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Qun Wang - One of the best experts on this subject based on the ideXlab platform.

  • b52 promotes alternative splicing of dscam in chinese mitten crab Eriocheir Sinensis
    Fish & Shellfish Immunology, 2019
    Co-Authors: Zhicheng Wan, Youting Zhu, Tianheng Gao, Qun Wang
    Abstract:

    B52 is a member of the classical serine/arginine (SR)-rich proteins, which are phylogenetically conserved and play significant roles in mRNA maturation, including alternative splicing. In the present study, the docking site, selector sequences and locus control region of the Chinese mitten crab (Eriocheir Sinensis) Down syndrome cell adhesion molecule (EsDscam) were identified. Alternative splicing of Dscam is essential to generate different isoforms. We also isolated and characterised the B52 gene from E. Sinensis (EsB52). The 876 bp open reading frame of EsB52 encodes a 291 amino acid residue polypeptide, and EsB52 has two RNA recognition motifs (RRMs) at the N-terminus and an arginine/serine-rich domain at the C-terminus. Each RRM contains two degenerate short submotifs, RNP-1 and RNP2. Analysis of tissue distribution revealed that EsB52 mRNA expression was widespread in all tested tissues, and especially high in brain and hemocytes. In hemocytes, EsB52 was upregulated significantly after stimulation with pathogen-associated molecular patterns and bacteria. Furthermore, EsB52 RNAi decreased the number of Ig7 inclusion in mRNA rather than Ig2 or Ig3. Taken together, these findings suggest that EsB52 acts as an alternative splicing activator of EsDscam.

  • p38 participates in spermatogenesis and acrosome reaction prior to fertilization in chinese mitten crab Eriocheir Sinensis
    Gene, 2015
    Co-Authors: Ming Zhu, Yuanli Wang, Wenjuan Sun, Hongdan Yang, Zelin Duan, Qun Wang
    Abstract:

    Abstract P38 mitogen-activated protein kinases (MAPKs) comprise a family of serine/threonine protein kinases that play important roles in cellular responses to inflammatory cytokines and environmental stresses. These kinases are involved in controlling cell division, differentiation and death in mammalian testes and therefore are critical to spermatogenesis. To explore their functions in male reproduction of Chinese mitten crabs, Eriocheir Sinensis p38 ( Es -p38) protein expression was determined in different tissues including testes at different developmental stages by Western blot. Es -p38 was expressed in various tissues, with higher levels in the heart, stomach, gills and testes. Total Es -p38 protein levels increased gradually during spermatogenesis, but phosphorylated Es -p38 was much higher in the spermatid (August–October) than the spermatocyte (July–August) and sperm (October–January) stages. Trypan blue staining and hematoxylin/eosin staining were both used to detect sperm motility and changes in sperm morphology during the acrosome reaction (AR) induced by pre-incubation with A23187 in vitro, activated Es -p38 proteins detected by fluorescent microscopy were translocated gradually to nuclear and apical cap regions, accumulating at the anterior of the acrosomal tubule. The results suggest the involvement of p38 MAPK in spermatogenesis and the AR in E. Sinensis .

  • morphological alterations of all stages of spermatogenesis and acrosome reaction in chinese mitten crab Eriocheir Sinensis
    Cell and Tissue Research, 2015
    Co-Authors: Yuanli Wang, Wenjuan Sun, Qun Wang
    Abstract:

    The Chinese mitten crab (Eriocheir Sinensis) is a seasonally breeding species and its reproductive system comprises paired symmetrical structures: testes, vasa deferentia, seminal vesicles, accessory glands and ejaculatory ducts. Histological examination of the testis of mature males reveals regression of the gonads and inhibition of the process of spermatogenesis during December to April of the following year, the regeneration of the gonads during June to July and the occurrence of the highest level of spermatogenesis during August to October. Microscopic assessments and hematoxylin and eosin (H&E) staining were used to describe all spermatogenic stages (spermatogonia, primary and secondary spermatocytes, spermatids and spermatozoids). To observe the morphological changes during spermiogenesis, we successfully initiated primary cell culture using testis tissue of E. Sinensis, which will lay a solid foundation for further work on the immortalization of crab cells. During the interaction between the sperm and oocyte, the fertilizing spermatozoon must undergo a series of terminal morphological changes, called the acrosome reaction (AR). This study also provides a detailed description of the structural alterations of the acrosome reaction of E. Sinensis. The acrosome complex and cup-shaped nucleus are located at the anterior and posterior of the spermatozoon, respectively. Male germ cell development involves a tightly controlled sequence of differentiation switches. The purpose of this study is to increase our knowledge of the morphological alterations during spermatogenesis and the acrosome reaction, whose changes are a fundamental requirement for fertilization of E. Sinensis.

  • two antibacterial c type lectins from crustacean Eriocheir Sinensis stimulated cellular encapsulation in vitro
    Developmental and Comparative Immunology, 2013
    Co-Authors: Shuang Li, Lin Cheng, Minhao Wu, Aiqing Yu, Weiwei Li, Qun Wang
    Abstract:

    The first step of host fighting against pathogens is that pattern recognition receptors recognized pathogen-associated molecular patterns. However, the specificity of recognition within the innate immune molecular of invertebrates remains largely unknown. In the present study, we investigated how invertebrate pattern recognition receptor (PRR) C-type lectins might be involved in the antimicrobial response in crustacean. Based on our previously obtained completed coding regions of EsLecA and EsLecG in Eriocheir Sinensis, the recombinant EsLectin proteins were produced via prokaryotic expression system and affinity chromatography. Subsequently, both rEsLecA and rEsLecG were discovered to have wide spectrum binding activities towards microorganisms, and their microbial-binding was calcium-independent. Moreover, the binding activities of both rEsLecA and rEsLecG induced the aggregation against microbial pathogens. Both microorganism growth inhibitory activities assays and antibacterial activities assays revealed their capabilities of suppressing microorganisms growth and directly killing microorganisms respectively. Furthermore, the encapsulation assays signified that both rEsLecA and rEsLecG could stimulate the cellular encapsulation in vitro. Collectively, data presented here demonstrated the successful expression and purification of two C-type lectins proteins in the Chinese mitten crab, and their critical role in the innate immune system of an invertebrate.

  • two novel toll genes estoll1 and estoll2 from Eriocheir Sinensis are differentially induced by lipopolysaccharide peptidoglycan and zymosan
    Fish & Shellfish Immunology, 2013
    Co-Authors: Xingkun Jin, Xiaonv Guo, Qun Wang
    Abstract:

    Tolls/Toll-like receptors (TLRs) play an essential role in initiating innate immune responses against pathogens and are found throughout the insect kingdom but have not yet been reported in the crustacean, Eriocheir Sinensis. For this purpose, we cloned two novel Toll genes from E. Sinensis, EsToll1 and EsToll2. The full-length cDNA of EsToll1 was 3963 bp with a 3042-bp open reading frame (ORF) encoding a 1013-amino acid protein. The extracellular domain of this protein contains 17 leucine-rich repeats (LRRs) and a 139-residue cytoplasmic Toll/interleukin-1 receptor (TIR) domain. The cDNA full-length of EsToll2 was 4419 bp with a 2667-bp ORF encoding an 888-amino acid protein with an extracellular domain containing 10 LRRs and a 139-residue cytoplasmic TIR domain. By phylogenetic analysis, EsToll1 and EsToll2 clustered into one group together with Tolls from other crustaceans. Quantitative RT-PCR analysis demonstrated that a) both EsToll1 and EsToll2 were constitutively expressed in all tested crab tissues; b) EsToll1 and EsToll2 were differentially induced after injection of lipopolysaccharides (LPS), peptidoglycan (PG) or zymosan (GLU). Importantly, EsToll2 expression was significantly upregulated at almost all time intervals post-challenge with LPS, PG and GLU. Our study indicated that EsToll1 and EsToll2 are differentially inducibility in response to various PAMPs, suggesting their involvement in a specific innate immune recognition mechanism in E. Sinensis.

Linsheng Song - One of the best experts on this subject based on the ideXlab platform.

  • a novel gata like zinc finger transcription factor involving in hematopoiesis of Eriocheir Sinensis
    Fish & Shellfish Immunology, 2018
    Co-Authors: Zhihao Jia, Lingling Wang, Xiaorui Song, Yu Liu, Yunke Jia, Linsheng Song
    Abstract:

    Abstract GATA transcription factor is a family of DNA-binding proteins that can recognize and bind to sequence of (A/T) GATA (A/G). In the present study, a GATA-like protein (named as EsGLP) was characterized from Eriocheir Sinensis, including an 834 bp full length open reading frame of EsGLP, encoding a polypeptide of 277 amino acids. The deduced amino acid sequence of EsGLP contained one conserved GATA-type zinc finger of the form Cys-X2-Cys-X17-Cys-X2-Cys, with four cysteine sites. The EsGLP mRNA transcripts were mainly detected in the hematopoietic tissue, hepatopancreas and gonad. The recombinant EsGLP protein was prepared for the antibody production. The EsGLP protein was mainly distributed in the edge of lobules in the HPT and the cytoplasm of hemocytes. The mRNA transcripts of EsGLP in hemocytes were significantly decreased at 24 h (0.39-fold and 0.27-fold, p

  • identification of a clip domain serine proteinase involved in immune defense in chinese mitten crab Eriocheir Sinensis
    Fish & Shellfish Immunology, 2018
    Co-Authors: Zhihao Jia, Huan Zhang, Lingling Wang, Mengqiang Wang, Xiudan Wang, Linsheng Song
    Abstract:

    Clip-domain serine proteinase is an important serine proteinase family involved in many biological processes, which is only found in invertebrates. In the present study, the full-length cDNA of a clip domain serine proteinase (designed as EsCDSP) gene was cloned from Chinese mitten crab Eriocheir Sinensis using rapid amplification of cDNA ends (RACE) technique. It was of 1488 bp with an open reading frame (ORF) of 1134 bp encoding a polypeptide of 377 amino acids. There were a signal peptide, a clip domain, and a Tryp_SPc domain in the deduced amino acid sequence of EsCDSP. Highly conserved cysteine residues were identified in the clip domain and Tryp_SPc domain. EsCDSP shared similarities of 40%-61% with CDSPs from Penaeus monodon (ACP19562.1), Scylla paramamosain (CCW43200.1), Drosophila melanogaster (NP_649734.2) and Delia antiqua (AAW57295.1). It was clustered with other CDSPs from crabs in the phylogenetic tree. EsCDSP transcript was highly expressed in hemocytes and it could response to the stimulations of Vibro anguillarum and Pichia pastoris. rEsCDSP could activate proPO system and significantly increase the PO activity of HLS. In addition, rEsCDSP could bond to Aeromonas hydrophila, Vibro anguillarum and Vibro alginolyticus, and reduced the mortality rate causing by pathogen infection. All the results suggested that EsCDSP was an important immune response participator involved in activation of the proPO system of crab.

  • functional characterization of hemocytes from chinese mitten crab Eriocheir Sinensis by flow cytometry
    Fish & Shellfish Immunology, 2017
    Co-Authors: Zhihao Jia, Lingling Wang, Shuai Jiang, Mingzhe Sun, Mengqiang Wang, Linsheng Song
    Abstract:

    Hemocytes comprise a diversity of cell types with functional and structural heterogeneity, and they play key roles in the host defense of invertebrates. In the present study, the hemocytes from Chinese mitten crab Eriocheir Sinensis were directly separated into two groups by flow cytometry. The hemocytes in P1 group were full of round and abundant granules with deeply staining cytoplasm, while P2 hemocytes were more diverse with a wide range of sizes and less granularity. Both P1 and P2 hemocytes exhibited phagocytic ability, but the phagocytic rate of P1 hemocytes increased which was significantly higher than that of P2 hemocytes after LPS stimulations. The levels of ROS production and intracellular Calcium as well as lysosome content were higher in P1 hemocytes than that in P2 hemocytes under both normal and immune-activated situations. The genes involved in phagocytosis, antimicrobial and antioxidant activities were mainly expressed in P1 hemocytes, while the genes involved in proPO activation system were highly expressed in P2 hemocytes. These results collectively suggested that P1 hemocytes were the main immunocompetent hemocytes in Chinese mitten crab and P2 hemocytes mainly participated in proPO activation system.

  • a galectin from Eriocheir Sinensis functions as pattern recognition receptor enhancing microbe agglutination and haemocytes encapsulation
    Fish & Shellfish Immunology, 2016
    Co-Authors: Mengqiang Wang, Huan Zhang, Lingling Wang, Yunchao Gai, Mengmeng Huang, Ying Guo, Hao Wang, Linsheng Song
    Abstract:

    Galectins are a family of β-galactoside binding lectins that function as pattern recognition receptors (PRRs) in innate immune system of both vertebrates and invertebrates. The cDNA of Chinese mitten crab Eriocheir Sinensis galectin (designated as EsGal) was cloned via rapid amplification of cDNA ends (RACE) technique based on expressed sequence tags (ESTs) analysis. The full-length cDNA of EsGal was 999 bp. Its open reading frame encoded a polypeptide of 218 amino acids containing a GLECT/Gal-bind_lectin domain and a proline/glycine rich low complexity region. The deduced amino acid sequence and domain organization of EsGal were highly similar to those of crustacean galectins. The mRNA transcripts of EsGal were found to be constitutively expressed in a wide range of tissues and mainly in hepatopancreas, gill and haemocytes. The mRNA expression level of EsGal increased rapidly and significantly after crabs were stimulated by different microbes. The recombinant EsGal (rEsGal) could bind various pathogen-associated molecular patterns (PAMPs), including lipopolysaccharide (LPS), peptidoglycan (PGN) and glucan (GLU), and exhibited strong activity to agglutinate Escherichia coli, Vibrio anguillarum, Bacillus subtilis, Micrococcus luteus, Staphylococcus aureus and Pichia pastoris, and such agglutinating activity could be inhibited by both d-galactose and α-lactose. The in vitro encapsulation assay revealed that rEsGal could enhance the encapsulation of haemocytes towards agarose beads. These results collectively suggested that EsGal played crucial roles in the immune recognition and elimination of pathogens and contributed to the innate immune response against various microbes in crabs.

  • the characterization of hematopoietic tissue in adult chinese mitten crab Eriocheir Sinensis
    Developmental and Comparative Immunology, 2016
    Co-Authors: Zhihao Jia, Lingling Wang, Sharath Kavungal, Shuai Jiang, Depeng Zhao, Mingzhe Sun, Linsheng Song
    Abstract:

    Invertebrates rely on the efficient innate immune mechanisms against invaders, in which the continuous production of hemocytes (hematopoiesis) is indispensable. In the present study, the hematopoietic tissue (HPT) from Chinese mitten crab Eriocheir Sinensis was identified and characterized. It was a thin and non-transparent sheet located at the dorsolateral side of the stomach, which was composed of a series of ovoid lobules. Each lobule was surrounded by connective tissue containing a large amount of spherical cells with big nucleus. In HPT, the cells were full of mitochondria and granules, and DNA replication was detected in some cells by EdU labeling technique. Cell proliferation was observed in HPT by transmission electron microscope (TEM). The distribution of two transcription factors, GATA1 and RUNX1, were examined by human GATA1 and RUNX1 antibodies, respectively. Three homologues of RUNX1 were detected in the HPT while no signal of RUNX1 was observed in hemocytes, and GATA1 was detected in both HPT and some hemocytes. The mRNA transcript of a novel hematopoiesis related cytokine EsAst was detected in hepatopancreas and hemocytes, but it was no detectable in HPT. The mRNA expression level of EsAst in hepatopancreas was 1.38-fold higher than that in hemocytes. Total hemocytes counts were related to the mRNA expression level of EsAst post Aeromonas hydrophila challenge. The results suggested that the stem cells in the hematopoietic tissue of Chinese mitten crab E. Sinensis were regulated by transcriptional and humoral factors to generate hemocytes.

Liqiao Chen - One of the best experts on this subject based on the ideXlab platform.

  • The metabolomics responses of Chinese mitten-hand crab (Eriocheir Sinensis) to different dietary oils
    Aquaculture, 2017
    Co-Authors: Qing Chen, Zhen-hua Shen, Tao Han, Jianguang Qin, Liqiao Chen
    Abstract:

    Abstract Nutritional values of animal diets depend on the type of oil in the feed ingredient, but the underlying metabolic mechanisms of dietary oil in animal feed have not been thoroughly studied in aquatic animals, especially in crustaceans. In the present study, we conducted assays on GC–MS-based metabolomics and nutritional parameters to investigate the metabolic mechanisms between juvenile Chinese mitten crabs ( Eriocheir Sinensis ) fed olive oil containing 69% oleic acid (OA) and perilla oil containing 56% linolenic acid (LNA). Crab fed OA displayed faster growth, lower concentrations of hepatic glycogen, triglycerides and peroxidation products than those fed LNA. In the metabolomics assay, among 222 peaks isolated, 69 peaks were identified in serum. Among 13 significantly different metabolites between OA and LNA groups, six metabolites related to glycolysis and TCA (tricarboxylic acid) cycle (pyruvate, succinic acid, lactose, malic acid, D-glyceric acid and threitol), methionine, 2-keto-isovaleric acid (intermediate for valine and leucine synthesis) and 2-hydroxybutanoic acid (intermediate for glutathione synthesis) were higher in the OA group than in the LNA group. Only glutaconic acid (intermediate of ketogenic amino acids breakdown) was higher in the LNA group. This study indicates that crab in the OA group increased degradation of glucose and lipids to provide energy for growth as compared with crab in the LNA group. This is the first metabolomics study to identify the key pathways and crucial metabolites as biomarkers to differentiate the metabolic mechanisms of crustaceans fed contrasting dietary oils.

  • transcriptome sequencing revealed the genes and pathways involved in salinity stress of chinese mitten crab Eriocheir Sinensis
    Physiological Genomics, 2014
    Co-Authors: Shaolin Wang, Xiaodan Wang, Ke Chen, Liqiao Chen
    Abstract:

    A total of 276.9 million reads were obtained and assembled into 206, 371 contigs with an average length of 614 bp and N50 of 1,470 bp. Comparison of digital gene expression between treatment and control group reveals 1,151 and 941 genes were significantly differentially expressed in crab gill and muscle, respectively. In gill and muscle, protein ubiquitination, ubiquinone biosynthesis, oxidative phosphorylation, and mitochondria dysfunction pathways were the top pathways differentially expressed following the challenge. EIF 2 signaling pathway and IGF-1 signaling pathway were the top ones among the signal-related pathways. Most of the amino acid metabolism pathways were found to be involved in this process. The expression patterns of 15 differentially expressed genes were validated by quantitative real-time RT-PCR (average correlation coefficient 0.80). This is the first report of expression analysis of genes and pathways involved in osmoregulation of Eriocheir Sinensis through transcriptome sequencing. The findings of this study will further promote the understanding of the underlying molecular mechanism of salinity stress adaptation for crustacean species.

  • functional annotation and analysis of expressed sequence tags from the hepatopancreas of mitten crab Eriocheir Sinensis
    Marine Biotechnology, 2009
    Co-Authors: Hui Jiang, Xiaowei Zhang, Liqiao Chen, Yimei Cai, Qun Wang
    Abstract:

    Due to its popularity as a traditional food, intensive harvesting of the mitten crab (Eriocheir Sinensis) is common and has lead to an increase in disease incidence, resulting in catastrophic losses to crab aquaculture. The hepatopancreas of E. Sinensis is not only an important digestive organ but also an indispensable immune organ. We constructed a nonnormalized cDNA library from the hepatopancreas of E. Sinensis and acquired 3,297 high-quality expressed sequence tags representing 1,178 unigenes. More than half of these unigenes were novel genes for this species; the remaining had homologs in public databases, which is of great importance for future functional research. We also investigated the association of these genes with immune processes for insight into one of the main functions of the hepatopancreas besides metabolism. Despite the relatively low sampling scalar of our cDNA library, we were able to demonstrate several important properties of the hepatopancreatic transcriptome and identified numerous genes that were closely associated with immune responses. These results might serve as the basis for an in-depth genomics study of E. Sinensis, including transcriptome analysis, physical mapping, and whole genome sequencing.

  • discovery of immune related genes in chinese mitten crab Eriocheir Sinensis by expressed sequence tag analysis of haemocytes
    Aquaculture, 2009
    Co-Authors: Daxian Zhao, Xiaowei Zhang, Qun Wang, Shuhui Song, Liqiao Chen
    Abstract:

    Abstract To identify distinctive genes associated with immunity in the Chinese mitten crab ( Eriocheir Sinensis ), an expressed sequence tag (EST) library was constructed from haemocytes of this economically important species. 3118 clones were unidirectionally sequenced and analyzed by homology searches against sequences in the GenBank, KEGG and Uniprot. Significant homology (BLASTX, e-values − 5 ) to known genes was found in 488 of the 1039 unique sequences. The automatic functional classification based on KEGG and Gene Ontology revealed 26 putative immune-related genes (2.5% of all unique sequences, 18.9% of all ESTs). These 26 genes coded for enzymes and proteins in the clotting and prophenoloxidase-activating system, antioxidative enzymes, antimicrobial peptides, and pattern recognition molecules. The existence of these molecular processes in the activation of cellular defense in crab has not been previously reported. According to EST abundance, the major immune-related genes were the Kazal-type proteinase inhibitor (337, 11.1% of all ESTs) and the C-type lectin (72, 2.4%). The EST sequences of mitten crab haemocytes provide important information for understanding the evolution of the immune system among crustaceans in general. This study lays the groundwork for development of molecular markers related to disease resistance in the crab.

  • discovery of immune related genes in chinese mitten crab Eriocheir Sinensis by expressed sequence tag analysis of haemocytes
    Aquaculture, 2009
    Co-Authors: Daxian Zhao, Xiaowei Zhang, Qun Wang, Shuhui Song, Liqiao Chen
    Abstract:

    Abstract To identify distinctive genes associated with immunity in the Chinese mitten crab (Eriocheir Sinensis), an expressed sequence tag (EST) library was constructed from haemocytes of this economically important species. 3118 clones were unidirectionally sequenced and analyzed by homology searches against sequences in the GenBank, KEGG and Uniprot. Significant homology (BLASTX, e-values

Lingling Wang - One of the best experts on this subject based on the ideXlab platform.

  • identification of a clip domain serine proteinase involved in immune defense in chinese mitten crab Eriocheir Sinensis
    Fish & Shellfish Immunology, 2018
    Co-Authors: Zhihao Jia, Huan Zhang, Lingling Wang, Mengqiang Wang, Xiudan Wang, Linsheng Song
    Abstract:

    Clip-domain serine proteinase is an important serine proteinase family involved in many biological processes, which is only found in invertebrates. In the present study, the full-length cDNA of a clip domain serine proteinase (designed as EsCDSP) gene was cloned from Chinese mitten crab Eriocheir Sinensis using rapid amplification of cDNA ends (RACE) technique. It was of 1488 bp with an open reading frame (ORF) of 1134 bp encoding a polypeptide of 377 amino acids. There were a signal peptide, a clip domain, and a Tryp_SPc domain in the deduced amino acid sequence of EsCDSP. Highly conserved cysteine residues were identified in the clip domain and Tryp_SPc domain. EsCDSP shared similarities of 40%-61% with CDSPs from Penaeus monodon (ACP19562.1), Scylla paramamosain (CCW43200.1), Drosophila melanogaster (NP_649734.2) and Delia antiqua (AAW57295.1). It was clustered with other CDSPs from crabs in the phylogenetic tree. EsCDSP transcript was highly expressed in hemocytes and it could response to the stimulations of Vibro anguillarum and Pichia pastoris. rEsCDSP could activate proPO system and significantly increase the PO activity of HLS. In addition, rEsCDSP could bond to Aeromonas hydrophila, Vibro anguillarum and Vibro alginolyticus, and reduced the mortality rate causing by pathogen infection. All the results suggested that EsCDSP was an important immune response participator involved in activation of the proPO system of crab.

  • a novel gata like zinc finger transcription factor involving in hematopoiesis of Eriocheir Sinensis
    Fish & Shellfish Immunology, 2018
    Co-Authors: Zhihao Jia, Lingling Wang, Xiaorui Song, Yu Liu, Yunke Jia, Linsheng Song
    Abstract:

    Abstract GATA transcription factor is a family of DNA-binding proteins that can recognize and bind to sequence of (A/T) GATA (A/G). In the present study, a GATA-like protein (named as EsGLP) was characterized from Eriocheir Sinensis, including an 834 bp full length open reading frame of EsGLP, encoding a polypeptide of 277 amino acids. The deduced amino acid sequence of EsGLP contained one conserved GATA-type zinc finger of the form Cys-X2-Cys-X17-Cys-X2-Cys, with four cysteine sites. The EsGLP mRNA transcripts were mainly detected in the hematopoietic tissue, hepatopancreas and gonad. The recombinant EsGLP protein was prepared for the antibody production. The EsGLP protein was mainly distributed in the edge of lobules in the HPT and the cytoplasm of hemocytes. The mRNA transcripts of EsGLP in hemocytes were significantly decreased at 24 h (0.39-fold and 0.27-fold, p

  • functional characterization of hemocytes from chinese mitten crab Eriocheir Sinensis by flow cytometry
    Fish & Shellfish Immunology, 2017
    Co-Authors: Zhihao Jia, Lingling Wang, Shuai Jiang, Mingzhe Sun, Mengqiang Wang, Linsheng Song
    Abstract:

    Hemocytes comprise a diversity of cell types with functional and structural heterogeneity, and they play key roles in the host defense of invertebrates. In the present study, the hemocytes from Chinese mitten crab Eriocheir Sinensis were directly separated into two groups by flow cytometry. The hemocytes in P1 group were full of round and abundant granules with deeply staining cytoplasm, while P2 hemocytes were more diverse with a wide range of sizes and less granularity. Both P1 and P2 hemocytes exhibited phagocytic ability, but the phagocytic rate of P1 hemocytes increased which was significantly higher than that of P2 hemocytes after LPS stimulations. The levels of ROS production and intracellular Calcium as well as lysosome content were higher in P1 hemocytes than that in P2 hemocytes under both normal and immune-activated situations. The genes involved in phagocytosis, antimicrobial and antioxidant activities were mainly expressed in P1 hemocytes, while the genes involved in proPO activation system were highly expressed in P2 hemocytes. These results collectively suggested that P1 hemocytes were the main immunocompetent hemocytes in Chinese mitten crab and P2 hemocytes mainly participated in proPO activation system.

  • a galectin from Eriocheir Sinensis functions as pattern recognition receptor enhancing microbe agglutination and haemocytes encapsulation
    Fish & Shellfish Immunology, 2016
    Co-Authors: Mengqiang Wang, Huan Zhang, Lingling Wang, Yunchao Gai, Mengmeng Huang, Ying Guo, Hao Wang, Linsheng Song
    Abstract:

    Galectins are a family of β-galactoside binding lectins that function as pattern recognition receptors (PRRs) in innate immune system of both vertebrates and invertebrates. The cDNA of Chinese mitten crab Eriocheir Sinensis galectin (designated as EsGal) was cloned via rapid amplification of cDNA ends (RACE) technique based on expressed sequence tags (ESTs) analysis. The full-length cDNA of EsGal was 999 bp. Its open reading frame encoded a polypeptide of 218 amino acids containing a GLECT/Gal-bind_lectin domain and a proline/glycine rich low complexity region. The deduced amino acid sequence and domain organization of EsGal were highly similar to those of crustacean galectins. The mRNA transcripts of EsGal were found to be constitutively expressed in a wide range of tissues and mainly in hepatopancreas, gill and haemocytes. The mRNA expression level of EsGal increased rapidly and significantly after crabs were stimulated by different microbes. The recombinant EsGal (rEsGal) could bind various pathogen-associated molecular patterns (PAMPs), including lipopolysaccharide (LPS), peptidoglycan (PGN) and glucan (GLU), and exhibited strong activity to agglutinate Escherichia coli, Vibrio anguillarum, Bacillus subtilis, Micrococcus luteus, Staphylococcus aureus and Pichia pastoris, and such agglutinating activity could be inhibited by both d-galactose and α-lactose. The in vitro encapsulation assay revealed that rEsGal could enhance the encapsulation of haemocytes towards agarose beads. These results collectively suggested that EsGal played crucial roles in the immune recognition and elimination of pathogens and contributed to the innate immune response against various microbes in crabs.

  • the characterization of hematopoietic tissue in adult chinese mitten crab Eriocheir Sinensis
    Developmental and Comparative Immunology, 2016
    Co-Authors: Zhihao Jia, Lingling Wang, Sharath Kavungal, Shuai Jiang, Depeng Zhao, Mingzhe Sun, Linsheng Song
    Abstract:

    Invertebrates rely on the efficient innate immune mechanisms against invaders, in which the continuous production of hemocytes (hematopoiesis) is indispensable. In the present study, the hematopoietic tissue (HPT) from Chinese mitten crab Eriocheir Sinensis was identified and characterized. It was a thin and non-transparent sheet located at the dorsolateral side of the stomach, which was composed of a series of ovoid lobules. Each lobule was surrounded by connective tissue containing a large amount of spherical cells with big nucleus. In HPT, the cells were full of mitochondria and granules, and DNA replication was detected in some cells by EdU labeling technique. Cell proliferation was observed in HPT by transmission electron microscope (TEM). The distribution of two transcription factors, GATA1 and RUNX1, were examined by human GATA1 and RUNX1 antibodies, respectively. Three homologues of RUNX1 were detected in the HPT while no signal of RUNX1 was observed in hemocytes, and GATA1 was detected in both HPT and some hemocytes. The mRNA transcript of a novel hematopoiesis related cytokine EsAst was detected in hepatopancreas and hemocytes, but it was no detectable in HPT. The mRNA expression level of EsAst in hepatopancreas was 1.38-fold higher than that in hemocytes. Total hemocytes counts were related to the mRNA expression level of EsAst post Aeromonas hydrophila challenge. The results suggested that the stem cells in the hematopoietic tissue of Chinese mitten crab E. Sinensis were regulated by transcriptional and humoral factors to generate hemocytes.

Zhaoxia Cui - One of the best experts on this subject based on the ideXlab platform.

  • large scale lysine crotonylation analysis reveals its potential role in spermiogenesis in the chinese mitten crab Eriocheir Sinensis
    Journal of Proteomics, 2020
    Co-Authors: Chenchang Bao, Chengwen Song, Yuan Liu, Yanan Yang, Zhaoxia Cui
    Abstract:

    Lysine crotonylation (Kcr) is a recently-discovered type of post-translational modification. Although Kcr has been reported in many species, little is known about this process in crustaceans. In this study, pan anti-lysine crotonylation antibody enrichment and high-resolution liquid chromatogram-mass spectrometry analysis were employed to characterize Kcr in testis of the Chinese mitten crab Eriocheir Sinensis testis. Overall, 2799 Kcr sites were identified on 908 proteins with 14 conserved motifs. Bioinformatics analysis showed that Kcr was predominant on proteins found in cytoplasm, mitochondria and nucleus, and those involved in ribosome, proteasome, carbon metabolism and protein processing in endoplasmic reticulum. In total, 83 up-regulated and 12 down-regulated non-histone crotonylated sites were identified during spermiogenesis. These differentially expressed proteins were enriched in protein processing in endoplasmic reticulum pathway during formation of acrosome. In contrast, histone Kcr associated with mammalian spermatogenesis. These results provide foundational knowledge on the role of non-histone Kcr in spermiogenesis of E. Sinensis. SIGNIFICANCE: Lysine crotonylation (Kcr) is a recently-identified post-translational modification, and histone Kcr was found to associate with mammalian spermatogenesis. However, crotonylation of non-histone proteins has not been reported in spermatogenesis regulation. Further, there is no information on crotonylation in crustaceans. This study was the first large-scale Kcr proteome characterization in crustaceans. A total of 2799 Kcr sites on 908 proteins with 14 conserved motifs were identified from Eriocheir Sinensis testis. Of which, 83 up-regulated and 12 down-regulated non-histone crotonylated sites were identified during spermiogenesis. Our results provide the basic information for further functional validation of Kcr proteins and revealed new roles of Kcr in spermiogenesis of E. Sinensis.

  • discovery of sex related genes from embryonic development stage based on transcriptome analysis in Eriocheir Sinensis
    Gene, 2019
    Co-Authors: Yuan Liu, Chengwen Song, Zhaoxia Cui
    Abstract:

    Investigation of sex determination system in Eriocheir Sinensis is important because of sex-dimorphism in its growth traits. However, little information about the sex-related genes in embryonic development stages were exposed. To obtain more information of sex determination in Chinese mitten crab, we performed the transcriptome analysis in embryonic development stage (fertilized egg stage, cleavage stage, blastula stage, gastrula stage and heartbeat stage) of Chinese mitten crab using nextgeneration sequencing technology. Thirty-one of 32,088 annotated unigenes were identified as sex-related genes including 16 genes involved in primary sex determination in model organisms and 8 genes of SOX family and 7 genes of DMRT gene family. Heatmap based on the RPKM value in five embryonic development stages indicated that these genes were clustered into two branches. Analysis of the differentially expressed 12 genes, including 3 genes of SOX family, 3 genes of DMRT gene family and 6 genes involved in primary sex determination in model organisms, showed significantly difference between the first three periods (fertilized egg stage-cleavage stage-blastula stage) and the last two periods (gastrula stage-heartbeat stage) and all 12 genes were up-regulated after blastula stage. In conclusion, we inferred that sex determination might be initiated after blastula stage in E. Sinensis. Transcriptome analysis from embryonic development stage could provide a background information for further investigation in sex determination of Eriocheir Sinensis.

  • comparative transcriptome analysis explores maternal to zygotic transition during Eriocheir Sinensis early embryogenesis
    Gene, 2019
    Co-Authors: Junhao Ning, Chengwen Song, Yuan Liu, Danli Luo, Hourong Liu, Zhaoxia Cui
    Abstract:

    The maternal genome directs almost all aspects of early animal development. As development proceeds, the elimination of maternal gene products and zygotic genome activation (ZGA) occur during the maternal to zygotic transition (MZT). To study the molecular mechanisms regulating this developmental event in Eriocheir Sinensis, RNA-Seq technology was applied to generate comprehensive information on transcriptome dynamics during early embryonic stages. In total, 32,088 annotated unigenes were obtained from the transcriptomes of fertilized eggs and embryos at the cleavage (2-4 cell) and blastula stage. A total of 566 maternal genes and 1165 zygotic genes were isolated, among which 103 and 266 genes were predicted conserved maternal transcripts (COMATs) and conserved zygotic transcripts (COZYTs), respectively. The COMATs performed housekeeping gene functions and may be essential for initiating early embryogenesis of the Bilateria. Furthermore, 87, 76 and 117 differentially expressed genes associated with the MZT, morphogenesis and immunity were identified when compared the three transcriptomic datasets. We also unmask that the MZT takes place around the cleavage stage, when the genes involved in the clearance of maternal gene products and the ZGA were significantly up-regulated. Taken together, these datasets provide a valuable resource for understanding the mechanisms of early developmental events in E. Sinensis, and facilitate further studies on molecular mechanisms of asynchronous development in crabs.

  • primary molecular basis of androgenic gland endocrine sex regulation revealed by transcriptome analysis in Eriocheir Sinensis
    Journal of Oceanology and Limnology, 2019
    Co-Authors: Chengwen Song, Min Hui, Yuan Liu, Hourong Liu, Lei Liu, Zhaoxia Cui
    Abstract:

    In crustaceans, the male sexual differentiation and maintenance are specially regulated by androgenic gland (AG). However, little is known about the genes involved in the regulation process. RNA-Seq was performed on AG with ejaculatory duct (AG_ED) and ejaculatory duct (ED) as control in Eriocheir Sinensis, one of the most important economic and fishery crabs with typically sex dimorphism. A total of 925 unigenes were identified as differentially expressed genes (DEGs) and the expression of nine genes randomly selected was confirmed by qRT-PCR. 667 unigenes were up-regulated in AG_ED, being supposed to be AG preferential genes. Among them, the full length of insulin-like androgenic gland factor (IAG) cDNA named as Es-IAG was obtained as a logo gene of AG, which together with the genes insulin-like receptor (INR), and single insulin binding domain protein (SIBD), might constitute the sex regulation pathway. Several sex related genes were identified, and their function will have to be investigated. Also, the identification of juvenile hormone epoxide hydrolase 1 (JHEH1), ecdysteroid 22-hydroxylase (DIB) and ecdysone receptor (ECR) preliminarily clarified the molecular regulation mechanism of eyestalk-AG-testis axis, which plays important roles in molting and reproduction. The results will enhance our understanding for the molecular basis of the AG involved in male sex regulation in crabs.

  • molecular characterization and expression profiles of four transformer 2 isoforms in the chinese mitten crab Eriocheir Sinensis
    Chinese Journal of Oceanology and Limnology, 2017
    Co-Authors: Danli Luo, Min Hui, Chengwen Song, Yuan Liu, Hourong Liu, Zhaoxia Cui
    Abstract:

    The transformer-2 (tra-2) gene plays a key role in the regulatory hierarchy of sexual differentiation in somatic tissues and in the germline of Drosophila melanogaster. In this study, sequences and expression profiles of tra-2 in the Chinese mitten crab Eriocheir Sinensis were characterized. Four tra-2 isoforms, designated as Estra-2a, Estra-2b, Estra-2c and Estra-2d, were isolated. They all contained an RNA-recognition motif (RRM) and a linker region, which shared high similarity with other reported tra-2s. Sequence analysis revealed that Estra-2a, Estra-2b and Estra-2c are encoded by the same genomic locus and are generated by alternative splicing of the pre-mRNA. Compared with the other three isoforms, Estra-2d lacks the RS2 domain. Quantitative real-time PCR showed that all four isoforms were highly expressed in the fertilized egg, and in the 2–4 cell and blastula stages compared with larval stages (P≤ 0.01), suggesting their maternal origin in early embryonic developmental stages. Notably, Estra-2a was highly expressed in male somatic tissues, while Estra-2c was significantly highly expressed in the ovary. These results suggest that Estra-2c is involved in sexual differentiation of the Chinese mitten crab. Our findings provide basic information for further functional studies of the tra-2 gene/protein in this species.