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

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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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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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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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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Effect of dietary supplement of inactivated Lactobacillus plantarum Ep-M 17 on growth performance,immune response,disease resistance,and intestinal microbiota in Penaeus vannamei

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Journal of Oceanology and Limnology 2024年第2期42卷 676-694页

作者： Ting WU Xiaoman HU Wenlong XU Yang DU Jiong CHEN State Key laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products Ningbo UniversityNingbo 315211China laboratory of Biochemistry and Molecular biology School of Marine SciencesNingbo UniversityNingbo 315211China Key laboratory of Applied marine biotechnology of Ministry of Education Ningbo UniversityNingbo 315211China

Our previous study found that feeding with Lactobacillus plantarum Ep-M17 could effectively affect the growth performance,immune response,and gut microbiota of Penaeus vannamei.However,high temperature and pressure during feed pelletizing is the main problem that can lead to a decrease in the activity of probiotics or cause their inactivation.Further investigation needs to investigate whether inactivated Ep-M17 can exert similar effects as live Ep-M17.Therefore,we evaluated the effects of inactivated L.plantarum Ep-M17 on growth performance,immune response,disease resistance,and gut microbiota in P.vannamei.Results show that adding inactivated Ep-M17 to the feed also promoted body weight gain and increased relative immune protection in shrimp.Also,histological examination revealed that the administration of inactivated Ep-M17 led to improvements in the density and distribution of microvilli in the intestines and enhancements in the abundance of B and R cells in the hepatopancreas.Additionally,the inactivated Ep-M17 supplementation resulted in increased activity levels of nutrient immune-related enzymes in both the shrimp hepatopancreas and intestines.Moreover,it stimulated the expression of Lvlec,PEN-3a,Crustin,LGBP,Lysozyme,and proPo genes in both the hepatopancreas and intestines.Furthermore,the inactivated Ep-M17 also increased bacterial diversity in the gut of shrimp and promoted the abundance of specific flora,facilitating the host organism’s metabolism and immunity to improve the disease resistance of shrimp.Therefore,supplementation of inactivated L.plantarum Ep-M17 in shrimp diets can exert similar effects as live L.plantarum Ep-M17 effectively improving growth performance,gut microbiota,immune response,and disease resistance in P.vannamei.

关键词： Penaeus vannamei Lactobacillus plantarum Ep-M17 inactivated probiotics immune response gut flora

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Spontaneous mutations and mutational responses to penicillin treatment in the bacterial pathogen Streptococcus pneumoniae D39

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marine Life Science & Technology 2024年第2期6卷 198-211页

作者： Wanyue Jiang Tongtong Lin Jiao Pan Caitlyn ERivera Clayton Tincher Yaohai Wang Yu Zhang Xiang Gao Yan Wang Ho‑Ching T.Tsui Malcolm E.Winkler Michael Lynch Hongan Long Institute of Evolution and marine Biodiversity KLMMEOcean University of ChinaQingdao 266003China laboratory for marine biology and biotechnology Laoshan LaboratoryQingdao 266237China Department of biology Indiana UniversityBloomingtonIN 47405USA School of Mathematics Science Ocean University of ChinaQingdao 266000China State Key laboratory of Microbial Technology Microbial Technology InstituteSchool of Life ScienceShandong UniversityQingdao 266237China Biodesign Center for Mechanisms of Evolution Arizona State UniversityTempeAZ 85281USA

Bacteria with functional DNA repair systems are expected to have low mutation rates due to strong natural selection for genomic stability.However,our study of the wild-type Streptococcus pneumoniae D39,a pathogen responsible for many common diseases,revealed a high spontaneous mutation rate of 0.02 per genome per cell division in mutation-accumulation(MA)lines.This rate is orders of magnitude higher than that of other non-mutator bacteria and is characterized by a high mutation bias in the A/T direction.The high mutation rate may have resulted from a reduction in the overall efficiency of selection,conferred by the tiny effective population size in nature.In line with this,S.pneumoniae D39 also exhibited the lowest DNA mismatch-repair(MMR)efficiency among bacteria.Treatment with the antibiotic penicillin did not elevate the mutation rate,as penicillin did not induce DNA damage and S.pneumoniae lacks a stress response pathway.Our findings suggested that the MA results are applicable to within-host scenarios and provide insights into pathogen evolution.

关键词： Neutral evolution Mutation spectrum Antibiotics DNA mismatch repair Oxidative damage repair

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RebL1 is required for macronuclear structure stability and gametogenesis in Tetrahymena thermophila

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marine Life Science & Technology 2024年第2期6卷 183-197页

作者： Huijuan Hao Yinjie Lian Chenhui Ren Sitong Yang Min Zhao Tao Bo Jing Xu Wei Wang Key laboratory of Chemical biology and Molecular Engineering of Ministry of Education Institute of BiotechnologyShanxi UniversityTaiyuan 030006China School of Life Science Shanxi UniversityTaiyuan 030006China Shanxi Key laboratory of biotechnology Taiyuan 030006China

Histone modification and nucleosome assembly play important roles in chromatin-related processes.Histone chaperones form different complexes and coordinate histone transportation and assembly.Various histone chaperone complexes have been identified in different organisms.The ciliate protozoa(ciliates)have various chromatin structures and different nuclear morphology.However,histone chaperone components and functions of different subunits remain unclear in ciliates.Tet-rahymema thermophila contains a transcriptionally active macronucleus(MAC)and a transcriptionally inactive micronu-cleus(MIC)which exhibit multiple replication and various chromatin remodeling progresses during vegetative growth and sexual developmental stages.Here,we found histone chaperone RebL1 not only localized evenly in the transcriptionally active MAC but also dynamically changed in the MIC during vegetative growth and sexual developmental stages.REBL1 knockdown inhibited cellular proliferation.The macronuclear morphology became bigger in growing mutants.The abnormal macronuclear structure also occurred in the starvation stage.Furthermore,micronuclear meiosis was disturbed during sexual development,leading to a failure to generate new gametic nuclei.RebL1 potentially interacted with various factors involved in histone-modifying complexes and chromatin remodeling complexes in different developmental stages.REBL1 knockdown affected expression levels of the genes involved in chromatin organization and transcription.Taken together,RebL1 plays a vital role in maintaining macronuclear structure stability and gametogenesis in T.thermophila.

关键词： Gametogenesis Histone chaperone Macronuclear structure RebL1 Tetrahymena thermophilaIntroduction

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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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From germline genome to highly fragmented somatic genome:genome-wide DNA rearrangement during the sexual process in ciliated protists

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marine Life Science & Technology 2024年第1期6卷 31-49页

作者： Liping Lyu Xue Zhang Yunyi Gao Tengteng Zhang Jinyu Fu Naomi A.Stover Feng Gao Key laboratory of Evolution&marine Biodiversity(Ministry of Education) and Institute of Evolution&Marine BiodiversityOcean University of ChinaQingdao 266003China Department of biology Bradley UniversityPeoriaIL 61625USA Laoshan laboratory Qingdao 266237China

Genomes are incredibly dynamic within diverse eukaryotes and programmed genome rearrangements(PGR)play important roles in generating genomic diversity.However,genomes and chromosomes in metazoans are usually large in size which prevents our understanding of the origin and evolution of PGR.To expand our knowledge of genomic diversity and the evolutionary origin of complex genome rearrangements,we focus on ciliated protists(ciliates).Ciliates are single-celled eukaryotes with highly fragmented somatic chromosomes and massively scrambled germline genomes.PGR in ciliates occurs extensively by removing massive amounts of repetitive and selfish DNA elements found in the silent germline genome dur-ing development of the somatic genome.We report the partial germline genomes of two spirotrich ciliate species,namely Strombidium cf.sulcatum and Halteria grandinella,along with the most compact and highly fragmented somatic genome for S.cf.sulcatum.We provide the first insights into the genome rearrangements of these two species and compare these features with those of other ciliates.Our analyses reveal:(1)DNA sequence loss through evolution and during PGR in S.cf.sulcatum has combined to produce the most compact and efficient nanochromosomes observed to date;(2)the compact,transcriptome-like somatic genome in both species results from extensive removal of a relatively large number of shorter germline-specific DNA sequences;(3)long chromosome breakage site motifs are duplicated and retained in the somatic genome,revealing a complex model of chromosome fragmentation in spirotrichs;(4)gene scrambling and alternative pro-cessing are found throughout the core spirotrichs,offering unique opportunities to increase genetic diversity and regulation in this group.

关键词： Alternative processing Ciliates Gene scrambling Genome rearrangement Germline genome Somatic genome

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