Time filter

Source Type

Zhao J.,CAS Yantai Institute of Coastal Zone Research | Qiu L.,Chinese Academy of Fishery Sciences | Ning X.,Yantai Oceanic Environmental Monitoring Central Station of SOA | Chen A.,CAS Yantai Institute of Coastal Zone Research | And 2 more authors.
Comparative Biochemistry and Physiology - B Biochemistry and Molecular Biology | Year: 2010

Lysozymes are key proteins to invertebrates in the innate immune responses against bacterial infections and providing nutrition as digestion enzymes. In the present study, an invertebrate type lysozyme (denoted as VpLYZ) was identified from Venerupis philippinarum haemocytes by cDNA library and RACE approaches. The full-length cDNA of VpLYZ consisted of 805 nucleotides with a canonical polyadenylation signal sequence AATAAA and a polyA tail, and an open-reading frame of 558. bp encoding a polypeptide of 185 amino acids with a calculated molecular mass of 20.87. kD and theoretical pI of 8.44. The high similarity of VpLYZ with other i-type lysozymes from mollusk indicated that VpLYZ should be a new member of i-type lysozyme family. Similar to most i-type lysozymes, VpLYZ possessed all conserved features critical for the fundamental structure and function of i-type lysozymes, such as three catalytic residues (Glu19, Asn72 and Ser75) and i-type specific motif CL(E/L/R/H)C(I/M)C. By semi-quantitative RT-PCR analysis, mRNA transcript of VpLYZ was found to be most abundantly expressed in the tissues of gills, hepatopancreas and haemocytes, weakly expressed in the tissues of muscle, foot and mantle. After clams were challenged by Vibrio anguillarum, the mRNA level of VpLYZ in overall haemocyte population was recorded by quantitative real-time RT-PCR. VpLYZ mRNA was down-regulated sharply from 6. h to 12. h post-infection. Then, the expression level increased to the peak at 72. h and recovered to the original level at 96. h. All these results indicated that VpLYZ was involved in the immune response against microbe infection and contributed to the clearance of bacterial pathogens. © 2010.


Li C.,CAS Yantai Institute of Coastal Zone Research | Wang L.,CAS Qingdao Institute of Oceanology | Ning X.,Yantai Oceanic Environmental Monitoring Central Station of SOA | Chen A.,CAS Yantai Institute of Coastal Zone Research | And 4 more authors.
Cell Stress and Chaperones | Year: 2010

Small heat shock proteins (sHSPs) encompass a widespread and diverse class of proteins with molecular chaperone activity. In the present study, two sHSP isoforms (VpsHSP-1 and VpsHSP-2) were cloned from Venerupis philippinarum haemocytes by Rapid Amplification of cDNA Ends (RACE) approaches. The expression profiles of these two genes under Vibrio anguillarum challenge and cadmium exposure were investigated by quantitative real-time reverse transcriptase polymerase chain reaction. The bacterial challenge could significantly up-regulate the mRNA expression of both VpsHSP-1 and VpsHSP-2, with the increase of VpsHSP-2 expression occurred earlier than that of VpsHSP-1. During the cadmium exposure experiment, the expression level of both VpsHSP-1 and VpsHSP-2 decreased significantly with larger amplitude in VpsHSP-2. As time progressed, the expression levels of both genes were up-regulated with more increment in the low-chemical exposure groups. The differences in the response to pathogen stimulation and cadmium exposure indicated that there were functional diversity between the two structurally different molecules, VpsHSP-1 and VpsHSP-2, and they probably played distinct roles in mediating the environmental stress and immune responses in calm. © 2010 Cell Stress Society International.


Li C.,CAS Yantai Institute of Coastal Zone Research | Qiu L.,CAS Qingdao Institute of Oceanology | Ning X.,Yantai Oceanic Environmental Monitoring Central Station of SOA | Chen A.,CAS Yantai Institute of Coastal Zone Research | And 3 more authors.
Fish and Shellfish Immunology | Year: 2010

Translationally controlled tumor protein (TCTP) is one of the abundant and ubiquitously expressed proteins in metazoans. In the present study, the first molluscan TCTP (denoted as VpTCTP) was identified from Venerupis philippinarum haemocytes by EST and RACE approaches. The full-length cDNA of VpTCTP consisted of 1148 nucleotides with an open-reading frame of 555 bp encoding 184 amino acids. The deduced amino acid sequence of VpTCTP shared high similarity with TCTPs from other species, indicating that VpTCTP should be a new member of TCTP family. Several highly conserved motifs, including 5'terminal ologopyrimidine (5'TOP) starting sequence and rich AU and AUUT elements in 3'UTR, were also identified in VpTCTP. The tissue and temporal expression of VpTCTP after Vibrio anguillarum challenge was recorded by quantitative real-time RT-PCR. VpTCTP transcript could be detected in all examined tissues with the highest expression level in haemocytes and the lowest in hepatopancreas. Concerning the time-course expression in haemocytes, the relative expression of VpTCTP mRNA was down-regulated sharply from 6 h to 12 h post-infection. Then, the expression level was obviously up-regulated and reached 3.4-fold to that in the control group at 48 h post challenge. As time progressed, the expression of VpTCTP recovered to the original level at 96 h. All these results indicated that VpTCTP was an acute-phase protein involved in the immune response of V. philippinarum. © 2010 Elsevier Ltd.


Wei L.,CAS Yantai Institute of Coastal Zone Research | Wei L.,University of Chinese Academy of Sciences | Wang Q.,CAS Yantai Institute of Coastal Zone Research | Ning X.,Yantai Oceanic Environmental Monitoring Central Station of SOA | And 9 more authors.
Comparative Biochemistry and Physiology - Part D: Genomics and Proteomics | Year: 2015

Ocean acidification (OA) has been found to affect an array of normal physiological processes in mollusks, especially posing a significant threat to the fabrication process of mollusk shell. In the current study, the impact of exposure to elevated pCO2 condition was investigated in mantle tissue of Crassostrea gigas by an integrated metabolomic and proteomic approach. Analysis of metabolome and proteome revealed that elevated pCO2 could affect energy metabolism in oyster C. gigas, marked by differentially altered ATP, succinate, MDH, PEPCK and ALDH levels. Moreover, the up-regulated calponin-2, tropomyosins and myosin light chains indicated that elevated pCO2 probably caused disturbances in cytoskeleton structure in mantle tissue of oyster C. gigas. This work demonstrated that a combination of proteomics and metabolomics could provide important insights into the effects of OA at molecular levels. © 2014 Elsevier Inc.


Li C.,Chinese Academy of Sciences | Li C.,CAS Yantai Institute of Coastal Zone Research | Li C.,Ningbo University | Li L.,Harbin University of Science and Technology | And 10 more authors.
Fish and Shellfish Immunology | Year: 2011

HSP40 was an understudied protein family with co-chaperone activity. In the present study, a HSP40 homology was cloned from Venerupis philippinarum haemocytes (designated as VpHSP40) by EST analysis and RACE approaches. The expression profiles of VpHSP40 under Vibrio anguillarum challenge and heavy metal exposure were investigated by quantitative real-time RT-PCR. The bacterial challenge could significantly up-regulate the mRNA expression, and the highest expression level was detected at 24 h post-infection with 6.0-fold increase compared with that in the control group. During heavy metal exposure experiment, the expression of VpHSP40 could also be induced by Cu2+ and Cd2+ at different concentration, where Cu2+ displayed more toxic effect on clam than that of Cd2+. Concerning the same heavy metal, varied effect on VpHSP40 expression was detected at different concentration of heavy metal. Taking together, these results suggested that VpHSP40 was perhaps involved in mediating the immune responses and environmental stresses in V. philippinarum. © 2010 Elsevier Ltd.


Wang Q.,CAS Yantai Institute of Coastal Zone Research | Cao R.,CAS Yantai Institute of Coastal Zone Research | Cao R.,University of Chinese Academy of Sciences | Ning X.,Yantai Oceanic Environmental Monitoring Central Station of SOA | And 7 more authors.
Fish and Shellfish Immunology | Year: 2016

Ocean acidification (OA), caused by anthropogenic CO2emissions, has been proposed as one of the greatest threats in marine ecosystems. A growing body of evidence shows that ocean acidification can impact development, survival, growth and physiology of marine calcifiers. In this study, the immune responses of the Pacific oyster Crassostrea gigas were investigated after elevated pCO2 exposure for 28 days. The results demonstrated that OA caused an increase of apoptosis and reactive oxygen species (ROS) production in hemocytes. Moreover, elevated pCO2 had an inhibitory effect on some antioxidant enzyme activities and decreased the GSH level in digestive gland. However, the mRNA expression pattern of several immune related genes varied depending on the exposure time and tissues. After exposure to pCO2 at ~2000 ppm for 28 days, the mRNA expressions of almost all tested genes were significantly suppressed in gills and stimulated in hemocytes. Above all, our study demonstrated that elevated pCO2 have a significant impact on the immune systems of the Pacific oyster, which may constitute as a potential threat to increased susceptibility of bivalves to diseases. © 2015 Elsevier Ltd.


Wang Q.,Chinese Academy of Sciences | Ning X.,Yantai Oceanic Environmental Monitoring Central Station of SOA | Chen L.,Ningbo University | Pei D.,Yantai University | And 6 more authors.
Comparative Biochemistry and Physiology - C Toxicology and Pharmacology | Year: 2011

Thioredoxin (abbreviated as Trx) is an important ubiquitous disulfide reductase, which can protect organisms against various oxidative stresses. In the present study, thioredoxin 1 (named as VpTrx1) and thioredoxin-related protein (named as VpTrp14) were identified from Venerupis philippinarum, respectively. Similar to most Trx1s, VpTrx1 possessed all conserved features critical for the fundamental structure and function of Trx1s, such as the conserved catalytic residues (C-G-P-C), but lacked the other cysteine residues, while VpTrp14 contained the conserved motif (C-P-D-C). Quantitative Real-time PCR assay showed that VpTrx1 and VpTrp14 transcripts were distributed in a wide array of tissues most abundantly expressed in the hepatopancreas. The expression of VpTrp14 mRNA in the hepatopancreas was significantly up-regulated after exposure to 10 and 40 μg/L Cd, while the VpTrx1 expression level was kept relatively constant. Both the expression levels of VpTrx1 and VpTrp14 in the hepatopancreas were induced after exposure to Cu, and increased to the peak value at 96 h under the 40 μg/L Cu exposure. These results showed that VpTrp14 transcripts responded to metal stress more acutely than VpTrx1, and both Trxs responded to Cu stress more sensitively than Cd. Together, it was suggested that VpTrx1 and VpTrp14 perhaps played important roles in the antioxidant responses against metal stress in V. philippinarum. © 2011 Elsevier Inc.


PubMed | CAS Yantai Institute of Coastal Zone Research, University of Chinese Academy of Sciences, Ningbo University and Yantai Oceanic Environmental Monitoring Central Station of SOA
Type: | Journal: Fish & shellfish immunology | Year: 2016

Ocean acidification (OA), caused by anthropogenic CO2emissions, has been proposed as one of the greatest threats in marine ecosystems. A growing body of evidence shows that ocean acidification can impact development, survival, growth and physiology of marine calcifiers. In this study, the immune responses of the Pacific oyster Crassostrea gigas were investigated after elevated pCO2 exposure for 28 days. The results demonstrated that OA caused an increase of apoptosis and reactive oxygen species (ROS) production in hemocytes. Moreover, elevated pCO2 had an inhibitory effect on some antioxidant enzyme activities and decreased the GSH level in digestive gland. However, the mRNA expression pattern of several immune related genes varied depending on the exposure time and tissues. After exposure to pCO2 at 2000ppm for 28 days, the mRNA expressions of almost all tested genes were significantly suppressed in gills and stimulated in hemocytes. Above all, our study demonstrated that elevated pCO2 have a significant impact on the immune systems of the Pacific oyster, which may constitute as a potential threat to increased susceptibility of bivalves to diseases.


Li C.,CAS Yantai Institute of Coastal Zone Research | Sun H.,CAS South China Sea Institute of Oceanology | Chen A.,CAS Yantai Institute of Coastal Zone Research | Ning X.,Yantai Oceanic Environmental Monitoring Central Station of SOA | And 4 more authors.
Fish and Shellfish Immunology | Year: 2010

Superoxide dismutase (SOD, EC 1.15.1.1) represents one kind of enzyme involved in scavenging the high level of reactive oxygen species (ROS) into molecular oxygen and hydrogen peroxide. In the present study, the intracellular Cu/Zn-SOD gene (icCu/Zn-SOD) of Venerupis philippinarum (denoted as VpSOD) was identified from haemocytes by homology cloning and RACR approaches. The full-length cDNA of VpSOD consisted of 910 nucleotides with a canonical polyadenylation signal sequence AATAAA, a polyA tail, and an open-reading frame of 465 bp encoding 154 amino acids. The deduced amino acid of VpSOD shared high similarity with the icCu/Zn-SODs from other species, indicating that VpSOD should be a new member of icCu/Zn-SOD family. Several highly conserved motifs including Cu, Zn binding sites (H46, H48, H63, H120 for Cu binding, and H63, H71, H80, D83 for Zn binding), intracellular disulfide bond and two Cu, Zn SOD signatures were also identified in VpSOD. The temporal expression of VpSOD in haemocytes after Vibrio anguillarum challenge was recorded by quantitative real-time RT-PCR. The relative expression level of VpSOD mRNA was up-regulated rapidly at 6 h post-infection and reached 18-fold of the control group. After a drastic decrease at 12 h, the expression level increased again and reached 22-fold to that in the control group at 96 h post-infection. All these results indicated that VpSOD was an acute-phase protein involved in the immune responses of V. philippinarum. © 2009 Elsevier Ltd. All rights reserved.


Wang Q.,Chinese Academy of Sciences | Ning X.,Yantai Oceanic Environmental Monitoring Central Station of SOA | Pei D.,Yantai University | Zhao J.,Chinese Academy of Sciences | And 3 more authors.
Chinese Journal of Oceanology and Limnology | Year: 2013

Thioredoxin (Trx) proteins are involved in many biological processes especially the regulation of cellular redox homeostasis. In this study, two Trx cDNAs were cloned from the mussel Mytilus galloprovincialis using rapid amplifi cation of cDNA ends-polymerase chain reaction (RACE-PCR). The two cDNAs were named MgTrx1 and MgTrx2, respectively. The open reading frames of MgTrx1 and MgTrx2 were 318 and 507 base pairs (bp) and they encoded proteins of 105 and 168 amino acids with estimated molecular masses of 11. 45 and 18. 93 kDa, respectively. Sequence analysis revealed that both proteins possessed the conserved active site dithiol motif Cys-Gly-Pro-Cys. In addition, MgTrx2 also possessed a putative mitochondrial targeting signal suggesting that it is located in the mitochondria. Quantitative real-time polymerase chain reaction (qPCR) revealed that both MgTrx1 and MgTrx2 were constitutively expressed in all tissues examined. The MgTrx1 transcript was most abundant in hemocytes and gills, whereas the MgTrx2 transcript was most abundant in gonad, hepatopancreas, gill and hemocytes. Following Vibrio anguillarum challenge, the expression of MgTrx1 was up-regulated and reached its peak, at a value 10-fold the initial value, at 24 h. Subsequently, expression returned back to the original level. In contrast, the expression level of MgTrx2 was down-regulated following bacterial stimulation, with one fi fth of the control level evident at 12 h post challenge. These results suggest that MgTrx1 and MgTrx2 may play important roles in the response of M. galloprovincialis to bacterial challenge. © 2013 Chinese Society for Oceanology and Limnology, Science Press and Springer-Verlag Berlin Heidelberg.

Loading Yantai Oceanic Environmental Monitoring Central Station of SOA collaborators
Loading Yantai Oceanic Environmental Monitoring Central Station of SOA collaborators