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Review
. 2013 Dec;51(4):271-6.
doi: 10.1016/j.bcmd.2013.07.009. Epub 2013 Aug 2.

Assaying hematopoiesis using zebrafish

Sonja Boatman  1 , Francesca Barrett, Sruthi Satishchandran, Lili Jing, Ilya Shestopalov, Leonard I Zon
Affiliations
Review

Assaying hematopoiesis using zebrafish

Sonja Boatman et al. Blood Cells Mol Dis. 2013 Dec.

Abstract

The zebrafish has become a commonly used model for studying hematopoiesis as a result of its unique attributes. Zebrafish are highly suitable for large-scale genetic and chemical screens compared to other vertebrate systems. It is now possible to analyze hematopoietic lineages in zebrafish and validate cell function via transplantation assays. Here, we review advancements over the past decade in forward genetic screens, chemical screens, fluorescence-activated cell sorting analysis, and transplantation assays. Integrating these approaches enables new chemical and genetic screens that assay cell function within the hematopoietic system. Studies in zebrafish will continue to contribute and expand our knowledge about hematopoiesis, and develop novel treatments for clinical applications.

Keywords: AGM; ALM; BM; CHT; ENU; FACS; FSC; GVHD; HCT; Hematopoiesis; ICM; MHC; PAF; PG; PLM; Pol II Associated Factor; SSC; Screens; Stem cell; TGFβ; TIF1γ; Transplantation; WKM; Zebrafish; anterior lateral mesoderm; aorta–gonad–mesonephros; bone marrow; caudal hematopoietic tissue; ethylnitrosourea; fluorescence-activated cell sorting; forward light scatter; graft-versus-host-disease; hCB; hematopoietic cellular transplantation; hours post fertilization; hpf; human cord blood; intermediate cell mass; major histocompatibility complex; posterior lateral mesoderm; prostaglandin; side light scatter; transcription intermediary factor1γ; transforming growth factor beta; whole kidney marrow.

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Figures

Figure 1
Figure 1. Tifγ regulates transcription elongation of blood genes
(A) In mon mutants, Pol II is paused by the DSIF and PAF and would not efficiently transcribe blood genes. (B) In wildtype cells, Tifγrecruits positive elongation factor p-TEFb and FACT to blood genes through the SCL complex and stimulates Pol II to proceed to finish transcription elongation. (C) In mon; cdc73−/− double mutants, the pausing factor PAF is lost, Pol II is no longer paused. Elongation continues without help from positive elongation factors.
Figure 2
Figure 2
In situ hybridization shows increased expression of HSC markers runx1 and cmyb in the AGM (indicated with arrowheads) of zebrafish embryos at 36hpf treated with dmPGE2. Lateral view with anterior to the left. nt = nueral tube, nc = notochord, agm = aorta-gonad-mesonephros, ys = yolk extension. Magnification = 40X. Figure panels are adapted from reference (Goessling et. al., 2007).
Figure 3
Figure 3
FACS light scatter profiles of the zebrafish kidney marrow, the site of adult hematopoiesis, and peripheral blood. FSC = forward light scatter. SSC = side light scatter.
See this image and copyright information in PMC

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