Metabolic Alterations of the Zebrafish Brain after Acute Alcohol Treatment by 1H Nuclear Magnetic Resonance Spectroscopy

Journal of Spectroscopy, May 2013

The purpose of this study is to investigate the metabolic alterations associated with acute alcohol treatment in zebrafish by 1H nuclear magnetic resonance spectroscopy (NMRS). The brain metabolism of zebrafish was investigated after acute alcohol treatment (one-hour long exposure of adult fish to 0.00%, 0.25%, 0.50%, or 1.00% ethyl alcohol) with whole brain extraction. The results of this study showed that glutamate (Glu) was significantly decreased, scyllo-inositol (sIns) showed a small apparent increase only in the highest acute treatment dose group, and myoinositol (mIns) showed a significant decrease. [Glu]/[tCr] and [mIns]/[tCr] levels were significantly reduced regardless of the alcohol dose, and [sIns]/[tCr] was increased in the highest alcohol treatment dose group. The present NMR study revealed that specific metabolites, such as Glu and mIns, were substantially decreased in case of acute alcohol exposed zebrafish brain.

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Metabolic Alterations of the Zebrafish Brain after Acute Alcohol Treatment by 1H Nuclear Magnetic Resonance Spectroscopy

Metabolic Alterations of the Zebrafish Brain after Acute Alcohol Treatment by 1H Nuclear Magnetic Resonance Spectroscopy Dong-Cheol Woo,1,2 Goo-Young Kim,3 Hyun-Ju Kim,4 Eunjung Bang,4 Hyang-Shuk Rhim,3 Sang-Young Kim,2 Do-Wan Lee,2 Chi-Bong Choi,2 Youl-Hun Seoung,2,5 and Bo-Young Choe2 1Department of Radiology, UT Southwestern Medical Center, Dallas, TX, USA 2Department of Biomedical Engineering, Research Institute of Biomedical Engineering, College of Medicine, The Catholic University of Korea, Seoul 137-701, Republic of Korea 3Department of Molecular Biology, College of Medicine, The Catholic University of Korea, Republic of Korea 4Environment & Metabolomics Research Team, Seoul Center, Korea Basic Science Institute, Seoul, Republic of Korea 5Department of Radiology, Cheongju University, Cheongju, Chungbuk, Republic of Korea Received 22 February 2013; Accepted 8 May 2013 Academic Editor: Tomasz Ruman Copyright © 2013 Dong-Cheol Woo et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Abstract The purpose of this study is to investigate the metabolic alterations associated with acute alcohol treatment in zebrafish by 1H nuclear magnetic resonance spectroscopy (NMRS). The brain metabolism of zebrafish was investigated after acute alcohol treatment (one-hour long exposure of adult fish to 0.00%, 0.25%, 0.50%, or 1.00% ethyl alcohol) with whole brain extraction. The results of this study showed that glutamate (Glu) was significantly decreased, scyllo-inositol (sIns) showed a small apparent increase only in the highest acute treatment dose group, and myoinositol (mIns) showed a significant decrease. [Glu]/[tCr] and [mIns]/[tCr] levels were significantly reduced regardless of the alcohol dose, and [sIns]/[tCr] was increased in the highest alcohol treatment dose group. The present NMR study revealed that specific metabolites, such as Glu and mIns, were substantially decreased in case of acute alcohol exposed zebrafish brain. 1. Introduction Alcohol (i.e., EtOH, ethyl alcohol, and ethanol) abuse and alcoholism are prevalent conditions that cause significant problems for both individual patients and society. Alcohol abuse and alcoholism involve neural processes distinct from other addictions [1]. Attempts to understand the common and unique aspects of alcohol addiction have spurred investigators to adopt new animal models and research methods that have not been widely used for the investigation of other addictions. Partially, alcohol in the brain reduces glutamate and it could be interpreted as a response to N-methyl-D-aspartate (NMDA) receptor blockade [2, 3]. However, the mechanisms associated with alcohol addiction have not been completely identified [1, 4–6], despite numerous reported studies on the topic during the past several decades. Animal models, including zebrafish, mice, rats, and monkeys, have been used to study alcohol addiction and the physiology of alcoholism [1]. Zebrafish have become a popular subject for embryology studies in genetic research, as well as neurobiology over the past three decades [7]. Attempts at in vivo MR imaging (MRI) and MR spectroscopy (MRS) of zebrafish have recently been reported [8, 9]. NMRS is a common modality for research of neurological diseases, such as Alzheimer and Parkinson disease, as well as epilepsy, depression, and schizophrenia in both animals and humans [10–14]. Previous human NMRS studies in alcoholics have shown brain metabolic changes, particularly in the hippocampus, frontal cortex, basal ganglia, and cerebellum [15–17]. The N-acetyl aspartate (NAA), choline (Cho), and glutamic acids were changed in the frontal lobe and cerebellum of alcohol dependents immediately after abstinence, which increased after a month of abstinence [16–22]. Although there have been several reports on alcohol related research using zebrafish, including behavioral tests [23, 24] and chromatography [4], in addition to in vivo MRS in both rats and humans, NMR study of the effects of alcohol in zebrafish has not yet been reported. However, NMRS information, which is included in the changes of various neuro-metabolites, can be useful to understand the neurobiology in the zebrafish brain. Furthermore, zebrafish and NMRS can be helpful to elucidate molecular mechanisms, such as NMDA in alcoholism. Therefore, the purpose of this study is to investigate the metabolic alterations associated with acute alcohol treatment in the zebrafish brain by in vitro NMRS. 2. Materials and Methods2.1. Animals Forty wild-type zebrafish naïve to ethanol (Danio rerio; 3~4 months old) were used in this study. In order to investigate the effects of alcohol in the zebrafish (males and females pooled), they were divided into four groups: EtOH 0.00% , EtOH 0.25% , EtOH 0.50% , and EtOH 1.00% , where the % EtOH represents the alcoho (...truncated)


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Dong-Cheol Woo, Goo-Young Kim, Hyun-Ju Kim, Eunjung Bang, Hyang-Shuk Rhim, Sang-Young Kim, Do-Wan Lee, Chi-Bong Choi, Youl-Hun Seoung, Bo-Young Choe. Metabolic Alterations of the Zebrafish Brain after Acute Alcohol Treatment by 1H Nuclear Magnetic Resonance Spectroscopy, Journal of Spectroscopy, 2013, 2013, DOI: 10.1155/2013/971914