饥饿及复投喂对金钱鱼肠型脂肪酸结合蛋白基因表达的影响
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摘要
为阐明肠型脂肪酸结合蛋白基因(ifabp)在金钱鱼(Scatophagus argus)脂肪代谢中的作用,从金钱鱼肝脏转录组中得到ifabp的unigene片段,设计特异引物克隆了金钱鱼2种亚型ifabp基因(ssifabp2a和ssifabp2b),并分析了这2种基因在雌、雄鱼中的组织分布以及饥饿及复投喂后肝脏和肠道中的表达变化。聚类结果表明,ssifabp2a与其他硬骨鱼类Ifabp2a、Ifabp或IfabpX1聚为一类, ssifabp2b则与其他鱼类的Ifabp2b或Ifabp-like聚为一类。同源性比较发现, ssifabp2a与其他硬骨鱼类Ifabp2a、Ifabp或IfabpX1的同源性为78.8%—87.9%;ssifabp2b与其他硬骨鱼类Ifabp2b或Ifabp-like的同源性为79.5%—87.9%; ssifabp2a与ssifabp2b的同源性为73.5%。RT-PCR发现:在雄鱼中, ssifabp2a在小肠中表达最强,在肾和肝脏等表达较弱; ssifabp2b也在小肠中表达最强,在肝脏、胃和下丘脑等较弱。在雌鱼中, ssifabp2a在胃中表达最强,在肾、肝脏和下丘脑组织表达较弱,在其他组织中有微弱表达,脑垂体中没有检测到表达。与ssifabp2a表达情况不同, ssifabp2b在下丘脑、卵巢、心脏、肠中表达较强,其他组织中有微弱表达,鳃中没有检测到表达。饥饿及复投喂结果表明:在肠中,饥饿2d后, ssifabp2a表达量显著降低, ssifabp2b无显著性变化;饥饿7d后, ssifabp2a表达量显著下降,但ssifabp2b无显著性变化;在复投喂后,与7d饥饿相比较, ssifabp2a和ssifabp2b的表达量均显著升高。在肝脏中,饥饿2d后, ssifabp2a表达量无显著变化,而ssifabp2b的表达量显著升高;饥饿7d后, ssifabp2a和ssifabp2b的表达量均显著升高;在复投喂后, ssifabp2a和ssifabp2b表达均显著下降,恢复到正常水平。结果表明,饥饿及复投喂对金钱鱼肝脏和肠道中的ssifabp2a和ssifabp2b的表达具有显著影响,表明两者都参与了金钱鱼的脂肪代谢调节。
To elucidate the role of intestinal fatty acid binding protein(Ifabp) in the regulation of lipids metabolism,unigenes were obtained from transcriptome of liver in spotted scat, Scatophagus argus. Two subtypes of ifabp genes(ssifabp2a and ssifabp2b) were isolated and analyzed in the female and male S. argus, respectively. Tissue distributions and changes of ssifabp2a and ssifabp2b in fast feeding and re-feeding were also observed. Phylogenetic tree results showed that ssIfabp2a was clustered with other Ifabp2a, Ifabp or IfabpX1, while ssIifabp2b was clustered with Ifabp2b or Ifabp-like in Osteichthyes. Homology analysis revealed that the sequence identity of ssifabp2a was78.8%—87.9% with other Osteichthyes Ifabp2a, Ifabp or IfabpX1. The sequence identity of ssifabp2b was79.5%—87.9% with other Osteichthyes Ifabp2b or Ifabp-like. The sequence identity was 73.5% between ssifabp2a and ssifabp2b. RT-PCR showed that ssifabp2a was the highest in intestine, and had a moderate level in kidney and liver.And ssifabp2b was also the highest in intestine, but had a moderate level in liver, stomach and hypothalamus in male.However, the expression of ssifabp2a was the highest in stomach, and moderate in kidney, liver and hypothalamus,with a weak expression level in other tissues and no expression in pituitary. The ssifabp2b was expressed strongly in hypothalamus, ovary, heart and intestine, and weakly in other tissues, but had no expression in gill of females. In the intestine, the expression of ssifabp2a decreased significantly, but there was no significant change of ssifabp2b after 2 d of food deprivation. The expression of ssifabp2a decreased significantly compared with the control group, but there was no significant difference on the expression of ssifabp2b within the 7 day fasting group. The expressions of ssifabp2a and ssifabp2b increased significantly with refeeding 3-h after the scheduled feeding time. In liver, the expression of ssifabp2a was not changed, but the expression of ssifabp2b increased significantly after 2-day of food deprivation.However, the ssifabp2a and ssifabp2b were all increased during the 7-day fasting, and decreased significantly with refeeding 3-h after the scheduled feeding time. In summary, ssIfabp2a and ssIfabp2b are involved in the regulation of lipids metabolism at liver and intestine in Scatophagus argus.
To elucidate the role of intestinal fatty acid binding protein(Ifabp) in the regulation of lipids metabolism,unigenes were obtained from transcriptome of liver in spotted scat, Scatophagus argus. Two subtypes of ifabp genes(ssifabp2a and ssifabp2b) were isolated and analyzed in the female and male S. argus, respectively. Tissue distributions and changes of ssifabp2a and ssifabp2b in fast feeding and re-feeding were also observed. Phylogenetic tree results showed that ssIfabp2a was clustered with other Ifabp2a, Ifabp or IfabpX1, while ssIifabp2b was clustered with Ifabp2b or Ifabp-like in Osteichthyes. Homology analysis revealed that the sequence identity of ssifabp2a was78.8%—87.9% with other Osteichthyes Ifabp2a, Ifabp or IfabpX1. The sequence identity of ssifabp2b was79.5%—87.9% with other Osteichthyes Ifabp2b or Ifabp-like. The sequence identity was 73.5% between ssifabp2a and ssifabp2b. RT-PCR showed that ssifabp2a was the highest in intestine, and had a moderate level in kidney and liver.And ssifabp2b was also the highest in intestine, but had a moderate level in liver, stomach and hypothalamus in male.However, the expression of ssifabp2a was the highest in stomach, and moderate in kidney, liver and hypothalamus,with a weak expression level in other tissues and no expression in pituitary. The ssifabp2b was expressed strongly in hypothalamus, ovary, heart and intestine, and weakly in other tissues, but had no expression in gill of females. In the intestine, the expression of ssifabp2a decreased significantly, but there was no significant change of ssifabp2b after 2 d of food deprivation. The expression of ssifabp2a decreased significantly compared with the control group, but there was no significant difference on the expression of ssifabp2b within the 7 day fasting group. The expressions of ssifabp2a and ssifabp2b increased significantly with refeeding 3-h after the scheduled feeding time. In liver, the expression of ssifabp2a was not changed, but the expression of ssifabp2b increased significantly after 2-day of food deprivation.However, the ssifabp2a and ssifabp2b were all increased during the 7-day fasting, and decreased significantly with refeeding 3-h after the scheduled feeding time. In summary, ssIfabp2a and ssIfabp2b are involved in the regulation of lipids metabolism at liver and intestine in Scatophagus argus.
引文
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[11]Cui D,Liu Z W,Liu N X,et al.Histological study on the gonadal development of Scatophagus argus[J].Journal of Fisheries of China,2013,37(5):696-704[崔丹,刘志伟,刘南希,等.金钱鱼性腺发育及其组织结构观察.水产学报,2013,37(5):696-704]
[12]Zhang M Z,Li G L,Zhu C H,et al.Effects of fish oil on ovarian development in spotted scat(Scatophagus argus)[J].Animal Reproduction Science,2013,141(1-2):90-97
[13]Li G L,Zhang M Z,Deng S P,et al.Effects of temperature and fish oil supplementation on ovarian development and foxl2 mRNA expression in spotted scat Scatophagus argus[J].Journal of Fish Biology,2015,86(1):248-260
[14]Cui X F,Zhao Y,Chen H P,et al.Cloning,expression and functional characterization on vitellogenesis of estrogen receptors in Scatophagus argus[J].General&Comparative Endocrinology,2017,246:37-45
[15]Deng S P,Wu B,Zhu C H,et al.Molecular cloning and dimorphic expression of growth hormone(gh)in female and male spotted scat Scatophagus argus[J].Fisheries Science,2014,80(4):715-723
[16]Wu B,Ye M,Chen L L,et al.Growth performance and digestive enzyme activity between male and female Scatophagus argus[J].Journal of Shanghai Ocean University,2013,22(4):545-551[吴波,叶满,陈娈娈,等.不同性别养殖金钱鱼生长性能及消化酶活性的比较.上海海洋大学学报,2013,22(4):545-551]
[17]Deng S P,Chen H P,Zhai Y,et al.Molecular cloning,characterization and expression analysis of spexin in spotted scat(Scatophagus argus)[J].General and Comparative Endocrinology,2018,266:60-66
[18]Wang M,Chen H P,Jiang D N,et al.Effect of hunger and refeeding on the expression of Scatophagus argus’s four igfbp genes[J].Journal of Hainan Tropical Ocean University,2017,24(5):1-7[王妹,陈华谱,邓思平,等.饥饿和复投喂对金钱鱼四种igfbp基因表达的影响.海南热带海洋学院,2017,24(5):1-7]
[19]Chen X W,Jiang S,Shi Z Y.Identification and expression analysis of fabp2 gene from common carp Cyprinus carpio[J].Journal of Fish Biology,2012,80(3):679-691
[20]Qin C J,Shao T,Yang J P,et al.The effect of starvation on lipid metabolism of darkbarbel catfish,Pelteobagrus vachelli[J].Acta Hydrobiologica Sinica,2015,39(1):58-65[覃川杰,邵婷,杨洁萍,等.饥饿胁迫对瓦氏黄颡鱼脂肪代谢的影响.水生生物学报,2015,39(1):58-65]
[21]Bay?r M,Bay?r A,Wright J M.Divergent spatial regulation of duplicated fatty acid-binding protein(fabp)genes in rainbow trout(Oncorhynchus mykiss)[J].Comparative Biochemistry and Physiology-Part D:Genomics and Proteomics,2015,14:26-32
[22]Cai Z P,Wang Y,Hu J W,et al.Sequence,linkage mapping and early developmental expression of the intestinaltype fatty acid-binding protein gene(fabp2)from zebrafish(Danio rerio)[J].Journal of Tropical Oceanography,2010,29(5):180-185[蔡泽平,王毅,胡家玮,等.金钱鱼繁殖生物学及诱导产卵试验.热带海洋学报,2010,29(5):180-185]
[23]Hughes A L,Piontkivska H.Evolutionary diversification of the avian fatty acid-binding proteins[J].Gene,2011,490(1):1-5
[24]Yamamoto T,Yamamoto A,Watanabe M,et al.Classification of FABP isoforms and tissues based on quantitative evaluation of transcript levels of these isoforms in various rat tissues[J].Biotechnology Letters,2009,31(11):1695
[25]Kaikaus R M,Bass N M,Ockner R K.Functions of fatty acid binding proteins[J].Experientia,1990,46(6):617-630
[26]Trevaskis N L,Lo C M,Li Y M,et al.An Acute and Coincident Increase in FABP expression and lymphatic lipid and drug transport occurs during intestinal infusion of lipid-based drug formulations to rats[J].Pharmaceutical Research,2006,23(8):1786-1796
[27]Pu L,Igbavboa U,Wood W G,et al.Expression of fatty acid binding proteins is altered in aged mouse brain[J].Molecular&Cellular Biochemistry,1999,198(1-2):69-78
[28]Shi Y B,Hayes W P.Thyroid hormone-dependent regulation of the intestinal fatty acid-binding protein gene during amphibian metamorphosis[J].Developmental Biology,1994,161(1):48-58
[29]Chmurzyńska A.The multigene family of fatty acid-binding proteins(FABPs):function,structure and polymorphism[J].Journal of Applied Genetics,2006,47(1):39-48
[30]Tian J,Wen H,Zeng L B,et al.Changes in the activities and mRNA expression levels of lipoprotein lipase(LPL),hormone-sensitive lipase(HSL)and fatty acid synthetase(FAS)of Nile tilapia(Oreochromis niloticus)during fasting and re-feeding[J].Aquaculture,2013,400-401(6):29-35
[2]Ockner R K,Manning J A,Poppenhausen R B,et al.Abinding protein for fatty acids in cytosol of intestinal mmucosa,liver,myocardium,and other tissues[J].Science,1972,177(4043):56-58
[3]Pierce M,Wang Y,Denovanwright E M,et al.Nucleotide sequence of a cDNA clone coding for an intestinaltype fatty acid binding protein and its tissue-specific expression in zebrafish(Danio rerio)[J].BBA-Gene Structure and Expression,2000,1490(1):175-183
[4]Zhang F,Lücke C,Baier L J,et al.Solution structure of human intestinal fatty acid binding protein:Implications for ligand entry and exit[J].Journal of Biomolecular NMR,1997,9(3):213-228
[5]Klapper M,B?hme M,Nitz I,et al.Transcriptional regulation of the fatty acid binding protein 2(FABP2)gene by the hepatic nuclear factor 1 alpha(HNF-1alpha)[J].Gene,2008,416(1):48-52
[6]Huang H L,Xue L Y,Wang J,et al.Effects of fasting and re-feeding on the expression of I-FABPb in Larimichthys crocea[J].Chinese Journal of Cell Biology,2016,38(4):397-404[黄红丽,薛良义,王俊.饥饿及复投喂对大黄鱼肠型脂肪酸结合蛋白b基因表达的影响.中国细胞生物学学报,2016,38(4):397-404]
[7]Xia Z L,Yu J H,Li H X,et al.Sequences analysis of jlFABP2 and the correlation between polymorphisms and body weight gain in Cyprinus carpio var.jian[J].Hereditas(Beijing),2013,35(5):628-636[夏正龙,俞菊华,李红霞,等.建鲤肠型脂肪酸结合蛋白基因的分离及其SNPs与增重的相关分析.遗传,2013,35(5):628-636]
[8]Sharma M K,Denovan-Wright E M,Degrave A,et al.Sequence,linkage mapping and early developmental expression of the intestinal-type fatty acid-binding protein gene(fabp2)from zebrafish(Danio rerio)[J].Comparative Biochemistry&Physiology Part B,2004,138(4):391-398
[9]Venold F F,Penn M H,Thorsen J.Intestinal fatty acid binding protein(fabp2)in Atlantic salmon(Salmo salar):Localization and alteration of expression during development of diet induced enteritis[J].Comparative Biochemistry&Physiology Part A,2013,164(1):229-240
[10]Lan G B,Yan B,Liao S M,et al.Biology of spotted scat Scatophagus argus:a review[J].Fisheries Science,2005,24(7):39-41[兰国宝,阎冰,廖思明,等.金钱鱼生物学研究及回顾.水产科学,2005,24(7):39-41]
[11]Cui D,Liu Z W,Liu N X,et al.Histological study on the gonadal development of Scatophagus argus[J].Journal of Fisheries of China,2013,37(5):696-704[崔丹,刘志伟,刘南希,等.金钱鱼性腺发育及其组织结构观察.水产学报,2013,37(5):696-704]
[12]Zhang M Z,Li G L,Zhu C H,et al.Effects of fish oil on ovarian development in spotted scat(Scatophagus argus)[J].Animal Reproduction Science,2013,141(1-2):90-97
[13]Li G L,Zhang M Z,Deng S P,et al.Effects of temperature and fish oil supplementation on ovarian development and foxl2 mRNA expression in spotted scat Scatophagus argus[J].Journal of Fish Biology,2015,86(1):248-260
[14]Cui X F,Zhao Y,Chen H P,et al.Cloning,expression and functional characterization on vitellogenesis of estrogen receptors in Scatophagus argus[J].General&Comparative Endocrinology,2017,246:37-45
[15]Deng S P,Wu B,Zhu C H,et al.Molecular cloning and dimorphic expression of growth hormone(gh)in female and male spotted scat Scatophagus argus[J].Fisheries Science,2014,80(4):715-723
[16]Wu B,Ye M,Chen L L,et al.Growth performance and digestive enzyme activity between male and female Scatophagus argus[J].Journal of Shanghai Ocean University,2013,22(4):545-551[吴波,叶满,陈娈娈,等.不同性别养殖金钱鱼生长性能及消化酶活性的比较.上海海洋大学学报,2013,22(4):545-551]
[17]Deng S P,Chen H P,Zhai Y,et al.Molecular cloning,characterization and expression analysis of spexin in spotted scat(Scatophagus argus)[J].General and Comparative Endocrinology,2018,266:60-66
[18]Wang M,Chen H P,Jiang D N,et al.Effect of hunger and refeeding on the expression of Scatophagus argus’s four igfbp genes[J].Journal of Hainan Tropical Ocean University,2017,24(5):1-7[王妹,陈华谱,邓思平,等.饥饿和复投喂对金钱鱼四种igfbp基因表达的影响.海南热带海洋学院,2017,24(5):1-7]
[19]Chen X W,Jiang S,Shi Z Y.Identification and expression analysis of fabp2 gene from common carp Cyprinus carpio[J].Journal of Fish Biology,2012,80(3):679-691
[20]Qin C J,Shao T,Yang J P,et al.The effect of starvation on lipid metabolism of darkbarbel catfish,Pelteobagrus vachelli[J].Acta Hydrobiologica Sinica,2015,39(1):58-65[覃川杰,邵婷,杨洁萍,等.饥饿胁迫对瓦氏黄颡鱼脂肪代谢的影响.水生生物学报,2015,39(1):58-65]
[21]Bay?r M,Bay?r A,Wright J M.Divergent spatial regulation of duplicated fatty acid-binding protein(fabp)genes in rainbow trout(Oncorhynchus mykiss)[J].Comparative Biochemistry and Physiology-Part D:Genomics and Proteomics,2015,14:26-32
[22]Cai Z P,Wang Y,Hu J W,et al.Sequence,linkage mapping and early developmental expression of the intestinaltype fatty acid-binding protein gene(fabp2)from zebrafish(Danio rerio)[J].Journal of Tropical Oceanography,2010,29(5):180-185[蔡泽平,王毅,胡家玮,等.金钱鱼繁殖生物学及诱导产卵试验.热带海洋学报,2010,29(5):180-185]
[23]Hughes A L,Piontkivska H.Evolutionary diversification of the avian fatty acid-binding proteins[J].Gene,2011,490(1):1-5
[24]Yamamoto T,Yamamoto A,Watanabe M,et al.Classification of FABP isoforms and tissues based on quantitative evaluation of transcript levels of these isoforms in various rat tissues[J].Biotechnology Letters,2009,31(11):1695
[25]Kaikaus R M,Bass N M,Ockner R K.Functions of fatty acid binding proteins[J].Experientia,1990,46(6):617-630
[26]Trevaskis N L,Lo C M,Li Y M,et al.An Acute and Coincident Increase in FABP expression and lymphatic lipid and drug transport occurs during intestinal infusion of lipid-based drug formulations to rats[J].Pharmaceutical Research,2006,23(8):1786-1796
[27]Pu L,Igbavboa U,Wood W G,et al.Expression of fatty acid binding proteins is altered in aged mouse brain[J].Molecular&Cellular Biochemistry,1999,198(1-2):69-78
[28]Shi Y B,Hayes W P.Thyroid hormone-dependent regulation of the intestinal fatty acid-binding protein gene during amphibian metamorphosis[J].Developmental Biology,1994,161(1):48-58
[29]Chmurzyńska A.The multigene family of fatty acid-binding proteins(FABPs):function,structure and polymorphism[J].Journal of Applied Genetics,2006,47(1):39-48
[30]Tian J,Wen H,Zeng L B,et al.Changes in the activities and mRNA expression levels of lipoprotein lipase(LPL),hormone-sensitive lipase(HSL)and fatty acid synthetase(FAS)of Nile tilapia(Oreochromis niloticus)during fasting and re-feeding[J].Aquaculture,2013,400-401(6):29-35
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