Quantitation of immune response gene expression and cellular localisation of interleukin-1ß mRNA in Atlantic salmon,
Salmo salar L., affected by amoebic gill disease (AGD)
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Bridle, AR and Morrison, RN and Cupit Cunningham, PM and Nowak, BF, Quantitation of immune response gene expression and cellular localisation of interleukin-1s mRNA in Atlantic salmon,
Salmo salar L., affected by amoebic gill disease (AGD), Veterinary Immunology and Immunopathology, 114, (1-2) pp. 121-134. ISSN 0165-2427 (2006) [Refereed Article]
The characterisation of selected immune response genes during amoebic gill disease (AGD) in Atlantic salmon, Salmo salar L., was performed using semi-quantitative RT-PCR, quantitative real-time RT-PCR (qRT-PCR), and in situ hybridisation (ISH). The immune response genes of interest were interleukin-1Î² (IL-1Î²), inducible nitric oxide synthase (iNOS), serum amyloid A (SAA), and serum amyloid P-like pentraxin (SAP). Atlantic salmon were inoculated with the ectoparasite Neoparamoeba sp., the causative agent of AGD, and gill, liver and anterior kidney tissue sampled at 0, 7 and 14 d post-inoculation (p.i.). Semi-quantitative RT-PCR was performed on the tissue samples to identify up/down-regulated mRNA expression relative to uninfected control fish and normalised to the housekeeping gene, Î²-actin. Interleukin-1Î² (IL-1Î²) was the only immune response gene of those investigated whose mRNA was differentially regulated in any of the tissues and was found to be up-regulated in the gills by semi-quantitative RT-PCR. Increased gill IL-1Î² mRNA expression was then accurately quantitated and confirmed using probe-based qRT-PCR. The cellular localisation of the IL-1Î² mRNA expression in the gills of uninfected and infected fish was then determined by ISH using an IL-1Î²-specific biotinylated cRNA probe. Expression of IL-1Î² mRNA was localised to filament and lamellar epithelium pavement cells in gills of uninfected and infected Atlantic salmon. These data implicate the involvement of IL-1Î² at the site of infection, the gills, of Atlantic salmon during AGD. This work supports previous studies that suggest IL-1Î² is important in the regulation of the fish immune response to parasitic infection but additionally shows the cellular localisation of fish IL-1Î² mRNA expression during infection. Â© 2006 Elsevier B.V. All rights reserved.
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