Document Type

Journal Article

Department/Unit

Department of Biology

Title

Assessments of the effects of nicotine and ketamine using tyrosine hydroxylase-green fluorescent protein transgenic zebrafish as biosensors

Language

English

Abstract

Transgenic zebrafish are a common vertebrate model system for the study of addictive behavior. In the present study, plasmid constructs containing green fluorescent protein (GFP) and the promoter of tyrosine hydroxylase (TH), a key synthetic enzyme for catecholamines, were produced. The TH-GFP constructs were microinjected into zebrafish embryonic cells. Three days post-fertilization, GFP began expressing in distinct catecholaminergic areas. The TH-GFP transgenic zebrafish were employed as live biosensors to test the effects of the commonly abused drugs nicotine and ketamine. First, locomotion assays were used to study the general excitatory effects of the drugs. Maximal locomotor activity was obtained after treatment with a high concentration of nicotine (10. μM), but with a much lower concentration of ketamine (0.1. μM). Second, TH protein levels in zebrafish brains were assessed by Western blot. TH protein levels were significantly increased, with maximal protein levels found after treatment with the same drug concentrations that gave maximal locomotor activity. Importantly, analysis of GFP in the zebrafish catecholaminergic areas revealed the same expression patterns as was obtained by Western blot. The present results indicate that increased locomotor activity can be correlated to TH protein expression, as indicated by Western blot and expression of TH-GFP. We have shown that TH-GFP expression is a reliable method to show the effects of drugs on TH expression that may be employed as a novel high-throughput live biosensor for screening drugs of abuse. © 2012 Elsevier B.V.

Keywords

Catecholamine, Dopamine, GFP, Ketamine, Nicotine, Tyrosine hydroxylase, Zebrafish

Publication Date

2013

Source Publication Title

Biosensors and Bioelectronics

Volume

42

Start Page

177

End Page

185

Publisher

Elsevier

DOI

10.1016/j.bios.2012.09.042

Link to Publisher's Edition

http://dx.doi.org/10.1016/j.bios.2012.09.042

ISSN (print)

09565663

ISSN (electronic)

18734235

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