PUBLICATION

Analysis of signaling pathways in zebrafish development by microinjection

Authors
Kinsey, W.H.
ID
ZDB-PUB-081218-34
Date
2009
Source
Methods in molecular biology (Clifton, N.J.)   518: 1-10 (Chapter)
Registered Authors
Kinsey, William H.
Keywords
Zebrafish, egg, oocyte, zygote, fertilization, Fyn, Yes, Src
MeSH Terms
  • Animals
  • Calcium/metabolism
  • Fertilization
  • Fertilization in Vitro
  • Fluorescence
  • Image Processing, Computer-Assisted
  • Microinjections/methods*
  • Ovum/metabolism
  • Signal Transduction*
  • Zebrafish/embryology*
  • Zebrafish/metabolism*
PubMed
19085131 Full text @ Meth. Mol. Biol.
Abstract
The zebrafish oocyte differs substantially from the zygote and cleavage-stage embryo with regard to the ease with which it can be microinjected with proteins or reagents that modify subsequent development. The objective of this chapter is to describe methods developed in this and other laboratories for microinjection and calcium imaging in the unfertilized zebrafish egg. Methods of immobilizing the oocyte include a holding chamber and a holding pipette. The holding chamber allows imaging of three or four oocytes simultaneously, while the holding pipette facilitates imaging of localized regions in the oocyte. Injection of calcium green dextran via holding chambers allowed detection of global changes in Ca(2+) release following fertilization and development through early blastula stages. Injection and imaging with the holding pipette method allowed discrimination of calcium changes in the egg cortex from that in the central regions of the cell. The results demonstrate the highly localized nature of calcium signaling in the zebrafish zygote and the implications of this signaling for embryonic development.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping