PUBLICATION
Biosensing Motor Neuron Membrane Potential in Live Zebrafish Embryos
- Authors
- Benedetti, L., Ghilardi, A., Prosperi, L., Francolini, M., Del Giacco, L.
- ID
- ZDB-PUB-170704-5
- Date
- 2017
- Source
- Journal of visualized experiments : JoVE (124): (Journal)
- Registered Authors
- Del Giacco, Luca, Ghilardi, Anna, Prosperi, Laura
- Keywords
- none
- MeSH Terms
-
- Animals
- Cell Communication
- Embryo, Nonmammalian
- Membrane Potentials/physiology*
- Motor Neurons/physiology*
- Zebrafish/embryology*
- PubMed
- 28671644 Full text @ J. Vis. Exp.
Citation
Benedetti, L., Ghilardi, A., Prosperi, L., Francolini, M., Del Giacco, L. (2017) Biosensing Motor Neuron Membrane Potential in Live Zebrafish Embryos. Journal of visualized experiments : JoVE. (124).
Abstract
The protocols described here are designed to allow researchers to study cell communication without altering the integrity of the environment in which the cells are located. Specifically, they have been developed to analyze the electrical activity of excitable cells, such as spinal neurons. In such a scenario, it is crucial to preserve the integrity of the spinal cell, but it is also important to preserve the anatomy and physiological shape of the systems involved. Indeed, the comprehension of the manner in which the nervous system-and other complex systems-works must be based on a systemic approach. For this reason, the live zebrafish embryo was chosen as a model system, and the spinal neuron membrane voltage changes were evaluated without interfering with the physiological conditions of the embryos. Here, an approach combining the employment of zebrafish embryos with a FRET-based biosensor is described. Zebrafish embryos are characterized by a very simplified nervous system and are particularly suited for imaging applications thanks to their transparency, allowing for the employment of fluorescence-based voltage indicators at the plasma membrane during zebrafish development. The synergy between these two components makes it possible to analyze the electrical activity of the cells in intact living organisms, without perturbing the physiological state. Finally, this non-invasive approach can co-exist with other analyses (e.g., spontaneous movement recordings, as shown here).
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping