Regulation of primitive hematopoiesis in zebrafish embryos by the death receptor gene
Copyright policy )Regulation of primitive hematopoiesis in zebrafish embryos by the death receptor gene
We investigated the regulatory mechanism of primitive hematopoiesis in zebrafish (Danio rerio) embryos with particular reference to the role of a death receptor (zDR) gene, based on a morpholino (MO) knockdown approach.MOs targeting the zDR and chordin (Chd) were injected into naturally spawned embryos at one- to four-cell stage. A random sequence (RS) MO was used as a control. Effects on hemoglobin formation (Hb), apoptosis, and lineage-specific gene expression were examined. Embryos injected with zDR, Chd, and RS-MOs were denoted zDR(mo), zChd(mo), and zRS(mo), respectively. Those co-injected with Chd+zDR-MOs and Chd+RS-MOs were abbreviated zChd+DR(mo) and zChd+RS(mo).zDR mRNA expression was restricted to the intermediate cell mass of wild-type (WT) and zChd(mo) embryos. At 48 hours postfertilization, zDR(mo) embryos showed increased Hb compared with WT or zRS(mo) embryos (2.36 x 10(-2) +/- 1.13 x 10(-3) vs 1.85 x 10(-2) +/- 5.60 x 10(-4) vs 1.79 x 10(-2) +/- 1.31 x 10(-3) U, p < 0.05). zChd+DR(mo) embryos also showed increased Hb compared with zChd(mo) or zChd+RS(mo) embryos (4.60 x 10(-2) +/- 2.79 x 10(-3) vs 3.17 x 10(-2) +/- 1.07 x 10(-3) vs 3.05 x 10(-2) +/- 1.25 x 10(-3) U, p < 0.05). zDR-MO reduced apoptosis, as shown by reduced terminal transferase-mediated dUTP nick end-labeling staining in zChd+DR(mo) compared with zChd+RS(mo) embryos and caspase-3 activity in zDR(mo) vs zRS(mo) (0.525 +/- 0.094 vs 0.953 +/- 0.113 U, p < 0.05), and zChd+DR(mo) vs zChd+RS(mo) embryos (0.247 +/- 0.121 vs 1.180 +/- 0.082, p < 0.05). zChd+DR(mo) embryos showed upregulation of erythroid-specific embryonic hemoglobin gene expression but not that of a myeloid-specific myeloperoxidase gene.Knockdown of zDR in zebrafish embryos decreased apoptosis and increased Hb, suggesting that zDR may regulate primitive hematopoiesis during development.
- University of California, Los Angeles United States
- KU Leuven Belgium
- University of Minnesota United States
- University of Minnesota Morris United States
- University of Minnesota System United States
Caspases - drug effects - metabolism, Embryo, Nonmammalian, Receptors, Tumor Necrosis Factor - administration & dosage - genetics - metabolism, Apoptosis, Research & Experimental Medicine, Receptors, Tumor Necrosis Factor, Hemoglobins, Receptors, Zebrafish - embryology - genetics - metabolism, BMP-4, Tumor Necrosis Factor - administration & dosage - genetics - metabolism, Morpholines - administration & dosage, 1102 Cardiorespiratory Medicine and Haematology, Zebrafish, Nonmammalian, Caspase 3, INDUCTION, Gene Expression Regulation, Developmental, Hematology, APOPTOSIS, Medicine, Research & Experimental, Embryo, Glycoproteins - administration & dosage, Caspases, Intercellular Signaling Peptides and Proteins, Intercellular Signaling Peptides and Proteins - administration & dosage, 3201 Cardiovascular medicine and haematology, Life Sciences & Biomedicine, Developmental - drug effects, Morpholines, Apoptosis - drug effects, Immunology, Hemoglobins - biosynthesis - drug effects, Gene Expression Regulation, Developmental - drug effects, Animals, Glycoproteins, Science & Technology, Animal, MESODERM, Hematopoiesis, Disease Models, Animal, Gene Expression Regulation, TRANSGENIC ZEBRAFISH, Disease Models, CELLS, ERYTHROPOIESIS, Hematopoiesis - drug effects - genetics - physiology
Caspases - drug effects - metabolism, Embryo, Nonmammalian, Receptors, Tumor Necrosis Factor - administration & dosage - genetics - metabolism, Apoptosis, Research & Experimental Medicine, Receptors, Tumor Necrosis Factor, Hemoglobins, Receptors, Zebrafish - embryology - genetics - metabolism, BMP-4, Tumor Necrosis Factor - administration & dosage - genetics - metabolism, Morpholines - administration & dosage, 1102 Cardiorespiratory Medicine and Haematology, Zebrafish, Nonmammalian, Caspase 3, INDUCTION, Gene Expression Regulation, Developmental, Hematology, APOPTOSIS, Medicine, Research & Experimental, Embryo, Glycoproteins - administration & dosage, Caspases, Intercellular Signaling Peptides and Proteins, Intercellular Signaling Peptides and Proteins - administration & dosage, 3201 Cardiovascular medicine and haematology, Life Sciences & Biomedicine, Developmental - drug effects, Morpholines, Apoptosis - drug effects, Immunology, Hemoglobins - biosynthesis - drug effects, Gene Expression Regulation, Developmental - drug effects, Animals, Glycoproteins, Science & Technology, Animal, MESODERM, Hematopoiesis, Disease Models, Animal, Gene Expression Regulation, TRANSGENIC ZEBRAFISH, Disease Models, CELLS, ERYTHROPOIESIS, Hematopoiesis - drug effects - genetics - physiology
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