Sep 12, 2015 - ESBRA 2015. EUROPEAN SOCIETY FOR BIOMEDICAL RESEARCH ON ALCOHOLISM. 12â15 SEPTEMBER 2015. VALENCIA, SPAIN. SY13.
Alcohol and Alcoholism, 2015, 50(S1) i1–i67
ESBRA 2015 EUROPEAN SOCIETY FOR BIOMEDICAL RESEARCH ON ALCOHOLISM 12–15 SEPTEMBER 2015 VALENCIA, SPAIN
SY13 NEW INSIGHTS INTO FETAL ALCOHOL SPECTRUM DISORDERS SY13-1 LONG-LASTING CHANGE ON ASTROCYTE SECRETOME FOLLOWING EARLY ALCOHOL EXPOSURE P. Trindade, B. Hampton, A. C. Manhães, and A. E. Medina School of Medicine, University of Maryland, Baltimore (USA) Glial cells are central players in the pathogenesis of neurodevelopmental disorders like Fetal Alcohol Spectrum Disorders (FASD). Ethanol can modulate gene expression in astrocytes imposing dramatic changes to the developing brain. We test the hypothesis that ethanol exposure in ferrets during a period equivalent to the last months of human gestation leads to a long-lasting change in astrocyte-secretome in vitro. Animals were treated every other day with ethanol (5g/kg) or saline between post-natal day (P)10-30. At P31, a time similar to birth in humans, animals were euthanized and puriﬁed astrocyte cultures
were made. Cells were submitted to stable isotope labeling by amino acids (SILAC). 24h-conditioned media of cells obtained from ethanolor saline-treated animals (ET-CM or SAL-CM) were collected and quantitative proteomics was performed through LC-MS/MS and bioinformatics analysis of differentially secreted proteins was done. We show that 65 out of 280 quantiﬁable proteins displayed signiﬁcant differences comparing ET-CM to SAL-CM. Among 59 proteins found reduced in ET-CM we highlight TIMP-1, Laminin subunits α2, α4, β1, β2 and γ1, Clusterin, PEDF, APP, HSPG2 and PA-1. Conversely, proteins like Thrombospondin-1, Hevin and Angiotensinogen were found increased in ET-CM. Gene ontology bioprocess analysis indicates that the majority of molecules decreased in ET-CM are constitutive components of the extracellular matrix and are related to differentiation, adhesion, neurogenesis, gliogenesis and negative modulation of catalytic activity. In contrast, proteins upregulated in ET-CM are positively associated with endopeptidase activity. Our results strongly suggest that early ethanol exposure leads to a longlasting change on astrocyte-secretome affecting the expression of trophic and matricellular proteins, which may underlie deﬁcits in cell growth and plasticity.
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