Cpsf1 is required for definitive HSC survival in zebrafish
ABSTRACT A comprehensive understanding of the genes and pathways regulating hematopoiesis is needed to identify genes causally related to bone marrow failure syndromes, myelodysplastic syndromes, and hematopoietic neoplasms. To identify novel genes involved in hematopoiesis, we performed an ethyl-nitrosourea mutagenesis screen in zebrafish (Danio rerio) to search for mutants with defective definitive hematopoiesis. We report the recovery and analysis of the grechetto mutant, which harbors an inactivating mutation in cleavage and polyadenylation specificity factor 1 (cpsf1), a gene ubiquitously expressed and required for 3' untranslated region processing of a subset of pre-mRNAs. grechetto mutants undergo normal primitive hematopoiesis and specify appropriate numbers of definitive HSCs at 36 hours postfertilization. However, when HSCs migrate to the caudal hematopoietic tissue at 3 days postfertilization, their numbers start decreasing as a result of apoptotic cell death. Consistent with Cpsf1 function, c-myb:EGFP(+) cells in grechetto mutants also show defective polyadenylation of snrnp70, a gene required for HSC development. By 5 days postfertilization, definitive hematopoiesis is compromised and severely decreased blood cell numbers are observed across the myeloid, erythroid, and lymphoid cell lineages. These studies show that cpsf1 is essential for HSC survival and differentiation in caudal hematopoietic tissue.
- SourceAvailable from: Jonathan Chernoff
[Show abstract] [Hide abstract]
- "Whole-Mount In Situ Hybridization, Immunohistochemistry, and Cartilage Staining Whole-mount in situ hybridization and immunohistochemistry was performed as described elsewhere (Bolli et al., 2011; Koshida et al., 2005; Yelon et al., 1999). p-Erk (Cell Signaling #9101) was used as primary antibody, and 3,3 0 diaminobenzidine was used for visualization. "
ABSTRACT: Proper neural crest development and migration is critical during embryonic development, but the molecular mechanisms regulating this process remain incompletely understood. Here, we show that the protein kinase Erk, which plays a central role in a number of key developmental processes in vertebrates, is regulated in the developing neural crest by p21-activated protein kinase 1 (Pak1). Furthermore, we show that activated Erk signals by phosphorylating the transcription factor Gata6 on a conserved serine residue to promote neural crest migration and proper formation of craniofacial structures, pigment cells, and the outflow tract of the heart. Our data suggest an essential role for Pak1 as an Erk activator, and Gata6 as an Erk target, during neural crest development.Developmental Cell 05/2014; 29(3):350-9. DOI:10.1016/j.devcel.2014.04.003 · 10.37 Impact Factor
[Show abstract] [Hide abstract]
- "Thus, the VDA, CHT and kidney of the zebrafish are respectively the functional analogs of the AGM, liver and bone marrow in mice (Chen and Zon, 2009). Using forward genetic analyses, three previously uncharacterized factors [rumba (znf574), haus3 and cpsf1] have recently been found to play an essential role in regulating fetal hematopoiesis in zebrafish (Bolli et al., 2011; Du et al., 2011), confirming that the employment of an unbiased forward genetic approach in zebrafish can complement our current knowledge of mammals. Sumoylation is a post-translational modification that participates in various cellular processes, such as regulation of gene transcription, the cell cycle, genomic integrity, stress responses and organogenesis (Hay, 2005). "
ABSTRACT: In vertebrates, establishment of the hematopoietic stem/progenitor cell (HSPC) pool involves mobilization of these cells in successive developmental hematopoietic niches. In zebrafish, HSPCs originate from the ventral wall of the dorsal aorta (VDA), the equivalent of the mammalian aorta-gonad-mesonephros (AGM). The HSPCs subsequently migrate to the caudal hematopoietic tissue (CHT) for transitory expansion and differentiation during the larval stage, and they finally colonize the kidney, where hematopoiesis takes place in adult fish. Here, we report the isolation and characterization of a zebrafish mutant, tango(hkz5), which shows defects of definitive hematopoiesis. In tango(hkz5) mutants, HSPCs initiate normally in the AGM and subsequently colonize the CHT. However, definitive hematopoiesis is not sustained in the CHT owing to accelerated apoptosis and diminished proliferation of HSPCs. Positional cloning reveals that tango(hkz5) encodes SUMO1-activating enzyme subunit 1 (Sae1). A chimera generation experiment and biochemistry analysis reveal that sae1 is cell-autonomously required for definitive hematopoiesis and that the tango(hkz5) mutation produces a truncated Sae1 protein (ΔSae1), resulting in systemic reduction of sumoylation. Our findings demonstrate that sae1 is essential for the maintenance of HSPCs during fetal hematopoiesis in zebrafish.Development 12/2012; 139(23):4321-9. DOI:10.1242/dev.081869 · 6.27 Impact Factor
[Show abstract] [Hide abstract]
- "These cells go on to populate the adult organs of haematopoiesis in the zebrafish, the kidney and the thymus. The precise timing of the move from primitive wave haematopoiesis to definitive wave haematopoiesis has yet to be fully established, but evidence from globin gene expression and mutants with normal primitive wave blood production suggests that the major contribution of haematopoiesis comes from definitive HSC derived cells by around 5 days post fertilization    "
ABSTRACT: Over the past ten years, studies using the zebrafish model have contributed to our understanding of vertebrate haematopoiesis, myelopoiesis, and myeloid leukaemogenesis. Novel insights into the conservation of haematopoietic lineages and improvements in our capacity to identify, isolate, and culture such haematopoietic cells continue to enhance our ability to use this simple organism to address disease biology. Coupled with the strengths of the zebrafish embryo to dissect developmental myelopoiesis and the continually expanding repertoire of models of myeloid malignancies, this versatile organism has established its niche as a valuable tool to address key questions in the field of myelopoiesis and myeloid leukaemogenesis. In this paper, we address the recent advances and future directions in the field of myelopoiesis and leukaemogenesis using the zebrafish system.Advances in Hematology 07/2012; 2012:358518. DOI:10.1155/2012/358518