As a member of mucins family,MUC2 is an important component of mucus,which is characterized by tandem and irregular repeat sequences rich in threonine and serine.It is strongly expressed by goblet cells that are invol...As a member of mucins family,MUC2 is an important component of mucus,which is characterized by tandem and irregular repeat sequences rich in threonine and serine.It is strongly expressed by goblet cells that are involved in innate immunity.Japanese flounder is an economically important marine flatfish.In this study,two homologous genes of MUC2 were identified,named MUC2-1 and MUC2-2.Depending on phylogenetic and structural analysis,MUC2-1 and MUC2-2 were clustered in two clades,respectively.Paralichthys olivaceus MUC2-1 showed a closer relationship with MUC2 in the higher vertebrates.Various healthy tissues were analyzed to determine the expression patterns,and both MUC2 genes showed high expression levels in the gills and intestines.Following Edwardsiella tarda challenge,P.olivaceus MUC2-1 and MUC2-2 both showed significant up-regulated expression in intestine and kidney.Moreover,MUC2-1 was significantly up-regulated in gill cell lines following PGN and polyI:C stimulation,and MUC2-2 was significantly up-regulated in gill cell lines following LPS stimulation.In addition,ISH results revealed that MUC2-1 and MUC2-2 showed the same tissue localization in intestine tissues,but displayed different localization in gill.The siRNA-mediated knockdown of MUC2-1 and MUC2-2 genes in the gill cell lines of Japanese flounder affected the expressions of galnt family genes and smad pathway members that are related to mucosal immunity.The results provided a valuable insight into understanding the functions of MUC2 on mucosal immune system in response to the invasion of bacterial pathogen.展开更多
MicroRNAs(miRNAs),22-nucleotide-long micromanagers that guide the post-transcriptional regulation of a wide range of target genes,can theoretically be used as a diagnostic or therapeutic target for inflammatory reacti...MicroRNAs(miRNAs),22-nucleotide-long micromanagers that guide the post-transcriptional regulation of a wide range of target genes,can theoretically be used as a diagnostic or therapeutic target for inflammatory reaction.In fish,miR-144-5p expression varies dramatically in response to the different bacterial infections and can regulate immunity-related genes to reduce the occurrence of inflammation.In this research,the regulation function of miR-144-5p to the intestinal innate immunity was udied in flounder Paralichthys olivaceus.The flounders were interfered by 2μg g^(-1) miR-144-5p antagomir and their tissues(intestine,liver and spleen)were harvested from the fish at 12 h post-injection.More than 60 million high-quality reads were collected.At 24 hours after miR-144-5p or miR-NC interference,a total of 2704 and 1823 different-expresion genes(DEGs)were identified in comparison with control group,respectively.The DEGs were enriched in a variety of immunity-related signaling pathways,including NOD-like receptor,Wnt and Toll-like receptor signaling pathways,according to GO and KEGG analyses.A total of 503 highly interacting DEGs engaged in 33 immunity-related signaling pathways were discovered using KEGG analyses.Additionally,5 hub genes were found by protein-protein interaction networks,which formed an intricate immune regulation network.Meanwhile,these hub genes were mostly involved in focal adhesion,Wnt signaling pathway,as well as the Intestinal Immune Network for IgT(IgA)Production Pathway.In conclusion,the loss of miR-144-5p can affect immunity-related genes and downstream signaling pathways.Our findings suggest that miR-144-5p is a modulator of gene networks and signaling pathways associated with intestinal innate immunity.展开更多
A library rich in CA/GT microsatellites was constructed from the Paralichthys olivaceus genome by combining biotin capture method and radioactive labeling hybridization. Five hundred and twenty six positive clones wer...A library rich in CA/GT microsatellites was constructed from the Paralichthys olivaceus genome by combining biotin capture method and radioactive labeling hybridization. Five hundred and twenty six positive clones were obtained through twice screens. Sequencing confirmed 133 microsatellite loci （number of repeats t〉 5） in 119 positive clones. Of these microsatellites, two （1.5%） had compound repeat motifs, 63 （47.37%） had perfect motifs and 68 （51.13%） had imperfect motifs. Primer pairs were designed in the flanking regions of 22 microsatelites and subjected to PCR amplification. In 8 artificial gynogenesis families, four pairs failed to amplification, one pair was monomorphic, and the rest were polymorphic with an average of 5.2 alleles per locus. Heterozygosities ranged between 0. 375 and 0. 846, PIC ranged between 0. 305 and 0. 823. The results suggested that most of the microsatellites we isolated were qualified to be applied to the population genetic studies of P. olivaceus.展开更多
Edwardsiellosis of flounder and turbot occurring in different mariculture farms during 2001～ 2004 was examined, including the conditions of disease occurrence, clinical signs and pathological changes. The results sho...Edwardsiellosis of flounder and turbot occurring in different mariculture farms during 2001～ 2004 was examined, including the conditions of disease occurrence, clinical signs and pathological changes. The results showed that all diseased fishes expressed bacterial septicaemia. A total of 148 strains were identified using a combination of traditional physiological and biochemical tests and partial 16S rRNA gene analysis. In addition, the mole fraction G ＋ C ratio of the DNA of representative strain of isolates and serum homology were detected, and pathogenicity tests of isolates were conducted by experimental infection. The results revealed that 148 strains were identified as E. tarda of genus Edwardsiella, all the isolates are of serologic similarity, and have strong pathogenicity to flounder and turbot.展开更多
基金supported by the Natural Science Foundation of Shandong Province (No.ZR2022QC037)the Shandong Key R&D Program for Academician Team in Shandong (No.2023ZLYS02)+3 种基金the High-Level Talents Research Fund of Qingdao Agricultural University (No.663/1120033)the Young Experts of Taishan Scholars (No.201909130)the Shandong Technical System of Fish Industry (No.SDAIT-12-03)the Science and Technology Support Plan for Youth Innovation of Colleges and Universities in Shandong Province (No.2019KJF003)。
文摘As a member of mucins family,MUC2 is an important component of mucus,which is characterized by tandem and irregular repeat sequences rich in threonine and serine.It is strongly expressed by goblet cells that are involved in innate immunity.Japanese flounder is an economically important marine flatfish.In this study,two homologous genes of MUC2 were identified,named MUC2-1 and MUC2-2.Depending on phylogenetic and structural analysis,MUC2-1 and MUC2-2 were clustered in two clades,respectively.Paralichthys olivaceus MUC2-1 showed a closer relationship with MUC2 in the higher vertebrates.Various healthy tissues were analyzed to determine the expression patterns,and both MUC2 genes showed high expression levels in the gills and intestines.Following Edwardsiella tarda challenge,P.olivaceus MUC2-1 and MUC2-2 both showed significant up-regulated expression in intestine and kidney.Moreover,MUC2-1 was significantly up-regulated in gill cell lines following PGN and polyI:C stimulation,and MUC2-2 was significantly up-regulated in gill cell lines following LPS stimulation.In addition,ISH results revealed that MUC2-1 and MUC2-2 showed the same tissue localization in intestine tissues,but displayed different localization in gill.The siRNA-mediated knockdown of MUC2-1 and MUC2-2 genes in the gill cell lines of Japanese flounder affected the expressions of galnt family genes and smad pathway members that are related to mucosal immunity.The results provided a valuable insight into understanding the functions of MUC2 on mucosal immune system in response to the invasion of bacterial pathogen.
基金supported by the National Natural Science Foundation of China(No.32002421)the Advanced Talents Foundation of QAU(No.6651118016)+5 种基金the Natural Science Foundation of Shandong Province(No.ZR2019BC009)the Fish Innovation Team of Shandong Agriculture Research System(No.SDAIT-12-06)the Aquatic Animal Immunologic Agents Engineering Research Center of Shandong Province,Key Research and Development Program in Shandong(No.2018YFJH0703)the Consulting Research Project of Shandong Research Institute,China Engineering Science and Technology Development Strategy(No.202103SDYB08)the‘First Class Fishery Discipline’Programme in Shandong Provincethe special top talent plan‘One Thing One Decision(YishiYiyi)’Programme in Shandong Province,China。
文摘MicroRNAs(miRNAs),22-nucleotide-long micromanagers that guide the post-transcriptional regulation of a wide range of target genes,can theoretically be used as a diagnostic or therapeutic target for inflammatory reaction.In fish,miR-144-5p expression varies dramatically in response to the different bacterial infections and can regulate immunity-related genes to reduce the occurrence of inflammation.In this research,the regulation function of miR-144-5p to the intestinal innate immunity was udied in flounder Paralichthys olivaceus.The flounders were interfered by 2μg g^(-1) miR-144-5p antagomir and their tissues(intestine,liver and spleen)were harvested from the fish at 12 h post-injection.More than 60 million high-quality reads were collected.At 24 hours after miR-144-5p or miR-NC interference,a total of 2704 and 1823 different-expresion genes(DEGs)were identified in comparison with control group,respectively.The DEGs were enriched in a variety of immunity-related signaling pathways,including NOD-like receptor,Wnt and Toll-like receptor signaling pathways,according to GO and KEGG analyses.A total of 503 highly interacting DEGs engaged in 33 immunity-related signaling pathways were discovered using KEGG analyses.Additionally,5 hub genes were found by protein-protein interaction networks,which formed an intricate immune regulation network.Meanwhile,these hub genes were mostly involved in focal adhesion,Wnt signaling pathway,as well as the Intestinal Immune Network for IgT(IgA)Production Pathway.In conclusion,the loss of miR-144-5p can affect immunity-related genes and downstream signaling pathways.Our findings suggest that miR-144-5p is a modulator of gene networks and signaling pathways associated with intestinal innate immunity.
文摘A library rich in CA/GT microsatellites was constructed from the Paralichthys olivaceus genome by combining biotin capture method and radioactive labeling hybridization. Five hundred and twenty six positive clones were obtained through twice screens. Sequencing confirmed 133 microsatellite loci （number of repeats t〉 5） in 119 positive clones. Of these microsatellites, two （1.5%） had compound repeat motifs, 63 （47.37%） had perfect motifs and 68 （51.13%） had imperfect motifs. Primer pairs were designed in the flanking regions of 22 microsatelites and subjected to PCR amplification. In 8 artificial gynogenesis families, four pairs failed to amplification, one pair was monomorphic, and the rest were polymorphic with an average of 5.2 alleles per locus. Heterozygosities ranged between 0. 375 and 0. 846, PIC ranged between 0. 305 and 0. 823. The results suggested that most of the microsatellites we isolated were qualified to be applied to the population genetic studies of P. olivaceus.
文摘Edwardsiellosis of flounder and turbot occurring in different mariculture farms during 2001～ 2004 was examined, including the conditions of disease occurrence, clinical signs and pathological changes. The results showed that all diseased fishes expressed bacterial septicaemia. A total of 148 strains were identified using a combination of traditional physiological and biochemical tests and partial 16S rRNA gene analysis. In addition, the mole fraction G ＋ C ratio of the DNA of representative strain of isolates and serum homology were detected, and pathogenicity tests of isolates were conducted by experimental infection. The results revealed that 148 strains were identified as E. tarda of genus Edwardsiella, all the isolates are of serologic similarity, and have strong pathogenicity to flounder and turbot.
