Abstract
Inhibition or ablation of neuronal activity combined with behavioral assessment is crucial in identifying neural circuits or populations essential for specific behaviors and to understand brain function. In the model vertebrate zebrafish, the development of genetic methods has allowed not only visualization but also targeted manipulation of neuronal activity, and quantitative behavioral assays allow precise measurement of animal behavior. Here, we describe a method to inhibit a specific neuronal population in adult zebrafish brain and assess their role in a learning behavior. We employed the Gal4–UAS system, gene trap and enhancer trap methods, and isolated transgenic zebrafish lines expressing Gal4FF transactivator in specific populations of neurons in the adult zebrafish brain. In these lines, a genetically engineered neurotoxin, botulinum toxin B light chain, was expressed and the fish were assessed in the active avoidance fear conditioning paradigm. The transgenic lines that showed impaired avoidance response were isolated and, in these fish, the Gal4-expressing neurons were analyzed to identify the neuronal circuits involved in avoidance learning.
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References
Wullimann MF, Rupp B, Reichert H (1996) Neuroanatomy of the zebrafish brain. Birkhäuser Verlag, Basel
Wullimann MF, Mueller T (2004) Teleostean and mammalian forebrains contrasted: evidence from genes to behavior. J Comp Neurol 475(2):143–162
Asakawa K, Suster ML, Mizusawa K, Nagayoshi S, Kotani T, Urasaki A et al (2008) Genetic dissection of neural circuits by Tol2 transposon-mediated Gal4 gene and enhancer trapping in zebrafish. Proc Natl Acad Sci U S A 105(4):1255–1260
Kawakami K, Asakawa K, Hibi M, Itoh M, Muto A, Wada H (2016) Gal4 driver transgenic zebrafish: powerful tools to study developmental biology, organogenesis, and neuroscience. Adv Genet 95:65–87
Lal P, Tanabe H, Suster ML, Ailani D, Kotani Y, Muto A et al (2018) Identification of a neuronal population in the telencephalon essential for fear conditioning in zebrafish. BMC Biol 16(1):45
Lal P, Kawakami K (2022) Integrated behavioral, genetic and brain circuit visualization methods to unravel functional anatomy of zebrafish amygdala. Front Neuroanat 16:837527
Sternberg JR, Severi KE, Fidelin K, Gomez J, Ihara H, Alcheikh Y et al (2016) Optimization of a neurotoxin to investigate the contribution of excitatory interneurons to speed modulation in vivo. Curr Biol 26(17):2319–2328
Kawakami K, Abe G, Asada T, Asakawa K, Fukuda R, Ito A et al (2010) zTrap: zebrafish gene trap and enhancer trap database. BMC Dev Biol 10:105
Wullimann MF, Rupp B, Reichert H (1996) Neuroanatomy of the zebrafish brain. Birkhauser, Basel
Portavella M, Torres B, Salas C (2004) Avoidance response in goldfish: emotional and temporal involvement of medial and lateral telencephalic pallium. J Neurosci 24(9):2335–2342
Acknowledgments
This work was supported by internal support from NORCE Norwegian Research Centre AS to P.L. and JSPS Kakenhi 21H02463 and NBRP from AMED to K.K.
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Lal, P., Tanabe, H., Kawakami, K. (2024). Genetic Identification of Neural Circuits Essential for Active Avoidance Fear Conditioning in Adult Zebrafish. In: Amatruda, J.F., Houart, C., Kawakami, K., Poss, K.D. (eds) Zebrafish. Methods in Molecular Biology, vol 2707. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3401-1_11
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DOI: https://doi.org/10.1007/978-1-0716-3401-1_11
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Publisher Name: Humana, New York, NY
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