Signal transducer and activator of transcription 3 (STAT3) is a major regulator of immune response and chronic inflammatory, which can be activated by interleukin-6 (IL-6). In mammals, STAT3 has multiple isoforms, and its function has been well studied. In teleost, a single stat3 has been cloned and identified in several species, but studies on its function are limited. In the present study, four stat3 isoforms including mastat3α1, mastat3α2, mastat3β1 and mastat3β2 were identified from blunt snout bream (Megalobrama amblycephala). The results of quantitative PCR (qPCR) showed that four mastat3 transcripts were ubiquitously expressed in all 10 tissues examined. After Aeromonas hydrophila challenge, the expression patterns of mastat3a1, mastat3a2 and mastat3β2 were similar, but significantly different from that of mastat3β1. In addition, western blot showed that rmaIL-6+rmasIL-6R (IL-6 trans-signaling) significantly up-regulated phosphorylation levels of the four maSTAT3 isoforms and mRNA levels of the il-10, il-11, tnf-a, socs3a and socs3b genes, while rmaIL-6 (IL-6 classical signaling) only significantly up-regulated phosphorylation levels of the two maSTAT3α isoforms and mRNA levels of the il-10, socs3a and socs3b genes. Meanwhile, overexpression or inhibition of JAK2 could significantly change the STAT3 phosphorylation. Finally, JAK2 and STAT3 inhibitors could significantly inhibit the up-regulation of il-10, il-11, tnf-a, socs3a and socs3b induced by rmaIL-6+rmasIL-6R or rmaIL-6. To sum up, this study reveals the functional distinctions and overlaps among the four maSTAT3 isoforms in blunt snout bream and reveals the differential regulation of IL-6 classical signaling and trans-signaling on downstream immune genes via the JAK2/STAT3 pathway, enriching our knowledge of fish's defense mechanisms against pathogens.
Keywords: Blunt snout bream; Classical signaling; IL-6; JAK2/STAT3; STAT3 isoforms; Trans-signaling.
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