Abstract
To elucidate cytolytic mechanisms in the channel catfish, lysates from catfish lymphoid and fibroblast cell lines were screened by Western blot analysis using a panel of antibodies reactive with components of the mammalian apoptotic pathway. Strong reactivity with three proteins (approximate Mr 70,000, 37,000, and 15,000) was seen using an antibody targeted to mammalian Fas ligand (FasL). The sizes of the two smaller proteins are consistent with their tentative designation as membrane-bound (37,000 Mr) and soluble (15,000 Mr) FasL. Treatments known to induce FasL in mammalian systems (e.g., PMA/calcium ionophore, UV-irradiation) induced expression of the 37,000-Mr protein in catfish T-cell lines. Moreover, expression of the 37,000-Mr protein in clonal T cells was up-regulated by increasing cell density. At the nucleotide level, homologues of Fas receptor (FasR), FADD, and caspase 8 were identified and characterized. These gene products likely constitute the teleost equivalent of the death-inducing signaling complex (DISC). FADD was constitutively expressed in all (T, B, macrophage, and fibroblast) cell lines examined as well as in peripheral blood lymphocytes (PBL), whereas FasR and caspase 8 were expressed in all cell lines except CCO, a FasL-positive fibroblast line. In contrast to FasL, expression of FasR and caspase 8 was inversely proportional to cell density. Collectively these studies identified four membrane-proximal proteins involved in the initiation of apoptosis in channel catfish and suggest that mechanisms of cell-mediated cytotoxicity in teleosts are similar to those used by mammals.
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Acknowledgements
We wish to thank Noel Hawke (Lineberger Cancer Center, University of North Carolina Medical School) for contributing the catfish FADD sequence. This work was supported by grants from the US Department of Agriculture (NRI/CGP 99-35204-7944 and 2002-35204-12211) and the National Institutes of Health (ROI-AI-19530). Nucleotide sequencing was performed in collaboration with Dr. Greg Warr and Darlene Middleton (Medical University of South Carolina).
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Long, S., Wilson, M., Bengtén, E. et al. Identification and characterization of a FasL-like protein and cDNAs encoding the channel catfish death-inducing signaling complex. Immunogenetics 56, 518–530 (2004). https://doi.org/10.1007/s00251-004-0701-2
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DOI: https://doi.org/10.1007/s00251-004-0701-2