Gene expression of pro- and anti-apoptotic proteins in rock bream (Oplegnathus fasciatus) infected with megalocytivirus (family Iridoviridae)

Abstract

Viruses belonging to the genus Megalocytivirus cause diseases in marine fishes primarily in East and Southeast Asian countries. Rock bream iridovirus (RBIV), which is a member of the Megalocytivirus genus, causes severe mass mortalities in rock beam (Oplegnathus fasciatus) in Korea. In this study, we assessed apoptosis-related gene expression patterns in Megalocytivirus-infected rock bream in high mortality and low mortality conditions to determine important apoptosis-related factors, which may affect fish survival/or death. In the high mortality group (100% mortality at 15 dpi), significantly high levels of perforin, granzyme, Fas ligand and caspase 9 expression (5.6-, 10.2-, 13.4- and 4.2-fold, respectively) were observed in the kidney at 8 dpi. Basal expression levels of Fas and caspase 3 were observed at 8 d (1.5-/0.7-fold) and 10 dpi (1.3-/0.6-fold), accompanied by heavy viral loads (8.12 × 106–2.21 × 107/μl). Inhibitor of apoptosis 1 (IAP1) was highly expressed (3.5- to 4.8-fold) at 1 d and 4 dpi; however, IAP1 was reduced when fish died at 8 d and 10 dpi (1.7- to 2.0-fold), which was not significantly different from that of the control group. A similar expression pattern was observed in the low mortality group (18% expected mortality at 30 dpi), which was characterised by a delayed lower magnitude of expression with lower viral loads than the high mortality group. Perforin, granzyme and Fas ligand expression was significantly higher in the low mortality group than in the control group at several sampling points until 30 dpi. Fas and caspases 8, 9 and 3 expression levels showed no statistical significance until 30 dpi. In the low mortality group, significantly higher IAP1 expression compared with the control was observed at 10 d (2.2-fold), 20 d (3.6-fold) and 22 dpi (2.0-fold). In summary, perforin- and granzyme-related apoptosis initiation signals were activated; however, the Fas-induced apoptosis pathway did not efficiently respond. Upregulated IAP1 in RBIV-infected rock bream, which was reported for the first time in this study, exhibited inhibited apoptotic responses in RBIV-infected fish. Although it remains unclear whether apoptosis inhibition aids or impedes fish survival, our data clearly show that the apoptotic response is inhibited in RBIV-infected rock bream.