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  • http://hdl.handle.net/10402/era.25357
  • Studies of the Mechanisms of Myelopoiesis in Goldfish (Carassius auratus L.)
  • Katzenback, Barbara A
  • English
  • Goldfish
    Macrophages
    Myelopoiesis
    Granulopoiesis
    Monopoiesis
    Transcription factors
    Stem cell factor
    Kit receptor
    Colony-stimulating factor-1
    Colony-stimulating factor-1 receptor
    Granulocyte colony-stimulating factor receptor
    Progenitor cells
  • Jan 31, 2012 12:28 PM
  • Thesis
  • English
  • Adobe PDF
  • 23192214 bytes
  • 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.
  • 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: 1165-1175
    Katzenback 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: 434-445
    Katzenback 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: 1224-1235
    Katzenback 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.
  • Doctoral
  • Doctor of Philosophy
  • Department of Biological Sciences
  • Physiology, Cell and Developmental Biology
  • Spring 2012
  • Belosevic, Miodrag (Biological Sciences)
  • Goss, Greg G (Biological Sciences)
    Field, Catherine (Agricultural, Food and Nutritional Science)
    Barreda, Daniel (Agricultural, Food and Nutritional Science)
    Bols, Niels (Biology)