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Characterization,expression dynamics,and potential function of OPA1 for regulation of mitochondrial morphology during spermiogenesis in Phascolosoma esculenta

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Journal of Oceanology and Limnology 2024年第1期42卷 187-200页

作者： Xinming GAO Binbin FENG Chen DU Congcong HOU Shan JIN Daojun TANG Junquan ZHU Key laboratory of Applied marine biotechnology by the Ministry of Education and Key laboratory of marine biotechnology of Zhejiang Province School of Marine SciencesNingbo UniversityNingbo 315211China College of Ecology Lishui UniversityLishui 323000China

Mitochondria undergo morphological changes during spermatogenesis in some animals.The mechanism and role of mitochondrial morphology regulation,however,remain somewhat unclear.In this study,we analyzed the molecular characteristics,expression dynamics and subcellular localization of optic atrophy protein 1(OPA1),a mitochondrial fusion and cristae maintenance-related protein,to reveal the possible regulatory mechanisms underlying mitochondrial morphology in Phascolosoma esculenta spermiogenesis.The full-length cDNA of the P.esculenta opa1 gene(Pe-opa1)is 3743 bp in length and encodes 975 amino acids.The Pe-OPA1 protein is highly conservative and includes a transmembrane domain,a GTPase domain,two helical bundle domains,and a lipid-interacting stalk.Gene and protein expression was higher in the coelomic fluid(a site of spermatid development)of male P.esculenta and increased first and then decreased from March to December.Moreover,their expression during the breeding stage was significantly higher than during the non-breeding stage,suggesting that Pe-OPA1 is involved in P.esculenta reproduction.The Pe-OPA1 protein was more abundant in components consisting of many spermatids than in components without,indicating that Pe-OPA1 mainly plays a role in the spermatid in coelomic fluid.Moreover,Pe-OPA1 was mainly detected in the spermatid mitochondria.Immunofluorescence experiments showed that the Pe-OPA1 are constitutively expressed and co-localized with mitochondria during spermiogenesis,suggesting its involvement in P.esculenta spermiogenesis.These results provide evidence for Pe-OPA1's involvement in the regulation of mitochondrial morphology during spermiogenesis.

关键词： optic atrophy protein 1(OPA1) spermiogenesis Phascolosoma esculenta mitochondrial dynamics spermatid

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Photoperiod Mediates the Effects of Temperature and Light Intensity on the Proliferation of Ulva prolifera

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Journal of Ocean University of China 2024年第1期23卷 255-263页

作者： JIANG Jianan YU Yanyan CHEN Yili LI Yahe XU Nianjun Key laboratory of marine biotechnology of Zhejiang Province School of Marine SciencesNingbo UniversityNingbo 315211China

In order to study the complex effects of photoperiod,temperature,and light intensity on the spore maturation and release number of Ulva prolifera,we cultured thalli segment(2–3 mm)under three different photoperiods(L:D=12:12,14:10 and 10:14),temperature(15℃(LT),25℃(MT)and 30℃(HT))and light intensity(100,200 and 400μmol m^(−2)s^(−1),noted as LL,ML and HL,respectively)conditions.Then the maturation time,spore release number and chlorophyll fluorescence were analyzed.The results suggested that:1)The spore maturation time was accelerated by higher temperature or higher light intensity from 62 h to 36 h,and changes in day length accelerated the spore maturation to a certain extent as compared with 12:12 light/dark cycle;2)Higher light intensity significantly decreased the chlorophyll fluorescence(Fv′/Fm′,NPQ,rETRmax andα)of the mature reproductive segment under 30℃with 12:12 light/dark cycle.But when in the other photoperiods(10:14 and 14:10 conditions),the inhibitory effects of high light intensity were alleviated significantly;3)The optimum condition for the spore maturation and release was 12:12 light/dark cycle,25℃,400μmol m^(−2)s^(−1),with both shorter and longer photoperiod reducing the spore release number;4)Higher light intensity significantly increased the spore release number under 25℃,but these effects were alleviated by 30℃treatment.This study is the first attempt to elucidate the coincidence effects of photoperiod,temperature and light intensity on the reproduction of Ulva,which would help to reveal the mechanism of the rapid proliferation of green tide.

关键词： light intensity maturation photoperiod reproduction temperature Ulva prolifera

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Full T-cell activation and function in teleosts require collaboration of first and co-stimulatory signals

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Zoological Research 2024年第1期45卷 13-24页

作者： Wei Liang Kang Li Haiyou Gao Kunming Li Jiansong Zhang Qian Zhang Xinying Jiao Jialong Yang Xiumei Wei State Key laboratory of Estuarine and Coastal Research School of Life SciencesEast China Normal UniversityShanghai 200241China laboratory for marine Biology and biotechnology Qingdao National Laboratory for Marine Science and TechnologyQingdaoShandong 266237China

Mammalian T-cell responses require synergism between the first signal and co-stimulatory signal.However,whether and how dual signaling regulates the T-cell response in early vertebrates remains unknown.In the present study,we discovered that the Nile tilapia(Oreochromis niloticus)encodes key components of the LAT signalosome,namely,LAT,ITK,GRB2,VAV1,SLP-76,GADS,and PLC-γ1.These components are evolutionarily conserved,and CD3εmAb-induced T-cell activation markedly increased their expression.Additionally,at least ITK,GRB2,and VAV1 were found to interact with LAT for signalosome formation.Downstream of the first signal,the NF-κB,MAPK/ERK,and PI3K-AKT pathways were activated upon CD3εmAb stimulation.Furthermore,treatment of lymphocytes with CD28 mAbs triggered the AKT-mTORC1 pathway downstream of the co-stimulatory signal.Combined CD3εand CD28 mAb stimulation enhanced ERK1/2 and S6 phosphorylation and elevated NFAT1,c-Fos,IL-2,CD122,and CD44 expression,thereby signifying T-cell activation.Moreover,rather than relying on the first or co-stimulatory signal alone,both signals were required for T-cell proliferation.Full T-cell activation was accompanied by marked apoptosis and cytotoxic responses.These findings suggest that tilapia relies on dual signaling to maintain an optimal T-cell response,providing a novel perspective for understanding the evolution of the adaptive immune system.

关键词： Oreochromis niloticus CD3 CD28 T cells Adaptive immunity Evolution

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Combined proteomic and metabolomic studies on the liver of Amur sturgeon Acipenser schrenckii under titanium dioxide nanoparticle exposure

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Journal of Oceanology and Limnology 2024年第3期42卷 1001-1015页

作者： Zhou ZHOU Ting LIU Jie KONG Zhenxing ZHAO Junquan ZHU Key laboratory of Aquacultural biotechnology Ministry of Educationand Key Laboratory of Marine Biotechnology of Zhejiang ProvinceCollege of Marine SciencesNingbo UniversityNingbo 315000China Guizhou Fisheries Research Institute Guizhou Academy of Agriculture SciencesGuiyang 550000China

Nanomaterials,particularly titanium dioxide nanoparticles(TiO_(2)-NPs),are extensively utilized across various industries.However,their environmental release has raised concerns regarding their potential ecological and environmental impacts.The reproductive toxicity of TiO_(2)-NPs in fish species has attracted considerable attention,yet conflicting research outcomes have been reported.We investigated the effects of TiO_(2)-NPs exposure on the liver of juvenile Amur sturgeon Acipenser schrenckii using label-free proteomic and untargeted metabolomic analyses.The experiment included a control group and three groups exposed to different concentrations of TiO_(2)-NPs(low,TL;medium,TM;high,TH).Compared to the control group,9,19,and 25 proteins and 35,73,and 158 metabolites were differentially expressed in the TH,TM,and TL TiO_(2)-NP-exposed groups,respectively.The differentially expressed genes(DEGs)were enriched in the Kyoto Encyclopedia for Genes and Genomes(KEGG)pathways related to glycolysis and gluconeogenesis.Moreover,among the 126 correlated proteins,the most enriched pathways were associated with endocytosis and protein processing in the endoplasmic reticulum.Notably,syringic acid was significantly downregulated across all three TiO_(2)-NP-exposed groups.To obtain a comprehensive overview of the TiO_(2)-NP-induced expression changes,a co-regulated network of proteins and metabolites associated with TiO_(2)-NPs exposure was constructed.Exposure to TiO_(2)-NPs led to enrichment and alteration of pathways related to immune responses,including endocytosis,protein processing in the endoplasmic reticulum,and peroxisome proliferator-activated receptor(PPAR)signaling.In conclusion,our findings indicate that exposure to TiO_(2)-NPs might disrupt glucose metabolism and induce immune responses,thus contributing to our understanding of the environmental impacts of nanomaterials and highlighting the need for further research and development of potential mitigation strategies.

关键词： TiO_(2)-nanoparticle exposure proteomics metabolomics glucose metabolism immune response

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Tbx 4 and Tbx 5 gene expression associated with appendage development and its relationship with the absence of the pelvic fin in Pampus argenteus

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Journal of Oceanology and Limnology 2024年第2期42卷 580-593页

作者： Shun ZHANG Xiaojing ZHU Lingzhu HU Kai LIAO Shanliang XU Danli WANG Chunyang GUO School of marine Science Ningbo UniversityNingbo 315211China Key laboratory of Applied marine biotechnology Ningbo UniversityChinese Ministry of EducationNingbo 315211China

The Tbx family is first known through the study of their functions in the body and limbs,and its members Tbx4 and Tbx5 genes are important factors in determining the characteristics of the appendages.Pampus argenteus is one of the important economical marine fishes widely distributed in offshore areas.Therefore,it is necessary to study the role of Tbx family genes in the deletion of pelvic fin in P.argenteus.In this study,we cloned Tbx4 and Tbx5 cDNA sequence of P.argenteus(GenBank:MH709128 and MH712458).The Western blot and real time PCR were used to detect the expressions of Tbx4 and Tbx5 in different developmental stages and tissues of P.argenteus.In addition,whole-mount in-situ hybridization was used to study the localization of Tbx4 and Tbx5 genes in different developmental stages of P.argenteus.Results show that the translation of Tbx4 mRNA was inhibited during the critical period of pelvic fin development.Among different tissues,Tbx4 protein levels were the lowest in the abdominal epithelium,and even lower than that in the pectoral fin,suggesting that the protein expression was also inhibited in the abdominal epithelium of adult P.argenteus.Therefore,the results indicated that upstream genes regulation led to the key stage-specific and low expression of Tbx4 during pelvic fin development and in the abdominal epithelium.

关键词： Stromateidae pelvic fin protein translation T-box aquaculture broodstock

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Findings on three endocommensal scuticociliates(Protista,Ciliophora)from freshwater mollusks,including their morphology and molecular phylogeny with descriptions of two new species

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marine Life Science & Technology 2024年第2期6卷 212-235页

作者： Tao Li Tengyue Zhang Mingjian Liu Zhe Zhang Jiachen Zhang Junhua Niu Xiangrui Chen Saleh A.Al‑Farraj Weibo Song Institute of Evolution and marine Biodiversity Ocean University of China and Key Laboratory of Evolution and Marine Biodiversity(Ministry of Education)Qingdao 266003China The Key laboratory of Zoological Systematics and Application College of Life SciencesHebei UniversityBaoding 071002China College of marine Life Sciences Ocean University of ChinaQingdao 266003China School of marine Sciences Ningbo UniversityNingbo 315800China Zoology Department College of ScienceKing Saud University11451 RiyadhSaudi Arabia laboratory for marine Biology and biotechnology Laoshan LaboratoryQingdao 266237China

Species of the ciliate genera Myxophyllum and Conchophthirus are found as endocommensals of terrestrial and freshwater mollusks,respectively.So far,there have been few studies of these genera and morphological data for most members are often incomplete.In the present work,two new species,Myxophyllum weishanense sp.***.and Conchophthirus paracurtus sp.nov.,and a known species,Conchophthirus lamellidens,were isolated from hosts in Lake Weishan Wetland,China.Taxonomic studies indicate that M.weishanense sp.***.can be recognized mainly by the combination of about 60 somatic kineties on both ventral and dorsal sides and the presence of caudal cilia.Conchophthirus paracurtus sp.***.differs from congeners in its body shape and size,having a glabrous area on the posterior right side,and having fewer somatic kineties.In addition,differences in their ITS2(Internally Transcribed Spacer 2)secondary structures support the discrimination of the two new species from their highly similar congeners.An improved diagnosis for the poorly known species,C.lamel-lidens is also provided.Phylogenetic analyses reveal that members of the genus Myxophyllum belong to a fully supported clade that is sister to a large,poorly supported clade consisting of Hemispeiridae,Ancistridae,and several lineages of the nonmonophyletic Cyclidiidae.The Myxophyllum clade also includes Protophyra ovicola JQ956552,a possible misidenti-fication.Sequences of the two new Conchophthirus species cluster with other congeners in a fully supported clade that is unrelated to either the‘typical’thigmotrichs or to pleuronematids,thus conflicting with the traditional classification,and may represent an orphan scuticociliate lineage.

关键词： Ciliates Endocommensal ITS2 secondary structure Mollusks New taxa Wetland

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Protein Disulfide Isomerase and Its Potential Function on Endoplasmic Reticulum Quality Control in Diatom Phaeodactylum tricornutum

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Phyton-International Journal of Experimental Botany 2024年第1期93卷 137-150页

作者： Yanhuan Lin Hua Du Zhitao Ye Shuqi Wang Zhen Wang Xiaojuan Liu Zhangzhou Institute of Technology ZhangzhouChina Guangdong Provincial Key laboratory of marine biotechnology Guangdong Provincial Key Laboratory of Marine Disaster Prediction and PreventionCollege of SciencesShantou UniversityShantouChina

PDI is a molecular chaperone and plays an important role in Endoplasmic Reticulum quality control (ERQC).PDI participates in the refolding of the misfolded/unfolded proteins to maintain cellular homeostasis under differentstresses. However, bioinformatic characteristics and potential functions of PDIs in diatom Phaeodactylumtricornutum (Pt) are still unknown so far. Hence, the genome-wide characteristics of PtPDI proteins in P. tricornutumwere first studied via bioinformatic and transcriptomic methods. 42 PtPDI genes were identified from thegenome of P. tricornutum. The motif, protein structure, classification, number of introns, phylogenetic relationship,and the expression level of 42 PtPDI genes under the tunicamycin stress were analyzed. A pair of tandemduplicated genes (PtPDI15 and PtPDI18) was observed in P. tricornutum. The 42 PtPDIs with different genecharacteristics were divided into three independent clades, indicating different evolutional relationships and functionsof these PtPDIs. The 14 up-regulated PtPDI genes under the tunicamycin treatment might have a positiveeffect on the ER quality control of the unfolded/misfolded proteins, while the 7 down-regulated PtPDIs mightnegatively affect the ERQC. The characteristics of all 42 PtPDIs and their proposed working model here providea comprehensive understanding of the PtPDIs gene family. The differential expression of 21 PtPDIs will be usefulfor further functional study in the ERQC.

关键词： Protein disulfide isomerase gene family Endoplasmic Reticulum quality control Phaeodactylum tricornutum

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Analysis and Characterization of the GABA Transaminase and Succinate Semialdehyde Dehydrogenase Genes in the Microalga Isochrysis zhanjiangensis in Response to Abiotic Stresses

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Journal of Ocean University of China 2024年第3期23卷 775-785页

作者： TIAN Jiaojiao ZHANG Lin LU Xinyue YE Lingzhi WU Yuanyuan CHEN Deshui CAO Jiayi JIANG Jiaxin XU Jilin YAN Xiaojun Key laboratory of marine biotechnology of Zhejiang Province School of Marine SciencesNingbo UniversityNingbo 315211China Key laboratory of Aquacultural biotechnology Ministry of Education Ningbo UniversityNingbo 315211China Laizhou Bay marine Technology Co. Ltd.Yantai 261400China Fujian Dalai Seed Science and Technology Co. Ltd.Ningde 352101China

Gamma-aminobutyric acid(GABA),widely existing in different organisms,is rapidly accumulated in plants in response to environmental stresses.The main biosynthesis and degradation pathways of GABA constitute the GABA shunt,which is tied to the tricarboxylic acid(TCA)cycle.GABA transaminase(GABA-T)and succinate semialdehyde dehydrogenase(SSADH)are two essential enzymes for the GABA degradation pathway.While there are abundant studies on GABA shunt in higher plants at the physiological and genetic levels,research on its role in microalgae remains limited.This study aimed at exploring the function of GABA-T and SSADH genes in Isochrysis zhanjiangensis,an important diet microalga,under different stresses.We cloned two GABA-T genes,IzGABA-T1 and IzGABA-T2,and one SSADH gene IzSSADH from Isochrysis zhanjiangensis and conducted heterologous expression experiments.The results showed that the overexpression of IzGABA-T1 or IzGABA-T2 enhanced the survival rates of yeast transformants under heat or NaCl stress,while the overexpression of IzSSADH improved yeast tolerance to NaCl stress but had no obvious effect on heat stress.Additionally,the results of quantitative real-time polymerase chain reaction(qPCR)showed that IzGABA-T1 transcription increased in the HT(salinity 25,35℃)and LS(salinity 15,25℃)groups.At 24 h,the IzGABA-T2 transcriptions increased in the HT,LS,and HS(salinity 35,25℃)groups,but their transcription levels decreased in all groups at 48 h.IzSSADH transcription increased in the LS group.These results suggest that IzGABA-T1,IzGABA-T2,and IzSSADH are associated with temperature and salinity stresses and possess a certain preference for different stresses.

关键词： abiotic stress GABA heterologous expression Isochrysis zhanjiangensis transcription

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Flow Field Characteristics of Multi-Trophic Artificial Reef Based on Computation Fluid Dynamics

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Journal of Ocean University of China 2024年第2期23卷 317-327页

作者： HUANG Junlin LI Jiao LI Yan GONG Pihai GUAN Changtao XIA Xu School of Engineering Ocean University of ChinaQingdao 266100China Yellow Sea Fisheries Research Institute Chinese Academy of Fishery SciencesKey Laboratory of Sustainable Development of Marine FisheriesMinistry of Agriculture and Rural AffairsQingdao Key Laboratory for Marine Fish Breeding and BiotechnologyQingdao 266071China laboratory for marine Fisheries Science and Food Production Processes Pilot National Laboratory for Marine Science and Technology(Qingdao)Qingdao 266071China

On the basis of computational fluid dynamics,the flow field characteristics of multi-trophic artificial reefs,including the flow field distribution features of a single reef under three different velocities and the effect of spacing between reefs on flow scale and the flow state,were analyzed.Results indicate upwelling,slow flow,and eddy around a single reef.Maximum velocity,height,and volume of upwelling in front of a single reef were positively correlated with inflow velocity.The length and volume of slow flow increased with the increase in inflow velocity.Eddies were present both inside and backward,and vorticity was positively correlated with inflow velocity.Space between reefs had a minor influence on the maximum velocity and height of upwelling.With the increase in space from 0.5 L to 1.5 L(L is the reef lehgth),the length of slow flow in the front and back of the combined reefs increased slightly.When the space was 2.0 L,the length of the slow flow decreased.In four different spaces,eddies were present inside and at the back of each reef.The maximum vorticity was negatively correlated with space from 0.5 L to 1.5 L,but under 2.0 L space,the maximum vorticity was close to the vorticity of a single reef under the same inflow velocity.

关键词： artificial reef flow field characteristics computation fluid dynamics multi-trophic structure

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Aptasensing biosynthesized phosphatidylserine with a AuNPs nanozyme-based colorimetric aptasensor

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Food Science and Human Wellness 2024年第2期13卷 823-829页

作者： Sai Wang Rui Ma Chengqiang Li Ling Zhang Haiyang Zhang Xuehan Li Xiangzhao Mao Qingdao Key laboratory of Food biotechnology College of Food Science and EngineeringOcean University of ChinaQingdao 266404China Key laboratory of Biological Processing of Aquatic Products China National Light IndustryQingdao 266404China laboratory for marine Drugs and Bioproducts of Qingdao National laboratory for marine Science and Technology Qingdao 266237China

Sensitive monitoring of the target products during the biosynthesis process is crucial,and facile analytical approaches are urgently needed.Herein,phosphatidylserine(PS)was chosen as the model target,a colorimetric aptasensor was developed for the rapid quantitation in biosynthesis samples.A chimeric aptamer was constructed with two homogeneous original PS aptamers.Specific recognition between the chimeric aptamer and PS results in the desorption of aptamer from the surface of the AuNPs nanozyme,and the peroxidase-like enzymatic activity of the AuNPs nanozyme was weakened in a relationship with the different concentrations.The developed aptasensor performed well when applied for analyzing PS in biosynthesis samples.The aptasensor offers good sensitivity and selectivity,under optimal conditions,achieving monitoring and quantitation of PS in the range of 2.5-80.0μmol/L,with a limit of detection at 536.2 nmol/L.Moreover,the aptasensor provides good accuracy,with comparison rates of 98.17%-106.40%,when compared with the HPLC-ELSD.This study provides a good reference for monitoring other biosynthesized products and promoting the development of aptamers and aptasensors in real-world applications.

关键词： Phosphatidylserine Chimeric aptamer AuNPs nanozyme Colorimetric aptasensor

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