Clone, identification and functional characterization of a novel toll (Shtoll1) from the freshwater crab Sinopotamon henanense in response to cadmium exposure and Aeromonas hydrophila infection. Issue 98 (March 2020)

• Record Type: Journal Article
• Authors: Lang, Lang; Bao, Minnan; Jing, Weixin; Chen, Wei; Wang, Lan
• Is Part Of: Fish & shellfish immunology. Issue 98(2020)
• Page Start: 401
• Page End: 413
• Publication Date: 2020-03
• Subjects: Sinopotamon henanense -- ShToll1 -- Aeromonas hydrophila -- Cadmium -- Innate immunity; Fishes -- Immunology -- Periodicals; Shellfish -- Immunology -- Periodicals; Poissons -- Immunologie -- Périodiques; Crustacés -- Immunologie -- Périodiques; 571.9617
• DOI: 10.1016/j.fsi.2020.01.012
• Languages: English
• ISSNs: 1050-4648

Abstract

Abstract: Toll is essential in innate immune system which is important for defense against bacterial, fungal and viral infections in invertebrates. Our previous study showed that cadmium (Cd) could change the expression pattern of ShToll3 in the epithelium (gills and midgut from the freshwater crab Sinopotamon henanense ) infected by Aeromonas hydrophila . To investigate the diverse innate immune roles of crustacean homolog Tolls, in this study, we cloned Shtoll1 from S. henanense . The full-length cDNA of Shtoll1 was 4746 bp, with an ORF of 3033 bp encoding a putative protein of 111 amino acids, a 5′-untranslated region of 255 bp and a 3′-untranslated region of 1713 bp. Phylogenetic analysis showed that ShToll1 was clustered into the group of DmToll1, DmToll 4 and DmToll 5. In addition, the tissue distribution results showed that Shtoll1 was expressed widely in different tissues, with the highest expression in heamocytes. Besides, Shtoll1 expressions were upregulated in heamocytes and hepatopancreas after A. hydrophila infection. At the same time, the increase of Shtoll1 expressions were examined in heamocytes in response to Cd exposure and A. hydrophila infection in combination. Through western blotting and immunohistochemical analysis, the ShToll1 expressions in heamocytes were increased in response to A. hydrophila and Cd independently as well as in combination. Moreover, the mRNA level of three antimicrobial peptides (AMPs) alf5, alf6, and c-lys, which possibly responded to Cd and A. hydrophila stimulation through Shtoll1, were analyzed. Thus, we conclude that Cd expand the susceptibility of ShToll1 to A. hydrophila infection in heamocytes. This suggest that ShToll1 may contribute to the innate immune defense of S. henanense against A. hydrophila and Cd in heamocytes. Highlights: A novel toll (Shtoll1) was identified from S. henanense. Shtoll1 expressed highest in heamocytes. Shtoll1 was up-regulated by A. hydrophila. The expressions of ShToll1 were increased in response to the combination of Cd exposure and A. hydrophila infection. alf5, alf6 and c-lys possibly responded to Cd and A. hydrophila stimulation through Shtoll1.