PUBLICATION
Optogenetic Signaling Activation in Zebrafish Embryos
- Authors
- Saul, A.J., Rogers, C.E., Garmendia-Cedillos, M., Pohida, T., Rogers, K.W.
- ID
- ZDB-PUB-231114-9
- Date
- 2023
- Source
- Journal of visualized experiments : JoVE (200): (Journal)
- Registered Authors
- Rogers, Katherine
- Keywords
- none
- MeSH Terms
-
- Animals
- Light
- Optogenetics*/methods
- Protein Domains
- Signal Transduction
- Zebrafish*/genetics
- Zebrafish Proteins/genetics
- Zebrafish Proteins/metabolism
- PubMed
- 37955383 Full text @ J. Vis. Exp.
Citation
Saul, A.J., Rogers, C.E., Garmendia-Cedillos, M., Pohida, T., Rogers, K.W. (2023) Optogenetic Signaling Activation in Zebrafish Embryos. Journal of visualized experiments : JoVE. (200):.
Abstract
Signaling pathways orchestrate fundamental biological processes, including development, regeneration, homeostasis, and disease. Methods to experimentally manipulate signaling are required to understand how signaling is interpreted in these wide-ranging contexts. Molecular optogenetic tools can provide reversible, tunable manipulations of signaling pathway activity with a high degree of spatiotemporal control and have been applied in vitro, ex vivo, and in vivo. These tools couple light-responsive protein domains, such as the blue light homodimerizing light-oxygen-voltage sensing (LOV) domain, with signaling effectors to confer light-dependent experimental control over signaling. This protocol provides practical guidelines for using the LOV-based bone morphogenetic protein (BMP) and Nodal signaling activators bOpto-BMP and bOpto-Nodal in the optically accessible early zebrafish embryo. It describes two control experiments: A quick phenotype assay to determine appropriate experimental conditions, and an immunofluorescence assay to directly assess signaling. Together, these control experiments can help establish a pipeline for using optogenetic tools in early zebrafish embryos. These strategies provide a powerful platform to investigate the roles of signaling in development, health, and physiology.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping