Visualization of Cellular Electrical Activity in Zebrafish Early Embryos and Tumors

Abstract

Bioelectricity, endogenous electrical signaling mediated by ion channels and pumps located on the cell membrane, plays important roles in signaling processes of excitable neuronal and muscular cells and many other biological processes, such as embryonic developmental patterning. However, there is a need for in vivo electrical activity monitoring in vertebrate embryogenesis. The advances of genetically encoded fluorescent voltage indicators (GEVIs) have made it possible to provide a solution for this challenge. Here, we describe how to create a transgenic voltage indicator zebrafish using the established voltage indicator, ASAP1 (Accelerated Sensor of Action Potentials 1), as an example. The Tol2 kit and a ubiquitous zebrafish promoter, ubi, were chosen in this study. We also explain the processes of Gateway site-specific cloning, Tol2 transposon-based zebrafish transgenesis, and the imaging process for early-stage fish embryos and fish tumors using regular epifluorescent microscopes. Using this fish line, we found that there are cellular electric voltage changes during zebrafish embryogenesis, and fish larval movement. Furthermore, it was observed that in a few zebrafish malignant peripheral nerve sheath tumors, the tumor cells were generally polarized compared to the surrounding normal tissues.

Generating Stable Knockout Zebrafish Lines by Deleting Large Chromosomal Fragments Using Multiple gRNAs

“…Zebrafish embryo microinjections were performed as we described previously (Silic and Zhang 2018). Briefly, adult fish were separated overnight in a fish mating tank by a divider, and 1-cell stage fish embryos were collected in the early morning of the second day immediately before injection.…”

Editorial: Interplay Between Ion Channels, the Nervous System, and Embryonic Development

“…Moreover, the zebrafish model has shown that ion channel effects can be exerted non-cell autonomously as well as cell autonomously (Pineda et al, 2006 ; Wright and Ribera, 2010 ). The paper forms a nice complement to past studies of developmental roles of channels in zebrafish morphogenesis and its disorders (Perathoner et al, 2014 ; Silic and Zhang, 2018 ; Lanni et al, 2019 ).…”

Revisiting Burr and Northrop’s “The Electro-Dynamic Theory of Life” (1935)

“…Burr had a remarkable ability to foresee advances in this field, and his work was no doubt an inspiration to the major figures in this area who advanced this science in the 20th century Jaffe 1975, 1976;Nuccitelli et al , 1977Nuccitelli et al , 1986Weisenseel et al 1975;Borgens et al 1977aBorgens et al , b, 1979aBorgens et al , b, c, 1980Borgens et al , 1981Jaffe and Nuccitelli 1977;Jaffe 1979;Jaffe and Poo 1979;Robinson and McCaig 1980;Stump et al 1980;Hinkle et al 1981;Kline et al 1981;Robinson et al 1981;Borgens 1982Borgens , 1983McCaig and Robinson 1982;McCaig 1986b;Nuccitelli 1986a). If he were alive today, Burr would have thrived in the context of the revolution in optogenetic reporters of bioelectric state and actuators that allow modulating tissue bioelectric patterns at will (Mutoh et al 2012;Adams et al 2014;Cohen and Venkatachalam 2014;Silic and Zhang 2018). Given his ability to see beyond the current mainstream and the willingness to embrace and extend the latest technology, we can only try to imagine the advances he would have made at the interface of molecular biology and developmental physiology.…”