Abstract
This chapter describes three fast and straightforward methods to introduce nucleic acids, dyes, and other molecules into small numbers of cells of zebrafish embryos, larvae, and adults using electroporation. These reagents are delivered through a glass micropipette and electrical pulses are given through electrodes to permeabilize cell membranes and promote uptake of the reagent. This technique allows the experimenter to target cells of their choice at a particular time of development and at a particular location in the zebrafish with high precision and facilitates long-term noninvasive measurement of biological activities in vivo. Applications include cell fate mapping, neural circuit mapping, neuronal activity measurement, manipulation of activity, ectopic gene expression, and genetic knockdown experiments.
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References
De Vry J, Martínez-Martínez P, Losen M, Temel Y, Steckler T, Steinbusch HWM, De Baets MH, Prickaerts J (2010) In vivo electroporation of the central nervous system: a non-viral approach for targeted gene delivery. Prog Neurobiol 92:227–244. doi:10.1016/j.pneurobio.2010.10.001
Concha ML, Russell C, Regan JC, Tawk M, Sidi S, Gilmour DT, Kapsimali M, Sumoy L, Goldstone K, Amaya E et al (2003) Local tissue interactions across the dorsal midline of the forebrain establish CNS laterality. Neuron 39:423–438
Rambabu KM, Rao SHN, Rao NM (2005) Efficient expression of transgenes in adult zebrafish by electroporation. BMC Biotechnol 5:29. doi:10.1186/1472-6750-5-29
Hendricks M, Jesuthasan S (2007) Electroporation-based methods for in vivo, whole mount and primary culture analysis of zebrafish brain development. Neural Dev 2:6. doi:10.1186/1749-8104-2-6
Bianco IH, Carl M, Russell C, Clarke JD, Wilson SW (2008) Brain asymmetry is encoded at the level of axon terminal morphology. Neural Dev 3:9. doi:10.1186/1749-8104-3-9
Kustermann S, Schmid S, Biehlmaier O, Kohler K (2008) Survival, excitability, and transfection of retinal neurons in an organotypic culture of mature zebrafish retina. Cell Tissue Res 332:195–209. doi:10.1007/s00441-008-0589-5
Pittman AJ, Gaynes JA, Chien C-B (2010) nev (cyfip2) is required for retinal lamination and axon guidance in the zebrafish retinotectal system. Dev Biol 344:784–794. doi:10.1016/j.ydbio.2010.05.512
Zou M, De Koninck P, Neve RL, Friedrich RW (2014) Fast gene transfer into the adult zebrafish brain by herpes simplex virus 1 (HSV-1) and electroporation: methods and optogenetic applications. Front Neural Circuits 8:41. doi:10.3389/fncir.2014.00041
Kitamura K, Judkewitz B, Kano M, Denk W, Hausser M (2008) Targeted patch-clamp recordings and single-cell electroporation of unlabeled neurons in vivo. Nat Methods 5:61–67
Haas K, Sin WC, Javaherian A, Li Z, Cline HT (2001) Single-cell electroporation for gene transfer in vivo. Neuron 29:583–591
Tawk M, Bianco IH, Clarke JD (2009) Focal electroporation in zebrafish embryos and larvae. Methods Mol Biol 546:145–151
Wullimann MF, Reichert H (1996) Neuroanatomy of the zebrafish brain: a topological atlas […] […]. Birkhäuser, Basel [u.a.]. doi:10.1007/978-3-0348-8979-7
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Zou, M., Friedrich, R.W., Bianco, I.H. (2016). Targeted Electroporation in Embryonic, Larval, and Adult Zebrafish. In: Kawakami, K., Patton, E., Orger, M. (eds) Zebrafish. Methods in Molecular Biology, vol 1451. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3771-4_17
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DOI: https://doi.org/10.1007/978-1-4939-3771-4_17
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