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Article: The role of phospholipase C gamma 1 in primitive hematopoiesis during zebrafish development

TitleThe role of phospholipase C gamma 1 in primitive hematopoiesis during zebrafish development
Authors
Issue Date2007
PublisherElsevier Inc. The Journal's web site is located at http://www.elsevier.com/locate/exphem
Citation
Experimental Hematology, 2007, v. 35 n. 3, p. 368-373 How to Cite?
AbstractObjectives: Phospholipase C (PLC) gamma 1 has been shown to mediate signal transduction of tyrosine kinases and affect function of hematopoietic cells. However, its role in hematopoiesis during embryonic development is currently unclear. In this study, we examined this issue using morpholino (MO) gene knockdown in zebrafish embryos. Methods: MO targeting at the exon-1-intron-1 junction of zebrafish PLC-γ1 was injected into embryos at the one- to four-cell stage (referred herein zPLC-γ1MO embryos). Primitive hematopoiesis was examined quantitatively by flow cytometry in Tg(gata1:GFP) embryos and by real-time quantitative polymerase chain reaction at 18 hours-post-fertilization (hpf), before the onset of circulation. The embryos were also treated with receptor inhibitors of vascular endothelial growth factor, fibroblast growth factor, and platelet-derived growth factor at 25, 1, and 30 μmol/L, respectively, from one cell until 48 hpf. Results: Erythropoiesis was reduced in zPLC-γ1MO embryos, as shown by the reduction in gata1+ cells (wild-type: 4.32% ± 0.10% vs zPLC-γ1MO: 2.38% ± 0.11%, p = 0.021) and gata1 and α-embryonic hemoglobin expression [0.47 ± 0.06-fold (p = 0.013) and 0.46 ± 0.04-fold (p = 0.013)]. Expression of scl, lmo-2 (early hematopoietic progenitors), pu.1, and l-plastin (myelomonocytic lineages) as well as fli1 (vascular progenitors) were unaffected. Fli1+ cells in Tg(fli1:GFP) embryos were also unaffected by zPLC-γ1MO. When embryos were incubated with receptor inhibitors of vascular endothelial growth factor (VEGFRTKI), fibroblast growth factor (SU5402), or platelet-derived growth factor (AG1296), only VEGFRTKI reduced erythropoiesis [VEGFRTKI: 2.10% ± 0.07% (p = 0.021) vs SU5402: 4.08% ± 0.12% (p = 0.248) vs AG1296: 4.12% ± 0.14% (p = 0.149)]. Conclusion: PLC-γ1 is involved in the regulation of primitive hematopoiesis in zebrafish embryos, which is distinct from its later effect on vascular formation. © 2007 International Society for Experimental Hematology.
Persistent Identifierhttp://hdl.handle.net/10722/163064
ISSN
2015 Impact Factor: 2.303
2015 SCImago Journal Rankings: 1.341
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorMa, ACHen_US
dc.contributor.authorLiang, Ren_US
dc.contributor.authorLeung, AYHen_US
dc.date.accessioned2012-09-05T05:27:07Z-
dc.date.available2012-09-05T05:27:07Z-
dc.date.issued2007en_US
dc.identifier.citationExperimental Hematology, 2007, v. 35 n. 3, p. 368-373en_US
dc.identifier.issn0301-472Xen_US
dc.identifier.urihttp://hdl.handle.net/10722/163064-
dc.description.abstractObjectives: Phospholipase C (PLC) gamma 1 has been shown to mediate signal transduction of tyrosine kinases and affect function of hematopoietic cells. However, its role in hematopoiesis during embryonic development is currently unclear. In this study, we examined this issue using morpholino (MO) gene knockdown in zebrafish embryos. Methods: MO targeting at the exon-1-intron-1 junction of zebrafish PLC-γ1 was injected into embryos at the one- to four-cell stage (referred herein zPLC-γ1MO embryos). Primitive hematopoiesis was examined quantitatively by flow cytometry in Tg(gata1:GFP) embryos and by real-time quantitative polymerase chain reaction at 18 hours-post-fertilization (hpf), before the onset of circulation. The embryos were also treated with receptor inhibitors of vascular endothelial growth factor, fibroblast growth factor, and platelet-derived growth factor at 25, 1, and 30 μmol/L, respectively, from one cell until 48 hpf. Results: Erythropoiesis was reduced in zPLC-γ1MO embryos, as shown by the reduction in gata1+ cells (wild-type: 4.32% ± 0.10% vs zPLC-γ1MO: 2.38% ± 0.11%, p = 0.021) and gata1 and α-embryonic hemoglobin expression [0.47 ± 0.06-fold (p = 0.013) and 0.46 ± 0.04-fold (p = 0.013)]. Expression of scl, lmo-2 (early hematopoietic progenitors), pu.1, and l-plastin (myelomonocytic lineages) as well as fli1 (vascular progenitors) were unaffected. Fli1+ cells in Tg(fli1:GFP) embryos were also unaffected by zPLC-γ1MO. When embryos were incubated with receptor inhibitors of vascular endothelial growth factor (VEGFRTKI), fibroblast growth factor (SU5402), or platelet-derived growth factor (AG1296), only VEGFRTKI reduced erythropoiesis [VEGFRTKI: 2.10% ± 0.07% (p = 0.021) vs SU5402: 4.08% ± 0.12% (p = 0.248) vs AG1296: 4.12% ± 0.14% (p = 0.149)]. Conclusion: PLC-γ1 is involved in the regulation of primitive hematopoiesis in zebrafish embryos, which is distinct from its later effect on vascular formation. © 2007 International Society for Experimental Hematology.en_US
dc.languageengen_US
dc.publisherElsevier Inc. The Journal's web site is located at http://www.elsevier.com/locate/exphemen_US
dc.relation.ispartofExperimental Hematologyen_US
dc.rightsExperimental Hematology. Copyright © Elsevier Inc.-
dc.subject.meshAnimalsen_US
dc.subject.meshErythropoiesis - Drug Effects - Physiologyen_US
dc.subject.meshFibroblast Growth Factor 1 - Administration & Dosageen_US
dc.subject.meshFlow Cytometry - Methodsen_US
dc.subject.meshGene Expression Profilingen_US
dc.subject.meshGene Expression Regulation, Developmentalen_US
dc.subject.meshGene Targeting - Methodsen_US
dc.subject.meshGreen Fluorescent Proteins - Geneticsen_US
dc.subject.meshHematopoiesis - Drug Effects - Genetics - Physiologyen_US
dc.subject.meshOligonucleotides, Antisense - Administration & Dosageen_US
dc.subject.meshPhospholipase C Gamma - Drug Effects - Genetics - Physiologyen_US
dc.subject.meshPlatelet-Derived Growth Factor - Administration & Dosageen_US
dc.subject.meshReverse Transcriptase Polymerase Chain Reaction - Methodsen_US
dc.subject.meshVascular Endothelial Growth Factor A - Administration & Dosageen_US
dc.subject.meshZebrafish - Embryology - Genetics - Physiologyen_US
dc.titleThe role of phospholipase C gamma 1 in primitive hematopoiesis during zebrafish developmenten_US
dc.typeArticleen_US
dc.identifier.emailLiang, R:rliang@hku.hken_US
dc.identifier.emailLeung, AYH:ayhleung@hku.hken_US
dc.identifier.authorityLiang, R=rp00345en_US
dc.identifier.authorityLeung, AYH=rp00265en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.doi10.1016/j.exphem.2006.11.010en_US
dc.identifier.pmid17309817-
dc.identifier.scopuseid_2-s2.0-33847030744en_US
dc.identifier.hkuros136885-
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-33847030744&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume35en_US
dc.identifier.issue3en_US
dc.identifier.spage368en_US
dc.identifier.epage373en_US
dc.identifier.isiWOS:000244605200004-
dc.publisher.placeUnited Statesen_US
dc.identifier.scopusauthoridMa, ACH=15849157500en_US
dc.identifier.scopusauthoridLiang, R=26643224900en_US
dc.identifier.scopusauthoridLeung, AYH=7403012668en_US
dc.identifier.citeulike10829715-

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