ZFIN ID: ZDB-PUB-171028-5
Using the Zebrafish as an Approach to Examine the Mechanisms of Vertebrate Erythropoiesis
Kafina, M.D., Paw, B.H.
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.
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.