In Vivo Analysis of Ifn-γ1 and Ifn-γ2 Signaling in Zebrafish

The zebrafish genome contains a large number of genes encoding potential cytokine receptor genes as judged by homology to mammalian receptors. The sequences are too divergent to allow unambiguous assignments of all receptors to specific cytokines, and only a few have been assigned functions by functional studies. Among receptors for class II helical cytokines-i.e., IFNs that include virus-induced Ifns (Ifn-phi) and type II Ifns (Ifn-gamma), together with Il-10 and its related cytokines (Il-20, Il-22, and Il-26)-only the Ifn-phi-specific complexes have been functionally identified, whereas the receptors for the two Ifn-gamma (Ifn-gamma 1 and Ifn-gamma 2) are unknown. In this work, we identify conditions in which Ifn-gamma 1 and Ifn-gamma 2 (also called IFNG or IFN-gamma and IFN-gammarel) are induced in fish larvae and adults. We use morpholino-mediated loss-of-function analysis to screen candidate receptors and identify the components of their receptor complexes. We find that Ifn-gamma 1 and Ifn-gamma 2 bind to different receptor complexes. The receptor complex for Ifn-gamma 2 includes cytokine receptor family B (Crfb)6 together with Crfb13 and Crfb17, whereas the receptor complex for Ifn-gamma 1 does not include Crfb6 or Crfb13 but includes Crfb17. We also show that of the two Jak2 paralogues present in the zebrafish Jak2a but not Jak2b is involved in the intracellular transmission of the Ifn-gamma signal. These results shed new light on the evolution of the Ifn-gamma signaling in fish and tetrapods and contribute toward an integrated view of the innate immune regulation in vertebrates. The Journal of Immunology, 2010, 185: 6774-6782.