RESUMEN
Fetal alcohol spectrum disorders (FASDs) are characterized by life-long changes in gene expression, neurodevelopment and behavior. What mechanisms initiate and maintain these changes are not known, but current research suggests a role for alcohol-induced epigenetic changes. In this study we assessed alterations to adult mouse brain tissue by assaying DNA cytosine methylation and small noncoding RNA (ncRNA) expression, specifically the microRNA (miRNA) and small nucleolar RNA (snoRNA) subtypes. We found long-lasting alterations in DNA methylation as a result of fetal alcohol exposure, specifically in the imprinted regions of the genome harboring ncRNAs and sequences interacting with regulatory proteins. A large number of major nodes from the identified networks, such as Pten signaling, contained transcriptional repressor CTCF-binding sites in their promoters, illustrating the functional consequences of alcohol-induced changes to DNA methylation. Next, we assessed ncRNA expression using two independent array platforms and quantitative PCR. The results identified 34 genes that are targeted by the deregulated miRNAs. Of these, four (Pten, Nmnat1, Slitrk2 and Otx2) were viewed as being crucial in the context of FASDs given their roles in the brain. Furthermore, ≈ 20% of the altered ncRNAs mapped to three imprinted regions (Snrpn-Ube3a, Dlk1-Dio3 and Sfmbt2) that showed differential methylation and have been previously implicated in neurodevelopmental disorders. The findings of this study help to expand on the mechanisms behind the long-lasting changes in the brain transcriptome of FASD individuals. The observed changes could contribute to the initiation and maintenance of the long-lasting effect of alcohol.
Asunto(s)
Metilación de ADN/genética , Trastornos del Espectro Alcohólico Fetal/genética , ARN no Traducido/genética , Envejecimiento/genética , Animales , Sitios de Unión/genética , Encéfalo/metabolismo , Encéfalo/patología , Factor de Unión a CCCTC , Análisis por Conglomerados , Biología Computacional , Epigénesis Genética , Femenino , Regulación del Desarrollo de la Expresión Génica , Redes Reguladoras de Genes/genética , Genoma/genética , Masculino , Exposición Materna/efectos adversos , Ratones , MicroARNs/genética , MicroARNs/metabolismo , Modelos Genéticos , Embarazo , Regiones Promotoras Genéticas/genética , Unión Proteica/genética , ARN no Traducido/metabolismo , Proteínas Represoras/metabolismo , Reproducibilidad de los Resultados , Factores de TiempoRESUMEN
The importance of understanding the effects of radiation exposure on wildlife is a critical responsibility of our stewardship of nuclear energy production. We tested the hypothesis that songbirds respond to exogenous radiation exposure with changes in plasma 8-hydroxy-2'-deoxyguanosine (8-OH-dG). We exposed two species of songbirds, house sparrows (Passer domesticus; n = 12) and song sparrows (Melospiza melodia; n = 12), to one of four acute whole-body radiation treatments: 75, 150, 300, or 600 mGy. We measured DNA damage by proxy as 8-OH-dG levels in the plasma at 0 hr (baseline), 36 hr, and 7 days post radiation. For both species, at all radiation treatments, 8-OH-dG levels increased significantly 36 hr following radiation exposure. However, songbird species differed significantly in response to treatment across time and between treatment groups. Song sparrows showed no significant changes in 8-OH-dG levels between 36 hr and Day 7. In contrast, house sparrows exposed to 300 and 600 mGy had significantly increased 8-OH-dG levels at Day 7 compared with 36 hr. This study demonstrates that in a controlled experiment, in isolation from other sources of genotoxicity, radiation exposure significantly affects songbirds. Our results suggest future research examining the effects of radiation on songbirds must consider using multiple species to assess the biological effects of radiation, as different species can show strikingly different responses to radiation dosage across time.