INVESTIGATION Developmental Ethanol Exposure Leads to Dysregulation of Lipid Metabolism and Oxidative Stress in Drosophila Theresa Logan-Garbisch, 1 Anthony Bortolazzo, 2 Peter Luu, Audrey Ford, David Do, Payam Khodabakhshi, and Rachael L. French 3 Department of Biological Sciences, San José State University, 1 Washington Square, San José, California 95192-0100 ABSTRACT Ethanol exposure during development causes an array of developmental abnormalities, both physiological and behavioral. In mammals, these abnormalities are collectively known as fetal alcohol effects (FAE) or fetal alcohol spectrum disorder (FASD). We have established a Drosophila melanogaster model of FASD and have previously shown that developmental ethanol exposure in flies leads to reduced expression of insulin-like peptides (dILPs) and their receptor. In this work, we link that observation to dysregulation of fatty acid metabolism and lipid accumulation. Further, we show that developmental ethanol exposure in Drosophila causes oxidative stress, that this stress is a primary cause of the developmental lethality and delay associated with ethanol exposure, and, finally, that one of the mechanisms by which ethanol increases oxidative stress is through abnormal fatty acid metabolism. These data suggest a previously uncharacterized mechanism by which ethanol causes the symptoms associated with FASD. KEYWORDS fetal alcohol syndrome reactive oxygen species lipid accumulation withered carnitine transporter Developmental exposure to ethanol causes a constellation of develop- mental and neurobehavioral problems in organisms from humans to Drosophila. These include slow growth, developmental delays (both physical and intellectual), reduced brain size, low birth weight, and a variety of behavioral and intellectual disabilities, including learning and memory deficits, sleep difficulties, impulse control problems, and reduced executive function (Hanson et al. 1976; Streissguth et al. 1980, 2004; Colangelo and Jones 1982; Kodituwakku 2007). In humans, these symptoms are referred to collectively as “fetal alcohol syndrome” or “fetal alcohol spectrum disorders” (FASD), reflecting the fact that the results of fetal ethanol exposure can vary substantially from individual to individual. In this article, we use the term “develop- mental alcohol exposure” (DAE) because it encompasses mamma- lian and invertebrate development. The variation in symptoms arising from DAE likely reflects a number of factors, including dose, timing, and duration of exposure (Guerri et al. 2009; Nunez et al. 2011). In addition, in twin studies of FASD in humans, greater concordance of phenotype was observed in identical twin pairs compared with fraternal twin pairs, indicating either a genetic effect on the susceptibility to the negative effects of DAE or effects of placental development on the amount of ethanol fetuses are exposed to (Warren and Li 2005). Ethanol affects the activity or expression of numerous molecules and pathways, including membrane lipids, a variety of transcription factors, the epidermal growth factor/mitogen-activated protein kinase (EGFR/MAPK) signaling pathway, 5-HT, and dopamine receptors, and insulin-like growth factors (IGFs) and their receptors (Harris et al. 2008; Corl et al. 2009; McClure et al. 2011) (our unpublished data). Although a few of these (MAPK and IGF signaling in particular) have been shown to be important for ethanol’s developmental effects in animal models, identification of the relevant targets and elucidation of ethanol’s mechanism(s) of action have been largely elusive. This is almost certainly due to the breadth of phenotypes associated with DAE, combined with the wide variety of potential targets. The combination of the widespread socioeconomic and health effects of DAE and the relative ineffectiveness of public awareness campaigns makes an understanding of the molecular mechanisms of Copyright © 2015 Logan-Garbisch et al. doi: 10.1534/g3.114.015040 Manuscript received October 15, 2014; accepted for publication November 7, 2014; published Early Online November 11, 2014. This is an open-access article distributed under the terms of the Creative Commons Attribution Unported License (http://creativecommons.org/licenses/ by/3.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Supporting information is available online at http://www.g3journal.org/lookup/ suppl/doi:10.1534/g3.114.015040/-/DC1 1 Present address: Department of Neurobiology, Stanford University, Stanford, CA 94305. 2 Present address: Laboratory of Genetics, University of Wisconsin, Madison, WI 53706. 3 Corresponding author: Department of Biological Sciences, San José State University, 1 Washington Square, San José, CA 95192-0100. E-mail: rachael.french@sjsu.edu Volume 5 | January 2015 | 49