PUBLICATION

FRAP Analysis of Extracellular Diffusion in Zebrafish Embryos

Authors
Soh, G.H., Müller, P.
ID
ZDB-PUB-181017-13
Date
2018
Source
Methods in molecular biology (Clifton, N.J.)   1863: 107-124 (Other)
Registered Authors
Müller, Patrick
Keywords
Developmental biology, Extracellular signaling molecules, FRAP, Fluorescence Recovery After Photobleaching, Morphogens, Zebrafish
MeSH Terms
  • Animals
  • Biological Transport
  • Diffusion
  • Embryo, Nonmammalian/cytology*
  • Embryo, Nonmammalian/physiology*
  • Fluorescence Recovery After Photobleaching/methods*
  • Image Processing, Computer-Assisted/methods*
  • Models, Biological
  • Morphogenesis*
  • Signal Transduction
  • Software
  • Zebrafish/embryology*
  • Zebrafish/physiology*
PubMed
30324594 Full text @ Meth. Mol. Biol.
Abstract
Morphogens are signaling molecules that provide positional information to cells during development. They must move through embryonic tissues in order to coordinate patterning. The rate of a morphogen's movement through a tissue-its effective diffusivity-affects the morphogen's distribution and therefore influences patterning. Fluorescence recovery after photobleaching (FRAP) is a powerful method to measure the effective diffusion of molecules through cells and tissues, and has been successfully employed to examine morphogen mobility and gain important insights into embryogenesis. Here, we provide detailed protocols for FRAP assays in vitro and in living zebrafish embryos, and we explain how to analyze FRAP data using the open-source software PyFRAP to determine effective diffusion coefficients.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping