Targeting the zebrafish nervous system using in vivo electroporation

Abstract

Zebrafish are a powerful model system for in vivo analysis of neural development. The degree of genetic characterization, the availability of transgenic animals, and the feasibility of in vivo imaging analysis makes zebrafish an excellent genetic model organism. In vivo electroporation has emerged as an important tool for studies examining the molecular mechanisms of neural development in zebrafish. In vivo electroporation offers precise temporal control over the induction of gain or loss of gene function because it can be used to target embryos at any developmental stage. Furthermore, in vivo electroporation allows for spatial targeting of different regions of the developing nervous system, and when combined with Gal4-based transgenic zebrafish can even target specific cell types. This chapter first outlines the basic protocol for targeting the zebrafish nervous system through electroporation, discussing the details concerning the choice of equipment, approach, and reagents. Then, a method utilizing a Gal4-based transgenic line to specifically target cells of the olfactory bulb is described in detail, and representative result are shown. Using this approach, in vivo electroporation can yield precise temporal control of genetic manipulations and can spatially target specific tissues or cell types.