Search

The feedback regulatory effects of estrogen (E2) and androgen (T) on the gonadotropin-releasing hormone (GnRH) and gonadotropin (GtH) within the brain–pituitary–gonad (BPG) axis in eels with undeveloped ovaries were investigated through in vivo studies. However, the regulatory role of the BPG axis only became apparent during ovary development in the migratory stage. To further elucidate the direct feedback regulation of the BPG axis, female Anguilla japonica underwent artificial induction of vitellogenesis, and the regulation of BPG axis tissues by GtH (human chorionic gonadotropin, hCG), E2, and T was explored through in vitro exposure. The mRNA expression levels of GnRH (mGnRH), GtH (fshb and lhb), and steroid biosynthesis enzymes (cyp11a1, hsd3b, cyp17a1, and cyp17a2) in the diencephalon, pituitary, and ovary, respectively, were determined. The results showed that the expression level of mGnRH in the diencephalon was significantly downregulated by 0.1 IU/mL hCG but upregulated by both 1 nM E2 and higher concentrations of T, suggesting a direct positive feedback regulation of E2 on mGnRH. In the pituitary, the expression levels of fshb and lhb were upregulated by E2, while fshb was suppressed by T. In the ovaries, the expression of cyp11a1 and hsd3b was upregulated by 1 nM E2, whereas T exposure resulted in an opposite effect. Cyp17a1 mRNA levels did not differ significantly with E2 treatment but were upregulated by 1 nM T. These findings suggest that low concentrations of E2 exhibited positive feedback regulation on all three levels (diencephalon, pituitary, and ovary) of the BPG axis, while T showed weaker and differential feedback regulation in BPG axis tissues. Overall, this study's results revealed the direct feedback regulation of hCG, E2, and T on the BPG axis in eels, a phylogenetic base of teleosts. [ABSTRACT FROM AUTHOR]

Simple Summary: In this study, we investigated the response mechanism of type I interferon receptor in the Japanese eel (Anguilla japonica) after an immune challenge, and we also explored the key role of type I interferon receptor in its immune response mechanism, which will provide new evidence for the study of type I interferon receptor in vertebrates, especially in teleost. IFNAR1, one of the type I IFN receptors, is crucial to mammalian host defense against viral invasion. However, largely unknown is the immunological role of the fish teleost protein IFNAR1, also known as CRFB5. We have successfully cloned the whole cDNA of the Japanese eel's (Anguilla japonica) CRFB5a homolog, AjCRFB5a. The two fibronectin-3 domains and the transmembrane region (238–260 aa) of AjCRFB5a are normally present, and it shares a three-dimensional structure with zebrafish, Asian arowana, and humans. According to expression analyses, AjCRFB5a is highly expressed in all tissues found, particularly the liver and intestine. In vivo, Aeromonas hydrophila, LPS, and the viral mimic poly I:C all dramatically increased AjCRFB5a expression in the liver. Japanese eel liver cells were reported to express AjCRFB5a more strongly in vitro after being exposed to Aeromonas hydrophila or being stimulated with poly I: C. The membranes of Japanese eel liver cells contained EGFP-AjCRFB5a proteins, some of which were condensed, according to the results of fluorescence microscopy. Luciferase reporter assays showed that AjCRFB5a overexpression strongly increased the expression of immune-related genes in Japanese eel liver cells, such as IFN1, IFN2, IFN3, IFN4, IRF3, IRF5, and IRF7 of the type I IFN signaling pathway, as well as one of the essential antimicrobial peptides LEAP2, in addition to significantly inducing human IFN-promoter activities in HEK293 cells. Additionally, RNA interference (RNAi) data demonstrated that knocking down AjCRFB5a caused all eight of those genes to drastically lower their expression in Japanese eel liver cells, as well as to variable degrees in the kidney, spleen, liver, and intestine. Our findings together showed that AjCRFB5a participates in the host immune response to bacterial infection by inducing antimicrobial peptides mediated by LEAP2 and favorably modulates host antiviral immune responses by activating IRF3 and IRF7-driven type I IFN signaling pathways. [ABSTRACT FROM AUTHOR]

Sox genes encode transcription factors with a high‐mobility group (HMG) box, playing critical roles in the initiation and maintenance of a variety of developmental processes, such as sex determination and differentiation. In the present study, we identified 26 Sox genes in the genome of spinyhead croaker Collichthys lucidus (Richardson, 1844) with homology‐based analysis of the HMG box. The transcriptome‐based expression profiles revealed that the expression of the Sox gene in gonads began to differ between sexes when the body length was 2.74 ± 0.24 cm. At that time, three Sox genes (Sox11b, Sox8a and Sox19) were significantly upregulated, accompanied by the downregulation of 12 Sox genes in the ovary, and six Sox genes were temporarily significantly upregulated in the testis. Afterwards, the expression profile of Sox genes changed only with a small amplitude in both the ovary and testis. For adult tissues, huge differences were observed in the expression profiles of Sox genes between ovaries and testes, as well as small differences in somatic tissues between sexes. These results provide clues to further decipher the role of Sox genes in the processes of sex determination and differentiation in spinyhead croaker and other teleosts. [ABSTRACT FROM AUTHOR]

Due to increased temperatures and aquaculture density, thermal and hypoxia stresses have become serious problems for the aquaculture of abalone Haliotis diversicolor. Stresses lead to immunosuppression, which can cause severe negative impacts on aquaculture farms. To study the mechanism of immunosuppression after hypoxia stress and bacterial challenge, transcriptomes of H. diversicolor hemocytes involved in immunity were profiled. A total of 307,395,572 clean reads were generated and assembled into 99,774 unigenes. KEGG analysis indicated that 225 unigenes with immunologic function were mapped into immune-related pathways. Expression of 41 unigenes measured by quantitative real-time PCR (qRT-PCR) showed consistent results with that of transcriptome analysis. When exposure challenge of Vibrio parahaemolyticus, it is indicated that the PI3K-AKT, MAPK, NF-κB and P53 signal pathways were involved in the hypoxia-induced immunosuppression of H. diversicolor. Furthermore, when the AKT gene (HdAKT) was inhibited by double-stranded RNA (dsRNA), expression levels of HdAKT was lower than the blank and control group in hemocytes at 4 h, 12 h and 24 h (p < 0.05). [ABSTRACT FROM AUTHOR]

The structure, evolution, and function of heat shock proteins 90 (Hsp90s) have been investigated in great detail in fungi and animals. However, studies on the Hsp90 genes in plants are generally limited. Brachypodium distachyon (L.) P. Beauv., as a model plant for cereal crops, has become a potential biofuel grass. During its long evolution, the Hsp90 gene family in Brachypodium has developed some strategies to cope with adverse environments. How the Hsp90 gene family in Brachypodium evolved in different plant lineages and what its role is in plant responses to drought and salt stresses remains to be elucidated. We used a set of different bioinformatics tools to identify 94 Hsp90 genes from 10 species representing four plant lineages and classified into three subgroups. Eight BdHsp90 genes were detected from B. distachyon. The number of exon-intron structures differed in each subgroup, and the motif analysis revealed that these genes were relatively conservative in each group. The fragments duplication and tandem duplication, which are the prime powers for functional diversity, generally occurred during the duplication of the whole plant genome. Transcriptional analysis of the BdHsp90 genes under salt and drought stress conditions indicated that the expression of these genes was delayed or increased at different stress time points; The expression was more affected in that of Bradi3g39630, Bradi4g06370, and Bradi1g30130. Our findings suggest the involvement of BdHsp90s in plant abiotic stress response, and further consolidate our views on the stress response mechanism of Hsp90 in general. [ABSTRACT FROM AUTHOR]

Vitellogenesis in crustaceans is controlled by several steroid hormones. In humans, the expression of SF-1 , a gene that regulates gonadal development and the synthesis of steroid hormones, is affected by DDX20. However, how the homologous gene FTZ-F1 is regulated by DDX20 and its association with vitellogenesis remains unknown in the mud crab Scylla paramamosain. In this study, SpDDX20 and SpFTZ-F1 were identified in the transcriptome of mature ovarian tissue from the mud crab. qRT-PCR results revealed that the expression levels of SpFTZ-F1 and SpVTG in the ovaries of crab in the experimental group injected with dsDDX20 (EO) were significantly higher (P < 0.05) than those in the negative control group injected with dsEGFP (NO) and the blank control group injected with SPSS (BO). The differentially expressed genes (DEGs) identified by comparative transcriptome analysis of the EO group and NO group were enriched into five pathways related to ovarian steroidogenesis. The expression of CYP17 , CYP3A4 , CYP1A1 and 3β-HSD were up-regulated in pathways related to steroid hormone production and biosynthesis. The expression of the INSR , IRS and PI3K genes in the insulin signaling pathway were significantly increased (P < 0.05). The expression level of the TGF-β gene was up-regulated (P < 0.05) in the transforming growth factor pathway, whereas the expression level of the Smad2 gene was down-regulated (P < 0.05). The expression of GnRHR , GS , AC and PKA genes in the gonadotropin-releasing hormone signaling pathway were up-regulated. Our data provide a foundation for investigating the relationship between DDX20 and FTZ-F1 in the regulation of vitellogenin expression in S. paramamosain. [Display omitted] • Full-length cDNA of SpDDX20 and SpFTZ-F1 were cloned and characterized from Scylla paramamosain. • The expression of SpDDX20 and SpFTZ-F1 had tissue and gender specificity. • The expression level of SpFTZ-F1 was up-regulated after SpDDX20 interference. • Five ovarian steroidogenesis pathways were differentially expressed after SpDDX20 knockdown. • The expression level of SpVTG was up-regulated after knockdown of SpDDX20. [ABSTRACT FROM AUTHOR]

Dermatopontin (DPT), a component of the extracellular matrix, plays important roles in cell-matrix interactions and matrix assembly. Some studies have revealed that it has more general functions in biological activities. However, its function in molluscs is poorly understood. In this study, a molluscan DPT gene, saDPT2, was cloned from small abalone Haliotis diversicolor. The full-length cDNA of saDPT2 sequence is 620 bp, with a 531 bp open reading frame encoding a protein of 177 amino acids (aa). Amino acid sequence analysis revealed that saDPT2 shares conserved signature motifs with other DPT proteins, including three repeats of 10-residue motif (S-X-H-X-N-X-Y-E-D-R), which is similar to mammalian 6-residue repeating sequence of D-R-E/Q-W-X-F/Y. Quantitative real-time PCR was employed to investigate the tissue distribution of saDPT2 mRNA, its expression at different developmental stages, and in abalone under bacteria challenge. The saDPT2 mRNA could be detected in all examined tissues and developmental stages. Moreover, the saDPT2 mRNA was up-regulated in haemocytes and gills after bacteria injection. The results indicate that the saDPT2 could respond to pathogenic infection and may play a role in adult abalone immune system. [ABSTRACT FROM AUTHOR]

Abstract: A total of 5160 high quality ESTs (expressed sequence tags) averaging 357bp were collected from normalized cDNA libraries created from testis, ovary and mixed organs of mud crab Scylla paramamosain. Clustering and assembly of these ESTs resulted in a total of 3837 unique sequences with 576 overlapping contigs and 3261 singletons. Comparisons with the GenBank non-redundant (Nr) protein database (BLASTx, e-values <10−5) revealed putative functions or matched homologs from other organisms for 847 (22%) of the ESTs. Several gonad development related genes such as cathepsin C, thioredoxin peroxidase, vitellogenin receptor precursor, 50S ribosomal protein L24 and ubiquitin-conjugating enzyme E2 isoform 2 were identified from this EST project and demonstrated as gonad differential expression genes by rqRT-PCR. Sixty five different types of SSRs (simple sequence repeats) were identified from the total 411 EST-SSR motifs. A home-made cDNA microarray containing 5664 spots was developed and the hybridization results indicated that 39 unique transcripts were differentially expressed in testis and ovaries (P<0.05). The expression levels of eleven unique transcripts examined by rqRT-PCR were matched with microarray fairly. These results will provide a useful resource for functional genomic studies on the biology of reproduction of mud crab. [Copyright &y& Elsevier]

Abstract: The analysis of expressed sequence tags (ESTs) is an efficient approach for gene discovery, expression profiling, and the development of resources for functional genomics. To analyze the transcriptome response to tributyltin of small abalone Haliotis diversicolor supertexta, a normalized cDNA library of hepatopancreas after exposure of animals to tributyltin was constructed. Three thousand and forty eight high quality ESTs were generated. After processing, a total of 2473 unigenes comprising 370 contigs and 2103 singlets were acquired. BLAST identified 1108 clones (45%) as known genes. Kyoto Encyclopedia of Genes and Genomes (KEGG) and Simple Sequence Repeat (SSR) identification were also carried out to acquire more information on pathway and EST–SSR. According to GO categories, the total ESTs distribute across the various functional categories. In addition, 43 potential toxicology-related clusters were identified. This work provides the first set of genetic data for small abalone which is of great value for further exploitation of this species in functional genomics and toxicogenomics. Our interesting results will be mainly useful to choose biomarkers for ecotoxicological studies. [Copyright &y& Elsevier]

Receptor-interacting protein 3 (RIP3) has been demonstrated to be a key regulator not only in cell death pathways including apoptosis and necroptosis but also in inflammation and host immune responses. In this study, a RIP3 ortholog named Lc-RIP3 is identified in large yellow croaker (Larimichthys crocea). The open reading frame (ORF) of Lc-RIP3 is 1524 bp long and encodes a protein of 507 amino acids (aa). The deduced Lc-RIP3 protein has an N-terminal kinase domain and a C-terminal RHIM domain, and the genome organization of Lc-RIP3 is conserved in teleosts with 12 exons and 11 introns but is different from that in mammals, which comprises 10 exons and 9 introns. Confocal microscopy revealed that Lc-RIP3 is a cytosolic protein. The expression analysis at the mRNA level indicated that Lc-RIP3 is ubiquitously distributed in various tissues/organs, and could be up-regulated under poly I:C, LPS, PGN, and Pseudomonas plecoglossicida stimulation in vivo. Notably, Lc-RIP3 could induce NF-κB but not IRF3 activation. In addition, Lc-RIP3 co-expression with Lc-TRIF, Lc-MAVS, or Lc-IRF3 significantly abolishes the activation of NF-κB but enhances the induction of IRF3 activity. Moreover, NF-κB activity could be up-regulated when Lc-RIP3 is co-expressed with Lc-RIP1 or Lc-IRF7. These results collectively indicate that Lc-RIP3 acts as an important regulator in host innate immune signaling in teleosts. [ABSTRACT FROM AUTHOR]

Abstract: In order to identify genes involved in oogenesis in shrimp, an ovarian cDNA library of Marsupenaeus japonicus was screened using a suppression-subtraction hybridization (SSH)-enriched probe. More than 20 genes were identified as differentially expressed genes between the ovary and the testis. Unexpectedly, one of these genes is a ribosomal protein that is normally considered a housekeeping gene. Northern blot shows that the shrimp ribosomal protein L24 gene (srpl24) is 0.6 kb in length. The expression level of srpl24 in the ovary is much higher than in the testis. Bioinformatics analyses show that srpl24 encodes a protein of 164 aa with a predicted molecular mass of 18.2 kDa, which is a cytoplasmic ribosomal protein. Real time PCR analyses demonstrated that the relative abundance of srpl24 mRNA in the different organs is: ovary ≫ testis, hepatopancreas, muscle and eye. The highest expression level of srpl24 in the ovary suggests that srpl24 has an important role in oogenesis. It is the first reported rpl24 in crustaceans and is the first reported rpl24 that is differentially expressed between the ovary and the testis in animals. [Copyright &y& Elsevier]

Heat-shock protein 70 (HSP70) is a molecular chaperone that plays critical roles in cell protein folding and metabolism, which helps to protect cells from unfavorable environmental stress. Haliotis diversicolor is one of the most important economic breeding species in the coastal provinces of south China. To date, the expression and transcriptional regulation of HSP70 in Haliotis diversicolor (HdHSP70) has not been well characterized. In this study, the expression levels of HdHSP70 gene in different tissues and different stress conditions were detected. The results showed that the HdHSP70 gene was ubiquitously expressed in sampled tissues and was the highest in hepatopancreas, followed by hemocytes. In hepatopancreas and hemocytes, the HdHSP70 gene was significantly up-regulated by Vibrio parahaemolyticus infection, thermal stress, and combined stress (Vibrio parahaemolyticus infection and thermal stress combination), indicating that HdHSP70 is involved in the stress response and the regulation of innate immunity. Furthermore, a 2383 bp of 5′-flanking region sequence of the HdHSP70 gene was cloned, and it contains a presumed core promoter region, a CpG island, a (TG)39 simple sequence repeat (SSR), and many potential transcription factor binding sites. The activity of HdHSP70 promoter was evaluated by driving the expression of luciferase gene in HEK293FT cells. A series of experimental results indicated that the core promoter region is located between −189 bp and +46 bp, and high-temperature stress can increase the activity of HdHSP70 promoter. Sequence-consecutive deletions of the luciferase reporter gene in HEK293FT cells revealed two possible promoter activity regions. To further identify the binding site of the key transcription factor in the two regions, two expression vectors with site-directed mutation were constructed. The results showed that the transcriptional activity of NF-1 site-directed mutation was significantly increased (p < 0.05), whereas the transcriptional activity of NF-κB site-directed mutation was significantly reduced. These results suggest that NF-1 and NF-κB may be two important transcription factors that regulate the expression of HdHSP70 gene. [ABSTRACT FROM AUTHOR]

In aquaculture, nutrigenomics or "nutritional genomics" is concerned with studying the impacts of nutrients and food ingredients on gene expressions and understanding the interactions that may occur between nutrients and dietary bioactive ingredients with the genome and cellular molecules of the treated aquatic animals at the molecular levels that will, in turn, mediate gene expression. This concept will throw light on or provide important information to recognize better how specific nutrients may influence the overall health status of aquatic organisms. In crustaceans, it is well known that the nutritional requirements vary among different species. Thus, studying the nutrigenomics in different crustacean species is of significant importance. Of interest, recognition of the actual mechanisms that may be associated with the effects of the nutrients on the immune responses of crustaceans will provide clear outstanding protection, build a solid immune system, and also decrease the possibilities of the emergence of infectious diseases in the culture systems. Similarly, the growth, molting, lipid metabolism, antioxidant capacity, and reproduction could be effectively enhanced by using specific nutrients. In the area of crustacean research, nutrigenomics has been rapidly grown for addressing several aspects related to the influences of nutrients on crustacean development. Several researchers have studied the relationships between several functional genes and their expression profile with several physiological functions of crustaceans. They found a close association between the effects of optimal feeding with efficient production, growth, reproduction development, and health status of several crustacean species. Moreover, they illustrated that regulation of the gene expression in individual cells by different nutrients and formulated feeds could improve the growth development and immunity-boosting of several crustacean species. The present review will spotlight on such relationships between the dietary nutrients and expression of genes linked with growth, metabolism, molting, antioxidant, reproduction, and immunity of several crustacean species. The literature included in this review article will provide references and future outlooks for the upcoming research plans. This will contribute positively for maintaining the sustainability of the sector of the crustacean industry. • This review evaluates the impacts of feed ingredients on gene expressions in tissues of different crustacean species. • The interactions between nutrients and bioactive components with the genome of treated crustaceans at the molecular levels were also studied. • There is a close association between the optimal feeding effects with efficient production, reproduction development, and health status in crustaceans. • This review throws light on the effects of nutrients on the expression of genes related to growth, metabolism, molting, antioxidant status, reproduction, and immunity of crustaceans. [ABSTRACT FROM AUTHOR]

Final oocyte maturation is the key step to successful spawning and fertilization. Quantitative real-time PCR (qPCR) is the technique of election to quantify the abundance of functional genes in such study. Reference gene is essential for correct interpretation of qPCR data. However, an ideal universal reference gene that is stable under all experimental circumstances has not been described. Researchers should validate their reference genes while performing qPCR analysis. The expression of 6 candidate reference genes: 18s rRNA , 28s rRNA , Cathepsin Z , Elongation factor 1-α , Glyceraldehyde-3-phosphate dehydrogenase and β-actin were investigated during final oocyte maturation induced by different compounds (DES and DEHP) in common carp ( Cyprinus carpio ). Four softwares (Bestkeeper, geNorm, NormFinder and RefFinder) were used to screen the most stable gene in order to evaluate their expression stability. The results revealed that EF1α was highly stable expressed when final oocyte maturation was induced by DES, while gapdh was the most stable gene when final oocyte maturation was induced by DEHP. Stable expressed reference gene selection is critical for all qPCR analysis to get accurate target gene mRNA expression information. [ABSTRACT FROM AUTHOR]

As a crucial member of pattern-recognition receptors (PRRs), the Tolls/Toll-like receptors (TLRs) gene family has been proven to be involved in innate immunity in crustaceans. In this study, nine members of TLR gene family were identified from the mud crab (Scylla paramamosain) transcriptome, and the structure and phylogeny of different Sp TLRs were analyzed. It was found that different Sp TLRs possessed three conserved structures in the TIR domain. Meanwhile, the expression patterns of different Sptlr genes in examined tissues detected by qRT-PCR had wide differences. Compared with other Sptlr genes, Sptlr-6 gene was significantly highly expressed in the hepatopancreas and less expressed in other tissues. Therefore, the function of Sptlr-6 was further investigated. The expression of the Sptlr-6 gene was up-regulated by Poly I: C, PGN stimulation and Vibrio parahaemolyticus infection. In addition, the silencing of Sptlr-6 in hepatopancreas mediated by RNAi technology resulted in the significant decrease of several conserved genes involved in innate immunity in mud crab after V. parahaemolyticus infection, including relish , myd88 , dorsal , anti-lipopolysaccharide factor (ALF), anti-lipopolysaccharide factor 2 (ALF-2) and glycine-rich antimicrobial peptide (glyamp). This study provided new knowledge for the role of the Sptlr-6 gene in defense against V. parahaemolyticus infection in S. paramamosain. • Nine TLRs were identified from the transcriptome data of mud crab Scylla paramamosain. • The novel Sptlr-6 had a high expression level in the hepatopancreas of S. paramamosain. • PGN, Poly I: C and Vibrio injection induced the expression of Sptlr-6 in the hepatopancreas. • Sptlr-6 played a crucial role via the Dorsal/Relish-AMPs pathway in response to Vibrio challenge. [ABSTRACT FROM AUTHOR]

In this study, a new continuous muscle cell line, LYCMS (l arge y ellow c roaker m u s cle cell line), derived from the muscle tissue of larva of large yellow croaker (Larimichthys crocea) was developed with modified DMEM/F12 medium at 27 °C. The muscle cell line could be passaged at different ratios for different growth rates. Karyotype analysis showed that a large proportion of LYCMS cells had 48 chromosomes. The proliferation of LYCMS cell line could be affected by mammalian growth factors such as human basic fibroblast growth factor (b-FGF), epidermal growth factor (EGF), and hepatocyte growth factor (HGF). GFP expression experiments indicated that the LYCMS cell line could be used for exogenous genes' expression. Different virus response-related genes tested in this study showed diverse change types in expression before and after (0–24 h) polycytidylic acid (poly I: C) challenge of LYCMS cells. This is the first study of virus response signaling pathways of large yellow croaker based on the muscle cell line. The results showed that compared with the in vivo experiments, the use of the LYCMS cell line for immune research is more convenient, efficient, and rapid. By using this model, we demonstrated that MDA5-IPS1-TRAF6-NFκB-cytokines, MDA5-IPS1-TRAF3-IRF3-interferon or TLR22-TRIF-IRF3-interferon, TLR8-MyD88-NFκB-cytokines, and TLR3-TRIF-IRF3-interferon pathways were able to response to poly I: C challenge in the muscle cell line of large yellow croaker. • A cell line from muscle tissue of larva of large yellow croaker was first developed. • Virus-responsive pathways in response to poly I: C challenge in large yellow croaker were evidenced. • The LYCMS is an in vitro model for antiviral study of large yellow croaker. • The use of LYCMS cell line for immune research is more convenient, efficient and rapid. [ABSTRACT FROM AUTHOR]

Aromatase is a key enzyme that catalyzes the biosynthesis of estrogens. Previous study indicated that putative tissue-specific promoters of the one aromatase gene (cyp19a1) may drive the differential regulatory mechanisms of cyp19a1 expression in Anguilla japonica. In the present study, for elucidating the transcription characteristics and the function of putative tissue-specific promoters of cyp19a1 in the brain-pituitary-gonad (BPG) axis during vitellogenesis, we investigated the transcriptional regulation of cyp19a1 by 17β-estrogen (E2), testosterone (T), or human chorionic gonadotropin (HCG) in A. japonica. The expression of estrogen receptor (esra), androgen receptor (ara), or luteinizing hormone receptor (lhr) was up-regulated as cyp19a1 in response to E2, T, or HCG, respectively in the telencephalon, diencephalon, and pituitary. The expression of cyp19a1 was also upregulated in the ovary by HCG or T in a dose-dependent manner. Unlike in the brain and pituitary, the expression of esra and lhr , rather than ara , was upregulated by T in the ovary. Subsequently, four primary subtypes of 5′-untranslated terminal regions of cyp19a1 transcripts and the corresponding two 5′ flanking regions (promoter P.I and P.II) were identified. The P.II existed in all BPG axis tissues, whereas the P.I with strong transcriptional activity was brain- and pituitary-specific. Furthermore, the transcriptional activity of promoters, the core promoter region, and the three putative hormone receptor response elements were validated. The transcriptional activity did not change when the HEK291T cells co-transfected with P.II and ar vector were exposed to T. These results suggested that the expression of cyp19a1 was upregulated indirectly through esra and lhr rather than ara by T in the ovary, whereas the expression of cyp19a1 was upregulated directly through androgen receptor and the downstream androgen response element of tissue-specific P.I in the brain and pituitary. The results of the study reveal the regulatory mechanisms of estrogen biosynthesis and provide a reference for optimizing the technology of artificially induced maturation in eels. • Promotor I with strong transcriptional activity was brain- and pituitary-specific. • esra and lhr , rather than ara , was upregulated as cyp19a1 by T in the ovary. • Transcriptional activity of Promoter II did not change by T and androgen receptor. • Promotor II mediated the upregulation of cyp19a1 by T indirectly in the ovary. [ABSTRACT FROM AUTHOR]

Toll-like receptors (TLRs) play essential roles in the innate immune system and have been extensively studied in mollusks. In this study, through a genome-wide search, TLR genes were identified as 29 in Haliotis discus hannai , 33 in H. rufescens , and 16 in H. laevigata. Domain analysis indicated that these TLR genes contain leucine-rich repeat (LRR) and Toll/IL-1 receptor (TIR) domains and exons ranging from 1 to 5. Polymorphism analysis showed that the TLRs in abalones did not have high diversities with 143 SNPs and no Indel in H. discus hannai , 92 SNPs and 3 Indels together with 6 missense mutations in H. rufescens , and no SNP or Indel in H. laevigata. The expression of 8 TLR genes in H. discus hannai was confirmed in the hepatopancreas, gill, hemolymph, gonads, intestine, muscle, and mantle. The expression of five TLR genes (out of 8) in gills (p < 0.05), three in hepatopancreas (p < 0.05), and three in hemolymph (p < 0.05) was upregulated separately in response to the infection caused by Vibrio parahaemolyticus. The findings in this study would contribute to a better understanding of the molecular immune mechanism of H. discus hannai against stimulation by V. parahaemolyticus and provide a basis for the study of TLRs in abalones. • 29, 33, 16 TLRs were identified from the genomes of Haliotis discus hannai H. rufescens and H. laevigata , separately. • Abalone TLR genes contain leucine-rich repeat (LRR) and Toll/IL-1 receptor (TIR) domains and exons ranging from 1 to 5. • 143 SNPs and no Indel in H. discus hannai , 92 SNPs and 3 Indels together with 6 missense mutations in H. rufescens , and no SNP or Indel in H. laevigata. • Vibrio exposure regulates the expression of TLRs in the gills, hepatopancreas, and hemolymph of H. discus hannai. [ABSTRACT FROM AUTHOR]

Octamer-binding transcription factor 4 (Oct4) is a crucial pluripotent transcription factor in controlling pluripotency of embryonic stem cells (ESCs), formation of primordial germ cells (PGCs), early embryonic development, and gonadal germ cells. To understand the roles of Oct4 in the large yellow croaker ( Larimichthys crocea ) ( Lc-Oct4 ), the full-length cDNA of Oct4 was cloned and analyzed. Lc-Oct4 includes a 104-bp 5′ untranslated region (UTR), a 567-bp 3′ UTR and a 1431-bp coding region encoding a protein of 476 amino acids (aa) with a predicted molecular mass (Mm) of 52.40 kDa and an isoelectric point (PI) of 6.34. Quantitative real time PCR (qRT-PCR) in tissues showed that Lc-Oct4 was only expressed in ovary. During ovarian development, the expression level in 635 days post hatching (635-dph) ovary was about 6.3-fold higher than in 270-dph and 1000-dph ovary. The in situ hybridization analysis showed that Lc-Oct4 had a high expression in the different developmental stages of oocytes with the highest level in stages II (later) and III oocytes. Lc-Oct4 was also expressed in spermatogonia and primary spermatocytes. Furthermore, the qRT-PCR analysis of Lc-Oct4 in different developmental embryos revealed that the expression from high to low was multiple-cell stage > mid-blastula stage = mid-gastrula stage, and the expression in multiple-cell stage was at less 1.6-fold higher than in other stages. Accordingly, the whole mount in situ hybridization (WISH) in embryos demonstrated that Lc-Oct4 was expressed only in early embryonic development from 2-cell stage to early-gastrula stage with the peak at 16-cell stage, but not detected in PGCs area. In conclusion, Lc- Oct4 participated in the oogenesis and early embryonic development in large yellow croaker. Overall, this study provided better understanding of Lc-Oct4 gene and lay the foundation for its function research in the large yellow croaker. [ABSTRACT FROM AUTHOR]

Sox genes play important roles in various developmental processes such as sex determination, embryogenesis, oogenesis, neurogenesis, and larval development. In order to clarify the roles of Sox genes in the developmental process of large yellow croaker, the full-length cDNA of the Sox11a gene ( Lc-Sox11a ) was cloned for the first time. Bioinformatics analysis indicated that Lc- Sox11a contains a protein of 366 amino acids with a Ser-rich region, a C-terminal conserved region, and a high mobility group box. The expression of Lc-Sox11a in different tissues of both sexes and in different developmental embryonic stages revealed that Lc-Sox11a were expressed with tissue and gender specificity, of which the expression level in female was ovary > brain > eye > gill; in male was brain > testis > gill. The gender differences occurred in the brain and eye with the male brain > female brain, female eye > male eye. Moreover, the expression of Lc-Sox11a in the gonad and brain at different growth stages was detected. Significant up-regulated expression of Lc-Sox11a was found in the ovary and the male brain at 1000 dph (days post hatching) compared with 270 dph and 635 dph. However, significant down-regulated expression of Lc-Sox11a occurred in the testis with growth. Besides, the expression of Lc-Sox11a in the female brain showed a trend of first rising then falling, with the highest peak in 635 dph. The results of in situ hybridization displayed that Lc-Sox11a was widely distributed only in cytoplasm of oocytes at each stage in oogenesis. In early stage of oocytes, Lc-Sox11a was expressed weakly and evenly. As the appearance of vacuoles and synthesis of yolks, positive signals of Lc-Sox11a distributed intensively in the residual cytoplasm. In spermatogenesis, Lc-Sox11a was distributed in cytoplasm of all male germ cells except spermatozoon with spermatogonium > spermatocyte > spermatid. During embryogenesis, Lc-Sox11a was expressed in most embryonic stages, the highest expression occurred in the formation-of-eye-lens stage, closely followed by the closure-of-blastopore stage, then the beginning-of-heart-pulsation stage. The results of whole mount in situ hybridization showed that the expression of Lc-Sox11a began to increase beginning with the multiple-cell stage, with the major distribution of Lc-Sox11a in the brain and eye areas in the pre-hatching stage. [ABSTRACT FROM AUTHOR]

Abstract: In this study, molecular characterization and expression of three heat shock responsive genes were analyzed as indicators to understand the mechanism of heat shock response of small abalone Haliotis diversicolor under stresses. The full length cDNA of heat shock transcriptional factor 1 (HdHSF1), heat shock factor binding protein 1(HSBP1), and heat shock protein 90 (HdHSP90) are 1548 bp, 809 bp, and 2592 bp respectively, encoding a protein of 515 aa, 75 aa, and 728 aa respectively. Real time quantitative PCR analysis revealed that these three genes are constitutively expressed in 7 selected tissues. The expression level of HdHSF1 in gills was higher than that in other tissues (p < 0.05). The highest expression level of HdHSBP1 was detected in hemocytes. The highest expression level of HdHSP90 was in the digestive tract and colleterial gland. The HdHSF1 expression level in the gills was up-regulated significantly (p < 0.05) after thermal stress and hypoxia exposure respectively. On the contrary, HdHSBP1 was down-regulated both in gills and hemocytes after thermal stress and the same as in gills after hypoxia stress. HdHSP90 expression level was also up-regulated in gills and hemocytes after both thermal and hypoxia stresses. These results indicated that these three heat shock responsive genes play important roles in response to thermal and hypoxia stress. [Copyright &y& Elsevier]

In this study, hypoxia inducible factor-1α (HIF-1α) and hypoxia inducible factor-1β (HIF-1β) from small abalone Haliotis diversicolor were cloned. The cDNA of H. diversicolor HIF-1α (HdHIF-1α) is 2833bp encoding a protein of 711aa and H. diversicolor HIF-1β (HdHIF-1β) is 1919bp encoding a protein of 590aa. Similar to other species' HIF-1, HdHIF-1 has one basic helix–loop–helix (bHLH) domain and two Per-Arnt-Sim (PAS) domains, and HdHIF-1α has a oxygen-dependent degradation domain (ODDD) with two proline hydroxylation motifs and a C-terminal transactivation domain (C-TAD) with an asparagine hydroxylation motif. Under normoxic conditions, HdHIF-1α and HdHIF-1β mRNAs were constitutively present in all examined tissues. Under hypoxia (2.0mg/L DO at 25°C) stress, HdHIF-1α expression was up-regulated in gills at 4h, 24h and 96h, and in hemocytes at 24h and 96h, while HdHIF-1β remained relatively constant. Under thermal stress (31°C), HdHIF-1α expression was significantly increased in gills at 4h, and hemocytes at 0h and 4h, while HdHIF-1β expression still remained relatively constant. These results suggested that HIF-1α may play an important role in adaption to poor environment in H. diversicolor. [ABSTRACT FROM AUTHOR]

Fish cell lines are important for the study in resource conservation, genetic breeding, diseases control, and environmental pollutants detection. The establishment of gonadal cell lines will provide a suitable in vitro model for studying the molecular regulation mechanisms underlying sex determination and sex-related gene expression and function. Large yellow croaker (Larimichthys crocea) is the unique marine economic fish with the highest production in China's coastal waters. In this study, the continuous gonadal cell lines, ovary cell line of large yellow croaker (LYCO) and testis cell line of large yellow croaker (LYCT), derived from the ovary and testis of juvenile fish were established with modified Leibovitz's L-15 medium at 27 °C. The medium was perfected to improve the growth rate and the increment ability of the LYCO and LYCT. The characteristics of the two cell lines were investigated, including the concentration of zeta-life fetal bovine serum (FBS), different passage ratios, cryopreservation, and recovery. The cell lines were successfully transfected with pEGFP-N1 and pNanog-N1 by electroporation, and the transfection efficiency of cells was high; these results showed that the cell lines could be used to study the expression of exogenous and endogenous genes. The LYCO expressed the marker gene of ovarian follicle cells, Foxl2 , while the LYCT expressed the marker gene of testicular sertoli cells, Dmrt1. However, LYCO and LYCT did not express Vasa , a marker gene of germ cells. Therefore, LYCO and LYCT appeared to consist of ovarian follicle cells and testicular sertoli cells, respectively. After the knockdown of Dmrt1 and Foxl2 in LYCO and LYCT, respectively, the results showed that the cell lines could be effective for RNA-mediated interference (RNAi) experiments and the study of the interactions among genes. As a first fish gonadal cell lines of L. crocea, which can provide a new model for the interactions between somatic cells and germ cells, and a basis for the further study of their reproduction and breeding in L.crocea. The results lay a foundation for the study of the gonadal cell culture in marine fish in the future. • Two cell lines from the ovary and testis of large yellow croaker (LYCO and LYCT) were first developed. • LYCO and LYCT can be used for high-efficiency and rapid electroporation to induce transformation to study gene function. • Overexpression of Nanog in LYCO and LYCT laid a foundation for the study of endogenous gene expression. • It is the first time to report the relationship of the sex-related genes by Dmrt1 and Foxl2 knockdown in LYCO and LYCT. [ABSTRACT FROM AUTHOR]

Abstract: The full-length (7816bp) cDNA of Vitellogenin (Vtg) encoding 2560 aa with an estimated molecular mass of 287.743kDa was cloned from the green mud crab Scylla paramamosain. Semi-quantitative PCR (sq-PCR) revealed a specific expression pattern of Sp-vtg gene in ovaries and hepatopancreas. With the development of ovaries, the expression level of Sp-vtg gene showed an increasing trend both in ovaries and hepatopancreas, and the expression level of Sp-vtg gene in hepatopancreas and ovary was stable after stage IV. By in situ hybridization, the positive signals of Sp-vtg gene were detected in the cytoplasm of oocytes in stage I, in the follicle cell and the surrounding of the nucleus in stage III, and in the nucleus in stage V. Furthermore, the signals become stronger with the later development stages of ovary. Moreover, in situ hybridization analysis revealed that positive signals of Sp-vtg gene are present in the hepatopancreatic tubule, and the signals increase during the development, becoming the strongest in stage V. Our results indicate that both ovaries and hepatopancreas are sites of Vitellogenin gene synthesis in S. paramamosain. [Copyright &y& Elsevier]

Abstract: The maturation promoting factor (MPF) is a key regulator of controlling G2/M phase transition in the meiotic maturation of oocyte and spermatocyte in animals, which is a complex of CDC2 (CDK1) and cyclin B. To better understand the molecular mechanism of oocyte and spermatocyte maturation in mud crab (Scylla paramamosain), the full length cDNA of cdc2 (Sp-cdc2) and cyclin B (Sp-cyclin B) were cloned and characterized. The full length cDNA of Sp-cdc2 gene is of 1593bp encoding a protein of 299 amino acids. Real-time quantitative PCR analysis revealed that the expression level of Sp-cdc2 in the ovary was higher than in other tissues (P<0.01); and its expression level was not significantly different in different stages of ovary development (P>0.05), meanwhile there was higher expression in T3 stage than in T1 and T2 stages (P<0.05). The full length cDNA of Sp-cyclin B is 1492bp encoding a protein of 391 amino acids. The real-time PCR results showed that its expression level in the ovary was the highest in all examined tissues (P<0.01), and the gonad expression level in O5 stage was significantly higher than in previous 4 stages and the testis (P<0.05), and was also significantly higher in T2 stage than in T1 stage (P<0.05). In situ hybridization analysis showed that the expressions of Sp-cdc2 and Sp-cyclin B transcripts were presented in similar distribution patterns in different developing stages of ovary and testis. The positive signals of Sp-cdc2 and Sp-cyclin B mRNA were detected in the oocytoplasm of oogonia and pre-vitellogenic and primary vitellogenic oocytes, while these two genes had higher expression level in the spermatid and secondary spermatocyte following primary spermatocyte. These results suggested that Sp-cdc2 and Sp-cyclin B may play essential roles in the oogenesis and spermatogenesis of the crab. [Copyright &y& Elsevier]

Abstract: Interleukin receptor‐associated kinase (IRAK)-1 binding protein 1 (IRAK1BP1) is a critical factor in preventing dangerous overproduction of proinflammatory cytokines by the innate immune system and in influencing the specificity of TLR responses. In this study, a first molluscan IRAK1BP1 gene, saIRAK1BP1, was cloned from the small abalone (Haliotis diversicolor). Its full-length cDNA sequence is 1047bp, with a 747bp open reading frame encoding a protein of 249 aa. The molecular mass of the deduced protein is approximately 28.1kDa with an estimated pI of 8.87, and shows highest identity (52%) to acorn worm Saccoglossus kowalevskii. Amino acid sequence analysis revealed that saIRAK1BP1 shares a conserved SIMPL domain. Quantitative real-time PCR was employed to investigate the tissue distribution of saIRAK1BP1 mRNA, and its expression in abalone under bacteria challenge and larvae at different developmental stages. The saIRAK1BP1 mRNA could be detected in all examined tissues, with the highest expression level in hemocytes, and was up-regulated in gills, kidneys and hemocytes after bacteria injection. Additionally, saIRAK1BP1 was constitutively expressed at all examined developmental stages. These results indicate that saIRAK1BP1 play an important role in the adult abalone immune system and might be essential in embryo and larval development in abalone. [Copyright &y& Elsevier]

Abstract: A selenium-dependent glutathione peroxidase cDNA was obtained from green mud crab Scylla paramamosain (SpGPx) by homology PCR technique and rapid amplification of cDNA ends (RACE) methods. The 1135 bp full-length cDNA contains a 9 bp 5′-untranslated region (UTR), an open reading frame (ORF) of 564 bp encoded a deduced protein of 187 amino acids (aa), and a 562 bp 3′-UTR with a 100 bp conserved eukaryotic selenocysteine insertion sequence (SECIS). It involves a putative selenocysteine (Sec40, or U40) residue which is encoded by an opal codon, 127TGA129, and forms an active site with residues Q74 and W142. Sequence characterization revealed that SpGPx contain a characteristic GPx signature motif 2 (64LAFPCNQF71), an active site motif (152WNFEKF157), a potential N-glycosylation site (76NTT78), and two residues (R90 and R168) which contribute to the electrostatic architecture by directing the glutathione donor substrate. Multiple sequence alignment and phylogenetic analysis showed that SpGPx share a high level of identities and closer relationship with other selected invertebrate GPxs and vertebrate GPx1 and GPx2. Molecular modelling analysis results also supported these observations. Real time quantitative PCR analysis revealed that SpGPx was constitutively expressed in 10 selected tissues, and its expression level in gill and testis was higher than that in the other tissues (p < 0.05). The SpGPx expression increased and then declined during ovarian and testicular development implying thatnscrpits yowed that SpGPx might play an important role in gonad development by protecting them from oxidative stress. The expression of SpGPx mRNA was induced by lipopolysaccharide (LPS) and hydrogen peroxide (H2O2) in hepatopancreas and haemocytes. The results suggested that SpGPx was implicated in the immune response induced by LPS and H2O2. [Copyright &y& Elsevier]

Abstract: Insulin-like growth factor binding protein 7 (IGFBP7), the only member of the IGFBP superfamily that binds strongly to insulin, may have different functions from other IGFBPs. Unlike other IGFBPs, there is no knowledge available on aquatic invertebrate IGFBP7. In this study, a molluscan IGFBP7 gene, saIGFBP7, was cloned for the first time from the small abalone Haliotis diversicolor. Its full-length cDNA sequence is 1812bp, with a 720bp open reading frame encoding a protein of 239 aa. The molecular mass of the deduced protein is approximately 25.37kDa with an estimated pI of 5.00, and it shares highest 41% identity to IGFBP7 of Amblyomma americanum. Analysis of conserved domains revealed the presence of an IGFBP N-terminal domain (IB), a kazal-type serine proteinase inhibitor domain (KI), and an immunoglobulin-like C2 domain (IgC2) in saIGFBP7. Furthermore, the 12 cysteine residues and the signature amino acid motif ‘xCGCCxxC’ which are characterized by the amino terminus region of the IGFBP superfamily are all presented in saIGFBP7. Quantitative real-time PCR and western blot were employed to investigate the tissue distribution of saIGFBP7, and its expression under bacterial challenge. The saIGFBP7 mRNA and protein could be detected in all examined tissues, with the highest expression level in hemocytes, higher expression level in gills, and was up-regulated in hemocytes and gills after bacterial injection. In addition, saIGFBP7 mRNA transcripts were observed in a subset of the branchial epithelium and the nucleus of hemocytes using the in situ hybridization method. Interestingly, saIGFBP7 was detected mainly in the goblet-like cell of the branchial epithelium by immunohistochemistry. These results suggested that saIGFBP7 was likely to be involved in a function associated with pathogenic infection and may play an important role in the adult abalone immune system. [Copyright &y& Elsevier]

Abstract: The small ubiquitin-related modifier-1 (SUMO-1) is a member of a family of ubiquitin-related proteins. SUMO pathway, which is involved in gene expression in eukaryotic posttranslational processing, plays important roles in gene expression, genomic stability and the occurrence of cells, development and other biological processes. Scylla paramamosain is one of the important economic breeding crabs in the southeast coast of China. To date, little is known about the distinct roles of SUMO in crustacean, especially in crabs. In the present study, we report the identification and characterization of mud crab, S. paramamosain SUMO-1 (SpSUMO-1) gene using an approach which combines expressed sequence tag (EST) and rapid amplification cDNA end (RACE). The full length cDNA of SpSUMO-1 gene (GenBank: HM581660) is of 732bp, including a 282bp open reading frame which encodes a protein of 93 amino acids. Tissue distribution analysis showed that SpSUMO-1 was expressed more abundantly in the ovary than in other tissues (P<0.01). And the expression profiles of SpSUMO-1 in the different gonad developing stages revealed that the highest expression of SpSUMO-1 occurred at proliferation stage, and then decreased gradually as the ovarian development progressed, while in the testis, the expression level of SpSUMO-1 was relatively stable at different stages of testis development. The distribution of SpSUMO-1 mRNA and its protein was observed in the crab gametogenesis by in situ hybridization and immunocytochemical method respectively. In oogenesis, SpSUMO-1 transcripts presented at the cytoplasm and nucleus of oocytes from proliferation stage to primary vitellogenesis stage, but only appeared in the nucleus of oocytes in secondary and tertiary vitellogenesis stages. Meanwhile, SpSUMO-1 protein was localized in the cytoplasm of oogonia and different developing oocytes. On the other hand, the SpSUMO-1 transcript was detected throughout the spermatogenesis, with the strong positive signals of SpSUMO-1 presented at the nuclei of primary and secondary spermatocytes, spermatids and spermatozoa. Interestingly, the positive signals of acrosomal tubules of spermatozoa were also detected. SpSUMO-1 protein was localized in spermatogonium, primary spermatocyte, secondary spermatocyte and spermatid, but the positive signal was only detected in the nucleus of spermatozoa. All these results suggested that SUMO-1 may play essential roles in the gametogenesis of the crustacea. [Copyright &y& Elsevier]

Abstract: Aquatic hypoxia caused by organic pollution and eutrophication is a pressing worldwide water pollution problem. Better methods for monitoring oxygen levels are needed to assist efforts to maintain and protect the health of natural aquatic environments. In this project, we used a Japanese ricefish (medaka, Oryzias latipes) 8K oligonucleotide array as a platform to identify potential hypoxic biomarkers in different organs (fin, gill, liver and brain) upon exposure to hypoxia. The microarray results were validated by qRT-PCR employing a subset of candidate biomarkers. Interestingly, the largest number and most significant of hypoxia responding array features were detected in hypoxia exposed fin tissues. We identified 173 array features that exhibited a significant response (over 2 fold change in expression) upon exposure to hypoxic conditions and validated a subset of these by quantitative RT-PCR. These gene targets were subjected to annotation and gene ontology mining. Positively identifiable gene targets that may be useful for development of a rapid and accurate biomarker test using fin clips are discussed in relation to previous reports on hypoxia responsive genes. [Copyright &y& Elsevier]

Abstract: In this study, we investigated the gene expression profiling of small abalone, Haliotis diversicolor by tributyltin (TBT) exposure using a cDNA microarray containing 2473 unique transcripts. Totally, 107 up-regulated genes and 41 down-regulated genes were found. For further investigation of candidate genes from microarray data and EST analysis, quantitative real-time PCR was performed at 6 h, 24 h, 48 h, 96 h and 192 h TBT exposure. 26 genes were found to be significantly differentially expressed in different time course, 3 of them were unknown. Some gene homologues like cellulose, endo-beta-1,4-glucanase, ferritin subunit 1 and thiolester containing protein II CG7052-PB might be the good biomarker candidate for TBT monitor. The identification of stress response genes and their expression profiles will permit detailed investigation of the defense responses of small abalone genes. [Copyright &y& Elsevier]

Abstract: Mammal interleukin-1 receptor-associated kinases (IRAKs) have been demonstrated to play important functions in TLRs (Toll-like receptor) signal pathway and T cell proliferation, but there is less knowledge available on mollusc IRAKs. In this study, a molluscan IRAK-4 gene, saIRAK-4, was cloned for the first time from the small abalone (Haliotis diversicolor). Its full-length cDNA sequence was 2062 bp, with a 1548 bp open reading frame encoding a protein of 516 aa. The molecular mass of the deduced protein was approximately 57.8 kDa with an estimated pI of 5.23, and showed highest identity (47%) to acorn worm Saccoglossus kowalevskii. Amino acid sequence analysis revealed saIRAK-4 shares conserved signature motifs with other IRAK-4 proteins, including the death domain (DD), serine/threonine/tyrosine protein kinase domain (STYKc), protein kinases ATP-binding region signature, serine/threonine protein kinases active-site signature and prokaryotic membrane lipoprotein lipid attachment site. Quantitative real-time PCR was employed to investigate the tissue distribution of saIRAK-4 mRNA, and its expression in abalone under bacteria challenge and larvae at different developmental stages. The saIRAK-4 mRNA could be detected in all examined tissues, with the highest expression level in gills, and was up-regulated in hemocytes and gills after bacteria injection. Additionally, saIRAK-4 was constitutively expressed at all examined developmental stages. These results indicate that saIRAK-4 could respond to pathogenic infection and may play an important role in the adult abalone immune system and early innate immunity in the process of abalone larval development. [Copyright &y& Elsevier]