ZFIN ID: ZDB-PUB-041022-1
Analysis of hematopoietic development in the zebrafish
Paffett-Lugassy, N.N., and Zon, L.I.
Date: 2005
Source: Methods in molecular medicine   105: 171-198 (Chapter)
Registered Authors: Paffett-Lugassy, Noelle, Zon, Leonard I.
Keywords: Zebrafish hematopoiesis; zebrafish blood mutants; whole-mount in situ hybridization; o-dianisidine staining; microinjection; morpholino-mediated knockdown; zebrafish transgenics; hematopoietic lineage separation
MeSH Terms:
  • Animals
  • Animals, Genetically Modified
  • Cell Lineage/genetics
  • Cell Lineage/physiology*
  • Gene Expression Regulation, Developmental/genetics
  • Gene Expression Regulation, Developmental/physiology*
  • Gene Transfer Techniques*
  • Hematopoiesis/genetics
  • Hematopoiesis/physiology*
  • Mutagenesis/genetics*
  • Zebrafish/embryology*
  • Zebrafish/genetics
PubMed: 15492396
The zebrafish (Danio rerio) has emerged as a powerful vertebrate genetic and developmental model that is particularly amenable to the study of hematopoiesis. The zebrafish embryo develops externally and its optical clarity allows the number and morphology of circulating blood cells to be visualized using a dissecting microscope. Both the morphology of the blood lineages and the expression of critical blood genes are highly conserved between zebrafish and mammals. The high fecundity and short generation time of zebrafish facilitate genetic analysis, and a number of large-scale mutagenesis screens have identified mutations in genes affecting blood development. The discovery of novel hematopoietic genes, as well as the cloning of zebrafish homologs of known hematopoietic genes, necessitates the use of efficacious and reliable methods for complete gene characterization. In this chapter, we illustrate frequently used techniques that are essential for evaluating hematopoiesis in the zebrafish, including whole-mount in situ hybridization, the detection of erythrocytes by o-dianisidine staining, and a description of the microinjection procedure, which has various applications, including overexpression of messenger ribonucleic acid, gene 'knockdown' by antisense technology, and the creation of transgenic zebrafish. Also included is an explanation of the use of flow cytometry to separate hema-topoietic lineages from the adult kidney and to isolate relatively pure populations of cell types from transgenic embryos based on the expression of fluorescent markers.