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Journal of Ocean University of China 2023年第6期22卷 1621-1634页

作者： MEI Yaoping HOU Zhishuai GAO Qinfeng DONG Shuanglin LI Xueqi XU Yuling Key Laboratory of Mariculture Ministry of EducationOcean University of ChinaQingdao 266003China Function Laboratory for Marine Fisheries Science and Food Production Processes Qingdao National Laboratory for Marine Science and TechnologyQingdao 266235China

Sea cucumber(apostichopus japonicus)is an excellent model for investigating effects of bottom-dwellers on carbon mig-ration and ***,the molecular mechanism of respiratory metabolism process variation caused by feeding rations is poorly *** this study,treatment groups set as 1%(about 0.63g),3%,and 7%of total body weight(named F1,F3 and F7 groups respectively).The potential molecular mechanisms behind the functions of respiratory tree and body wall were investigated by RNA-Seq.A total of 52411 expressed genes were identified from 89342 expressed *** results showed 759,254 and 334 genes were up-regulated,and 334,445 and 992 genes were down-regulated in respiratory tree of F1 vs.F3,F1 vs.F7 and F3 vs.F7,***,2070,1601 and 896 genes were up-regulated,and 1303,1337 and 1144 genes were down-regulated in body wall between F1 vs.F3,F1 vs.F7 and F3 vs.F7,*** expressed genes were enriched in salivary secretion and ECM-receptor interaction pathways in respiratory tree,and in various types of N-glycan biosynthesis,ribosome and sphingolipid metabolism pathways in body *** results suggested respiratory tree and body wall were involved in activation of respiratory metabolisms in response to different feeding *** research provided valuable knowledge for physiological differences in res-piratory metabolism.

关键词： transcriptomics apostichopus japonicus feeding ration respiration metabolism

Analysis of differentially expressed genes in the sea cucumber apostichopus japonicus under heat stress

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Acta Oceanologica Sinica 2023年第11期42卷 117-126页

作者： Dongxue Xu Jingjing Zhang Wenqi Song Lina Sun Ji Liu Yuanxue Gu Yanru Chen Bin Xia School of Marine Science and Engineering Qingdao Agricultural UniversityQingdao 266109China Key Laboratory of Mariculture of Ministry of Education Ocean University of ChinaQingdao 266003China CAS Key Laboratory of Marine Ecology and Environmental Sciences Institute of OceanologyChinese Academy of SciencesQingdao 266071China

The sea cucumber apostichopus japonicus plays important roles in marine benthic ecosystem as environmental cleaners,and it is the important aquaculture species in *** water temperature poses critical threat for the survival of ***,which has resulted in extensive death in *** explore the genes expression profiles under different levels of heat stress,the high-throughput RNA-seq was applied in this *** results revealed a total of 1371,1225 and 1408 differentially expressed genes(DEGs)in 26℃for 6 h,26℃for 48 h and 30℃for 6 h respectively in comparison with Control *** pathway analysis suggested“Protein processing in endoplasmic reticulum(ER)”was significantly enriched in all these heat stress(HS)treatment *** expression results of key DEGs in this pathway(Hsp70,Derlin,NEF,PDI,GPR94 and ERP57)by qRT-PCR was in accordance with the RNA-seq *** subcluster analysis of DEGs revealed that a variety of heat shock proteins(HSPs)and calcium ion binding proteins had an obvious up-regulated expression in 26℃for 6 h,comparatively low expression in 26℃for 48 h,and the highest expression in 30℃for 6 *** other DEGs subcluster,consisting of critical components of extracellular matrix(ECM)and a subset of peptidases and proteases,showed significantly rising tendency in 30℃for 6 ***,the expression of matrix metalloproteases(MMP1,MMP16 and MMP19)was prominently affected by HS,and peaked in 30℃for 6 *** study provides a series of candidate genes for further study about heat shock response in ***,especially genes associated with protein processing in ER and regulation of ECM,which also offers new insights into cellular homeostasis under stressful conditions in marine invertebrates.

关键词： apostichopus japonicus heat stress RNA-seq protein processing extracellular matrix

Isolation, Identification, and Quantitative Determination of Saponin in apostichopus japonicus by HPLC-DAD

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Journal of Ocean University of China 2022年第2期21卷 473-478页

作者： LIU Guiying WANG Xuda GE Kun WU Jinhao WANG Zhaohui DU Jing SONG Lun ZHOU Zunchun Liaoning Ocean and Fisheries Science Research Institute Dalian 116023China Dalian Ocean School Dalian 116023China

apostichopus japonicus is an important invertebrate that is widely used as a tonic food in Asian *** purpose of this study is to purify and identify a class of compound,the saponins,from the body wall of ***,and to establish a new me-thod to determine the quantity of saponins in the sea *** this study,the saponins of ***,cladoloside A(CA),were ob-tained from 80%ethanol extract by column chromatography for the first time and were characterized using the spectral *** resulting purified saponins were then profiled using ^(1)HNMR,^(13)CNMR,and ESI-MS,which revealed the CA molecular formula of C_(53)H_(82)O_(2) and contained a triterpenoid backbone,a methylglucopyranosyl moiety,a quinovopyranosyl,and two xylopyranosyls.A me-thod for the quantitative determination of CA,comprising microwave-assisted extraction,high-performance liquid chromatography,and diode array detector method,was *** efficiency was optimized by changing microwave power,extraction sol-vent,volume,time,and *** showed that under the optimum conditions(extraction time of 10 min,temperature of 45℃,and solvent of 25 mL 70%ethanol under 400 W),the detection limit of CA was 0.0015 mg mL^(−1),and the recoveries of CA from samples at spiking levels of 10,20,and 50μg g^(−1) ranged from 90.1%-104%.The proposed method was successfully applied to ana-lyze the saponins in different tissues of *** collected in different *** method developed in this study can provide quantitative technical support for the quality control of ***.

关键词： cladoloside A apostichopus japonicus saponin microwave-assisted extraction HPLC-DAD

Choline dehydrogenase interacts with SQSTM1 to activate mitophagy and promote coelomocyte survival in apostichopus japonicus following Vibrio splendidus infection

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动物学研究(英文) 2023年第5期44卷 905-918页

作者： Lian-Lian Sun Ying-Fen Dai Mei-Xiang You Cheng-Hua Li State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products Ningbo UniversityNingboZhejiang 315211China Laboratory for Marine Fisheries Science and Food Production Processes Qingdao National Laboratory for Marine Science and TechnologyQingdaoShandong 266071China

Previous studies have shown that Vibrio splendidus infection causes mitochondrial damage in apostichopus japonicus coelomocytes,leading to the production of excessive reactive oxygen species(ROS)and irreversible apoptotic cell *** evidence suggests that mitochondrial autophagy(mitophagy)is the most effective method for eliminating damaged mitochondria and ROS,with choline dehydrogenase(CHDH)identified as a novel mitophagy receptor that can recognize non-ubiquitin damage signals and microtubule-associated protein 1 light chain 3(LC3)in ***,the functional role of CHDH in invertebrates is largely *** this study,we observed a significant increase in the mRNA and protein expression levels of *** CHDH(AjCHDH)in response to *** infection and lipopolysaccharide(LPS)challenge,consistent with changes in mitophagy under the same ***,AjCHDH was localized to the mitochondria rather than the cytosol following *** ***,AjCHDH knockdown using si RNA transfection significantly reduced mitophagy levels,as observed through transmission electron microscopy and confocal *** investigation into the molecular mechanisms underlying CHDH-regulated mitophagy showed that AjCHDH lacked an LC3-interacting region(LIR)for direct binding to LC3 but possessed a FB1 structural domain that binds to *** interaction between AjCHDH and SQSTM1 was further confirmed by immunoprecipitation ***,laser confocal microscopy indicated that SQSTM1 and LC3 were recruited by AjCHDH in coelomocytes and HEK293T *** contrast,AjCHDH interference hindered SQSTM1 and LC3 recruitment to the mitochondria,a critical step in damaged mitochondrial ***,AjCHDH interference led to a significant increase in both mitochondrial and intracellular ROS,followed by increased apoptosis and decreased coelomocyte ***,these findings indicate that AjCHDH-mediated mitophagy plays a

关键词： Choline dehydrogenase Mitophagy SQSTM1 Microtubule-associated protein 1 light chain 3 apostichopus japonicus

ROS-mediated BNIP3-dependent mitophagy promotes coelomocyte survival in apostichopus japonicus in response to Vibrio splendidus infection

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Zoological Research 2022年第2期43卷 285-300页

作者： Lian-Lian Sun Yi-Na Shao Mei-Xiang You Cheng-Hua Li State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products Ningbo UniversityNingboZhejiang 315211China Laboratory for Marine Fisheries Science and Food Production Processes Qingdao National Laboratory for Marine Science and TechnologyQingdaoShandong 266071China State-Province Joint Laboratory of Marine Biotechnology and Engineering Ningbo UniversityNingboZhejiang 315211China

Organisms produce high levels of reactive oxygen species(ROS)to kill pathogens or act as signaling molecules to induce immune responses;however,excessive ROS can result in cell *** maintain ROS balance and cell survival,mitophagy selectively eliminates damaged mitochondria via mitophagy receptors in *** marine invertebrates,however,mitophagy and its functions remain largely *** the current study,Vibrio splendidus infection damaged mitochondrial morphology in coelomocytes and reduced mitochondrial membrane potential(ΔΨm)and mitophagosome *** colocalization of mitochondria and lysosomes further confirmed that lipopolysaccharide(LPS)treatment increased mitophagy *** explore the regulatory mechanism of mitophagy,we cloned Bcl2/adenovirus E1 B 19 kDa protein-interacting protein 3(BNIP3),a common mitophagy receptor,from sea cucumber apostichopus japonicus(Aj BNIP3)and confirmed that Aj BNIP3 was significantly induced and accumulated in mitochondria after *** infection and LPS *** the mitochondrial membrane,Aj BNIP3 interacts with microtubule-associated protein 1 light chain 3(LC3)on phagophore membranes to mediate *** Aj BNIP3 interference,mitophagy flux decreased ***,Aj BNIP3-mediated mitophagy was activated by ROS following the addition of exogenous hydrogen peroxide(H2 O2),ROS scavengers,and ROS ***,inhibition of BNIP3-mediated mitophagy by Aj BNIP3 small interfering RNA(si RNA)or high concentrations of lactate increased apoptosis and decreased coelomocyte *** findings highlight the essential role of Aj BNIP3 in damaged mitochondrial degradation during *** mitophagy activity is required for coelomocyte survival in *** against *** infection.

关键词： apostichopus japonicus Mitophagy Bcl2/adenovirus E1B 19 kDa protein-interacting protein 3 Reactive oxygen species Microtubule-associated protein 1 light chain 3

Fine‑mapping and association analysis of candidate genes for papilla number in sea cucumber,apostichopus japonicus

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Marine Life Science & Technology 2022年第3期4卷 343-355页

作者： Xinghai Zhu Ping Ni Marc Sturrock Yangfan Wang Jun Ding Yaqing Chang Jingjie Hu Zhenmin Bao Ministry of Education Key Laboratory of Marine Genetics and Breeding College of Marine Life SciencesOcean University of ChinaQingdao 266003China Department of Physiology and Medical Physics Royal College of Surgeons in IrelandDublin D02 YN77Ireland College of Fisheries and Life Science Dalian Ocean UniversityDalian 116023China Laboratory of Tropical Marine Germplasm Resources and Breeding Engineering Sanya Oceanographic InstitutionOcean University of ChinaSanya 572000China

The papilla number is one of the most economically important traits of sea cucumber in the China marketing ***,the genetic basis for papilla number diversity in holothurians is still *** the present study,we conducted genomewide association studies(GWAS)for the trait papilla number of sea cucumbers utilizing a set of 400,186 high-quality SNPs derived from 200 sea *** significant trait-associated SNPs that passed Bonferroni correction(P<1.25E−7)were located in the intergenic region near PATS1 and the genic region of EIF4G,which were reported to play a pivotal role in cell growth and *** fine-mapping regions around the top two lead SNPs provided precise causative loci/genes related to papilla formation and cellular activity,including PPP2R3C,GBP1,and *** SNPs with P<1E−4 were acquired for the following GO and KEGG enrichment ***,the two lead SNPs were verified in another population of sea cucumber,and the expressive detection of three potential candidate genes PATS1,PPP2R3C,and EIF4G that near or cover the two lead SNPs was conducted in papilla tissue of TG(Top papilla number group)and BG(Bottom papilla number group)by *** found the significantly higher expression profile of PATS1(3.34-fold),PPP2R3C(4.90-fold),and EIF4G(4.23-fold)in TG,implying their potential function in papilla *** present results provide valuable information to decipher the phenotype differences of the papilla trait and will provide a scientific basis for selective breeding in sea cucumbers.

关键词： Genome-wide association study apostichopus japonicus Molecular breeding Candidate genes Papilla number

Ultrastructural variation and key ER chaperones response induced by heat stress in intestinal cells of sea cucumber apostichopus japonicus

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Journal of Oceanology and Limnology 2021年第1期39卷 317-328页

作者： Shasha WANG Yingqiu ZHENG Muyan CHEN Kenneth B.STOREY2 Key Laboratory of Mariculture(Ocean University of China) Ministry of EducationQingdao 266003China Institute of Biochemistry Carleton UniversityOttawa ON K1S 5B6Canada

Abstract The unfolded protein response(UPR)is an important protective and compensatory strategy used during endoplasmic reticulum stress caused by factors including glucose starvation,low pH,or heat ***,there is very little information on the possible role(s)of the UPR under adverse conditions experienced by marine *** observed that rough endoplasmic reticulum(ER)was dramatically expanded and numerous autophagosomes were accumulated in the intestinal cells of sea cucumbers,apostichopus japonicus,under heat stress(4 h at 25°C compared with 15°C controls).Moreover,heat stress led to sharp increases in the relative transcript and protein expression levels of two primary ER chaperones:the endoplasmic reticulum resident protein 29-like(ERP29)and protein disulfi de-isomerase A6-like(PDIA6).These results suggest a potential adaptive mechanism to deal with heat-induced stress in sea cucumber intestine.

关键词： apostichopus japonicus endoplasmic reticulum(ER) unfolded protein response(UPR) endoplasmic reticulum resident protein 29-like(ERP29) protein disulfi de-isomerase A6-like(PDIA6)

Comparative analysis of transcriptomes from albino and control sea cucumbers, apostichopus japonicus

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Acta Oceanologica Sinica 2014年第8期33卷 55-61页

作者： MA Deyou YANG Hongsheng SUN Lina XU Dongxue Institute of Oceanology Chinese Academy of Sciences University of Chinese Academy of Sciences

The sea cucumberapostichopus japonicus is an important economic species in China. Its dorsal body wall color is commonly tawny, whereas its ventral surface is fawn. Albino sea cucumbers are rarely observed. In order to profile gene expression and screen albinism-related genes, we compared the transcriptome of albino samples with a control by 454 cDNA sequencing. We found that 6 539 identified genes on the basis of sequence similarity to known genes were expressed in the albino A. japonicus. The gene ontology analysis indicated that the transcription of genes associated with the terms of biological regulation and pigmenta-tion was non-abundant in the albino library compared to the control. Based on an analysis using the Kyoto Encyclopedia of Genes and Genomics （KEGG） database, we identified 14 important genes that were in-volved in major intercellular signaling pathways related to melanin synthesis, such as tyrosine metabolism, the mitogen-activated protein kinase （MAPK） pathway, and melanogenesis. The expressions of fibroblast growth factor receptor 4 （FGFR4）, protein kinase C （PKC）, protein kinase A （PKA）, and Ras genes were sig-nificantly down-regulated in the albino transcriptome compared with the control, while the expressions of homogentisate 1, 2-dioxygenase gene （HGO）, cAMP-responsive element binding protein （CREB）, transcrip-tion factor AP-1（c-jun）, and calmodulin （CaM） were significantly up-regulated （Fisher＇s exact test,p 〈 0.05）. These differentially expressed genes could be candidate genes for revealing the mechanism of albinism and investigating regulation of melanin synthesis inA. japonicus.

关键词： 454 cDNA sequencing albinism apostichopus japonicus transcriptome

Effects of rearing temperature and density on growth,survival and development of sea cucumber larvae,apostichopus japonicus(Selenka)

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Chinese Journal of Oceanology and Limnology 2010年第4期28卷 842-848页

作者：刘广斌杨红生刘石林 Institute of Oceanology Chinese Academy of Sciences Graduate University of Chinese Academy of Sciences

In laboratory conditions,effects of rearing temperature and stocking density were examined on hatching of fertilized egg and growth of auricularia larvae of apostichopus japonicus *** series like larval length and density,metamorphic time,and survival rate of the larvae were *** showed that for ***,survival rate(from fertilized egg to late auricularia) decreased significantly with the increasing rearing temperature(P26℃).Hatching rate was significantly different between 0.2-5 ind./ml groups and 20-50 ind./ml *** larvae at the higher density had the smaller maximal-length,whereas needed longer time to complete *** study suggested that 21℃ and 0.4 ind./ml can be used as the most suitable rearing temperature and stocking density for large-scale artificial breeding of ***'s larvae.

关键词： apostichopus japonicus egg hatch larvae growth metamorphism

Differences in MITF gene expression and histology between albino and normal sea cucumbers (apostichopus japonicus Selenka)

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Chinese Journal of Oceanology and Limnology 2012年第1期30卷 80-91页

作者： ZHAO Heling YANG Hongsheng ZHAO Huan LIU Shilin WANG Tianming Graduate University of Chinese Academy of Sciences Beijing 100049China Key Laboratory of Marine Ecology and Environmental Sciences Institute of OceanologyChinese Academy of SciencesQingdao 266071China

Albino apostichopus japonicus occur both in the wild and in captivity. The offspring of albino A. japonicus also suffer from albinism. The formation of melanin in the melanocytes is dependant on microphthalmia-associated transcription factor (MITF). To investigate the role of MITF in controlling albinism, we cloned the full-length MITF cDNA from A. japonicus and compared MITF mRNA expression in albino and normal A. japonicus. In addition, we used light and electron microscopy to compare histological samples of normal and albino A. japonicus. The body wall of albino adults was characterized by significantly lower levels of MITF expression and lower numbers of epidermal melanocytes, which also contained less melanin. In albino juvenile offspring, MITF expression levels were significantly lower 32 d after fertilization and there were fewer, and less developed, epidermal melanocytes. Thus, we conclude that albino A. japonicus have fewer melanocytes and a reduced ability to synthesize melanin, likely because of lower expression of MITF.

关键词： microphthalmia-associated transcription factor (MITF) melanocyte melanosome melanin apostichopus japonicus