PUBLICATION
Real-time monitoring of endogenous Fgf8a gradient attests to its role as a morphogen during zebrafish gastrulation
- Authors
- Harish, R.K., Gupta, M., Zöller, D., Hartmann, H., Gheisari, A., Machate, A., Hans, S., Brand, M.
- ID
- ZDB-PUB-230905-52
- Date
- 2023
- Source
- Development (Cambridge, England) 150(19): (Journal)
- Registered Authors
- Brand, Michael, Gupta, Mansi, Hans, Stefan, Machate, Anja, Zöller, Daniela
- Keywords
- Diffusion, Fgf8a, Morphogen, Neural plate, Patterning, Zebrafish
- MeSH Terms
-
- Animals
- Body Patterning/genetics
- Fibroblast Growth Factors*/genetics
- Fibroblast Growth Factors*/metabolism
- Gastrulation*/genetics
- Morphogenesis/genetics
- Signal Transduction/genetics
- Zebrafish*
- Zebrafish Proteins*/genetics
- Zebrafish Proteins*/metabolism
- PubMed
- 37665167 Full text @ Development
Citation
Harish, R.K., Gupta, M., Zöller, D., Hartmann, H., Gheisari, A., Machate, A., Hans, S., Brand, M. (2023) Real-time monitoring of endogenous Fgf8a gradient attests to its role as a morphogen during zebrafish gastrulation. Development (Cambridge, England). 150(19):.
Abstract
Morphogen gradients impart positional information to cells in a homogenous tissue field. Fgf8a, a highly conserved growth factor, has been proposed to act as a morphogen during zebrafish gastrulation. However, technical limitations have so far prevented direct visualization of the endogenous Fgf8a gradient and confirmation of its morphogenic activity. Here, we monitor Fgf8a propagation in the developing neural plate using a CRISPR/Cas9-mediated EGFP knock-in at the endogenous fgf8a locus. By combining sensitive imaging with single-molecule Fluorescence Correlation Spectroscopy (FCS), we demonstrate that Fgf8a, produced at the embryonic margin, propagates by diffusion through the extracellular space and forms a graded distribution towards the animal pole. Overlaying the Fgf8a gradient curve with expression profiles of its downstream targets determines the precise input-output relationship of Fgf8a-mediated patterning. Manipulation of the extracellular Fgf8a levels alters the signaling outcome, thus establishing Fgf8a as a bona fide morphogen during zebrafish gastrulation. Furthermore, by hindering Fgf8a diffusion, we demonstrate that extracellular diffusion of the protein from the source is critical for it to achieve its morphogenic potential.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping