DNA methylation at the DAT promoter and risk for psychopathology. Intergenerational transmission between school-age youths and their parents in a community sample

Background: The effect of gene polymorphisms and promoter methylation, associated with maladaptive developmental outcomes, vary depending on environmental factors (e.g., parental psychopathology). Most studies have focused on 0- to 5-year-old children, adolescents, or adults, whereas there is dearth of research on school-age youths and pre-adolescents. Methods: In a sample of 21 families recruited at schools, we addressed parents' psychopathological symptoms (through SCL-90-R); offspring emotional-behavioral functioning (through CBCL-6-18); dopamine transporter gene (DAT1) for epigenetic status of the 5'-untranslated region (UTR) and for genotype, i.e., variable number of tandem repeats polymorphism at the 3'-UTR. Possible associations were explored between bio-genetic and psychological characteristics within the same individual and between triplets of children, mothers, and fathers. Results: DAT methylation of CpG at positions M1, M6, and M7 in mothers was correlated with maternal (phobic) anxiety, whereas in fathers' position M6 was related to paternal depression, anxiety, hostility, psychoticism, and higher Global Severity Index (GSI). No significant correlations were found between maternal and offspring DAT methylation. Significant correlations were found between fathers' methylation at CpG M1 and children's methylation at CpG M6. Linear regressions showed that mothers and fathers' GSI predicted children's methylation at CpG sites M2, M3, and M6, whereas fathers' GSI predicted children's methylation at CpG sites, particularly M1, M2, and M6. Moreover, offspring methylation of DAT at CpG M2 predicted somatic complaint, internalizing and attention problems; methylation of DAT at CpG M6 predicted withdraw. Conclusion: This study may have important clinical implication for the prevention and treatment of emotional-behavioral difficulties in children, as it adds to previous knowledge about the role of genetic and environmental factors in predicting psychopathological symptoms within non-clinical population

Role of serotonin on cocaine-mediated effects on prodynorphin gene expression in the rat brain

The effect of the selective serotonin uptake inhibitor fluoxetine was examined on prodynorphin gene expression. Fluoxetine or vehicle was infused continuously for 7 d via osmotic minipumps into male rats. Northern blot analysis showed significant increases in prodynorphin gene expression in the hypothalamus (171% of controls) and significant decreases in the caudate putamen and nucleus accumbens (62% and 70% of controls, respectively). There were no significant changes in the hippocampus. Thus, chronic inhibition of serotonin uptake can regulate prodynorphin gene expression in the hypothalamus, caudate putamen, and nucleus accumbens. Fluoxetine effects were also evaluated in rats treated with p-chloroamphetamine (PCA), a neurotoxin that depletes serotonin. Because we previously reported that continuous infusion of cocaine for 7 d (which inhibits dopamine, serotonin, and norepinephrine uptake), or GBR 12909 (a selective dopamine uptake inhibitor), produced significant decreases in the hypothalamus and cocaine also produced a significant increase in prodynorphin gene expression in caudate putamen, regulation of prodynorphin gene expression by fluoxetine is suggested to be different from that by cocaine. Because neither a selective dopamine uptake inhibitor nor a selective serotonin uptake inhibitor produced the same effect as cocaine in the caudate putamen, this effect is likely regulated by the inhibition of norepinephrine uptake, by a combination of effects on two or three neurotransmitter transporters, or by a mechanism unrelated to transporter inhibition

Selective alterations in endogenous opioid system genes expression in rats selected for high ethanol intake during adolescence

Historically, the roots of alcoholism have been linked to either environment or heredity. However, the interaction between these factors is still largely unexplored. The evidence supports a link between alcohol consumption and the endogenous opioid system. We here studied the opioid genes expression in male and female Wistar rats derived from a short-term breeding program which selected -- at adolescence -- for high (ADHI line) or low (ADLO line) ethanol drinking. Specifically, in this work we analyzed central opioid gene expression in the rats of the second filial generation (S2-ADLO and S2-ADHI). Selective downregulation of pronociceptin (Pnoc) and its receptor (Oprl1) mRNA levels were observed in the prefrontal cortex of male S2-ADHI rats when compared to S2-ADLO, and for Oprl1 also in the nucleus accumbens. An increase in gene expression was instead observed for pro-opiomelanocortin (Pomc) in the nucleus accumbens of S2-ADHI males when compared to S2-ADLO, as well as for mu opioid receptor (Oprm1) but in females. The differences in mRNA levels may be due to the different alcohol consumption between the two groups of rats or may represent pre-existing differences between them. Moreover, we show a sex-specific modulation of the expression of these genes, thus pointing out the importance of sex on ethanol responses. The results might lead to more specific and effective pharmacological treatments for alcoholism.Fil: Bellia, Fabio. Università degli Studi di Teramo; ItaliaFil: Fernandez, Macarena Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; ArgentinaFil: Fabio, Maria Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; Argentina. Universidad Nacional de Córdoba. Facultad de Psicología; ArgentinaFil: Pucci, Mariangela. Università degli Studi di Teramo; ItaliaFil: Pautassi, Ricardo Marcos. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; Argentina. Universidad Nacional de Córdoba. Facultad de Psicología; ArgentinaFil: D’Addario, Claudio. Università degli Studi di Teramo; Italia. Karolinska Huddinge Hospital; Sueci

OXTR Gene DNA Methylation Levels Are Associated with Discounting Behavior with Untrustworthy Proposers

Individual differences in temporal and probabilistic discounting are associated with a wide range of life outcomes in literature. Traditional approaches have focused on impulsiveness and cognitive control skills, on goal-oriented personality traits as well as on the psychological perception of time. More recently, literature started to consider the role of social and contextual factors in discounting behavior. Between others, higher generalized trust in human beings and specific trust in people who will deliver the future/probabilistic rewards have been related to a stronger willingness to wait and to assume risk. Moreover, the tendency to trust others has been associated with the oxytocin receptor gene regulation that can be modified by life experiences. In this perspective, we hypothesized that differences in the tendency to wait and to take risks for a more desirable reward according to the proposer’s trustworthiness could be related to a different level of DNA methylation at the oxytocin receptor gene. Findings confirmed that participants are less willing to wait and to risk when the proposer is considered highly untrustworthy and revealed how higher oxytocin receptor gene DNA methylation is associated with a stronger effect due to the presence of an untrustworthy proposer. Limits and future directions are outlined

OCT-angiography follow-up of choroidal neovascularization treated with treat-and- extend aflibercept regimen to avoid over-treatment

Purpose To propose optical coherence tomography angiography (OCT-A) for the follow-up of neovascular age-related macular degeneration (nAMD) treated with a treat-and-extend (T&E) aflibercept regimen to avoid overtreatment. Methods Retrospective, cohort, pilot study. We analysed 16 consecutive-treatment naive nAMD eyes following up 2-years at the Eye Clinic, Bari, Italy. Intravitreal aflibercept injections in the T&E regimen for no less than 12 months, during which the macula was dry without any sign of intraretinal or subretinal fluid (SRF) at each visit, were performed. Parametric data were evaluated using an analysis of variance (ANOVA); any non-parametric statistical calculations were performed using the Wilcoxon and Mann-Whitney test. Results The average number of injections during follow-up was: 8.8 +/- 1. Treatment regimen adjustments were 4 weeks (W), 8W (4 + 4), 10W (8 + 2), 12W (8 + 4, or 10 + 2). No significant CNV size change from 4 to 8W (-0.027 +/- 0.22 mm(2), p = 0.088), and from 8 to 12W (-0.04 +/- 0.11 mm(2), p = 0.065) were found. Likewise, no significant decrease in choriocapillaris flow (CF) was detected (p = 0.056). Conclusion We suggest that OCT-A may be useful in the evaluation of dry macula to decide the best approach for perchance adjusting injection intervals based on changes of CNV size

Iron modulates the membrane-binding and the intracellular trafficking of 5-lipoxygenase: functional implications in neurodegeneration

Iron modulates the membrane-binding and the intracellular trafficking of 5-lipoxygenase: functional implications in neurodegeneration Beatrice Dufrusine1, Andrea Di Francesco1, Annalaura Sabatucci1, Clotilde Beatrice Angelucci2, Sergio Oddi2,3, Claudio D’Addario1,3, Dieter Steinhilber4, Mauro Maccarrone3,5* and Enrico Dainese1,3* 1Faculty of Biosciences, University of Teramo, Teramo, 64100, Italy; 2 Faculty of Veterinary Medicine, University of Teramo, Teramo, 64100, Italy; 3 European Center for Brain Research (CERC)/Santa Lucia Foundation, Rome, Italy; 4 Institute of Pharmaceutical Chemistry, Goethe University Frankfurt, Germany; 5 Center of Integrated Research, Campus Bio-Medico University of Rome, Italy. *Equally senior authors. Introduction 5-lipoxygenase (5-LOX) is a non-heme iron-containing enzyme catalyzing the initial steps in the biosynthesis of leukotrienes, inflammatory eicosanoids that are capable of promoting neurodegeneration. Iron accumulation has been demonstrated in Alzheimer’s disease (AD). Here we present the structure of 5-LOX in solution and a new molecular mechanism throughout the iron content of the enzyme is able to influence its membrane binding and subcellular distribution in THP-1 macrophages. Methods 3D structure in solution and membrane binding experiments were done using Small angle X-ray scattering (SAXS) and fluorescence resonance energy transfer (FRET) approaches, as reported [1]. Confocal fluorescence microscopy, subcellular fractionation and western blot (WB) analysis were carried out as already described [1]. Late Onset AD (LOAD) patients and non-demented healthy subjects (CT) were recruited as described [2]. Results SAXS analysis revealed that 5-LOX in solution is mostly organized as a homodimer. Iron removal from the recombinant human 5-LOX altered the catalytic activity of the enzyme, and impaired its membrane-binding ability. THP-1 cells exposed to increasing amounts of iron showed a redistribution of the cytosolic 5-LOX to the nuclear compartment. Additionally, a significant correlation between plasma levels of the 5-LOX end-product LTB4 and hemoglobin was observed in a population of healthy subjects and AD patients. Conclusions These results suggest that exogenous iron modulates 5-LOX activity by increasing its ability to bind to nuclear membranes, further supporting a new role for iron in inflammation-based neurological diseases where its homeostasis is altered. Acknowledgements. M.M. and E.D. wish to thank EU for granting the Biostruct-X project within the FP VII programme. [1] E. Dainese et al., FASEB J. 24 (2010) 1725-1736. [2] A. Di Francesco et al., JAD 37 (2013) 3-8

Peripheral Biomarkers in DSM-5 Anxiety Disorders: An Updated Overview

Anxiety disorders are prevalent and highly disabling mental disorders. In recent years, intensive efforts focused on the search for potential neuroimaging, genetic, and peripheral biomarkers in order to better understand the pathophysiology of these disorders, support their diagnosis, and characterize the treatment response. Of note, peripheral blood biomarkers, as surrogates for the central nervous system, represent a promising instrument to characterize psychiatric disorders, although their role has not been extensively applied to clinical practice. In this report, the state of the art on peripheral biomarkers of DSM-5 (Diagnostic and Statistical Manual of Mental Disorders, 5th edition) Anxiety Disorders is presented, in order to examine their role in the pathogenesis of these conditions and their potential application for diagnosis and treatment. Available data on the cerebrospinal fluid and blood-based biomarkers related to neurotransmitters, neuropeptides, the hypothalamic–pituitary–adrenal axis, neurotrophic factors, and the inflammation and immune system are reviewed. Despite the wide scientific literature and the promising results in the field, only a few of the proposed peripheral biomarkers have been defined as a specific diagnostic instrument or have been identified as a guide in the treatment response to DSM-5 Anxiety Disorders. Therefore, further investigations are needed to provide new biological insights into the pathogenesis of anxiety disorders, to help in their diagnosis, and to tailor a treatment

On the role of adenosine a2a receptor gene transcriptional regulation in parkinson’s disease

Adenosine A2A receptors (A2ARs) have attracted considerable attention as an important molecular target for the design of Parkinson’s disease (PD) therapeutic compounds. Here, we studied the transcriptional regulation of the A2AR gene in human peripheral blood mononuclear cells (PBMCs) obtained from PD patients and in the striatum of the well-validated, 6-hydroxydopamine (6-OHDA)-induced PD mouse model. We report an increase in A2AR mRNA expression and protein levels in both human cells and mice striata, and in the latter we could also observe a consistent reduction in DNA methylation at gene promoter and an increase in histone H3 acetylation at lysine 9. Of particular relevance in clinical samples, we also observed higher levels in the receptor gene expression in younger subjects, as well as in those with less years from disease onset, and less severe disease according to clinical scores. In conclusion, the present findings provide further evidence of the relevant role of A2AR in PD and, based on the clinical data, highlight its potential role as disease biomarker for PD especially at the initial stages of disease development. Furthermore, our preclinical results also suggest selective epigenetic mechanisms targeting gene promoter as tool for the development of new treatments