PUBLICATION

Using the Zebrafish as an Approach to Examine the Mechanisms of Vertebrate Erythropoiesis

Authors
Kafina, M.D., Paw, B.H.
ID
ZDB-PUB-171028-5
Date
2018
Source
Methods in molecular biology (Clifton, N.J.)   1698: 11-36 (Chapter)
Registered Authors
Paw, Barry
Keywords
Danio rerio, Flow cytometry, Hemoglobin, In situ hybridization, Microinjection, o-dianisidine staining
MeSH Terms
  • Animals
  • Biomarkers
  • Cell Differentiation
  • Cell Lineage
  • Drosophila Proteins/genetics
  • Drosophila Proteins/metabolism
  • Erythrocytes/metabolism
  • Erythropoiesis*
  • Flow Cytometry
  • GATA1 Transcription Factor/genetics
  • GATA1 Transcription Factor/metabolism
  • GATA2 Transcription Factor/genetics
  • GATA2 Transcription Factor/metabolism
  • Gene Expression Regulation, Developmental
  • Hemoglobins
  • Humans
  • In Situ Hybridization*/methods
  • Microinjections
  • Models, Animal
  • Mutation
  • Phenotype
  • Vertebrates*
  • Zebrafish*
PubMed
29076082 Full text @ Meth. Mol. Biol.
Abstract
The zebrafish, Danio rerio, is a powerful model for the study of erythropoiesis and defining the genetic basis of hematological diseases. The mechanisms of erythroid differentiation are highly conserved in the zebrafish, permitting translational research studies and the modeling of erythropoiesis in higher vertebrates. An advantage of the system is the ability to manipulate gene expression and observe the effect on erythroid development in vivo, with relative ease and rapidity. The production of optically transparent embryos also makes it an attractive tool for visual analysis of circulating erythrocytes that can be used to study erythropoiesis. Through large-scale chemical mutagenesis screens, a variety of zebrafish blood mutants have been identified that are used for gene discoveries and the recapitulation of human diseases. Experimental techniques including in situ hybridization, o-dianisidine staining, flow cytometry, and microinjection are now commonly employed to study red blood cell biochemistry and erythropoiesis in the zebrafish. These techniques have been applied for identifying novel genes required for the hemoglobin synthesis, isolating blood cell lineages, visualizing genetic expression within erythroid tissues, and characterizing the phenotype of blood disorders. The applications of zebrafish methodology to the study of erythropoiesis and optimized step-by-step protocols are discussed in this chapter.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping