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Studies of the Mechanisms of Myelopoiesis in Goldfish (Carassius auratus L.) Open Access
- Other title
Granulocyte colony-stimulating factor receptor
Stem cell factor
Colony-stimulating factor-1 receptor
- Type of item
- Degree grantor
University of Alberta
- Author or creator
Katzenback, Barbara A
- Supervisor and department
Belosevic, Miodrag (Biological Sciences)
- Examining committee member and department
Barreda, Daniel (Agricultural, Food and Nutritional Science)
Goss, Greg G (Biological Sciences)
Bols, Niels (Biology)
Field, Catherine (Agricultural, Food and Nutritional Science)
Department of Biological Sciences
Physiology, Cell and Developmental Biology
- Date accepted
- Graduation date
Doctor of Philosophy
- Degree level
Development of progenitor cells into myeloid cells (i.e. macrophages and neutrophils) is critical to the survival of metazoans for maintenance of homeostasis and defense against pathogens. While much is known about myeloid cell development in mammals, less is known about this process in fish. The objective of my thesis was to characterize the myeloid progenitor cell subpopulations of goldfish, their growth factors, receptors and transcription factors (TFs) involved in the process of myeloid cell development (myelopoiesis).
Goldfish kita receptor and its ligand, kitla, were cloned, expressed and characterized. The kita was highly expressed in progenitor cells, compared to monocytes and mature macrophages. The recombinant KITLA was glycosylated, formed non-covalent homodimers, induced a progenitor cell chemotactic response and promoted the survival of progenitor cells. These results suggest a role for KITA/KITLA in the retention and survival of progenitor cells.
Colony stimulating factor-1 receptor (CSF-1R) is important for macrophage development. Antibody to goldfish CSF-1R was shown to specifically identify cells of the macrophage lineage, including a subpopulation of progenitor cells. The CSF-1R+ population of progenitor cells decreased with time of culture, coinciding with the generation of monocytes and macrophages.
The assessment of mRNA levels of a panel of TFs in progenitor cells showed that this cell population was committed to the macrophage lineage by day 2 of cultivation. Addition of rgKITLA and rgCSF-1 to progenitor cells modulated specific myeloid TF mRNA levels consistent with the functional characterization studies.
A procedure for the isolation of highly purified primary neutrophils was developed in this study. Primary neutrophils, isolated from the goldfish kidney produced, reactive oxygen intermediates and degranulated after exposure to mitogens and the fish pathogen, Aeromonas salmonicida, in a dose- and time-dependent manner. Goldfish granulocyte colony stimulating factor receptor (gcsfr) was identified and expressed by neutrophils. gcsfr mRNA levels were found to be significantly higher after exposure of neutrophils to mitogens or A. salmonicida, suggesting a role for GCSFR in neutrophil survival or activation during inflammation.
Taken together, my results highlight the importance of growth factors, receptors and transcription factors in the modulation of goldfish myelopoiesis.
- Permission is hereby granted to the University of Alberta Libraries to reproduce single copies of this thesis and to lend or sell such copies for private, scholarly or scientific research purposes only. Where the thesis is converted to, or otherwise made available in digital form, the University of Alberta will advise potential users of the thesis of these terms. The author reserves all other publication and other rights in association with the copyright in the thesis and, except as herein before provided, neither the thesis nor any substantial portion thereof may be printed or otherwise reproduced in any material form whatsoever without the author's prior written permission.
- Citation for previous publication
Katzenback BA, and Belosevic M. 2009 Molecular and functional characterization of kita and kitla of the goldfish (Carassius auratus L.). Developmental and Comparative Immunology 33: 1165-1175Katzenback BA and Belosevic M. 2012. Colony stimulating factor-1 receptor protein expression is a specific marker for goldfish (Carassius auratus L.) macrophage progenitors and their differentiated cell types. Fish and Shellfish Immunology 32: 434-445Katzenback BA, Karpman M, and Belosevic M. 2011 Distribution and expression analysis of transcription factors in tissues and progenitor cell populations of the goldfish (Carassius auratus L.) in response to growth factors and pathogens. Molecular Immunology 48: 1224-1235Katzenback BA and Belosevic M. 2009. Isolation and functional characterization of neutrophil-like cells from goldfish (Carassius auratus L.) kidney. Developmental and Comparative Immunology 33: 601-611.Katzenback BA and Belosevic M. 2012. Characterization of granulocyte colony stimulating factor receptor of the goldfish (Carassius auratus L.). Developmental and Comparative Immunology 36: 199-207.
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