Zebrafish has emerged as an important model for the study of embryonic hematopoiesis. It is a well-characterized model with numerous advantages. Large amount of embryos can be produced by a single pair of zebrafish and the optically transparent embryos allow direct visualization and manipulation of embryonic processes. Large-scale chemical screening using zebrafish embryos can be developed for robust screening of chemical libraries.
The zebrafish hematopoiesis resembles that of mammals and occurs in two successive waves, primitive and definitive hematopoiesis. High-throughput read-outs are available to study the effects of different chemicals and genetic modifications on hematopoiesis.
In first part of this study, an initial screening using O-dianisidine staining and whole-mount in-situ hybridization as read-out for chemicals that might perturb the regulation of hematopoiesis was conducted. Positive hit was further evaluated by flow cytometry of dissociated transgenic Tg(gata1:GFP) zebrafish embryos. A total of 50 compounds were screened from the "Mechanistic set" chemical libraries obtained from Developmental Therapeutics Program of the National Cancer Institute. One compound, "NSC 643834" was shown to reduce O-dianisidine staining at different concentrations tested.
The second part of this study was performed to investigate the role of inca2 in zebrafish hematopoiesis. inca2 was found to be upregulated in chordin morphant zebrafish in which primitive hematopoiesis was expanded. The spatial expression of inca2 was examined by whole mount in-situ hybridization of embryos at different developmental stages. Furthermore the function of inca2 was investigated by gene knockdown using inca2 anti-sense morpholino. Primitive hematopoiesis was perturbed, suggesting that inca2 might play an important role in the regulation of this process.
In conclusion, the present study demonstrated the distinct advantages and feasibility of using zebrafish as a platform of high throughput chemical screening and genetic manipulation. The result provided important ground to investigate the regulatory mechanisms of embryonic hematopoiesis. / published_or_final_version / Medicine / Master / Master of Research in Medicine
| Identifer | oai:union.ndltd.org:HKU/oai:hub.hku.hk:10722/192789 |
| Date | January 2013 |
| Creators | Ng, Koon-kiu., 吳官橋. |
| Publisher | The University of Hong Kong (Pokfulam, Hong Kong) |
| Source Sets | Hong Kong University Theses |
| Language | English |
| Detected Language | English |
| Type | PG_Thesis |
| Source | http://hub.hku.hk/bib/B5071322X |
| Rights | The author retains all proprietary rights, (such as patent rights) and the right to use in future works., Creative Commons: Attribution 3.0 Hong Kong License |
| Relation | HKU Theses Online (HKUTO) |
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