Functions In Crustaceans Research Articles

Polystyrene nanoplastics induce apoptosis, histopathological damage, and glutathione metabolism disorder in the intestine of juvenile East Asian river prawns (Macrobrachium nipponense)

Nanoplastics (NPs) are widely distributed in the aquatic environment and have become a global concern as a new type of pollutant. Many researchers have studied the physiological effects of NPs on aquatic organisms, but relatively little is known about their effects on intestinal immune function in crustaceans. Therefore, we used NPs concentrations of 0, 5, 10, 20, 40 mg/L for 28 days of stress, evaluated the effects of NPs exposure on intestinal cell apoptosis, histopathological damage, and glutathione (GSH) metabolism of juvenile East Asian river prawns (Macrobrachium nipponense). As NPs concentration increased, the contents of total GSH and oxidized glutathione decreased gradually (P < 0.05), the concentration of GSH first increased and then decreased (P < 0.05), and the activities of lysozyme, acid phosphatase, phenoloxidase, and alkaline phosphatase first increased and then decreased (P < 0.05). Additionally, intestinal tissue structure was damaged, and the apoptosis rate significantly increased (P < 0.05). The expression of intestinal autophagy genes (CTL, ALF, Crustin, ATG8, and BCL-2) increased at first and then decreased, the expression levels of TNF and Wnt4 significantly decreased, and the expression of Beclin significantly increased with increasing NPs concentration. We also found that AP-1 and PTEN were highly expressed in the hepatopancreas and were involved in intestinal immune responses. Our results showed that exposure to NPs may induce apoptosis of intestinal tissue cells, induce autophagy, and inhibit GSH metabolism, thereby reducing intestinal immune function of M. nipponense. These findings provide a reference for healthy aquaculture and ecological risk assessment of prawns.

Soluble trehalase responds to heavy metal stimulation by regulating apoptosis in Neocaridina denticulata sinensis

Trehalase plays an important role in insect metabolism and development by hydrolyzing blood sugar trehalose, but it seems to perform primarily an immunomodulatory function in crustaceans whose blood sugar is glucose. Metal ions as pollutants seriously affecting crustacean health, but studies on trehalase in metal immunity are still limited. In this study, a soluble trehalase (NdTre1) that could bind to multiple metals was identified from Neocaridina denticulata sinensis for investigating metal resistance. Expression profiling revealed that NdTre1 was mainly expressed in the gill and was significantly decreased following stimulation with copper (Cu²⁺) and cadmium (Cd²⁺). Transcriptomic analysis of gills revealed an increase in ecdysone synthesis after interference with NdTre1. Increased ecdysone activated the endogenous mitochondrial pathway and the mitogen activated protein kinase (MAPK) pathway to further induced apoptosis. In vitro, Escherichia coli overexpressing recombinant NdTre1 had higher survival and faster growth rates to better adapted the metal-containing medium. Overall, NdTre1 exercises an important immune function in shrimp resistance to metal stimulation by regulating apoptosis and molting. Further investigation can further explore specific response mechanisms of NdTre1 to multiple metals.

Chitinases: expanding the boundaries of knowledge beyond routinized chitin degradation.

Chitinases, enzymes that degrade chitin, have long been studied for their role in various biological processes. They play crucial roles in the moulting process of invertebrates, the digestion of chitinous food, and defense against chitin-bearing pathogens. Additionally, chitinases are involved in physiological functions in crustaceans, such as chitinous food digestion, moulting, and stress response. Moreover, chitinases are universally distributed in organisms from viruses to mammals and have diverse functions including tissue degradation and remodeling, nutrition uptake, pathogen invasion, and immune response regulation. The discovery of these diverse functions expands our understanding of the biological significance and potential applications of chitinases. However, recent research has shown that chitinases possess several other functions beyond just chitin degradation. Their potential as biopesticides, therapeutic agents, and tools for bioremediation underscores their significance in addressing global challenges. More importantly, we noted that they may be applied as bioweapons if ethical regulations regarding production, engineering and application are overlooked.

Changes in Hemolymph Microbiota of Chinese Mitten Crab (Eriocheir sinensis) in Response to Aeromonas hydrophila or Staphylococcus aureus Infection.

The Chinese mitten crab (Eriocheir sinensis) has significant economic potential in both the Chinese domestic and global markets. The hemolymph microbiota is known to play a critical role in regulating physiological and biochemical functions in crustaceans. However, the study of the hemolymph microbiota of E. sinensis in response to infections has not been undertaken. In this study, changes in the composition and function of the hemolymph microbiota in E. sinensis infected with either Staphylococcus aureus (Sa) or Aeromonas hydrophila (Ah) were investigated using 16S rRNA sequencing, with a phosphate buffer saline (PBS) injection serving as the control. Results showed that the dominant hemolymph microbiota of E. sinensis were Proteobacteria, Bacteroidota, and Firmicutes. The relative abundance of the phyla Firmicutes, Bdellovibrionota, and Myxococcota was significantly reduced in both Sa and Ah groups compared to the PBS group. At the genus level, compared to the PBS group, a significant increase in the abundance of Flavobacterium and Aeromonas was found in both Ah and Sa groups. The analysis of the functional profile showed that pathways related to 'cell growth and death', 'metabolism of terpenoids and polyketides', 'cancers', 'lipid metabolism', 'neurodegenerative diseases', 'metabolism of other amino acids', 'xenobiotics biodegradation and metabolism', and 'circulatory system and endocrine system' were predominant in the Ah group. Meanwhile, pathways related to 'metabolism or genetic information progressing', such as 'translation', 'metabolic diseases', and 'cellular processes and signaling', were enriched in the Sa group. This study revealed the effects of pathogens (S. aureus or A. hydrophila) on the maintenance of the hemolymph microbiota in E. sinensis. It shed light on the mechanisms employed by the hemolymph microbiota of E. sinensis under pathogen stimulation.

Comparative transcriptome analysis reveals the effects of different feeding times on the hepatopancreas of Chinese mitten crabs

ABSTRACT Feeding rhythms affect a range of physiological functions in crustaceans. To investigate their effect on the physiological functions of Eriocheir sinensis, herein, we analyzed the influence of different feeding times on the hepatopancreas transcriptome via high-throughput sequencing. We harvested the hepatopancreas of crabs at 12:00 on day 11 of the experiment. We weighted the crabs before and after the experiment and found that those in the 06:00 group had the highest weight gain rate. In addition, 512 differentially expressed genes (DEGs) were grouped into nine distinct clusters. Functional enrichment analysis of DEGs showed that E. sinensis metabolic and immune processes were affected by the feeding time. Furthermore, we mapped the DEGs involved in retinol metabolism and the lysosome pathway. To our knowledge, this is the first comparative transcriptomic analysis of the hepatopancreas of E. sinensis based on different feeding times, which provides multi-level information to reveal the mechanism underlying the regulation of feeding rhythms in E. sinensis.

Nemo-like kinase (NLK) gene regulates apoptosis via the p53 signaling pathway in Litopenaeus vannamei under low-temperature stress

The Nemo-like kinase (NLK) is an important serine/threonine-protein kinase in many signaling pathways. However, its function in crustaceans, such as shrimps, is still poorly understood and needs to be further explored. In the present study, the full-length cDNA of NLK from Litopenaeus vannamei (LvNLK) was cloned. The full-length LvNLK cDNA has 2497 bp, including an open reading frame (ORF) of 1524 bp encoding a protein with 507 amino acids and a predicted molecular mass of 56.1 kDa. Phylogenetic analysis revealed that LvNLK shared high similarities with NLK from other known species. Low-temperature stress markedly upregulated the expression of LvNLK. Its overexpression in hemocytes suppressed the expression of BCL2-associated X (Bax) and tumor protein P53 (p53) in vitro. Meanwhile, the BCL2 apoptosis regulator (Bcl-2), MDM2 proto-oncogene (MDM2), and Yin Yang 1 (YY1) were upregulated. Moreover, LvNLK silencing in vivo increased the susceptibility of shrimps to low-temperature stress. The generation of ROS and the rate of hemocyte apoptosis also increased when LvNLK was silenced. Additionally, qPCR results indicated that LvNLK might participate in apoptosis via the p53 signaling pathway in vitro and in vivo. These results suggested that LvNLK is indispensable for the environmental adaptation of L. vannamei. Our current findings also demonstrated that NLK is evolutionarily conserved in crustaceans and provided insights into the environmental adaptation of invertebrates.

Understanding antimicrobial peptides and combatting antimicrobial resistance problems

Understanding antimicrobial peptides and combatting antimicrobial resistance problems

Comparative transcriptome analysis reveals the impact of the daily rhythm on the hemolymph of the Chinese mitten crab (Eriocheir sinensis)

ABSTRACT The daily rhythm affects a series of physiological functions in crustaceans. To study its effect on the physiological function in Eriocheir sinensis, a crustacean species of high economic value, we analyzed the hemolymph transcriptome during the daily rhythm by high-throughput sequencing. We sampled the hemolymph from crabs at four time points in a single day (06:00, 12:00, 18:00, and 24:00 h) and identified 3,01,661 and 1,03,998 transcripts and unigenes, respectively; some of the unigenes were annotated as core clock genes. Moreover, 15,564 differentially expressed genes (DEGs) were divided into nine different clusters. Functional enrichment analysis of DEGs indicated that the molting, metabolism, and immunity processes in E. sinensis were impacted by its daily rhythm. In addition, we mapped the DEGs involved in the daily entrainment pathway. To the best of our knowledge, this is the first comparative transcriptome analysis of crustacean hemolymph during the day–night cycle, and provides multi-level information for unraveling the finer regulatory effects of the daily cycle in crustaceans.

FADD regulates antibacterial immune responses via the immune deficiency signaling pathway in the Chinese mitten crab

In invertebrates, innate immune responses are the only defense against invading pathogens. The immune deficiency (IMD) signaling pathway protects invertebrates from bacterial infection by secreting antimicrobial peptides (AMPs). Fas-associated protein with death domain (FADD) activates AMPs and triggers apoptosis. However, FADD's function in crustaceans is unclear. Herein, the full-length FADD cDNA (EsFADD) was cloned from the Chinese mitten crab, Eriocheir sinensis. Vibrio parahaemolyticus infection upregulated EsFADD expression markedly. Knockdown of EsFADD in hemocytes suppressed the cytoplasm-to-nucleus translocation of transcription factor Relish under V. parahaemolyticus stimulation, which in turn reduced the expression of several AMPs. In vivo, silencing of EsFADD rendered crabs susceptible to bacterial infection and impaired their bacterial clearance. The results suggest that EsFADD is indispensable in IMD signal transduction in E. sinensis. In contrast to Drosophila, EsFADD barely promoted apoptosis. Our findings revealed the evolutionary conservation of FADD in crustaceans and provided insights into IMD signaling in invertebrates.

Comparative analysis of gut bacterial community composition during a single day cycle in Chinese mitten crab (Eriocheir sinensis)

Many physiological functions in crustaceans adapt to daily environmental change through circadian changes. However, the daily variation of the gut bacterial community composition has not been reported for crustaceans. In this study, we aimed to identify and elaborate on 24 h dynamic variations in the gut bacterial community of the Chinese mitten crab (Eriocheir sinensis). To this end, we processed gut samples from crabs collected at four time points (6:00, 12:00, 18:00, and 24:00) in a 24 h period and applied 16 S rRNA high-throughput gene sequencing. The gut bacterial community alpha-diversity and richness were highest in the samples collected at 18:00. Vagococcus salmoninarum was the most abundant species in the 12:00 samples, and Vibrio rumoiensis was the most abundant species in the 18:00 samples. Thus, our findings suggest that the daily variation may affect the response of the gut bacterial community to oxidative stress, and even affect the innate immune response of E. sinensis. Firmicutes and Bacteroidetes were identified as the dominant phyla in the gut bacterial community network. To the best of our knowledge, this is the first report describing the daily change in the gut microbiome composition of a crustacean species, E. sinensis. Our findings provide new insight into the 24 h dynamic variations of gut bacteria in crustaceans and may be useful for improving E. sinensis aquaculture.

Transcriptome Profiling Analysis of the Testis After Eyestalk Ablation for Selection of the Candidate Genes Involved in the Male Sexual Development in Macrobrachium nipponense.

The eyestalk of crustacean species secretes many hormones, affecting the process of reproduction, molting, metabolism of glucose, and other functions in crustaceans. In this study, important metabolic pathways and candidate genes involved in the male sexual development were identified through performing the transcriptome profiling analysis of the testis after the ablation of eyestalk from Macrobrachium nipponense. The histological observations revealed that the testis development became vigorous after eyestalk ablation, indicating that the hormones secreted by the eyestalk have negative effects on the testis development in M. nipponense. Transcriptome profiling analysis revealed that 1,039, 1,226, and 3,682 differentially expressed genes (DEGs) were identified between normal prawns (CG) vs single-side eyestalk ablation prawns (SS), SS vs double-side eyestalk ablation prawns (DS), and CG vs DS, respectively, indicating that the ablation of double-side eyestalk has more significant regulatory roles on male sexual development than that of single-side ablation, which was consistent with the histological observations. Lysosome, Apoptosis, Glycolysis/Gluconeogenesis, and Insulin signaling pathway were the main enriched metabolic pathways in all of these three comparisons, and the important genes from these metabolic pathways were also selected. The qPCR verifications of 10 DEGs from these metabolic pathways were the same as those of RNA-seq. The qPCR, in situ hybridization, and RNA interference analysis of Mn-NFkBα revealed that NFkBα has a positive regulatory effect on testis development. This study provided new insights on male sexual development in M. nipponense, promoting the studies on male sexual development in other crustaceans as well.

Biochemical characterization of the cyclooxygenase enzyme in penaeid shrimp.

Cyclooxygenase (COX) is a two-step enzyme that converts arachidonic acid into prostaglandin H2, a labile intermediate used in the production of prostaglandin E2 (PGE2) and prostaglandin F2α (PGF2α). In vertebrates and corals, COX must be N-glycosylated on at least two asparagine residues in the N-(X)-S/T motif to be catalytically active. Although COX glycosylation requirement is well-characterized in many species, whether crustacean COXs require N-glycosylation for their enzymatic function have not been investigated. In this study, a 1,842-base pair cox gene was obtained from ovarian cDNA of the black tiger shrimp Penaeus monodon. Sequence analysis revealed that essential catalytic residues and putative catalytic domains of P. monodon COX (PmCOX) were well-conserved in relation to other vertebrate and crustacean COXs. Expression of PmCOX in 293T cells increased levels of secreted PGE2 and PGF2α up to 60- and 77-fold, respectively, compared to control cells. Incubation of purified PmCOX with endoglycosidase H, which cleaves oligosaccharides from N-linked glycoproteins, reduced the molecular mass of PmCOX. Similarly, addition of tunicamycin, which inhibits N-linked glycosylation, in PmCOX-expressing cells resulted in PmCOX protein with lower molecular mass than those obtained from untreated cells, suggesting that PmCOX was N-glycosylated. Three potential glycosylation sites of PmCOX were identified at N79, N170 and N424. Mutational analysis revealed that although all three residues were glycosylated, only mutations at N170 and N424 completely abolished catalytic function. Inhibition of COX activity by ibuprofen treatment also decreased the levels of PGE2 in shrimp haemolymph. This study not only establishes the presence of the COX enzyme in penaeid shrimp, but also reveals that N-glycosylation sites are highly conserved and required for COX function in crustaceans.

Function analysis and molecular characterization of cyclin A in ovary development of oriental river prawn, Macrobrachium nipponense.

Macrobrachium nipponense has the characteristics of fast ovarian development cycle, which leads to the coexistence of multiple generations, the reduction of commodity specifications and the low economic benefit. Therefore, the study on the mechanism of ovarian development is of great significance to the development of industry. Cyclin A (CycA)is a key gene regulating ovarian development in vertebrates, but little information was available for its function in crustaceans. In this study, the full-length cDNA of Mn-CycA was obtained from the ovary. The full-length cDNA (2033bp) with an open reading frame of 1368bp, encoded a 456-amino acid protein. qRT-PCR revealed tissue-specific expression pattern of Mn-CycA, with abundant expression in the ovary. Results in different developmental stages of ovary indicated that Mn-CycA expression is positively correlated with ovarian maturation. qRT-PCR In different developmental stages, the expression of Mn-CycA mRNA gradually increased during the embryonic stage and decreased significantly on the first day of the hatching stage. At the 25th day of the metamorphosis stage, the expression level of Mn-CycAmRNA in female shrimp was 3.5 times higher than that in male shrimp, which may be related to the proliferation of oogonia and the formation of oocytes. In situ hybridization (ISH) of ovary showed Mn-CycA was examined in all stages and was mainly located in oogonia and oocytes. Compared with the control group, the obvious change of gonad somatic index (GSI) proved that injection of Mn-CycA dsRNA could delay the ovarian development cycle, which provided strong evidence for the involvement of Mn-CycA in ovarian maturation and oogenesis, and expanded a new perspective for studying the fast ovarian development cycle in M. nipponense.

The presence of an insulin-like peptide-binding protein (ILPBP) in the ovary and its involvement in the ovarian development of the red deep-sea crab, Chaceon quinquedens

Invertebrate insulin-like peptide-binding proteins (ILPBPs) are structurally homologous to vertebrate insulin-like growth factor binding protein 7 (IGFBP7). One of the invertebrate ILPBPs is considered as a potential binding protein of insulin-like androgenic gland factor (IAG) in males of crayfish, Cherax quadricarinatus. However, the ILPBP expression is not limited in the androgenic gland and found in most examined tissues, implicating that ILPBP may have additional functions in crustaceans. Here, the full-length cDNA sequence of ILPBP (termed ChqILPBP) is isolated from the ovary of the red deep-sea crab, Chaceon quinquedens. ChqILPBP transcripts are present in the various tissues, as similar to other crab species. The crustacean ILPBPs have their putative amino acid sequences conserved much less than vertebrate IGFBP7s. To understand if ChqILPBP is involved in ovarian development, examined are levels of ChqILPBP, together with vitellogenin (ChqVTG) in the same ovary and hepatopancreas of adult females at the different ovarian stages: 2, 3, and 5. Chaceon hepatopancreas exhibits as the primary VTG synthesis site, while VTG transcript levels do not differ by the ovarian stages. The ovary contains ChqILPBP transcripts ~10-fold higher than hepatopancreas that changes significantly from stage 2 to 3. Such an expression pattern mirrors that of ovarian ChqVTG. In hepatopancreas, ChqILPBP transcripts are similar at stages 2 and 3 and increase significantly at stage 5. The data indicate that ovarian ILPBP may function differently from that of the hepatopancreas and may play a role in ovarian development. ChqAK transcripts are ~six folds higher in the ovary than the hepatopancreas. While they do not differ by ovarian stages, suggesting that AK may not be involved in vitellogenesis of the cold water crustacean species.

Stimulatory roles of epidermal growth factor receptor (EGFR) in ovarian development of mud crab Scylla paramamosain

The epidermal growth factor receptor (EGFR) is a pleiotropic glycoprotein which plays a role in regulating cell proliferation, migration and differentiation. However, to date little is known about its functions in crustaceans. In this study, we successfully identified SpEGFR from mud crab Scylla paramamosain. RT-PCR result showed that SpEGFR was widely distributed in all tested tissues and highly expressed in ovary. In situ hybridization revealed that SpEGFR mainly localized in oocyte perinuclear region with notably obvious signals. In vitro experiments showed that the expression of SpVgR and SpCyclin B in ovary explants from late vitellogenic stage crabs (summer) were significantly increased when treated with 1 nM human EGF (hEGF) for 1 h, while there was no obvious change towards SpEGFR. Interestingly, as for winter crab at the same vitellogenic stage, the expression of SpVgR and SpCyclin B in ovary explants did not show significant increase until treated with higher concentration of 10 nM hEGF and longer incubation time of 12 h. In addition, the hEGF-induced effect could be suppressed by pre-treated with EGFR inhibitor AG1478 and PD153035, respectively, which further indicated that EGF-EGFR pathway played a vital role in ovarian development in mud crab. In conclusion, SpEGFR might promote ovarian development by stimulating the expression of SpVgR and SpCyclin B under hEGF-induced treatment. The different physiological response to hEGF in the same vitellogenic stage crabs between summer and winter might be attributed to the changes in metabolism and physiological sensitivity.

Molecular characteristic of activin receptor IIB and its functions in growth and nutrient regulation in Eriocheir sinensis.

Activin receptor IIB (ActRIIB) is a serine/threonine-kinase receptor binding with transforming growth factor-β (TGF-β) superfamily ligands to participate in the regulation of muscle mass in vertebrates. However, its structure and function in crustaceans remain unknown. In this study, the ActRIIB gene in Eriocheir sinensis (Es-ActRIIB) was cloned and obtained with a 1,683 bp open reading frame, which contains the characteristic domains of TGF-β type II receptor superfamily, encoding 560 amino acids. The mRNA expression of Es-ActRIIB was the highest in hepatopancreas and the lowest in muscle at each molting stage. After injection of Es-ActRIIB double-stranded RNA during one molting cycle, the RNA interference (RNAi) group showed higher weight gain rate, higher specific growth rate, and lower hepatopancreas index compared with the control group. Meanwhile, the RNAi group displayed a significantly increased content of hydrolytic amino acid in both hepatopancreas and muscle. The RNAi group also displayed slightly higher contents of saturated fatty acid and monounsaturated fatty acid but significantly decreased levels of polyunsaturated fatty acid compared with the control group. After RNAi on Es-ActRIIB, the mRNA expressions of five ActRIIB signaling pathway genes showed that ActRI and forkhead box O (FoxO) were downregulated in hepatopancreas and muscle, but no significant expression differences were found in small mother against decapentaplegic (SMAD) 3, SMAD4 and mammalian target of rapamycin. The mRNA expression s of three lipid metabolism-related genes (carnitine palmitoyltransferase 1β (CPT1β), fatty acid synthase, and fatty acid elongation) were significantly downregulated in both hepatopancreas and muscle with the exception of CPT1β in muscles. These results indicate that ActRIIB is a functionally conservative negative regulator in growth mass, and protein and lipid metabolism could be affected by inhibiting ActRIIB signaling in crustacean.

Differential expression of microRNAs in mud crab Scylla paramamosain in response to white spot syndrome virus (WSSV) infection

Till date numerous microRNAs (miRNAs) have been discovered from various organisms, including fish, shellfish and crustaceans. The miRNAs are known to regulate immune functions in crustaceans, but little is known about the role of miRNAs against viral infection in crab. We performed small RNA sequencing to characterize the differentially expressed miRNAs in WSSV infected Scylla paramamosain, in comparison to that in uninfected crab, at 2 h and 12 h post infection. In total, 24 host miRNAs were up-regulated and 25 host miRNAs were down-regulated in response to WSSV infection at 2 h post infection. And 27 host miRNAs were up-regulated and 30 host miRNAs were down-regulated in response to WSSV infection at 12 h post infection. Further, the gene ontology analysis revealed that many signaling pathways were mediated by these miRNAs. The integral component of membrane is the most important biological process and endocytosis pathway is the most important pathway, which indicates that endocytosis is very important for WSSV infection. This study is one important attempt at characterizing crab miRNAs that response to WSSV infection, and will help unravel the miRNA pathways involved in antiviral immunity of crab.

Identification and characterization of hemocyte microRNAs in mud crab Scylla paramamosain in response to Vibrio parahemolyticus infection

The microRNAs (miRNAs) are known to regulate immune functions in crustaceans, but little is known about the role of miRNAs against bacterial infection. We performed small RNA sequencing to characterize the differentially expressed microRNAs in V. parahemolyticus infected crab, in comparison to that in control uninfected crab, at 12 h and 24 h post infection. In total, 23 host miRNAs were up-regulated in response to the infection and 23 host miRNAs were down-regulated at both the time-points. Among these miRNAs, miR-125, PC-3p-11,483, miR-281-5p, miR-10-3p and PC-5p-50,686 were significantly up-regulated by above 10 folds, and stem-loop RT-qPCR confirmed this result. Further, gene ontology analysis revealed that many signaling pathways, especially phagocytosis, were mediated by these miRNAs. Golgi apparatus and Magnesium ion binding are important biological processes, endocytosis and phagosome are important pathways in the immune response to V. parahemolyticus. This study is one important attempt at characterizing crab miRNAs that response to V. parahemolyticus infection, and will help unravel the miRNA pathways involved in antibacterial immunity of crab.

Cloning and expression analysis of Serum Amyloid A (SAA) from Exopalaemon carinicauda (Holthuis, 1950) (Caridea, Palaemonidae)

Abstract Serum amyloid A (SAA), an acute-phase protein, has been demonstrated to play a critical role in the inflammatory response. However, data regarding its function in crustaceans are still scarce. In the present study, cDNA of SAA from Exopalaemon carinicauda (Holthuis, 1950) (designated as EcSAA) was cloned and characterized. The full-length cDNA was 683 bp, including an open reading frame (ORF) of 267 bp that encoded 88 amino acid residues. The predicted amino acid sequence of EcSAA contained characteristic motifs of the SAA family. The EcSAA was widely expressed in all tissues, with the highest expression in gill and hepatopancreas. In response to the Vibrio anguillarum Bergeman, 1909 challenge, EcSAA mRNA was up-regulated in gill, hepatopancreas, and haemolymph. These results suggest thatEcSAA may be a constitutive and inducible acute-phase protein involved in anti-pathogen responses in E. carinicauda.

Characterization and expression patterns of wnt4 in Exopalaemon carinicauda (Holthuis, 1950) (Caridea, Palaemonidae) during embryonic and larval development

Abstract Wnt4 (Wingless-type MMTV integration site family member 4) has been demonstrated to play critical roles in a wide variety of biological processes, including embryonic development, cell proliferation, and differentiation in vertebrates, but its function in crustaceans is still not clear. In the present study, the full-length wnt4 cDNA sequence was cloned and characterized for the ridgetail white prawn Exopalaemon carinicauda. The expression patterns of the wnt4 mRNA in embryos and larvae at different stages were investigated. The tissue distribution showed that wnt4 was obviously expressed in eyestalk and hepatopancreas. During embryonic development, the wnt4 was highly expressed in all developmental stages except the zygote, two-cell stage, and late zoaea stage. The wnt4 mRNA was expressed in Z1-Z5 and post-larval stages. Taken together, the present study indicates that the wnt4 gene may be involved in the regulation of embryonic and larval development in the ridgetail white prawn.