PUBLICATION
A 3-D Tail Explant Culture to Study Vertebrate Segmentation in Zebrafish
- Authors
- Simsek, M.F., Özbudak, E.M.
- ID
- ZDB-PUB-210720-11
- Date
- 2021
- Source
- Journal of visualized experiments : JoVE (172): (Journal)
- Registered Authors
- Ozbudak, Ertugrul, Simsek, Muhammed
- Keywords
- none
- MeSH Terms
-
- Animals
- Body Patterning
- Embryonic Development
- Gene Expression Regulation, Developmental
- Mesoderm/metabolism
- Somites*
- Zebrafish*/genetics
- Zebrafish Proteins/metabolism
- PubMed
- 34279500 Full text @ J. Vis. Exp.
Citation
Simsek, M.F., Özbudak, E.M. (2021) A 3-D Tail Explant Culture to Study Vertebrate Segmentation in Zebrafish. Journal of visualized experiments : JoVE. (172):.
Abstract
Vertebrate embryos pattern their major body axis as repetitive somites, the precursors of vertebrae, muscle, and skin. Somites progressively segment from the presomitic mesoderm (PSM) as the tail end of the embryo elongates posteriorly. Somites form with regular periodicity and scale in size. Zebrafish is a popular model organism as it is genetically tractable and has transparent embryos that allow for live imaging. Nevertheless, during somitogenesis, fish embryos are wrapped around a large, rounding yolk. This geometry limits live imaging of PSM tissue in zebrafish embryos, particularly at higher resolutions that require a close objective working distance. Here, we present a flattened 3-D tissue culture method for live imaging of zebrafish tail explants. Tail explants mimic intact embryos by displaying a proportional slowdown of axis elongation and shortening of rostrocaudal somite lengths. We are further able to stall axis elongation speed through explant culture. This, for the first time, enables us to untangle the chemical input of signaling gradients from the mechanistic input of axial elongation. In future studies, this method can be combined with a microfluidic setup to allow time-controlled pharmaceutical perturbations or screening of vertebrate segmentation without any drug penetration concerns.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping