Identification of natural substances with antioxidant properties is ongoing research for addressing issues related to oxidative stress especially attributed to environmental effects.Our previous study demonstrated tha...Identification of natural substances with antioxidant properties is ongoing research for addressing issues related to oxidative stress especially attributed to environmental effects.Our previous study demonstrated that Lateolabrax japonicus peptides(LPH),rich in Glu,Gly,and hydrophobic amino acids,exhibited remarkable antioxidant activity in vitro,with though its action mechanism yet to be revealed.Therefore,to assess the in vivo antioxidative properties of LPH,we employed H_(2)O_(2) to generate oxidative stress in Drosophila melanogaster model.Results indicated that LPH significantly prolonged the lifespan of Drosophila subjected to oxidative stress mostly mediated via LPH’s enhancement of the antioxidant defense system and intestinal functions.Antioxidant effects were manifested by a decrease in malondialdehyde(MDA)levels,elevated superoxide dismutase(SOD),catalase(CAT),and glutathione peroxidase(GSH-Px)activities,decreased levels of reactive oxygen species(ROS)in intestinal epithelial cells,and the preservation of intestinal length.LPH effectively controlled the excessive proliferation and differentiation of oxidative stress-induced Drosophila intestinal stem cells.At the gene level,LPH upregulated the expression of antioxidant-related Nrf2 genes while concurrently downregulated mTOR expression level.Furthermore,high-throughput 16S rDNA sequencing revealed that the addition of LPH significantly influenced the diversity and abundance of the intestinal microbiota in H_(2)O_(2)-induced Drosophila.These findings provide a deeper understanding of the antioxidative mechanism of LPH,suggesting its potential applications in food industry and to be assessed using other in vivo oxidative stress models.展开更多
The Chinese seabass(Lateolabrax maculatus)is one of the most popular and valuable aquaculture species in China.Recently,the disease caused by Vibrio anguillarum has brought huge economic losses in the L.maculatus indu...The Chinese seabass(Lateolabrax maculatus)is one of the most popular and valuable aquaculture species in China.Recently,the disease caused by Vibrio anguillarum has brought huge economic losses in the L.maculatus industry.However,the immune response of L.maculatus after V.anguillarum infection remains unknown.In this study,the blood homeostasis,gut microbiota and transcriptomic profiling of L.maculatus after V.anguillarum infection were investigated.Our results indicated that the levels of superoxide dismutase(SOD),alanine aminotransferase(ALT)and total bilirubin(TBIL)increased,while the levels of blood glucose(BG),total protein(TP)and albumin(ALB)decreased after V.anguillarum infection.The analysis of the gut microbiota composition revealed that the dominant phyla was Firmicutes and Proteobacteria,and the relative abundance of genus Vibrio increased after V.anguillarum infection.Subsequently,the differentially expressed genes(DEGs)in the kidney and spleen after V.anguillarum infection were analyzed by transcriptome sequencing.The results indicated that immunity-related genes like TLR5,TLR8,TLR9,IL-1β,CCL3,IFNγ,CXCL11 and TNFαwere affected and the NOD-like receptor signaling pathway,cytokine-cytokine receptor interaction and Toll-like receptor signaling were activated.Thus,an effective immune and pro-flammatory response can help resist V.anguillarum infection.Our results provide a theoretical support for improving the disease resistance ability of L.maculatus.展开更多
生长激素受体(GHR)作为GH/IGF轴的中心环节,在内分泌调控中发挥重要作用。本实验采用c DNA末端快速扩增法(RACE)技术克隆出花鲈GHR1和GHR2的c DNA全长序列。急性低盐度调控实验设为海水组、半海水组和淡水组。测定了急性低盐度调控24、4...生长激素受体(GHR)作为GH/IGF轴的中心环节,在内分泌调控中发挥重要作用。本实验采用c DNA末端快速扩增法(RACE)技术克隆出花鲈GHR1和GHR2的c DNA全长序列。急性低盐度调控实验设为海水组、半海水组和淡水组。测定了急性低盐度调控24、48、96、144、192h后,花鲈肝脏中GHRs、IGF-1及垂体GH表达情况。结果表明,GHR1 c DNA全长序列2436bp,编码637个氨基酸;GHR2 c DNA全长序列2940bp,编码582个氨基酸。GHR1与GHR2由信号肽、胞外区、跨膜区、胞内区组成,且结构存在差异。脑、肾、鳃中GHR1表达明显高于GHR2;而在肌肉、垂体、肝脏、盲肠、胸腺、心脏中,GHR2表达明显高于GHR1。24h时,各组GHR1表达不变,GHR2、GH、IGF-1显著下降。之后,相对于海水组,淡水组和半海水组GHRs和IGF-1表达升高,而GH下降,GH与GHR负相关。据结果推测,花鲈GHR2可能为SL受体,GH/IGF轴参与低渗调控可能是通过增加GHRs,进而激活下游IGF-1表达而实现。展开更多
A homologue of the lower vertebrates translationally controlled tumor protein (TCTP) was cloned from the marine fish Japanese sea perch (Lateolabrax japonicus) by the technology of homology cloning. The full-length cD...A homologue of the lower vertebrates translationally controlled tumor protein (TCTP) was cloned from the marine fish Japanese sea perch (Lateolabrax japonicus) by the technology of homology cloning. The full-length cDNA sequence of the sea perch TCTP gene contained a 5' untranslated region (UTR) of 47 bp, a 3' UTR of 433 bp, and a putative open reading frame (ORF) of 510 bp encoding a polypeptide of 170 amino acids. The deduced amino acid sequence of the sea perch TCTP gene showed a high similarity to that of zebrafish, rohu, rabbit, chicken and human. Sequence analysis revealed there were a signature sequence of TCTP family, an N-glycosylation site, and five Casein kinase phosphorylation sites in the sea perch TCTP. The temporal expression of TCTP genes in healthy and lipopolysaccharide (LPS) challenged fishes was measured by semi-quantitative reverse transcription-PCR (RT-PCR). The results indicated that LPS could up-regulate the expression of sea perch TCTP in the examined tissues, including head-kidney, spleen and liver.展开更多
The Na^+/K^+/2Cl^-cotransporter(NKCC)and the cystic fibrosis transmembrane conductance regulator(CFTR)proteins play crucial roles in the transportation of Na^+and Cl^-.In this study,we identified cftr,nkcc1 a,nkcc1 b ...The Na^+/K^+/2Cl^-cotransporter(NKCC)and the cystic fibrosis transmembrane conductance regulator(CFTR)proteins play crucial roles in the transportation of Na^+and Cl^-.In this study,we identified cftr,nkcc1 a,nkcc1 b and nkcc2 in spotted sea bass(Lateolabrax maculatus)genomic and transcriptomic databases.We also characterized these genes via phylogenetic and structural analyses.The results showed that both cftr and nkcc were highly conservative in L.maculatus.Quantitative real-time polymerase chain reaction(qRT-PCR)analysis in ten tissues showed that cftr,nkcc1 a and nkcc2 highly express in osmoregulatory organs such as gill,kidney and intestine.Furthermore,the expressions of cftr and nkcc1 a in gill as well as nkcc2 in intestine were up-regulated by high salinity,indicating that these genes function potentially in osmoregulation.Our findings provided the insights into the cftr and nkcc functions in euryhaline teleost.展开更多
The t-SNARE protein SNAP-25 (synaptosome-associated protein of 25 kDa) plays an essential role in regulating fusion between the vesicle and plasma membranes during exocytosis. To clone and characterize SNAP-25 gene, t...The t-SNARE protein SNAP-25 (synaptosome-associated protein of 25 kDa) plays an essential role in regulating fusion between the vesicle and plasma membranes during exocytosis. To clone and characterize SNAP-25 gene, the first step in the functional study of SNARE proteins in marine teleostean, was to obtain the cDNA of sea perch SNAP-25 (SPsn25) by RT-PCR and RACE-PCR amplification of a Japanese sea perch. The full-length cDNA of 831bp contains a CDS of 615 bp, coding 204 amino acid residues, and a 5′UTR of 219bp. Bioinformatic analysis revealed that SPsn25 corresponds with SNAP-25a isoform and shares 91.1% identity with SNAP-25a of a goldfish and a zebrafish. The SPsn25 expression in both mRNA and protein levels in the Japanese sea perch had been identified through semi-quantitative RT-PCR and Western Blot assay. Together, these data again confirmed the nerve tissue specificity of the fish SNAP-25 gene expression.展开更多
基金supported by National Key R&D Program of China(2023YFD2100205)Fuzhou Science&Technology Project,China(2022-Y-0022022-P-023).
文摘Identification of natural substances with antioxidant properties is ongoing research for addressing issues related to oxidative stress especially attributed to environmental effects.Our previous study demonstrated that Lateolabrax japonicus peptides(LPH),rich in Glu,Gly,and hydrophobic amino acids,exhibited remarkable antioxidant activity in vitro,with though its action mechanism yet to be revealed.Therefore,to assess the in vivo antioxidative properties of LPH,we employed H_(2)O_(2) to generate oxidative stress in Drosophila melanogaster model.Results indicated that LPH significantly prolonged the lifespan of Drosophila subjected to oxidative stress mostly mediated via LPH’s enhancement of the antioxidant defense system and intestinal functions.Antioxidant effects were manifested by a decrease in malondialdehyde(MDA)levels,elevated superoxide dismutase(SOD),catalase(CAT),and glutathione peroxidase(GSH-Px)activities,decreased levels of reactive oxygen species(ROS)in intestinal epithelial cells,and the preservation of intestinal length.LPH effectively controlled the excessive proliferation and differentiation of oxidative stress-induced Drosophila intestinal stem cells.At the gene level,LPH upregulated the expression of antioxidant-related Nrf2 genes while concurrently downregulated mTOR expression level.Furthermore,high-throughput 16S rDNA sequencing revealed that the addition of LPH significantly influenced the diversity and abundance of the intestinal microbiota in H_(2)O_(2)-induced Drosophila.These findings provide a deeper understanding of the antioxidative mechanism of LPH,suggesting its potential applications in food industry and to be assessed using other in vivo oxidative stress models.
基金received funding from the Natural Science Foundation of Shandong Province(No.ZR2023MC141)the Innovation and Entrepreneurship Training Program for College Students(No.202210435003)financial support was also provided by the‘First Class Fishery Discipline’Program and the Special Talent Program‘One Thing One Decision(Yishi Yiyi)’in Shandong Province,China。
文摘The Chinese seabass(Lateolabrax maculatus)is one of the most popular and valuable aquaculture species in China.Recently,the disease caused by Vibrio anguillarum has brought huge economic losses in the L.maculatus industry.However,the immune response of L.maculatus after V.anguillarum infection remains unknown.In this study,the blood homeostasis,gut microbiota and transcriptomic profiling of L.maculatus after V.anguillarum infection were investigated.Our results indicated that the levels of superoxide dismutase(SOD),alanine aminotransferase(ALT)and total bilirubin(TBIL)increased,while the levels of blood glucose(BG),total protein(TP)and albumin(ALB)decreased after V.anguillarum infection.The analysis of the gut microbiota composition revealed that the dominant phyla was Firmicutes and Proteobacteria,and the relative abundance of genus Vibrio increased after V.anguillarum infection.Subsequently,the differentially expressed genes(DEGs)in the kidney and spleen after V.anguillarum infection were analyzed by transcriptome sequencing.The results indicated that immunity-related genes like TLR5,TLR8,TLR9,IL-1β,CCL3,IFNγ,CXCL11 and TNFαwere affected and the NOD-like receptor signaling pathway,cytokine-cytokine receptor interaction and Toll-like receptor signaling were activated.Thus,an effective immune and pro-flammatory response can help resist V.anguillarum infection.Our results provide a theoretical support for improving the disease resistance ability of L.maculatus.
文摘生长激素受体(GHR)作为GH/IGF轴的中心环节,在内分泌调控中发挥重要作用。本实验采用c DNA末端快速扩增法(RACE)技术克隆出花鲈GHR1和GHR2的c DNA全长序列。急性低盐度调控实验设为海水组、半海水组和淡水组。测定了急性低盐度调控24、48、96、144、192h后,花鲈肝脏中GHRs、IGF-1及垂体GH表达情况。结果表明,GHR1 c DNA全长序列2436bp,编码637个氨基酸;GHR2 c DNA全长序列2940bp,编码582个氨基酸。GHR1与GHR2由信号肽、胞外区、跨膜区、胞内区组成,且结构存在差异。脑、肾、鳃中GHR1表达明显高于GHR2;而在肌肉、垂体、肝脏、盲肠、胸腺、心脏中,GHR2表达明显高于GHR1。24h时,各组GHR1表达不变,GHR2、GH、IGF-1显著下降。之后,相对于海水组,淡水组和半海水组GHRs和IGF-1表达升高,而GH下降,GH与GHR负相关。据结果推测,花鲈GHR2可能为SL受体,GH/IGF轴参与低渗调控可能是通过增加GHRs,进而激活下游IGF-1表达而实现。
基金supported by the“863"Prijetof China under contract Nos 2001AA628180 and 2002AA626020.
文摘A homologue of the lower vertebrates translationally controlled tumor protein (TCTP) was cloned from the marine fish Japanese sea perch (Lateolabrax japonicus) by the technology of homology cloning. The full-length cDNA sequence of the sea perch TCTP gene contained a 5' untranslated region (UTR) of 47 bp, a 3' UTR of 433 bp, and a putative open reading frame (ORF) of 510 bp encoding a polypeptide of 170 amino acids. The deduced amino acid sequence of the sea perch TCTP gene showed a high similarity to that of zebrafish, rohu, rabbit, chicken and human. Sequence analysis revealed there were a signature sequence of TCTP family, an N-glycosylation site, and five Casein kinase phosphorylation sites in the sea perch TCTP. The temporal expression of TCTP genes in healthy and lipopolysaccharide (LPS) challenged fishes was measured by semi-quantitative reverse transcription-PCR (RT-PCR). The results indicated that LPS could up-regulate the expression of sea perch TCTP in the examined tissues, including head-kidney, spleen and liver.
基金supported by the China Agriculture Research System (No. CARS-47)the National Natural Science Foundation of China (No. 31602147)
文摘The Na^+/K^+/2Cl^-cotransporter(NKCC)and the cystic fibrosis transmembrane conductance regulator(CFTR)proteins play crucial roles in the transportation of Na^+and Cl^-.In this study,we identified cftr,nkcc1 a,nkcc1 b and nkcc2 in spotted sea bass(Lateolabrax maculatus)genomic and transcriptomic databases.We also characterized these genes via phylogenetic and structural analyses.The results showed that both cftr and nkcc were highly conservative in L.maculatus.Quantitative real-time polymerase chain reaction(qRT-PCR)analysis in ten tissues showed that cftr,nkcc1 a and nkcc2 highly express in osmoregulatory organs such as gill,kidney and intestine.Furthermore,the expressions of cftr and nkcc1 a in gill as well as nkcc2 in intestine were up-regulated by high salinity,indicating that these genes function potentially in osmoregulation.Our findings provided the insights into the cftr and nkcc functions in euryhaline teleost.
基金the NSFC (No.40476060)Hi-Tech Research and Development Program of China (No. 2002AA629120)
文摘The t-SNARE protein SNAP-25 (synaptosome-associated protein of 25 kDa) plays an essential role in regulating fusion between the vesicle and plasma membranes during exocytosis. To clone and characterize SNAP-25 gene, the first step in the functional study of SNARE proteins in marine teleostean, was to obtain the cDNA of sea perch SNAP-25 (SPsn25) by RT-PCR and RACE-PCR amplification of a Japanese sea perch. The full-length cDNA of 831bp contains a CDS of 615 bp, coding 204 amino acid residues, and a 5′UTR of 219bp. Bioinformatic analysis revealed that SPsn25 corresponds with SNAP-25a isoform and shares 91.1% identity with SNAP-25a of a goldfish and a zebrafish. The SPsn25 expression in both mRNA and protein levels in the Japanese sea perch had been identified through semi-quantitative RT-PCR and Western Blot assay. Together, these data again confirmed the nerve tissue specificity of the fish SNAP-25 gene expression.
