Characterization and functional analysis of pl-miR-2188 in melanin synthesis in leopard coral grouper (Plectropomus leopardus)

Abstract

MicroRNAs (miRNAs) are known to regulate gene expression and play a role in body color formation in fish. However, the molecular mechanisms underlying miRNA involvement in the body color of leopard coral grouper (Plectropomus leopardus) remain largely unexplored. In this study, we investigated the expression levels of miR-2188 in red and black P. leopardus (pl-miR-2188) and found significantly higher expression levels in red fish samples compared to those in black fish samples. Silencing pl-miR-2188 in vivo using a pl-miR-2188 antagomir resulted in increased melanin concentration. Following pl-miR-2188 silencing, the expression levels of melanin-related genes, such as tyrosinase (tyr), TYR-related protein 1 (tyrp1–1 and tyrp1–2) and TYR-related protein 2 (tyrp2), and microphthalmia-associated transcription factor (mitf), were elevated. RNAhybrid predictions and dual-luciferase reporter assays identified sox5 as a target mRNA of pl-miR-2188. Following pl-miR-2188 antagomir injection, sox5 expression was significantly upregulated in the injection group compared to that in control groups (P < 0.05). These results suggest that pl-miR-2188 may regulate melanin synthesis in P. leopardus by targeting sox5. This study provides new insights into the miRNA–mRNA interactions involved in melanin synthesis and body color formation in the leopard coral grouper.