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• W4387375172 abstract "Background Apoptosis regulates normal development, homeostasis, immune tolerance and response to environmental stress by eliminating unwanted or diseased cells, and plays a key role in non-specific immunity of invertebrates. The exogenous pathway mediated by death receptors and death ligands is a very important pathway for cell apoptosis. Death ligands are mainly members of the tumour necrosis factor (TNF) family, of which FasL is an important member. The deep involvement of FasL in vertebrates cell apoptosis and immunity has been reported many times, but there is limited research on the FasL gene in shellfish, and its functional importance in oyster cell apoptosis and immunity remains unclear. Methods The full length of Ch FasL was identified and cloned based on the genome of Crassostrea hongkongensis . Quantitative PCR was used to detect the relative expression of Ch FasL in different developmental stages and tissues, as well as the changes of relative expression in hemocytes after bacterial infection. The expression position of Ch FasL in HEK293T cells was also located by subcellular localization, and the effect of increased recombinant protein content on the activity of reporter genes p53 and p21 was studied by dual-fluorescence reporter gene. Finally, the changes of apoptosis rate in hemocytes after Ch FasL silencing was identified by RNA interference technology. Results We identified a novel FasL gene from C. hongkongensis and named it Ch FasL. We found that Ch FasL has potential N-linked glycosylation site, a transmembrane domain and a TNF region, which was a typical characteristics of TNF family. Ch FasL was expressed in all developmental stages of larvae and in all tissues of oysters. After stimulation by V. alginolyticus or S. haemol yticus, its relative expression in hemocytes increased significantly, suggesting that Ch FasL was deeply engaged in the immune response process of C. hongkongensis to external microbial stimulation. The results of subcellular localization showed that Ch FasL was mainly distributed in the cytoplasm of HEK293T cells. With the overexpression of the recombinant protein pcDNA3 1- Ch FasL, the activity of p53 and p21 significantly increased, showing a positive regulatory effect. Moreover, after dsRNA successfully reduced the relative expression of Ch FasL, the apoptosis rate of hemocytes was significantly lower than that the dsGFP group. Conclusion These results comprehensively confirmed the important role of Ch FasL in the apoptosis process of C. hongkongensis , which provided the basis and premise for the in-depth understanding of the immune function of apoptosis in molluscs, and also contributed to the research on the pathogenic death mechanism and disease resistance breeding of marine bivalves." @default.
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• W4387375172 date "2023-10-05" @default.
• W4387375172 modified "2023-10-16" @default.
• W4387375172 title "A novel Fas ligand plays an important role in cell apoptosis of Crassostrea hongkongensis: molecular cloning, expression profiles and functional identification of ChFasL" @default.
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• W4387375172 doi "https://doi.org/10.3389/fimmu.2023.1267772" @default.
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