灌喂氧化鱼油后黄颡鱼胃肠道黏膜黑色素细胞分化和黑色素合成途径基因的差异表达
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摘要
以黄颡鱼(Pelteobagrus fulvidraco)为实验对象,灌喂氧化鱼油、鱼油7d后,提取胃肠道黏膜总RNA,采用RNA-seq测序并做转录组分析,分析了黑色素生物合成途径关键酶(酪氨酸酶)及其相关蛋白基因、黑素体运动的3个蛋白基因、α黑素细胞刺激激素途径和WNT/β-catenin、EDN3和EDNRB、KIT及其配体KITL3个信号通路的主要蛋白基因的差异表达活性。结果显示,黄颡鱼胃肠道黏膜中存在黑色素细胞分化和发育过程、黑色素合成及其调控途径的代谢网络,通过绘制代谢网络得到了关键性酶或蛋白质的基因信息。在灌喂氧化鱼油后,控制黑色素合成途径主要基因的表达活性显著下调,可能导致黑色素合成量的不足;α-MSH激素途径主要基因差异表达上调,具备促进黑色素细胞分化和发育的调控基础;而调控黑色素细胞分化和发育的3个信号通路主要基因也有差异表达。因此,黄颡鱼受灌喂氧化鱼油的影响,黑色素细胞分化和发育过程受到较大影响,会影响到鱼体成熟的黑色素细胞的数量,同时,黑色素的生物合成量不足将导致引起黄颡鱼体色的变化。
Transcriptome analysis of yellow catfish(Pelteobagrus fulvidraco) by RNA-seq method extracting gastrointestinal mucosa Total RNA was studied in this paper. The fish were fed with oxidized fish oil and fish oil respectively for 7 d. Differential expression of genes related to melanin synthesis, melanosomes movement, melanocyte stimulating hormone-α pathway and three signal pathways controlling melanocyte differentiation were analyzed. The results showed that there were metabolic networks of melanocyte differentiation and development, melanin synthesis and regulation in the gastrointestinal mucosa. The genetic information of key enzymes and protein of melanin synthesis were introduced with plotting metabolic networks graph. The melanin content was deficient because of the genes expression level were significantly down-regulation related to controlling melanin synthesis after fed with oxidized fish oil. While melanocyte differentiation and development were improved because of the genes expression level were significantly up-regulation related to melanocyte stimulating hormone-α pathway(α-MSH). In the same time, genes expression level of WNT/β-catenin, EDN3/EDNRB and KIT/KITL signal pathways were different. In conclusion, feeding oxidized fish oil affected melanocyte differentiation and development and reduced melanocyte number which benefited to maturity in yellow catfish. Insufficient melanin biosynthesis also led to body color variation of yellow catfish.
Transcriptome analysis of yellow catfish(Pelteobagrus fulvidraco) by RNA-seq method extracting gastrointestinal mucosa Total RNA was studied in this paper. The fish were fed with oxidized fish oil and fish oil respectively for 7 d. Differential expression of genes related to melanin synthesis, melanosomes movement, melanocyte stimulating hormone-α pathway and three signal pathways controlling melanocyte differentiation were analyzed. The results showed that there were metabolic networks of melanocyte differentiation and development, melanin synthesis and regulation in the gastrointestinal mucosa. The genetic information of key enzymes and protein of melanin synthesis were introduced with plotting metabolic networks graph. The melanin content was deficient because of the genes expression level were significantly down-regulation related to controlling melanin synthesis after fed with oxidized fish oil. While melanocyte differentiation and development were improved because of the genes expression level were significantly up-regulation related to melanocyte stimulating hormone-α pathway(α-MSH). In the same time, genes expression level of WNT/β-catenin, EDN3/EDNRB and KIT/KITL signal pathways were different. In conclusion, feeding oxidized fish oil affected melanocyte differentiation and development and reduced melanocyte number which benefited to maturity in yellow catfish. Insufficient melanin biosynthesis also led to body color variation of yellow catfish.
引文
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[13]Fabien M,Sabrina G,Friedrich B.The tyrosinase enhancer is activated by Sox10 and Mitf in mouse melanocytes[J].Pigment Cell Research,2007,20(3):173-184
[14]Bondurand N,Pingault V,Goerich D E,et al.Interaction among Sox10,Pax3 and Mitf three genes altered in waardenburg syndrome[J].Human Molecular Genetics,2000,9(13):1907-1917
[15]Mascarenhas J B,Littlejohn E L,Wolsky R J,et al.PAX3and SOX10 activate Mitf receptor expression in melanoma[J].Pigment Cell&Melanoma Research,2010,23(2):225-237
[16]Vachtenheim J,Borovansky J.“Transcription physiology”of pigment formation in melanocytes:central role of MITF[J].Experimental Dermatology,2010,19(7):617-627
[2]Masazumi S.Morphological color changes in fish:regulation of pigment cell density and morphology[J].Microscopy Research and Technique,2002,58(6):496-503
[3]Cheng W X,Xu G H,Xiong D,et al.Primary culturing of melanoma cells and cloning analysis of migration-related gene in Pelteobagrus fulvidraco[J].Asian Journal of Ecotoxicology,2014,9(6):1035-1040[程炜轩,许国焕,熊达,等.黄颗鱼黑色素细胞原代培养及迁移相关基因克隆分析.生态毒理学报,2014,9(6):1035-1040]
[4]Zhu J,Zhang X M,Gao T Y,et al.The metamorphosis of turbot Scophthamus maximus and morphological observation on melanophores in larval skin[J].Journal of Fisheries of China,2002,26(3):193-200[朱杰,张秀梅,高天翔,等.大菱坪早期变态发育和体表黑色素细胞形态学观察.水产学报,2002,26(3):193-200]
[5]Wu D W,Shui D Z,Cai C F,et al.Nutrition changes associated with anchovy enzymolysis process[J].Feed Industry,2015,36(24):25-30[吴代武,税典章,蔡春芳,等.鳀鱼鱼浆的酶解过程与营养成分的变化.饲料工业,2015,36(24):25-30]
[6]Ye Y T,Cai C F,Xu F,et al.Feeding grass carp(Ctenopharyngodon idellus)with oxidized fish oil up-regulates the gene expression in the cholesterol and bile acid synthesis pathway in intestinal mucosa[J].Acta Hydrobiologica Sinica,2015,39(1):90-100[叶元土,蔡春芳,许凡,等.灌喂氧化鱼油使草鱼肠道黏膜胆固醇胆汁酸合成基因通路表达上调.水生生物学报,2015,39(1):90-100]
[7]Ye Y T,Cai C F,Xu F,et al.Effects of oxidized fish oil on oxidative stress pathways of intestinal mucosa of grass carp(Ctenopharyngodon idellus)[J].Acta Hydrobiologica Sinica,2016,40(4):758-766[叶元土,蔡春芳,许凡,等.氧化鱼油对草鱼肠道钻膜抗氧化应激通路基因表达水平的影响.水生生物学报,2016,40(4):758-766]
[8]Zhu G Y,YE Y T,Gao Y L,et al.The comparison of pignent and tryosinase activity in seven secies of freshwater fishes[J].Journal of Shanghai Fisheries Univerdity,2007,16(5):431-436[诸葛燕,叶元土,高艳玲,等.七种淡水鱼类色素含量和酪氨酸酶活力的比较研究.上海水产大学学报,2007,16(5):431-436]
[9]Victoria A,Kdmler,John D,et al.Morphological studies on the mechanisms of pigmentary organelle transport in fish xanthophores and melanophores[J].Microscopy Research and Technique,2002,58(6):470-480
[10]Yang C L,Hao T T,Li Q F,et al.Signal regulation for melanoblast cytopoiesis process:a review[J].China Animal Husbandry and Veterinary Medicine,2013,40(9):187-194[杨春玲,郝甜甜,李强飞,等.成黑色素细胞生成过程中信号调节的研究进展.中国畜牧兽医,2013,40(9):187-194]
[11]Lan X M,Yang X C.Wnt/β-catenin signaling pathways and hair follicle morphogenesis[J].Journal of Practical Dermatology,2015,8(3):205-207[兰雪梅,杨希川.Wnt/β-catenin信号通路与毛囊的生长.实用皮肤病学杂志,2015,8(3):205-207]
[12]Xue C Y,Li L.The regulation ofα-melanocyte-stimulating hormone to melanogenesis of skin[J].Journal of Medical Postgraduates,2004,17(9):823-828[薛春雨,李蠢.α黑素细胞刺激素对皮肤黑素合成的调控作用.医学研究生学报,2004,17(9):823-828]
[13]Fabien M,Sabrina G,Friedrich B.The tyrosinase enhancer is activated by Sox10 and Mitf in mouse melanocytes[J].Pigment Cell Research,2007,20(3):173-184
[14]Bondurand N,Pingault V,Goerich D E,et al.Interaction among Sox10,Pax3 and Mitf three genes altered in waardenburg syndrome[J].Human Molecular Genetics,2000,9(13):1907-1917
[15]Mascarenhas J B,Littlejohn E L,Wolsky R J,et al.PAX3and SOX10 activate Mitf receptor expression in melanoma[J].Pigment Cell&Melanoma Research,2010,23(2):225-237
[16]Vachtenheim J,Borovansky J.“Transcription physiology”of pigment formation in melanocytes:central role of MITF[J].Experimental Dermatology,2010,19(7):617-627