ALBERT

Description: 〈p〉Publication date: March 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 98〈/p〉 〈p〉Author(s): Lang Lang, Minnan Bao, Weixin Jing, Wei Chen, Lan Wang〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉 〈p〉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 〈em〉Sinopotamon henanense〈/em〉) infected by 〈em〉Aeromonas hydrophila〈/em〉. To investigate the diverse innate immune roles of crustacean homolog Tolls, in this study, we cloned 〈em〉Shtoll1〈/em〉 from 〈em〉S. henanense〈/em〉. The full-length cDNA of 〈em〉Shtoll1〈/em〉 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 〈em〉Shtoll1〈/em〉 was expressed widely in different tissues, with the highest expression in heamocytes. Besides, 〈em〉Shtoll1〈/em〉 expressions were upregulated in heamocytes and hepatopancreas after 〈em〉A. hydrophila〈/em〉 infection. At the same time, the increase of 〈em〉Shtoll1〈/em〉 expressions were examined in heamocytes in response to Cd exposure and 〈em〉A. hydrophila〈/em〉 infection in combination. Through western blotting and immunohistochemical analysis, the ShToll1 expressions in heamocytes were increased in response to 〈em〉A. hydrophila〈/em〉 and Cd independently as well as in combination. Moreover, the mRNA level of three antimicrobial peptides (AMPs) 〈em〉alf5, alf6,〈/em〉 and 〈em〉c-lys〈/em〉, which possibly responded to Cd and 〈em〉A. hydrophila〈/em〉 stimulation through 〈em〉Shtoll1,〈/em〉 were analyzed.〈/p〉 〈p〉Thus, we conclude that Cd expand the susceptibility of ShToll1 to 〈em〉A. hydrophila〈/em〉 infection in heamocytes. This suggest that ShToll1 may contribute to the innate immune defense of 〈em〉S. henanense〈/em〉 against 〈em〉A. hydrophila〈/em〉 and Cd in heamocytes.〈/p〉 〈/div〉 〈/div〉