The Role Of As-rarres3 In The Immune Process Of Artemia Sinica

Artemia sinica is a small crustacean living in heavy salt water.As an initial feeding,it plays an important role in the seedling production of fish,shrimp and crabs.Since invertebrates such as Artemia do not have a specific immune system,the innate immune system is their only defense system.The innate immunity of invertebrates can be triggered by PRRs,which indirectly cause a defense response through intracellular immune signal pathways,or directly kill pathogens through encapsulation and phagocytosis.Retinoic acid receptor responder 3(RARRES3)gene is located in the cytoplasm and is the main pattern recognition receptors for recognizing RNA of the viruses in the cytoplasm and activating downstream signal cascades.MAVS is activated as an important adaptor protein,which recruits downstream TRAF3 and TRADD to form a signal complex,and then activates the transcription factor IRF3 through phosphorylation.Finally,IRF3 enters the nucleus,induces host cells to release pro-inflammatory cytokines and type Ⅰ interferons to inhibits pathogen replication and infection.In previous studies,it was found that RARRES3 is involved in the innate immunity of Artemia sinica,but its molecular regulation in the innate immune response of Artemia sinica is not clear.In this study,the full length c DNA sequence of As-rarres3 gene was obtained for the first time(Gen Bank Access No.MW965463),which encodes 274 amino acids.The domain contains a typical Nlp C/P60 family domain,which proves that it belongs to the Nlp C/P60 family.Multiple sequence alignment verified the conservation of its domain.The expression site of As-rarres3 was analyzed by immunofluorescence technique.The results showed that As-RARRES3 was mainly located in the cytoplasm and widely distributed in all stages of development.Real-time q PCR method was used to detect the m RNA expression level of As-rarres3 gene of Artemia sinica in different developmental stages and under the conditions of Gram-positive bacteria or Gram-negative bacteria stress.Western blotting was used to detect the protein expression pattern and expression level of As-RARRES3,As-MAVS,AsTRAF3 and As-IRF3 during the embryonic development and bacterial stress of Artemia sinica.The results showed that the expression of As-rarres3 was high in the early stage of Artemia sinica embryo development(0-5 h),and the level of transcription and translation was low in the following periods.Under the stimulation of Gram-negative bacteria,the level of transcription and translation of As-rarres3 increased at first and then decreased,but higher than that of the control group,which indicated that As-rarres3 played a certain role in the response process of Artemia sinica to the attack of Gram-positive and Gram-negative bacteria.This study aims to clarify the role of RARRES3 in the immune process of Artemia sinica,and has important theoretical significances for studying the molecular mechanism of RARRES3 in the immune process of Artemia sinica and the immune process of other invertebrates.It has potential application value in the development and utilization of Artemia sinica bait protein sources.