Characterization of granulocyte colony stimulating factor receptor of the goldfish ( Carassius auratus L.)

Abstract

Granulocyte colony stimulating factor receptor (GCSFR) is a member of the class I cytokine receptor superfamily and signaling through this receptor is important for the proliferation, differentiation and activation of neutrophils and their precursors. In this study we report on the cloning and molecular characterization of goldfish GCSFR. The identified goldfish GCSFR sequence possesses the conserved Ig-like domain, the cytokine receptor homology domain (CRH), three fibronectin domains as well as several intracellular signaling motifs characteristic of other vertebrate GCSFRs. Goldfish gcsfr mRNA was highly expressed in kidney and spleen, and in primary kidney neutrophils. The neutrophils have significantly higher mRNA levels of the transcription factors pu.1 and cebpα, and down-regulated levels of transcription factors important for macrophage development such as egr1 and cjun, compared to progenitor cells from the kidney. The gcsfr mRNA was present in the kidney progenitor cells, albeit at much lower levels compared to the neutrophils, and the expression of gcsfr in progenitor cells was not affected by duration of cultivation. Furthermore, gcsfr mRNA levels were up-regulated in neutrophils after treatment with heat-killed Aeromonas salmonicida A449 or with mitogens. Our results indicate that GCSFR may be a useful marker for fish neutrophils.