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The ceRNA Network Indicates Its Immune Function in Peripheral Blood Leukocytes of half-smooth tongue sole(Cynoglossus semilaevis)After Stimulation with Chitosan Oligosaccharide

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Journal of Ocean University of China 2024年第6期23卷 1615-1629页

作者： WANG Wenwen WEI Shu CHEN Yadong TAN Suxu SHI Kunpeng ZANG Shaoqing WANG Minmin YANG Teng YANG Yingming CUI Zhongkai SHA Zhenxia Institute of Aquatic Biotechnology College of Life SciencesQingdao UniversityQingdao 266071China State Key Laboratory of Mariculture Biobreeding and Sustainable Goods Yellow Sea Fisheries Research InstituteChinese Academy of Fishery SciencesQingdao 266071China Laboratory for Marine Fisheries Science and Food Production Processes Qingdao Marine Science and Technology CenterQingdao 266237China

As an efficient immunostimulant,chitosan oligosaccharide(COS)can enhance the immunity of teleosts;however,the underlying molecular mechanisms still require *** endogenous RNAs(ceRNAs)are vital regulators in the immune response,but their roles in half-smooth tongue sole(Cynoglossus semilaevis)remain *** this study,for the first time,we studied whole-transcriptome expression profiles and analyzed ceRNA networks in peripheral blood leukocytes of half-smooth tongue sole treated with COS.A total of 19 circRNAs(DE-circRNAs),18 miRNAs(DE-miRNAs)and 50 previously identified lncRNAs(DElncRNAs)were differentially expressed after COS *** DE-lncRNAs and DE-miRNAs targeted numerous immunity-related genes,and were enriched in important pathways,including MAPK and Toll-like receptor signaling pathways,suggesting the immunoregulatory roles of ***,we constructed circRNA-miRNA-mRNA and lncRNA-miRNA-mRNA regulatory networks using DE-circRNAs,DE-miRNAs,DE-lncRNAs,and ***,a ceRNA network with immunity-related DEmRNAs was constructed,showing that 3 DE-circRNAs,12 DE-lncRNAs,and 29 DEGs exhibited crosstalk through 9 ***,a DE-miRNA in the ceRNA network,miR-144-3p,was targeted by DE-lncRNA tnrc6a,and negatively regulated the genes of inhibitor of nuclear factor kappa B kinases(IKKs)(ikbkg,ikbkb,and ikbip)and ***,ikbkb,and c3ar1 were significantly up-regulated in macrophages stimulated by *** could be inferred that ncRNAs participated in the immune and inflammatory response by acting as ceRNAs after COS stimulation in *** findings indicate that COS could enhance the immunity of teleosts by regulating ncRNAs,and lay the foundation for further practical application of COS in aquaculture.

关键词： ceRNA circRNA miRNA half-smooth tongue sole peripheral blood leukocytes chitosan oligosaccharide

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Application of AFLP markers for population genetic study on half-smooth tongue sole Cynoglossus semilaevis

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Chinese Journal of Oceanology and Limnology 2011年第3期29卷 609-615页

作者：刘云国李俊峰叶乃好 Inspection & Quarantine Technology Center Shandong Entry-Exit Inspection & Quarantine Bureau Jiangsu Key Laboratory of Marine Biotechnology Huaihai Institute of Technology College of Chemical Engineering Qingdao University of Science & Technology Yellow Sea Fisheries Research Institute Chinese Academy of Fishery Sciences

The genetic diversity of wild and hatchery populations of half-smooth tongue sole Cynoglossus semilaevis, based on observation of amplified fragment length polymorphism (AFLP) was described. Two hundred individuals from four wild populations, Laizhou (LZ), Weihai (WH), Qingdao (QD), Rizhao (RZ), and one hatchery population, Mingbo (MB), were screened using eight different AFLP primer combinations. A total of 384 loci were screened in the five studied populations. 48.4%, 51.3%, 50.7%, 49.3% and 45.8% of these loci were polymorphic among the individuals tested in the LZ, WH, QD, RZ and MB populations, respectively. The number of polymorphic loci detected by single primer combinations ranged from 17 to 35. The average heterozygosity of the LZ, WH, QD, RZ and MB populations was 0.072, 0.093, 0.092, 0.090 and 0.063, respectively. The WH population showed the highest genetic diversity in terms of total number of AFLP bands, total number of polymorphic bands, average heterozygosity and percentage of low frequency (0-0.2) polymorphic loci among all the populations, while the LZ population was the lowest among the wild populations. Compared with the wild populations, the hatchery population showed a low genetic viability.

关键词： half-smooth tongue sole Cynoglossus semilaevis AFLP marker genetic characteristics

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Molecular characterization and expression analysis of Patched 1 gene in the half-smooth tongue sole(Cynoglossus semilaevis)

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Acta Oceanologica Sinica 2016年第6期35卷 19-28页

作者： DONG Zhongdian QI Qian ZHANG Ning SHAO Changwei ZHANG Liyan WEN Haishen CHEN Songlin Fisheries College Ocean University of China Yellow Sea Fisheries Research Institute Chinese Academy of Fishery Sciences Function Laboratory for Marine Fisheries Science and Food Production Processes National Laboratory for Marine Science and Technology

The signaling molecule hedgehog（Hh） is essential for cellular signaling required for body pattern formation in embryonic and individual development in vertebrate and *** 1（Ptc1）,a receptor of Hh,mediates the activity of Hh by binging the Hh *** this study,the Ptc1 of half-smooth tongue sole Cynoglossus semilaevis（CsPtc1） was cloned and *** full-length of CsPtc1 cDNA is 5 212 nucleotides（nt）,and encodes a protein with 1 543 amino acid ***1 shared many features with Ptc1 of other telesot ***-time quantitative PCR revealed CsPtc1 mRNA expressed in brain,liver,heart,gill,intestines,spleen,gonad and kidney,with expression level in testis significantly higher than in *** testis,hybridization signals were mainly detected in primary spermatocytes,secondary spermatocytes,and sertoli cells,while weak signals were found in ***,the degree of methylation is higher in female than in male and *** indicate that CsPtc1 may be involved in Desert Hedgehog（DHH） maintenance of the male and pseudo-male germ line and spermatogenesis.

关键词： half-smooth tongue sole male and female Patched 1 RACE qRT-PCR methylation

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Phenotypic and genetic parameter estimation of juvenile growth and bottom color traits in half-smooth tongue sole,Cynoglossus semilaevis

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Acta Oceanologica Sinica 2016年第10期35卷 83-87页

作者： LIU Feng Key Laboratory for Sustainable Development of Marine Fisheries of Ministry of Agriculture Yellow Sea Fisheries Research InstituteChinese Academy of Fishery SciencesQingdao 266071China Laboratory for Marine Fisheries Science and Food Production Processes Qingdao National Laboratory for Marine Science and TechnologyQingdao 266237China College of Fisheries and Life Science Shanghai Ocean UniversityShanghai 201306China Marine and Fishery Research Institute Zhejiang Ocean UniversityZhoushan 316021China

half-smooth tongue sole（Cynoglossus semilaevis） is a commercially important fish species. There were no reports about accurate genetic parameters of growth traits in C. semilaevis, despite efforts aimed at the culture of this species. Therefore, the main goal of this study was to estimate the parameters for the early growth traits（total length, body depth and body weight） of C. semilaevis based on the data obtained from 80 families. The trait of fish with pure white bottom or not（named as the trait of bottom color） was also analyzed. All the heritabilities estimated for the traits were medium, which varied from 0.210 to 0.362. Genetic correlations among growth traits were highly positive, varying from 0.913 to 0.959, indicating that selection of one of the three traits would result in correlated increase in the others. Also, genetic correlations between bottom color and growth traits were positive and varying from 0.241 to 0.353, suggesting that selection for the bottom color can be utilized to enhance the selection of growth traits. In addition, fish from the top performing 16 full-sib families were selected as broodstock, based on the above analysis and the comparison of family breeding value. This research provides an important basic material to implement selective breeding in C. semilaevis.

关键词： half-smooth tongue sole Cynoglossus semilaevis growth trait bottom color genetic parameter family selection

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Characterization of the cyp19a1a gene from a BAC sequence in half-smooth tongue sole (Cynoglossus semilaevis) and analysis of its conservation among teleosts

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Acta Oceanologica Sinica 2013年第8期32卷 35-43页

作者： SHAO Changwei LIU Geng LIU Shanshan LIU Changlin CHEN Songlin Ocean University of China Yellow Sea Fisheries Research Institute Chinese Academy of Fisheries Sciences BGI-Shenzhen

The cyp19ala gene encodes an aromatase that plays a key role in sex differentiation of the gonad. The first bacterial artificial chromosome （BAC） sequence of half-smooth tongue sole （Cynoglossus sernilaeuis） containing the intact cyp19ala gene was reported and the conservation and synteny of the cyp19ala gene among teleosts were analyzed in the study. The BAC is 107 367 bp in size, with an overall guanine-cytosine （GC） content of 43.44%, and contains 4.38% transposable elements. Nine genes were predicted, including seven functional genes and two hypothetical genes. The cyp19ala gene of all tested teleosts has nine exons and eight introns, and potential binding sites flanking the transcriptional start site are conserved. The ex- pression pattern among teleosts is also similar during ovarian differentiation. Synteny analysis revealed a conserved gene cluster PKH4B-SL9A5-FHOD3-CEBPG-CEBPA among teleosts. These findings suggest that, among teleosts, cyp19ala genes not only have similar genomic structures, but also have conserved function- s. The genomic environment of cyp19ala in tongue sole is not universal in teleosts, reflecting the particular evolution of tongue sole cyp19ala after it diverged from the other teleosts.

关键词： BAC cypl9ala conservation gene synteny half-smooth tongue sole

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Molecular cloning,characterization and expression analysis of Sox9a and Foxl2 genes in half-smooth tongue sole(Cynoglossussemilaevis)

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Acta Oceanologica Sinica 2011年第1期30卷 68-77页

作者： DONG Xiaoli CHEN Songlin JI Xiangshan SHAO Changwei College of Fisheries Ocean University of China Qingdao 266003 China Key Laboratory for Sustainable Utilization of Marine Fisheries Resources of Ministry ofAgriculture Yellow Sea Fisheries Research Institute Chinese Academy of Fishery SciencesQingdao 266071 China College of Animal Science and Technology Shandong Agricultural University Taian 271018 China

Two transcription factors, Sox9a and Foxl2 were cloned from half-smooth tongue sole （Cynoglossus semilaevis）. Sox9a is a new duplication of *** Sox9 gene. The complete cDNA of Sox9a gene was 1 842 bp long coding for 487 amino acids and Foxl2 gene was 1 817 bp coding for 308 amino acids. Sox9a was expressed higher in male brain, pituitary and gonad and Foxl2 were higher in female brain, pituitary and gonad. The expression of Sox9a gene in gonads of neo-males was higher than that of normal females. Sox9a and Foxl2 were expressed higher in gastrula stage than in other stages. In the period of sex differentiation, the expression of Sox9a was first going up and then going down and Foxl2 was higher expressed at 37 dph. The highest expressions of Sox9a and Foxl2 genes occurred in nine-month and 12-month old gonad tissues, respectively. Sox9a gene was considered to have inevitable links with sex reversal, sex differentiation and cell differentiation of embryos and formation of spermatogenic ceils. Foxl2 was considered to play a role in sex differentiation, cell differentiation of embryos but not to be necessary for sex determination and sex reversal.

关键词： half-smooth tongue sole Cynoglossus semilaevis Sox9 Foxl2 real-time PCR sexreversal neo-male

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Isolation and Characterization of Pathogenic Listonella anguillarum of Diseased half-smooth tongue sole(Cynoglossus semilaevis Günther)

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Journal of Ocean University of China 2008年第3期7卷 343-351页

作者： TANG Xiaoqian ZHOU Li ZHAN Wenbin LaboratoO qf Patholog） and Immunology of Aquatic Animals LMMEC Ocean University of ChinaQingdao 266003 P. R. China

Studies were conducted to determine the cause of the acute mortality of half-smooth tongue sole Cynoglossus semilaevis Günther juveniles in a fish farm in Jimo, Shandong Province, China, in June 2006. Gross signs of the diseased tongue sole included several petechiae and ecchymoses on the body and fin necrosis and hemorrhagic lesion at the base of the fin. Bacteria were isolated from kidney, liver and hemorrhagic lesions of the diseased tongue sole. Among 14 strains, SJ060621 was proved to be highly virulent to juvenile tongue sole with LD50 value of 〈1.0×10^5 colony forming units （CFU）mL^-1, while the remaining 13 were avirulent. Among the 16 antibiotics tested, SJ060621 was sensitive to gentamicin and nitrofurantoin. It was identified as Listonella anguillantm with conventional plate and tube tests in combination with API 20E analysis. 16S rRNA gene and partial HSP60 gene sequenceing analysis revealed that the strain was highly homologous with L. anguillarum. Examination of the infected musculature by electron microscopy indicated numerous bacteria and lots of macrophages containing phagocytosed bacteria. Histopathological investigations revealed severe necrotic degenerative changes in the infected organs. Indirect immunofluorescence assay （IFA） was employed to detect the location of occurrence of bacteria, and bacteria were found in aggregations in the inflammatory areas in musculature.

关键词： half-smooth tongue sole Cynoglossus semilaevis Günther Listonella anguillarum 16S rRNA gene HSP60 gene histopathology IFA

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Analysis of SSRs Derived from an EST Library of half-smooth tongue sole Cynoglossus semilaevis

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Journal of Ocean University of China 2008年第3期7卷 339-342页

作者： LIU Yunguo BAO Baolong CHEN Songlin SUN Xiuqin College of Fisheries Ocean Universitv of China Qingdao 266003 P. R. China Key Laboratory of Marine and Estuarine Fisheries Ministry of Agriculture East China Sea Fisheries ResearchInstitute Shanghai 200090 P. R. China College of Aqua-Life Science and Technology Shanghai Ocean University Shanghai 200090 P. R. China Yellow Sea Fisheries Research Institute Chinese Academy of Fishery Sciences Qingdao 266071 P. R. China The First Institute of Oceanography State Oceanic Administration Qingdao 266061 P. R. China

A complementary DNA(cDNA) library was constructed from half-smooth tongue sole spleen. A long-read expressed sequence tag(EST) database was generated,containing 3100 cDNA clones,of which 220 clones were fully sequenced. A total of 1060 non-redundant simple sequence repeats(SSRs) were obtained from the cDNA library. An average of 5 kb sequence generates 1 SSR in the half-smooth tongue sole spleen cDNA library. The proportion of the SSR unit size was different in the cDNA library. The monomeric repeats(51.4%) are the most abundant class of SSR in the dataset. The dimeric,trimeric,tetrameric and hexameric re-peats are represented in decreasing proportions of 27.2%,16.0%,2.8% and 1.9%,respectively. The frequency of pentameric repeats was observed the least(only 0.7%) . Most of the monomeric and dimeric repeats are distributed in 3' and 5' un-translation region. If translation regions are considered merely,trimeric repeats are the highest,accounting for 57% of the total microsatellites.

关键词： half-smooth tongue sole Cynoglossus semilaevis EST microsatellite

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Development of 57 Novel Polymorphic Microsatellite Markers in half-smooth tongue sole(Cynoglossus semilaevis)

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Journal of Ocean University of China 2011年第3期10卷 241-245页

作者： MIAO Guidong XU Ying WANG Di CHEN Songlin FAN Tingjun TIAN Yongsheng College of Marine Life Sciences Ocean University of China Qingdao 266003 P. R. China Key Lab for Sustainable Utilization of Marine Fisheries Resources Ministry of Agriculture Yellow Sea Fisheries Research Institute Chinese Academy of Fishery Sciences Qingdao 266071P. R. China Zibo Entry-Exit Inspection and Quarantine Bureau of P. R. China Zibo 255031 P. R. China

half-smooth tongue sole (Cynoglossus semilaevis) is a promising species for aquaculture in *** wild population of C. semilaevis is under threat from environmental factors. Microsatellite markers are very suitable for assessing genetic diversity. Four microsatellite-enriched libraries of half smooth tongue sole (Cynoglossus semilaevis) were constructed,from which 57 polymorphic microsatellites were isolated and *** polymorphism of these microsatellites was assessed by genotyping in 30 individual *** number of alleles ranged from 2 to 11, with an average of 4.614 alleles per *** values of observed and expected heterozygosities ranged from 0.1000 to 1.0000 and from 0.0966 to 0.8847 respectively. Polymorphism information content (PIC) ranged from 0.0905 to *** markers would be useful for population structure assessment,genetic linkage map construction and parentage analysis for this species.

关键词： Cynoglossus semilaevis microsatellite polymorphic marker half-smooth tongue sole

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Construction of a microsatellite-based genetic linkage map for half-smooth tongue sole Cynoglossus semilaevis

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Current Zoology 2013年第1期59卷 99-108页

作者： Wentao SONG Guidong MIAO Yongwei ZHAO Yuze NIU Renyi PANG Xiaolin LIAO Changwei SHAO Songlin CHEN Yellow Sea Fisheries Research Institute Chinese Academy of Fishery Sciences Qingdao 266071 China College of Fisheries and Life Science Shanghai Ocean University Shanghai 201306 China

The half-smooth tongue sole Cynoglossus semilaevis is an important cultured marine fish and a promising model fish for the study of sex determination. Sex-specific genetic linkage maps of half-smooth tongue sole were developed with 567 mark- ers （565 microsatellite markers and two SCAR markers）. The parents and F1 progeny （92 individuals） were used as segregating populations. The female map was composed of 480 markers in 21 linkage groups, covering a total of 1388.1 cM, with an average interval 3.06 cM between markers. The male map consisted of 417 markers in 21 linkage groups, spanning 1480.9 cM, with an average interval of 3.75 cM. The female and male maps had 474 and 416 unique positions, respectively. The genome length of half-smooth tongue sole was estimated to be 1522.9 cM for females and 1649.1cM for males. Based on estimations of map length, the female and male maps covered 91.1% and 89.8% of the genome, respectively. Furthermore, two female-specific SCAR mark- ers, f-382 and f-783, were mapped on LG15f （linkage group 15 in female maps）. The present study presents a mid-density genetic linkage map for half-smooth tongue sole. These improved genetic linkage maps may facilitate systematic genome searches to identify quantitative trait loci （QTL）, such as disease resistance, growth and sex-related traits, and are very useful for marker-assisted selection breeding programs for economically important traits in half-smooth tongue sole [Current Zoology 59 （1）： 99-108, 2013].

关键词： half-smooth tongue sole Cynoglossus semilaevis Microsatellite Genetic linkage map MAS

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