Basic Study
Copyright ©The Author(s) 2015. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Biol Chem. Nov 26, 2015; 6(4): 389-408
Published online Nov 26, 2015. doi: 10.4331/wjbc.v6.i4.389
DNA microarray unravels rapid changes in transcriptome of MK-801 treated rat brain
Yuka Kobayashi, Sofya P Kulikova, Junko Shibato, Randeep Rakwal, Hiroyuki Satoh, Didier Pinault, Yoshinori Masuo
Yuka Kobayashi, Hiroyuki Satoh, Department of Biomolecular Science, Faculty of Science, Toho University, Chiba 274-8510, Japan
Yuka Kobayashi, Junko Shibato, Randeep Rakwal, Yoshinori Masuo, Health Technology Research Center, National Institute of Advanced Industrial Science and Technology, Tsukuba 305-8569, Japan
Sofya P Kulikova, Didier Pinault, INSERM U1114, Neuropsychologie Cognitive et Physiopathologie de la Schizophrénie, FMTS (Fédération de Médecine Translationnelle de Strasbourg), Faculté de Médecine, Université de Strasbourg, 67400 Strasbourg, France
Junko Shibato, Randeep Rakwal, Department of Anatomy, School of Medicine, Showa University, Shinagawa, Tokyo 142-8555, Japan
Junko Shibato, Laboratory of Exercise Biochemistry and Neuroendrocrinology, Institute of Health and Sport Sciences, University of Tsukuba, Tsukuba 305-8574, Japan
Junko Shibato, Randeep Rakwal, Global Research Center for Innovative Life Science, Peptide Drug Innovation, School of Pharmacy and Pharmaceutical Sciences, Hoshi University, Shinagawa, Tokyo 142-8501, Japan
Randeep Rakwal, Organization for Educational Initiatives, University of Tsukuba, Tsukuba 305-8577, Japan
Randeep Rakwal, Faculty of Health and Sport Sciences and Tsukuba International Academy for Sport Studies, University of Tsukuba, Tsukuba 305-8574, Japan
Yoshinori Masuo, Laboratory of Neuroscience, Department of Biology, Faculty of Science, Toho University, Funabashi, Chiba 274-8510, Japan
Author contributions: Kobayashi Y, Shibato J, Rakwal R and Masuo Y performed the majority of experiments; Kulikova SP and Pinault D performed the electrophysiology experiments and contributed to the design and the writing and revision; Masuo Y and Satoh H co-ordinated and provided financial support for this work; Kobayashi Y, Masuo Y, Shibato J and Rakwal R designed the study and wrote and edited/revised the manuscript.
Institutional animal care and use committee statement: All animal care procedures were achieved in accordance with European Union Guidelines (Directive 2010/63/EU) and with CREMEAS, the National and Regional Ethics Committee. The experiments were also conducted in accordance with the Guidelines for the Care and Use of Laboratory Animals at the National Institute of Advanced Industrial Science and Technology (AIST), Japan.
Conflict-of-interest statement: The authors declare no conflict of interest.
Data sharing statement: The obtained gene expression data deposited at the NCBI, Gene Expression Omnibus (GEO) site (accession number GSE63639) is freely available to the scientific community (http://www.ncbi.nlm.nih.gov/geo/).
Open-Access: This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Correspondence to: Dr. Randeep Rakwal, Professor, Faculty of Health and Sport Sciences and Tsukuba International Academy for Sport Studies, University of Tsukuba, 1-1-1 Tennoudai, Tsukuba 305-8574, Japan. plantproteomics@gmail.com
Telephone: +81-90-18537875
Received: November 26, 2014
Peer-review started: November 29, 2014
First decision: December 26, 2014
Revised: July 21, 2015
Accepted: August 30, 2015
Article in press: August 31, 2015
Published online: November 26, 2015
Abstract

AIM: To investigate the impact of MK-801 on gene expression patterns genome wide in rat brain regions.

METHODS: Rats were treated with an intraperitoneal injection of MK-801 [0.08 (low-dose) and 0.16 (high-dose) mg/kg] or NaCl (vehicle control). In a first series of experiment, the frontoparietal electrocorticogram was recorded 15 min before and 60 min after injection. In a second series of experiments, the whole brain of each animal was rapidly removed at 40 min post-injection, and different regions were separated: amygdala, cerebral cortex, hippocampus, hypothalamus, midbrain and ventral striatum on ice followed by DNA microarray (4 × 44 K whole rat genome chip) analysis.

RESULTS: Spectral analysis revealed that a single systemic injection of MK-801 significantly and selectively augmented the power of baseline gamma frequency (30-80 Hz) oscillations in the frontoparietal electroencephalogram. DNA microarray analysis showed the largest number (up- and down- regulations) of gene expressions in the cerebral cortex (378), midbrain (376), hippocampus (375), ventral striatum (353), amygdala (301), and hypothalamus (201) under low-dose (0.08 mg/kg) of MK-801. Under high-dose (0.16 mg/kg), ventral striatum (811) showed the largest number of gene expression changes. Gene expression changes were functionally categorized to reveal expression of genes and function varies with each brain region.

CONCLUSION: Acute MK-801 treatment increases synchrony of baseline gamma oscillations, and causes very early changes in gene expressions in six individual rat brain regions, a first report.

Keywords: Dizocilpine, Dye-swap, Gene expression, Microarray, MK801, N-Methyl-D-aspartate receptors

Core tip: N-Methyl-D-aspartate receptors (NMDAr) are involved in multiple physiological functions and neuropsychiatric disorders. Dizocilpine (commonly referred to as MK-801) is a well-known non-competitive NMDAr antagonist with psychotomimetic properties. A combination of electrophysiological and molecular analyses reveals not only the increased synchrony of baseline cortical gamma oscillations by MK-801, but also more importantly new insight into differential gene expressions in the cerebral cortex, midbrain, hippocampus, ventral striatum, amygdala, and hypothalamus regions after acute low-dose (0.08 mg/kg) MK-801 treatment; only the ventral striatum showed increased gene expression at a high dose (0.16 mg/kg) of MK-801. We believe that our present study will contribute in the understanding of the pathogenic mechanisms of neuropsychiatric disorders.