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Sci Rep. 2017 Jul 12;7(1):5230. doi: 10.1038/s41598-017-04657-x.

Genetic targeting and anatomical registration of neuronal populations in the zebrafish brain with a new set of BAC transgenic tools.

Author information

1
Max Planck Institute of Neurobiology, Department Genes - Circuits - Behavior, Am Klopferspitz 18, D-82152, Martinsried, Germany.
2
Max-Delbrück Center for Molecular Medicine, Berlin, Germany.
3
Department of Biological Sciences, Purdue University, West Lafayette, USA.
4
Department of Biological Sciences, University of Toronto Scarborough, Toronto, Canada.
5
Max Planck Institute of Neurobiology, Department Genes - Circuits - Behavior, Am Klopferspitz 18, D-82152, Martinsried, Germany. hbaier@neuro.mpg.de.
6
Max Planck Institute of Neurobiology, Department Genes - Circuits - Behavior, Am Klopferspitz 18, D-82152, Martinsried, Germany. fumikubo@neuro.mpg.de.

Abstract

Genetic access to small, reproducible sets of neurons is key to an understanding of the functional wiring of the brain. Here we report the generation of a new Gal4- and Cre-driver resource for zebrafish neurobiology. Candidate genes, including cell type-specific transcription factors, neurotransmitter-synthesizing enzymes and neuropeptides, were selected according to their expression patterns in small and unique subsets of neurons from diverse brain regions. BAC recombineering, followed by Tol2 transgenesis, was used to generate driver lines that label neuronal populations in patterns that, to a large but variable extent, recapitulate the endogenous gene expression. We used image registration to characterize, compare, and digitally superimpose the labeling patterns from our newly generated transgenic lines. This analysis revealed highly restricted and mutually exclusive tissue distributions, with striking resolution of layered brain regions such as the tectum or the rhombencephalon. We further show that a combination of Gal4 and Cre transgenes allows intersectional expression of a fluorescent reporter in regions where the expression of the two drivers overlaps. Taken together, our study offers new tools for functional studies of specific neural circuits in zebrafish.

PMID:
28701772
PMCID:
PMC5507991
DOI:
10.1038/s41598-017-04657-x
[Indexed for MEDLINE]
Free PMC Article

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