A calmodulin targeted by miRNA scaffold659_26519 regulates IL-17 expression in the early immune response of oyster Crassostrea gigas. (November 2021)

• Record Type: Journal Article
• Authors: Han, Zirong; Li, Jialuo; Wang, Weilin; Li, Jiaxin; Zhao, Qi; Li, Meijia; Wang, Lingling; Song, Linsheng
• Is Part Of: Developmental and comparative immunology. Volume 124(2021)
• Publication Date: 2021-11
• Subjects: Calmodulin (CaM) -- miRNA -- Immune response -- IL-17 expression -- Crassostrea gigas; Immunology -- Periodicals; Developmental immunology -- Periodicals; 616.079
• DOI: 10.1016/j.dci.2021.104180
• Languages: English
• ISSNs: 0145-305X

Abstract

Abstract: Calmodulin (CaM) is a highly conserved second messenger protein transducing calcium signals by binding and modulating intracellular calcium ions (Ca 2+ ), and involves in the Ca 2+ -dependent physical processes including host defense in vertebrates. In the present study, a CaM homologue (designated as Cg CaM) was identified from Pacific oyster Crassostrea gigas . The open reading frame of Cg CaM cDNA was of 471 bp encoding a polypeptide of 156 amino acid residues. There were four EFh domains predicted in Cg CaM, which shared high homologies with those in CaMs from oyster C. virginica and other invertebrates . The mRNA transcripts of Cg CaM were constitutively expressed in all the tested tissues including labellum, mantle, gonad, gills, adductor muscle, haemocytes and hepatopancreas, with the highest expression level in haemocytes. The mRNA expression level of Cg CaM in haemocytes decreased significantly (0.31-fold of that in blank, p < 0.05) at 3 h after LPS stimulation, while the intracellular Ca 2+ (1.57-fold of that in blank, p < 0.05) and the mRNA expression of cytokine Cg IL17-1 (4.87-fold of that in blank, p < 0.05) both increased in haemocytes. Meanwhile, an oyster miRNA scaffold659_26519 was identified, and it was proved to target the 3′-untranslated regions (3′-UTR) of Cg CaM mRNA by luciferase reporter assay. The expression of scaffold659_26519 increased significantly at 3 h (43.523-fold of that of blank, p < 0.05) and 6 h (55.91-fold of that of blank, p < 0.05) after LPS stimulation. When the expression of scaffold659_26519 was inhibited by transfection with its inhibitor in vitro, the expression of Cg IL17-1 declined significantly to 0.58-fold of that in LPS stimulation group. These findings indicated that the miRNA scaffold659_26519 targeted CaM was involved in the early inflammatory response of oyster immunity, and provided a new evidence for CaM-mediated immune mechanism in molluscs. Highlights: A CaM homologue was identified from oyster which was highly expressed in hemocytes. Cg CaM was modulated by scaffold659_26519 and involved in immune response . Sscaffold659_26519 regulated Cg IL17-1 expression by targeting Cg CaM.