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Generation and Characterization of Neurogeninl-GFP Transgenic Medaka for High Throughput Developmental Neurotoxicity Screening
Citation:
Fan, C., S. SIMMONS, S. Law, K. Jensen, J. COWDEN, D. Hinton, S. J. PADILLA, AND T. V. RAMABHADRAN. Generation and Characterization of Neurogeninl-GFP Transgenic Medaka for High Throughput Developmental Neurotoxicity Screening. AQUATIC TOXICOLOGY. Elsevier Science Ltd, New York, NY, 105(1-2):127-135, (2011).
Impact/Purpose:
This paper describes the creation of a transgenic line of medaka fish that can be used to monitor the progress of neuronal development
Description:
Fish models such as zebrafish and medaka are increasingly used as alternatives to rodents in developmental and toxicological studies. These developmental and toxicological studies can be facilitated by the use of transgenic reporters that permit the real-time, noninvasive observation of the fish. Here we report the construction and characterization of transgenic medaka lines expressing green fluorescent protein (GFP) under the control of the zebrafish neurogenin J (ngnJ) gene promoter. Neurogenin (ngnl) is a helix-loop-helix transcription factor expressed in proliferating neuronal progenitor cells early in neuronal differentiation and plays a crucial role in directing neurogenes. GFP expression was detected from 24 hours post-fertilization until hatching, in a spatial pattern consistent with the previously reported zebrafish ngnJ expression. Temporal expression of the transgene parallels the expression profile of the endogenous medaka ngnJ transcript. Further, we demonstrate that embryos from the transgenic line permit the screening of ngnJ promoter-directed GFP expression in a 96-well format, enabling higher throughput studies of developmental neurotoxicants. This strain has been deposited with the National BioResource Project (http://www.nbrp.jp/report/reportProject.jsp?project=medaka) and is available on request.
