PUBLICATION
Ontogenetic development of erythropoiesis can be studied non-invasively in GATA-1:DsRed transgenic zebrafish
- Authors
- Yaqoob, N., Holotta, M., Prem, C., Kopp, R., and Schwerte, T.
- ID
- ZDB-PUB-090716-5
- Date
- 2009
- Source
- Comparative biochemistry and physiology. Part A, Molecular & integrative physiology 154(2): 270-278 (Journal)
- Registered Authors
- Kopp, Renate, Schwerte, Thorsten
- Keywords
- DsRed, Erythropoiesis, GATA-1, Hypoxia, Zebrafish
- MeSH Terms
-
- Animals
- Animals, Genetically Modified
- Aorta/ultrastructure
- Blood Cell Count
- Blood Volume/physiology*
- Erythrocyte Volume
- Erythropoiesis/physiology*
- Erythropoietin/metabolism
- Fluorescent Dyes
- GATA1 Transcription Factor/genetics
- GATA1 Transcription Factor/metabolism
- Gene Expression Regulation
- Genes, Reporter
- Larva/growth & development
- Microscopy, Interference
- Oxygen/physiology
- Promoter Regions, Genetic
- RNA, Messenger/metabolism
- Vasodilation/physiology
- Zebrafish/embryology
- Zebrafish/growth & development*
- Zebrafish/physiology
- Zebrafish Proteins/genetics
- Zebrafish Proteins/metabolism
- PubMed
- 19584002 Full text @ Comp. Biochem. Physiol. A Mol. Integr. Physiol.
Citation
Yaqoob, N., Holotta, M., Prem, C., Kopp, R., and Schwerte, T. (2009) Ontogenetic development of erythropoiesis can be studied non-invasively in GATA-1:DsRed transgenic zebrafish. Comparative biochemistry and physiology. Part A, Molecular & integrative physiology. 154(2):270-278.
Abstract
For the erythroid cell lineage development in vertebrates, GATA-1 transcription factor is essential. In our report, we have demonstrated that the approximate developmental status of erythrocytes and the progression of blood formation can be studied non-invasively in GATA-1:DsRed transgenic zebrafish (Danio rerio) embryo and larva by characterization of fluorescence luminance spectra. Study was carried out for the animals maintained under normoxic and hypoxic (152 and 20torr PO(2) respectively) conditions up to 10days post-fertilization (dpf) and total blood cells concentrations and fluorescent cells percentage were determined for this purpose. The erythroids were classified into five intensity stages (IS) on the basis of their fluorescence intensity. The luminescent cells with medium intensities (IS3) in normoxic animals were found throughout 2 to 10dpf although in lower quantity while in hypoxic group they appeared from 5dpf till 10dpf showing a maximum of 15% of the total luminescent cells at 8dpf. The total blood cell concentration dropped after 8dpf in contrast to hypoxic group which showed further increasing trend. The fluorescent cells percentage in normoxic group was generally higher as compared to the hypoxic ones. Our method successfully defined various stages of erythroid development. An effort was also made to correlate our luminance data (GATA-1 expression) and total blood cells concentrations with Epo mRNA production. Quantitative RT-PCR of 2-15dpf old zebrafish was carried out for this purpose. Normoxic animals showed 1-3 Epo mRNA copies per ng RNA in contrast to the hypoxic larvae that showed remarkable fluctuation of 1 to 12 Epo mRNA copies per ng RNA during development. The blood volume (aortic diameter) and production time scale proved to be important factors to define the relationship of Epo mRNA with total blood cells concentration and GATA-1 protein expression respectively.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping