grant Agreement No 634143, the Spanish Ministry of Economy,Innovation and Competitiveness (SAF2016-75966-R), the SpanishMinistryof Education and Science and Innovation and EuropeanRegional Development Fund (PSI2013-45924-P, PSI2015-73111-EXP, PSI2017-83038-P, PSI2017-83893-R, the Plan Nacional sobre Drogas(#2014/020), and the Plan de Ciencia, Tecnología e Innovación delPrincipado de Asturias (FC-15-GRUPIN14-088). Department ofExperimental and Health Sciences is a“Unidad de Excelencia María deMaeztu”funded by the MINECO (MDM-2014-0370). The authors de-clare no potential conflicts of interest