ABSTRACT
Toll-like receptors (TLRs) are one of the extensively studied pattern recognition receptors (PRRs) and play crucial roles in the immune responses of vertebrates and invertebrates. In this study, 14 TLR genes were identified from the genome-wide data of Octopus sinensis. Protein structural domain analysis showed that most TLR proteins had three main structural domains: extracellular leucine-rich repeats (LRR), transmembrane structural domains, and intracellular Toll/IL-1 receptor domain (TIR). The results of subcellular localization prediction showed that the TLRs of O. sinensis were mainly located on the plasma membrane. The results of quantitative real-time PCR (qPCR) showed that the detected TLR genes were differentially expressed in the hemolymph, white bodies, hepatopancreas, gills, gill heart, intestine, kidney, and salivary gland of O. sinensis. Furthermore, the present study investigated the expression changes of O. sinensis TLR genes in hemolymph, white bodies, gills, and hepatopancreas in different phases (6 h, 12 h, 24 h, 48 h) after stimulation with PGN, poly(I: C) and Vibrio parahaemolyticus. The expression of most of the TLR genes was upregulated at different time points after infection with pathogens or stimulation with PAMPs, a few genes were unchanged or even down-regulated, and many of the TLR genes were much higher after V. parahaemolyticus infection than after PGN and poly(I:C) stimulation. The results of this study contribute to a better understanding of the molecular immune mechanisms of O. sinensis TLRs genes in resistance to pathogen stimulation.
Subject(s)
Gene Expression Regulation , Immunity, Innate , Octopodiformes , Toll-Like Receptors , Vibrio parahaemolyticus , Animals , Toll-Like Receptors/genetics , Toll-Like Receptors/immunology , Toll-Like Receptors/chemistry , Vibrio parahaemolyticus/physiology , Octopodiformes/genetics , Octopodiformes/immunology , Immunity, Innate/genetics , Gene Expression Regulation/immunology , Phylogeny , Gene Expression Profiling/veterinary , Poly I-C/pharmacology , Peptidoglycan/pharmacology , Arthropod Proteins/genetics , Arthropod Proteins/immunology , Arthropod Proteins/chemistry , Pathogen-Associated Molecular Pattern Molecules/pharmacologyABSTRACT
In the present study, the next-generation sequencing technology was used to develop a transcriptome database of gonad and liver from 3-year-old male and female Amur sturgeons (Acipenser schrenckii). A total of 139,406 unigenes were generated after the Illumina Hiseq. 2500 sequence and assembled by Trinity. The differential expression analysis between male and female obtained 5,199 differentially expressed genes (DEGs) in gonad and 457 DEGs in liver. Gene Ontology enrich analysis showed that the specific DEGs of gonad play a dominant role in reproductive processes. Although the specific DEGs of liver indicated their primary responsibility for energy metabolism, the DEGs of liver and gonad co-own enriched in terms associated with reproduction suggested that liver also plays a role in sex-related differences in Amur sturgeon. Furthermore, genes related to sex-related differences were selected to validate among the four different tissues by real-time quantitative polymerase chain reaction (qRT-PCR). In addition, by trans-acting analysis, a total of 5,206 putative long noncoding RNAs (lncRNAs) and 3,490 target genes of lncRNAs were predicted from gonad and liver. Moreover, several lncRNAs targeting Mea1, Piwil1, Tdrd1, Nanos2, Ankrd49, and ZP3 may have potential regulatory effect related to gametogenesis and gonadal differentiation were identified and validated by qRT-PCR. These results suggested for the first time that lncRNAs might be one of the effect factors in regulating the differential expression of messenger RNAs associated with sex-related differences in Amur sturgeon.
