Marsupenaeus Japonicas Research Articles

Integrative analysis of transcriptomic profile reveals potential roles of miRNAs in regulating development of Marsupenaeus japonicas

Integrative analysis of transcriptomic profile reveals potential roles of miRNAs in regulating development of Marsupenaeus japonicas

Dietary β-Hydroxy-β-Methylbutyrate Supplementation Affects Growth Performance, Digestion, TOR Pathway, and Muscle Quality in Kuruma Shrimp (Marsupenaeus japonicas) Fed a Low Protein Diet.

An 8-week feeding trial was performed to evaluate the effects of dietary β-hydroxy-β-methylbutyrate (HMB) supplementation on growth performance and muscle quality of kuruma shrimp (Marsupenaeus japonicas) (initial weight: 2.00 ± 0.01 g) fed a low protein diet. The positive control diet (HP) with 490 g/kg protein and negative control diet (LP) with 440 g/kg protein were formulated. Based on the LP, 0.25, 0.5, 1, 2 and 4 g/kg β-hydroxy-β-methylbutyrate calcium were supplemented to design the other five diets named as HMB0.25, HMB0.5, HMB1, HMB2 and HMB4, respectively. Results showed that compared with the shrimp fed LP, the HP, HMB1 and HMB2 groups had significantly higher weight gain and specific growth rate, while significantly lower feed conversion ratio (p < 0.05). Meanwhile, intestinal trypsin activity was significantly elevated in the above three groups than that of the LP group. Higher dietary protein level and HMB inclusion upregulated the expressions of target of rapamycin, ribosomal protein S6 kinase, phosphatidylinositol 3-kinase, and serine/threonine-protein kinase in shrimp muscle, accompanied by the increases in most muscle free amino acids contents. Supplementation of 2 g/kg HMB in a low protein diet improved muscle hardness and water holding capacity of shrimp. Total collagen content in shrimp muscle increased with increasing dietary HMB inclusion. Additionally, dietary inclusion of 2 g/kg HMB significantly elevated myofiber density and sarcomere length, while reduced myofiber diameter. In conclusion, supplementation of 1-2 g/kg HMB in a low protein diet improved the growth performance and muscle quality of kuruma shrimp, which may be ascribed to the increased trypsin activity and activated TOR pathway, as well as elevated muscle collagen content and changed myofiber morphology caused by dietary HMB.

Shrimp miR-965 transfers tumoricidal mitochondria

BackgroundMicro RNA of Marsupenaeus japonicas has been known to promote apoptosis of tumor cells. However, the detailed mechanisms are not well understood.ResultsUsing tomographic microscope, which can detect the internal structure of cells, we observed breast tumor cells following treatment of the miRNA. Intriguingly, we found that mitochondria migrate to an adjacent tumor cells through a tunneling nanotube. To recapitulate this process, we engineered a microfluidic device through which mitochondria were transferred. We show that this mitochondrial transfer process released endonuclease G (Endo G) into tumor cells, which we referred to herein as unsealed mitochondria. Importantly, Endo G depleted mitochondria alone did not have tumoricidal effects. Moreover, unsealed mitochondria had synergistic apoptotic effects with subtoxic dose of doxorubicin thereby mitigating cardiotoxicity.ConclusionsTogether, we show that the mitochondrial transfer through microfluidics can provide potential novel strategies towards tumor cell death.

Identification and Taxonomic Study of Shrimps in Bardawil Lagoon, North Sinai, Egypt

Taxonomic study of shrimp on the Bardawil lagoon, were studied in specimens of shrimp collected from Bardawil lagoon fishers from Seasonal occurrence of shrimp During, 2019. Identification of species was based on the morphological characters of rostrum (dorsal and ventral teeth), remarks of carapace, antenna, sub apical spines on telson and the colored pattern of the whole body based on standard keys and diagnoses available from the current literature. Modified Key identify shrimp species from Bardawil lagoon, was prepared after morphometric analysis. A total of 5 species from belong to three genus; Penaeus, Melicertus and Metapenaeus. Among these, two species belong to the genus Penaeus (Penaeus semisulcatus (De Haan, 1844) and Penaeus (Marsupenaeus) japonicas (Bate, 1888). The species which taken from genus Metapenaeus namely Metapenaeus stebbingi (Nobili, 1904) and Metapenaeus monoceros. (Fabricus, 1798), One species belong to the genus Melicertus kerathurus. (Forskal, 1775). Additional research is needed to more clearly define the distribution of shrimp species in Bardawil lagoon.

The role of Nrf2 in mitigating cadmium-induced oxidative stress of Marsupenaeus japonicus.

Nuclear factor-erythroid 2-related factor-2 (Nrf2) is an important modulator of cellular responses against Cd in mammalian cells. However, whether such modulation is conserved in Marsupenaeus japonicas remains unknown.In our study, the shrimps were injected with dsRNA targeting Nrf2 at 4μgg-1 body weight (b.w.) or sulforaphane (SFN) at 5μgg-1 b.w., and then were exposed to 40mgL-1 CdCl2 for 48h. After Nrf2 knockdown, the Cd content increased, but decreased in the SFN group. This suggested that Nrf2 could promote Cd excretion. A terminal deoxynulceotidyl transferase nick-end-labeling (TUNEL) assay revealed that the Nrf2 knockdown increased the number of apoptotic cells in M.japonicas, while SFN decreased the number of apoptotic cells. After Nrf2 knockdown, the total antioxidant capacity (T-AOC), superoxide dismutase (Sod) activity, and related gene expression decreased significantly, while the malondialdehyde (MDA) content increased remarkably. By contrast, SFN injection alleviated the oxidative stress, as evidenced by increased T-AOC, Sod activity, sod mRNA expression and a reduced MDA content. Similarly, detoxification related enzyme activities (ethoxyresorufin O-deethylase and glutathione-S-transferase (GST)) and their corresponding gene expressions (cyp3a (cytochrome P450 family 3 subfamily A) and gst) were suppressed in the ds-Nrf2 injection group, while they were elevated in the SFN group. In addition, ds-Nrf2 activated mitochondrial apoptotic pathway, as evidenced the mRNA and protein levels of caspase-3, Bcl2 associated X protein (Bax), and p53, while SFN treatment suppressed them. These results displayed that in M.japonicus Cd-induced cellular oxidative damage probably acts via the Nrf2 pathway.

Impacts of microplastics on three different juvenile shrimps: Investigating the organism response distinction

The effects of microplastics (MPs) on aquaculture animals have raised increasing concern, but studies on MPs contamination in cultured shrimp are still limited. Therefore, the responses of three widely farmed shrimp species to MPs, including Penaeus monodon (P. monodon), Marsupenaeus japonicas (M. japonicus) and Litopenaeus vannamei (L. vannamei), were investigated in this study. The results showed that the mortality of P. monodon, M. japonicus and L. vannamei were 47%, 53% and 20% respectively after 48 h of 300 mg/L MPs exposure. After 48 h of 100 mg/L MPs exposure, for P. monodon, the MPs content in water and excreta were significantly different from that in M. japonicus and L. vannamei. For genes expressions, the expression of catalase (Cat) was significantly increased and the expression of apoptosis protein (IAP) was inhibited in these three shrimps, but only the expression of Lysozyme (Lys) was increased in L. vannamei after MPs exposure. After 48 h of depuration, the Cat and IAP expression of P. monodon and M. japonicus was significant decreased while the IAP and Lys expression of L. vannamei still maintained at a high level. The results suggested that the metabolic rate of MPs in P. monodon was significantly higher than that in M. japonicus and L. vannamei. The tolerance of L. vannamei to MPs was higher than that of P. monodon and M. japonicas and their different responses in anti-microbial gene might be one of the reasons for the difference of their mortality. This study provides the first report comparing the organism response distinction in cultured shrimp and enriching to the understanding of the impact of MPs on ecosystem.

Clinical signs of naturally white spot syndrome virus (WSSV)-infected kuruma shrimp Marsupenaeus japonicas, based on their physiological and behavioural states

White spot syndrome virus (WSSV) proliferates rapidly and causes a high fatality rate in shrimp. Additionally, there is a lack of effective prevention and treatment methods for this disease. Although WSSV pathology is well-studied by molecular biology techniques, little research has have focused on the clinical signs of this disease based on individual shrimp performances. In this study, we recorded the clinical signs of diseased shrimp by observing their physiological and behavioural states. The physiological state of shrimp, including their heart rate (HR), heart rate variability (HRV), and ventilatory rate (VR), was recorded with a high-speed camera, which was used for crustacean disease diagnosis for the first time. The behavioural state of the shrimp consisted of their behaviour in the pond before sampling and the action demonstrated by the shrimp while falling into the water. Quantitative of WSSV in the shrimp was performed after measuring their physiological and behavioural characteristics. The virus was detected in both shrimp with and without white spots. The amount of virus in shrimp with or without white spots in the sand was relatively low and that of shrimp with white spots swimming in the water was high. The HR and VR both first increased and then decreased as the amount of virus increased gradually. There was a significant negative correlation between HRV and virus quantity in diseased shrimp. The swimming ability of shrimp with high virus quantities was weaker than that of shrimp with low virus quantities. Based on the data, we infer that the burrowing behaviour of shrimp is a disease resistance strategy; the slow swimming or knocking at the wall are symptoms of shrimp suffering from severe infection stage; WSSV destroys the heart, gill and muscle tissue of shrimp, causing damage to their basic motor function. Using the HRV index, we uncovered a fast and effective method with which to determine the virus quantity in diseased shrimp. On the basis of the above symptoms, WSSV is demonstrated as a shrimp disease not only related to the immune system, but also to the motor nervous system and respiratory circulatory systems.

Rab9 defense against white spot syndrome virus by participation in autophagy in Marsupenaeus japonicas

White spot syndrome virus (WSSV) is the main pathogen of shrimp and has led to considerable economic losses to the shrimp industry around the world. However, so far there are still no effective strategies to address this problem. In this paper, the tissue distribution of Rab9 as well as its defense mechanism against WSSV in Japanese shrimp (Marsupenaeus japonicas) was investigated. The results revealed that Rab9 had a higher expression in hemocyte and gill while expression was lower in heart, muscle, intestine, liver, indicating Rab9 was involved in the innate immune process. The results showed that the Rab9 expression increased when shrimp was challenged with WSSV compared with that of control, while the silence of Rab9 led to the increase of WSSV copies. In order to explore the antiviral mechanism of Rab9, it was demonstrated that the expression level of Rab9 changed during autophagy process, which indicated that Rab9 is participated in the autophagy procedure of shrimp. The fact that autophagy decreased after Rab9 silenced, may also suggest that Rab9 protein could affect autophagy. In short, the results showed Rab9 played a key role in antivirus through regulating autophagy. The results not only enlarge the limited views about molecular mechanism of Rab in invertebrate, but also help to enrich the immunological content in marine invertebrate.

Comparative toxicity of seven neonicotinoid insecticides and their two metabolites to juveniles of the marine crustacean kuruma prawn (Marsupenaeus japonicas)

Comparative toxicity of seven neonicotinoid insecticides and their two metabolites to juveniles of the marine crustacean kuruma prawn (Marsupenaeus japonicas)

Chemotaxis Assay for Marsupenaeus japonicas Hemocytes and Application for the Development of an Oral Immunostimulant Against White Spot Syndrome Virus.

The chemotactic activity of vertebrate leukocytes is an important host-defense mechanism. However, chemotaxis of invertebrate immune cells, particularly those of shrimp species, is incompletely understood and critically understudied. In this study, we aimed to optimize the conditions for a Boyden chamber chemotaxis assay using hemocytes (granulocytes) from cultured kuruma shrimp, Marsupenaeus japonicas (Mj) and the optimal conditions were: 5 μm-pore-size Polyvinylpyrrolidone membrane; culture buffer at pH 7.0; and chemotactic factor N-formyl-methionyl-leucyl-phenylalanine (fMLP) 10–8 mol/L; 4 h incubation time. We then applied the chemotaxis assay to develop an oral immunostimulant against white spot syndrome virus (WSSV), which results in high mortality rates in several shrimp species worldwide. We focused on the kelp Laminaria japonica, as this species contains immunostimulative molecules such as β-glucan. We prepared Heat Extracts (HE) and Crude Laminarans (CL) from kelp using hot water and hydrochloric acid extraction methods, respectively. HE and CL ware mixed with normal shrimp feed. Kelp extracts were orally administered for 7 days, and hematocyte chemotaxis toward fMLP was compared. No difference was detected between control and kelp extracts on day 3, but HE stimulated chemotaxis 2-fold and CL stimulated chemotaxis 3-fold relative to control on day 7 after initiating administration. Kelp extract administration protected against WSSV exposure. Finally, we identified that Kelp extracts stimulated hematocyte superoxide production on days 3 and 7, and increased hematocyte phagocytosis and phenol oxidase activity on day 7 after administration. We concluded that the chemotaxis assay is informative in assessment of shrimp hemocyte immunological activity, and is applicable to the development of immunostimulants against shrimp infectious diseases.

Comparative proteomic investigation of Marsupenaeus japonicus hepatopancreas challenged with Vibrio parahaemolyticus and white spot syndrome virus

This study aimed to use isobaric tags (IBTs) to investigate the immune response of the hepatopancreas of Marsupenaeus japonicas infected with Vibrio parahaemolyticus or white spot syndrome virus (WSSV). Liquid chromatography-tandem mass spectrometry and protein sequencing identified 1005 proteins. Among them, 109 proteins were upregulated and 94 were downregulated after V. parahaemolyticus infection. After WSSV infection, 130 proteins were identified as differentially abundant, including 88 that were upregulated and 42 were downregulated. Fifty-four proteins were identified as differentially abundant after both V. parahaemolyticus and WSSV infection. A number of proteins related to cytoskeletal processes, including actin and myosin, and apoptosis-related proteins were upregulated in shrimp after V. parahaemolyticus and WSSV infection, indicating that phagocytosis and apoptosis may be involved in the response to in V. parahaemolyticus or WSSV infection. Quantitative real-time PCR was carried out to verify the reliability of the proteomic data. These data provide a basis to characterize the immunity-related processes of shrimp in response to infection with WSSV or V. parahaemolyticus.

Molecular characterization of the dual oxidase (LvDuox) gene from the pacific white shrimp Litopenaeus vannamei

The reactive oxygen species（ROS） generated by dual oxidases (Duox) play a role in innate immunity in many organisms. In this study, a 4,735 bp full-length cDNA of the Pacific white shrimp dual oxidase (LvDuox) gene was cloned; the sequence included an open reading frame of 4,497 bp, encoding a protein of 1,498 aa with a calculated mass of 173 kDa. Structural analysis revealed that LvDuox contains several domains. Homology analysis revealed that LvDuox exhibits 96.1%, 67.3% and 67.3% sequence identity with Marsupenaeus japonicas, Drosophila melanogaster and Acyrthosiphon pisum Duox, respectively. The mRNA transcripts of LvDuox were detected in all tested tissues. The mRNA expression of LvDuox was significantly induced in the midgut after Vibrio parahaemolyticus E1 (VPE1) stimulation. After the level of H2O2 in the midgut increased, expression of the superoxide dismutase and catalase genes in the midgut increased significantly. These results suggested that the LvDuox gene was upregulated in the midgut after the challenge by VPE1, and antioxidant genes were involved in the regulation of ROS in the shrimp midgut. LvDuox may therefore be a new target for intestinal disease research in the Pacific white shrimp.

Heat and mass transfer, shrinkage, and thermal protein denaturation of kuruma prawn (Marsupenaeus japonicas) during water bath treatment: A computational study with experimental validation

Excessive shrinkage of processed food lowers its perceived quality by consumers, and therefore should be avoided. This study aims to clarify the multiphysics involved in the shrinkage of prawn during heating. A model that describes changes in moisture content of prawn due to pressure-driven water transport was reported according to Darcy's law. The transport model for the heating process included a stress-strain analysis (a structural mechanics model) coupled to a virtual work principle, which is applicable to a body undergoing shrinkage in two dimensions. Simultaneous calculation of changes in internal pressure and thermal protein denaturation (TPD, using a non-isothermal kinetics method) was used to describe the physics behind shrinkage. Temperature, moisture, pressure, as well as TPD profiles and distributions were calculated, and were validated with measured results. Results indicated that shrinkage was delayed due to slow rate of water release, which promoted the increment in internal pressure. This phenomenon, in combination with actin denaturation, resulted in dramatic water release and volumetric shrinkage. The proposed model improves the understanding of the shrinkage phenomena of prawn during heating and has the potential to contribute in the reduction of food quality losses associated with shrinkage and water release.

Deep eutectic solvent-based extraction and fabrication of chitin films from crustacean waste

In this study, chitin was exclusively extracted from shrimp shells (Marsupenaeus japonicas) through a green solvent called deep eutectic solvent (DES), and various types of DES were utilized to extract chitin. The physicochemical properties of the obtained chitin were compared with the conventional method. A high purity of chitin was obtained while using DES-8 (choline chloride-malonic acid) with a yield of 19.41% ± 1.35%, and purity was confirmed using 13C nuclear magnetic resonance. The DES-produced chitin was utilized to produce chitin films and was compared with standard chitin films. The obtained films were characterized by SEM, AFM, TGA, DSC, FTIR, mechanical properties, moisture sorption, swelling behavior, and biodegradation. The DES film showed similar properties to the standard film, while the mechanical properties, swelling behavior, and biodegradation of the DES chitin films proved to be similar to standard chitin film. These chitin films can be used as wound healing resources.

A C-type lectin (MrLec) with high expression in intestine is involved in innate immune response of Macrobrachium rosenbergii

C-type lectins (CTLs) are pattern-recognition proteins that play an important role in innate immunity of vertebrates and invertebrates. In this study, a lectin cDNA named MrLec was cloned and characterized from giant freshwater prawns (Macrobrachiun rosenbergii). The full-length cDNA of MrLec was 1431 bp, which contained an open reading frame of 1041 bp that encoded a protein with 346 amino acids. MrLec was found to contain a typical signal peptide of 18 amino acids and a single carbohydrate-recognition domain with 121 amino acids. The phylogenetic analysis showed that MrLec was grouped with vertebrates and had 57% identity with C-type lectin 3 from Marsupenaeus japonicas. Tissue expression analysis showed that MrLec was ubiquitously distributed at a high level in the intestine, with lower expression levels in the hemocytes, heart, hepatopancreas, gill and stomach. Vibrio parahaemolyticus infection induced the upregulation of MrLec in the gills and intestine. For the white spot syndrome virus (WSSV) challenge, MrLec in gills was upregulated at 24, 36 and 48 h. In intestine, MrLec also went up at 36 and 48 h WSSV challenge. Recombinant MrLec can agglutinate (Ca2+-dependent) and bind both Gram-negative and Gram-positive bacteria. rMrLec could attach to lipopolysaccharide and peptidoglycan in a dose-dependent manner. These results indicated possible MrLec involvement in the immune response of giant freshwater prawns.

Identification of a novel class B scavenger receptor homologue in Portunus trituberculatus: Molecular cloning and microbial ligand binding

Class B scavenger receptors (SRBs), which are present in mammals and insects, have been implicated in a wide range of functions. Herein, a novel SRB homologue, PtSRB, was cloned from the swimming crab, Portunus trituberculatus. PtSRB has 538 amino acid residues, and it consists of two transmembrane regions, a large extracellular loop, and two intracellular tails. A phylogenetic analysis showed that PtSRB distinctly clustered with Marsupenaeus japonicas SRB-1 and most Drosophila SRB homologues, including Croquemort, Peste, NinaD, and Santa Maria, but was separate from the Drosophila sensory neuron membrane protein, MjSRB-2, and all vertebrate SRBs. Real-time quantitative PCR analyses showed that the PtSRB gene was constitutively expressed in all tissues tested. When PtSRB was overexpressed in human embryonic kidney 293T cells, it was distributed in the membrane and cytoplasm. Moreover, in vitro assays showed that rPtSRB bound microbial lipopolysaccharide with low affinity, and lipoteichoic acid and peptidoglycan with high affinity. PtSRB transcripts were down-regulated after challenge with Vibrio alginolyticus or white spot syndrome virus, but not after a Candida lusitaniae challenge. This study provides valuable data for understanding the role of SRBs in the host defense against microbial pathogens, which will facilitate future studies of host–pathogen interactions in crabs.

Minichromosome maintenance protein 7 regulates phagocytosis in kuruma shrimp Marsupenaeus japonicas against white spot syndrome virus.

Minichromosome maintenance protein (MCM7) belongs to the MCM protein family and participates in the MCM complex by playing a role in the cell replication cycle and chromosome initiation of eukaryotes. Previously, we found that several genes, including MCM7, were over-expressed in Drosophila melanogaster after white spot syndrome virus (WSSV) infection. In this study, we aimed to further research the MCM7 of kuruma shrimp, Marsupenaeus japonicus (mjMCM7) and determine its role in the innate immune system. To this end, we cloned the entire 2307-bp mjMCM7 sequence, including a 1974-bp open reading frame (ORF) encoding a 658-aa-long protein. Real-time PCR showed that the gene was primarily expressed in the hemolymph and hepatopancreas and over-expressed in shrimp challenged with WSSV. Gene function study was carried out by knocking down the expression of MCM7 using small interference RNA (siRNA). The results revealed that β-actin, hemocyanin, prophenoloxidase (proPO) and tumor necrosis factor-α (TNF-α) were up-regulated while the cytoskeleton proteins such as myosin and Rho were significantly down-regulated at 24h after treatment. The results indicate a possible relationship between mjMCM7 and the innate immune system, and suggest that mjMCM7 may play a role in phagocytosis. After WSSV challenge, WSSV copies and mortality count were both higher in the MCM7-siRNA-treated groups at 60h after treatment, and the mortality count approached that of the control groups over time. The phagocytosis rate was significantly lower in the MCM7-siRNA-treated group than in the WSSV group. The findings of this study confirm that mjMCM7 positively regulates phagocytosis and plays an important role against WSSV. These results could help researchers to further understand the function of the MCM7 protein and reveal its potential role in the innate immunity of invertebrates.

Studies on molecular markers for hemocytes in kuruma shrimp Marsupenaeus japonicas

Major shrimp immune response such as phagocytosis, encapsulation and prophenoloxidase activity take place in the hemocytes. It is said that different kinds of hemocytes play different roles in the immune system therefore their classification are very important for understanding hemocytes function, as well as the entire immune system. Generally, hemocytes have three types of cell, hyaline cell, semigranular cell, and granular cell with reference to morphological features. However, morphological features are ambiguous and results may vary depending on the method used so that an accurate method is needed to establish classification of hemocytes. To resolve this problem, classification up to molecular level using molecular marker should be done. This study aims to develop an accurate classification method through identifying candidate molecular markers of hemocytes. Enable to do this, hemocyte samples were separated into two layers by density gradient centrifugation (upper layer and lower layer). Morphological features of hemocytes in each layer were analyzed by May Giemsa staining and flow cytometry analysis. In the result, hemocytes in upper layer showed larger size of cell and nucleus and less complexity of cell than lower layer. Then, gene expression analysis in each layer was conducted by transcriptome analysis and qPCR. Sixteen genes which showed more than eight times difference in total number of reads with p-value less than 1×10 were selected from transcriptome analysis. Fourteen genes showed significantly larger number of reads in upper layer and two genes showed larger number of reads in lower layer which was further confirmed by the mRNA levels of these sixteen genes were then analyzed by qPCR, where all genes indicated higher level of mRNA in same layers. These results imply that hemocytes can be separated into two types through morphological features, where sixteen candidate genes were identified to be possible markers, which may further classify hemocytes to different types. These results may provide technical basis for the classification of shrimp hemocytes.

Differential expression of microRNAs in shrimp Marsupenaeus japonicus in response to Vibrio alginolyticus infection

Till date numerous microRNAs (miRNAs) have been discovered from various organisms, including mammals, plants, insects, nematodes and viruses. They are known to have antiviral functions in crustaceans such as shrimp Marsupenaeus japonicas. However, little is known about the role of miRNAs against bacterial infection in this shrimp caused by Vibrio alginolyticus. We performed small RNA sequencing to characterize the differentially expressed microRNAs in V. alginolyticus challenged shrimp, in comparison to that in control uninfected shrimp, at 24 h and 48 h. In total, 55 host miRNAs were differentially expressed in response to the infection and most of these were downregulated at both the time-points. TargetScan and miRanda algorithms showed that the target genes of these down-regulated miRNAs were related to innate immune functions such as production of phenoloxidase enzyme, apoptosis and phagocytosis. Further, gene ontology analysis revealed that many immune signaling pathways were mediated by these miRNAs. This study is one of the earliest attempts at characterizing shrimp miRNAs that respond to V. alginolyticus infection, and will help unravel the miRNA pathways involved in antibacterial action in shrimp.

CDNA cloning, expression, and immunolocalization of gonad-inhibiting hormone (GIH) in Litopenaeus vannamei

In this study, the full-length GIH cDNA sequence from Litopenaeus vannamei was cloned from the eyestalk by reverse transcriptase polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends. The full-length GIH cDNA was 865 bp with a 288 bp open-reading frame, which encoded a 96 amino acid prepro-GIH with 17 amino acid signal peptide. L. vannamei GIH (LvGIH) can be classified as a member of type-II crustacean hyperglycemic hormone polypeptide family. LvGIH shares 93.8 and 66.7 % amino acid sequence identity with GIH from Penaeus monodon, and the molt-inhibiting hormone from Marsupenaeus japonicas, respectively. By quantitative real-time PCR (qPCR), LvGIH mRNA transcripts were detected in fertilized eggs, nauplius, zoea, mysis and juveniles of 25, 35 and 40 days old. LvGIH transcript levels increased significantly with the development from fertilized eggs to juveniles. LvGIH transcript levels were highest in juveniles at 35 days old. By RT-PCR, LvGIH mRNA transcripts were detected only in the eyestalks and brains but not in the muscles, intestines, gills, heart, hepatopancreas, ovaries and testes of adults, and there was no difference in the expression level of LvGIH between males and females. Using the P. monodon anti-GIH antibody, we showed that LvGIH was located mainly in the XO-SG and slightly in axon, with similar fluorescence intensity found in XO and SG. To summarize, we have cloned and characterized the GIH of the shrimp L. vannamei. In addition to the GIH properties described in other crustaceans, a peak of LvGIH expression was identified at the time of sexual differentiation (i.e., day 35 larvae) suggesting that LvGIH may also be involved in the control of this process.