Development and assessment of new risk assessment methods

Title of Research:
13_PT01-01

Development of in vivo fluorescent imaging of neuronaldifferentiation in zebrafish for developmental neurotoxicity testing
Principal Investigator:
Yuhei Nishimura (Department of Pharmacogenomics, Mie University Graduate School ofMedicine)Collaborators:
Toshio Tanaka and Reiko Kawase (Department of Systems Pharmacology, Mie University
Graduate School of Medicine)
Summary of Research:
Various chemicals may affect the differentiation of neurons, oligodendrocytes and astrocytes,
which may cause neurodevelopmental disorders such as autism and attention deficit hyperactivedisorder. In vivo models that can be used in both visualization of the neuronal differentiation andquantitative assessment of behavior are highly valuable for developmental neurotoxicity testing.
Recently, zebrafish has emerged as an alternative non-mammalian animal model that allows
testing of large numbers of subjects while reducing expenses and minimizing the use ofmammalian subjects. In this study, we have developed three-color zebrafish exhibiting blue, yellow
and red fluorescence in neurons, oligodendrocytes and astrocytes, respectively. Using thezebrafish, we evaluated the developmental neurotoxicity of four positive control chemicals,
valproic acid, methyl mercury, nicotine, and bisphenol A. We were able to detect the toxicity ofthese chemicals on the neuronal differentiation. We will use more positive and negative chemicals
to examine the sensitivity and specificity of the in vivo fluorescent imaging of neuronaldifferentiation in zebrafish for developmental neurotoxicity testing.
Timeline:
Mar 2016 ~ Feb 2017
Publications:1) Activation of Sterol Regulatory Element Binding Factors by Fenofibrate and Gemfibrozil
Stimulates Myelination in Zebrafish. Ashikawa, Y., Nishimura, Y., Okabe, S., Sasagawa, S.,Murakami, S., Yuge, M., Kawaguchi, K., Kawase, R., and Tanaka, T. Frontiers in pharmacology 7,206 (2016)2) EP300 protects from light-induced retinopathy in zebrafish. Kawase, R., Nishimura, Y.,
Ashikawa, Y., Sasagawa, S., Murakami, S., Yuge, M., Okabe, S., Kawaguchi, K., Yamamoto, H.,Moriyuki, K., Yamane, S., Tsuruma, K., Shimazawa, M., Hara, H., and Tanaka, T. Frontiers inpharmacology 7, 126 (2016).
3) E2F4 promotes neuronal regeneration and functional recovery after spinal cord injury inzebrafish. Sasagawa, S., Nishimura, Y., Hayakawa, Y., Murakami, S., Ashikawa, Y., Yuge, M.,Okabe, S., Kawaguchi, K., Kawase, R., and Tanaka, T. Frontiers in pharmacology 7, 119 (2016).
4) Downregulation of GSTK1 Is a Common Mechanism Underlying Hypertrophic
Cardiomyopathy. Sasagawa, S., Nishimura, Y., Okabe, S., Murakami, S., Ashikawa, Y., Yuge, M.,Kawaguchi, K., Kawase, R., Okamoto, R., Ito, M., and Tanaka, T. Frontiers in pharmacology 7, 162(2016).
5) Comparative transcriptome analysis identifies CCDC80 as a novel gene associated withpulmonary arterial hypertension. Sasagawa, S., Nishimura, Y., Sawada, H., Zhang, E., Murakami,S., Ashikawa, Y., Yuge, M., Okabe, S., Kawaguchi, K., Kawase, R., Mitani, Y., Maruyama, K., andTanaka, T. Frontiers in pharmacology 7, 142 (2016).
6) Integrated Approaches to Drug Discovery for Oxidative Stress-Related Retinal Diseases.
Nishimura, Y., and Hara, H. Oxid Med Cell Longev. 2370252 (2016).