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DNA Methylation of pacap Gene Is Involved in Growth Superiority of Female Half Smooth Tongue Sole(Cynoglossus semilaevis)in Different Salinities

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Journal of Ocean University of China 2021年第1期20卷 169-180页

作者： si yufeng HE Feng WEN Haishen LI siping HE Huiwen The Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources Ministry of EducationShanghai Ocean UniversityShanghai 201306China The Key Laboratory of Mariculture Ministry of EducationOcean University of ChinaQingdao 266003China

Pituitary adenylate cyclase activating polypeptide(PACAP)and growth hormone-releasing hormone(GHRH)play important roles in the GH/IGF growth axis in *** determine whether epigenetic change is involved in the regulation of pacap and ghrh responses to low salinity stress in Cynoglossus semilaevis,the correlation between growth traits,DNA methylation status and gene expression level in low salinity(15,S15)and optimal salinity(30,S30)at day 7(D7)and day 60(D60)were *** showed that exposure to low salinity for 60 days attenuated *** growth *** low salinity,DNA methylation level of pacap promoter increased in females and decreased in males at day 7,but inverted at day ***,pacap expression was up-regulated in both males and *** pacap promoter methylation level was higher and expression level was lower in female than in *** results suggest that pacap promoter methylation level is negatively correlated to mRNA level and positively correlated to body weight,while gene expression level is negatively related with body *** the decrease of salinity,DNA methylation level of ghrh promoter and exon1,as well as its gene expression displayed minor ***,pacap gene seems to play an important role in fish growth,contributing to female growth superiority,while ghrh gene seems not pertinent under salinity *** results indicate that low salinity potentially affects fish growth through regulating DNA methylation in pacap *** study expands the understanding of the molecular mechanism of how salinity modulates fish growth from the epigenetic perspective.

关键词： DNA methylation Cynoglossus semilaevis salinity stress pacap growth difference

Half Smooth Tongue Sole(Cynoglossus semilaevis) Under Low Salinity Stress Can Change Hepatic igf2 Expression Through DNA Methylation

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Journal of Ocean University of China 2020年第1期19卷 171-182页

作者： LI siping HE Feng WEN Haishen si yufeng LIU Mingyuan HUANG Yajuan WU Shuxian Fishery College the Key Laboratory of Mariculture of Ministry of EducationOcean University of ChinaQingdao 266003China

Salinity is a crucial environmental stress that severely affects fish growth and *** environmental stress,DNA methylation plays an important role in gene expression and genome *** better understand the epigenetic regulation mechanism of igf2 under low salinity stress,we analyzed the DNA methylation at 5’UTR,exon1,intron1,and exon2,and analyzed the relationship of DNA methylation with mRNA abundance as well as the special single CpG sites methylation patterns of igf2 in the liver of half smooth tongue sole under low salinity(15)for 0,7,and 60 *** exposed to low salinity,DNA methylation at 5’UTR and exon2 remained stable,while it was up-regulated firstly and then down-regulated at exon1 and *** single CpG sites of igf2 associated with low salinity,and most of these sites with significantly changed methylation levels(Psingle CpG sites methylation levels and igf2 expression further revealed that females and males showed different response to low ***,the 38-CpG site of intron1 servers as a sexual ***,our integrative analysis demonstrated that regional DNA of igf2 methylation had highly complex interplay on gene *** single CpG sites in intron1 were indispensable epigenetic markers under external environmental *** all,to resist the low salinity stress,half smooth tongue sole liver can regulate the expression of igf2 through methylation of CpG sites in intron1.

关键词： salinity hepatic igf2 DNA methylation gene expression half smooth tongue sole

Expression Analysis of the Insulin-Like Growth Factors Ⅰ and Ⅱ During Embryonic and Early Larval Development of Turbot(Scophthalmus maximus)

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Journal of Ocean University of China 2015年第2期14卷 309-316页

作者： WEN Haishen QI Qian HU Jian si yufeng HE Feng LI Jifang Fisheries College Ocean University of China

The insulin-like growth factors Ⅰ and Ⅱ （IGF-Ⅰ and IGF-Ⅱ） are important proteins involved in fish growth and develop- ment. Here, we report the isolation of IGF-Ⅱ and expression analysis of IGFs in turbot Scophthalmus maximus, aiming to clarify their function in embryonic and larval development of fish. The deduced IGF-Ⅱ gene is 808 bp in full length, which encodes a protein of 219 amino acids and is 93% similar with that ofParalichthys olicaceus in amino acid sequence. The tissue abundance and the ex- pression pattern of IGFs in a turbot at early development stages were investigated via reverse transcription-polymer chain reaction. Result showed that the IGF-Ⅰ and IGF-Ⅱ genes were widely expressed in tissues of S. maximus. IGF-Ⅰ was detected in all tissues ex- cept intestines with the highest level in liver, while IGF-Ⅱ transcript presented in all tissues except muscle. At the stages of embry- onic and larval development, the mRNA levels of IGFs sharply increased from the stage of unfertilized egg to post larva, followed by a decrease with larval development. However, there was an increase in IGF-Ⅰ at the embryonic stage and IGF-Ⅱ at the gastrula stage, respectively. These results suggested that IGFs play important roles in cell growth and division of the turbot. Our study provides reference data for further investigation of growth regulation in turbot, which can guarantee better understanding of the physiological role that IGFs play in fish.

关键词： Scophthalmus maximus insulin-like growth factor cloning expression analysis

Analysis of DNA Methylation Level by Methylation-Sensitive Amplification Polymorphism in Half Smooth Tongue Sole(Cynoglossus semilaevis)Subjected to Salinity Stress

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Journal of Ocean University of China 2017年第2期16卷 269-278页

作者： LI siping HE Feng WEN Haishen LI Jifang si yufeng LIU Mingyuan HE Huiwen HUANG Zhengju The Key Laboratory of Mariculture of Ministry of Education Fishery CollegeOcean University of ChinaQingdao 266003P.R.China

Increasingly arisen environmental constraints may contribute to heritable phenotypic variation including methylation changes,which can help the animals with development,growth and *** this study,we assessed the DNA methylation levels in three tissues(gonad,kidney and gill) of half smooth tongue sole under the salinity *** methylation-sensitive amplification polymorphism(MSAP) technique was applied to illustrate the regulation of epigenetic mechanism in environmental *** were subjected to 15 salinity treatment for 7 and 60 days,respectively.A total of 11259 fragments were amplified with 8 pairs of selective *** levels of methylated DNA in different tissues of females and males without salinity stress were analyzed,which were 32.76% and 47.32% in gonad;38.13% and 37.69% in kidney;37.58% and 34.96% in gill,*** addition,the significant difference was observed in gonad between females and males,indicating that discrepant regulation in gonadal development and differentiation may involve sex-related *** analysis showed that total and hemi-methylation were significantly decreased under 15 salinity for 7 days,probably resulting in up-regulating salt-tolerance genes expression to adjust salt *** the adjustment for 60 days,total and hemi-methylation prominently went back to its normal levels to obtain ***,full methylation levels were steady along with salinity stress to maintain the stability of gene ***,the data showed that gonads in females and gills in males were superior in *** a result,DNA methylation regulates tissue-specific epiloci,and may respond to salinity stress by regulating gene expression to maintain animal survival and activity.

关键词： DNA methylation diversity half smooth tongue sole salinity stress MSAP

The double mutations of acvr2aa and acvr2ba leads to muscle hypertrophy in zebrafish

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Aquaculture and Fisheries 2023年第6期8卷 706-712页

作者： Jinyuan Che Cunjie Hu Qi Wang Chunxin Fan yufeng si Xiaoling Gong Baolong Bao Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources(Shanghai Ocean University) Ministry of EducationInternational Research Center for Marine Biosciences at Shanghai Ocean UniversityMinistry of Science and TechnologyNational Demonstration Center for Experimental Fisheries Science EducationShanghai Ocean UniversityShanghai201306China

Activin A receptor,type II(Acvr2)is a member of the transforming growth factor beta receptor family and can function as a negative regulator of skeletal muscle ***2 plays an important role in regulating muscle development that can inhibit skeletal muscle growth in ***,there is very little research reported on the function of acvr2 in muscle development of *** this study,we analyzed the effect of acvr2aa and acvr2ba on muscle development in *** rates of WT and acvr2a^(-/-b-/-)􀀀were measured from juvenile stage to adult *** addition,effects of acvr2 on skeletal muscle were tested in histological,protein and molecular *** a result,acvr2a^(-/-b-/-)􀀀exhibited a wider body trunk than WT and showed a significant increase in body weight and width from two months *** analysis of skeletal muscle indicated that the size of muscle fiber in acvr2a^(-/-b-/-)􀀀(female:1809±123μm^(2);male:2261±130μm^(2))was larger than that in WT(709.8±49μm^(2);815±53μm^(2)).In addition,western blot of fast MyLc protein showed the protein synthesis of acvr2a^(-/-b-/-)􀀀are ***,Histological analysis of heart showed the ventricle area is aslo increased in acvr2a^(-/-b-/-)􀀀.Our results demonstrated acvr2 attends the development of muscle fiber and will cause muscle hypertrophy when they were knocked out in *** conclusion,acvr2 in zebrafish can control the development of muscle fibers during posthatch growth.

关键词： acvr2 Muscle fiber Hypertrophy Zebrafish Skeletal muscle growth

Methylation Status of the Follistatin Gene at Different Development Stages of Japanese Flounder(Paralichthys olivaceus)

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Journal of Ocean University of China 2018年第5期17卷 1243-1252页

作者： HUANG Yajuan HU Nan si yufeng LI siping WU Shuxian ZHANG Meizhao WEN Haishen LI Jifang LI Yun HE Feng The Key Laboratory of Mariculture of Ministry of Education Fishery CollegeOcean University of China

Follistatin （Fst） is a hyperplasia factor that plays a crucial role in muscle development. DNA methylation, a significant process, regulates gene expression. The aim of our study is to examine the DNA methylation and expression patterns ofFst gene at five different development stages of Japanese flounder （stage A, 7 dph; stage B, 90 dph; stage C, about 180 dph; stage D, about 24 months; stage E, about 36 months）. The muscle tissue of Japanese flounder was obtained at different development stages in dais experiment. DNA methylation levels in the promoter and exon 2 of Fst were determined by bisulfite sequencing, and the relative expression of the Fst gene at the five stages was measured by quantitative PCR. The results showed that the lowest methylation level was at stage A and the highest methylation level was at stage B. Moreover, the highest expression level of the Fst gene was observed at stage A. The mRNA abundance was negatively correlated with DNA methylation level. Three CpG islands in the promoter region and three CpG islands in exon 2 of Fst were found in the binding sequence of the putative transcription factor. These results offered a theoretical basis for the mechanism of Fst gene regulation to muscle development at different development stages.

关键词： Japanese flounder Follistatin DNA methylation gene expression muscle growth

Decoding the fish genome opens a new era in important trait research and molecular breeding in China

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Science China(Life Sciences) 2024年第10期67卷 2064-2083页

作者： Qian Zhou Jialin Wang Jiongtang Li Zhangfan Chen Na Wang Ming Li Lei Wang yufeng si Sheng Lu Zhongkai Cui Xuhui Liu Songlin Chen State Key Laboratory of Mariculture Biobreeding and Sustainable Goods Yellow Sea Fisheries Research InstituteChinese Academy of Fishery SciencesQingdaoShandong 266071China Laboratory for Marine Fisheries Science and Food Production Processes Qingdao Marine Science and Technology CenterQingdaoShandong 266237China Key Laboratory of Aquatic Genomics Ministry of Agriculture and Rural AffairsBeijing Key Laboratory of Fishery BiotechnologyChinese Academy of Fishery SciencesBeijing 100041China

Aquaculture represents the fastest-growing global food production sector,as it has become an essential component of the global food *** has the world's largest aquaculture industry in terms of production ***,the sustainable development of fish culture is hindered by several concerns,including germplasm degradation and disease *** practice of genomic breeding,which relies heavily on genome information and genotypephenotype relationships,has significant potential for increasing the efficiency of aquaculture *** 2014,the completion of the genome sequencing and annotation of the Chinese tongue sole signified the beginning of the fish genomics era in *** then,domestic researchers have made dramatic progress in functional genomic *** date,the genomes of more than 60 species of fish in China have been assembled and *** on these reference genomes,evolutionary,comparative,and functional genomic studies have revolutionized our understanding of a wide range of biologically and economically important traits of fishes,including growth and development,sex determination,disease resistance,metamorphosis,and ***,genomic tools and breeding techniques such as SNP arrays,genomic selection,and genome editing have greatly accelerated genetic improvement through the incorporation of functional genomic information into breeding *** review aims to summarize the current status,advances,and perspectives of the genome resources,genomic study of important traits,and genomic breeding techniques of fish in *** review will provide aquaculture researchers,fish breeders,and farmers with updated information concerning fish genomic research and breeding *** summary will help to promote the genetic improvement of production traits and thus will support the sustainable development of fish aquaculture.

关键词： fish genomics economic traits genomic selection genome editing genomic breeding

钇对Ti-23Al-25Nb合金热处理组织及性能的影响

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稀有金属材料与工程 2006年第4期35卷 655-658页

作者：陈玉勇司玉锋孔凡涛陈子勇哈尔滨工业大学黑龙江哈尔滨150001

采用DTA分析Ti-23Al-25Nb合金和Ti-23Al-25Nb-0.36Y合金的相变特征,并根据DTA曲线特征制定热处理工艺,用SEM,XRD分析热处理后合金试样的显微组织及相组成,用TEM分析原始组织中钇的存在形式,用Instron-5569万能材料试验机进行室温压缩试... 详细信息

采用DTA分析Ti-23Al-25Nb合金和Ti-23Al-25Nb-0.36Y合金的相变特征,并根据DTA曲线特征制定热处理工艺,用SEM,XRD分析热处理后合金试样的显微组织及相组成,用TEM分析原始组织中钇的存在形式,用Instron-5569万能材料试验机进行室温压缩试验。结果表明,钇在原始组织中以Y2O3形式存在;钇没有改变Ti-23Al-25Nb合金的相变温度;加入钇后,在1320℃淬火,B2相晶粒比较粗大,随着淬火温度的降低,O相增加,并且O相板条变细而杂乱,有利于提高合金的塑性和抗蠕变性能;加入钇后,合金强度提高,原因是第二相粒子Y2O3弥散强化的结果。

关键词： Ti-23Al-25Nb合金钇差热分析(DTA) 显微组织性能

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水流对斑马鱼体色和肥满度的影响

上海海洋大学学报 2024年第5期33卷 1064-1073页

作者：丁长晨黄亚娟司玉凤鲍宝龙上海海洋大学水产种质资源发掘与利用教育部重点实验室上海201306 上海海洋大学水产科学国家级实验教学示范中心上海201306

为了探究水流对鱼类体色的影响,比较了不同流速水流养殖下斑马鱼皮肤黑色素细胞数量和基因的表达,结果表明:相对于低流速(0.010 m/s),在高流速(0.022 m/s)下养殖斑马鱼3个月后,皮肤黑色素细胞数量显著增加,尤其是鱼体尾部区域;实时荧光... 详细信息

为了探究水流对鱼类体色的影响,比较了不同流速水流养殖下斑马鱼皮肤黑色素细胞数量和基因的表达,结果表明:相对于低流速(0.010 m/s),在高流速(0.022 m/s)下养殖斑马鱼3个月后,皮肤黑色素细胞数量显著增加,尤其是鱼体尾部区域;实时荧光定量PCR(RT-qPCR)结果显示高流速组斑马鱼皮肤黑色素细胞标记基因kita、mitfa和tyrp1a相对表达量显著升高,黄色素细胞标记基因csf1ra显著降低,虹彩细胞标记基因pnp4a无显著变化,表明高流速水流诱导黑色素细胞而抑制黄色素细胞的形成。RT-qPCR结果显示,相比于低流速组,高流速组斑马鱼皮肤中黑色素细胞形成抑制因子基因asip1的相对表达量显著减少,与黑色素细胞形成相关的黑皮质素受体基因mc1r、阿黑皮素原基因pomca无显著性变化,视黄酸合成相关的视黄醛脱氢酶基因raldh2和raldh3显著下调,提示水流刺激可通过降低asip1的表达来诱导黑色素细胞形成;水流刺激瞬时受体通道蛋白基因trpv4和压电式机械敏感通道蛋白基因piezo2的表达上调,而piezo1无显著性变化。以上结果表明,水流能诱导斑马鱼黑色素细胞的形成,可能是通过皮肤机械力感受蛋白Trpv4或Piezo2介导水流刺激减少asip1表达的结果。Asip1除了调控黑色素细胞形成以外还调控脂肪的积累,相比于低流速组,高流速组斑马鱼肥满度也显著下降,GO富集分析显示差异表达基因主要富集到甘油三酯分解代谢、高密度脂蛋白颗粒、脂质结合等条目,差异表达基因显著富集的KEGG通路有:氨基糖和核苷酸糖的代谢、PPAR信号通路、亚油酸代谢、花生四烯酸代谢等。RT-qPCR检测与脂肪代谢相关的脂肪酸过饱和酶2基因fads2显著上调,瘦素基因lep无显著性差异,提示水流可能通过Asip1影响Fads2进而影响脂肪降解。本研究首次报道了水流对鱼类体色的影响,为理解目前越来越普遍的流水养殖对鱼类体色和肥满度的影响提供了初步的理论基础。

关键词：水流斑马鱼体色肥满度机械力感受蛋白