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Int J Mol Sci
2014 Oct 27;1511:19472-86. doi: 10.3390/ijms151119472.
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Expression analysis of immune related genes identified from the coelomocytes of sea cucumber (Apostichopus japonicus) in response to LPS challenge.
Dong Y, Sun H, Zhou Z, Yang A, Chen Z, Guan X, Gao S, Wang B, Jiang B, Jiang J.
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The sea cucumber (Apostichopus japonicus) occupies a basal position during the evolution of deuterostomes and is also an important aquaculture species. In order to identify more immune effectors, transcriptome sequencing of A. japonicus coelomocytes in response to lipopolysaccharide (LPS) challenge was performed using the Illumina HiSeq™ 2000 platform. One hundred and seven differentially expressed genes were selected and divided into four functional categories including pathogen recognition (25 genes), reorganization of cytoskeleton (27 genes), inflammation (41 genes) and apoptosis (14 genes). They were analyzed to elucidate the mechanisms of host-pathogen interactions and downstream signaling transduction. Quantitative real-time polymerase chain reactions (qRT-PCRs) of 10 representative genes validated the accuracy and reliability of RNA sequencing results with the correlation coefficients from 0.88 to 0.98 and p-value <0.05. Expression analysis of immune-related genes after LPS challenge will be useful in understanding the immune response mechanisms of A. japonicus against pathogen invasion and developing strategies for resistant markers selection.
Figure 1. Hypothetical diagram of LPS-triggered inflammation and apoptosis pathways summarized in A. japonicus coelomocytes. Genes listed here play important roles in these potential pathways. On the left, three pathways will promote the expression of inflammation factors. The existence of the Type I type I interferon (IFN) pathway was unclear for the absence of IFN homologues in invertebrates. On the right, the apoptosis pathway will result in the degradation of DNA. The abbreviations: LPS, lipopolysaccharide; CCP, Complement control proteins; TRIF, TLR and interleukin-1 receptor (TIR) domain-containing adaptor inducing IFN-β; Cyto C, Cytochrome C; TRAF, tumor necrosis factor (TNF)-receptor-associated factor; TBK1, TRAF family member-associated NF-κB activator (TANK)-binding kinase 1; IRF3/7, interferon regulatory factor 3/7; CD, cluster of differentiation; LBP, lipopolysaccharide binding protein; TLR, toll-like receptor; MyD88, myeloid differentiation primary response gene 88; FADD, Fas-associated death domain protein; IRAK1/4, interleukin-1 receptor-associated kinase 1/4; NF-κB, nuclear factor-κ-B.
Figure 2. Validation of RNA-Seq results using qRT-PCR. The relative fold changes of 10 genes expressed in A. japonicus coelemocytes at 4 h (A); 24 h (B) and 72 h (C) after LPS challenge. Gene abbreviations are: Bf, complement factor B; C3-2, complement component 3-2; Cat B, cathepsin B; CD36, Cluster of differentiation 36; Hsp90, heat shock protein 90; LBP, lipopolysaccharide binding protein; MyD88, myeloid differentiation primary response gene 88; Mys, Myosin; Rel, NF-κB transcription factor Rel; Thy, Thymosin β.
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