种蛋孵化早期蛋黄蛋白质组学比较分析
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摘要
卵黄为发育胚胎提供脂肪、蛋白质等营养物质,直接影响胚胎发育。研究旨在探索胚胎发育早期卵黄蛋白质变化,采用2-DE结合MALDI-TOF MS/MS分析孵化0和10 d种蛋卵黄蛋白质组成。共鉴定出代表10种蛋白质的57个蛋白点丰度存在显著差异(P<0.05),其中8个蛋白点仅存在于0 d卵黄;24个蛋白点仅存在于孵化10 d卵黄;另有25个蛋白点共同存在于0和10 d卵黄中,其中9个点丰度上调、16个点丰度下调。首次检测出卵黄膜外层蛋白1存在于胚胎孵化0和10 d卵黄中。2-DE凝胶图像分析表明,39个蛋白质相对分子质量低于其理论值。生物信息学富集分析表明,4种蛋白质与脂质转运和脂质定位有关,2种蛋白质与抗原结合有关。孵化10 d后卵黄差异蛋白主要参与脂质转运以维持胚胎发育。研究结果有助于进一步了解卵黄蛋白质对胚胎发育的调控机理。
The egg yolk provides nutrients such as fat and protein to the developing embryo, and can directly affect embryo development. This study was designed to explore the changes of egg yolk protein during the early embryonic development. The egg yolk protein composition at hatching for 0 and10 days was analyzed by 2-DE combined with MALDI-TOF MS/MS. A significant difference in the abundance of 57 protein spots representing 10 proteins was identified(P<0.05). Among them, eight protein spots existed only on 0 day, 24 protein spots existed only on hatching for 10 days, and another25 protein spots co-existed in hatching for 0 and 10 days, in which 9 spots exhibited up regulation and16 spots was down regulated. The vitelline membrane outer layer protein 1 was detected in the yolk at embryonic days for the first time. Two-DE gel image analysis showed that the relative molecular mass of 39 proteins were lower than that of the theoretical value. Bioinformatics enrichment analysis showed that four proteins were involved in lipid transport and lipid localization, and two proteins were involved in antigen binding. The differentially expressed proteins in egg yolk at 10 embryonic days were mostly involved in lipid transportation for embryo development. The results will help to further understand the regulation mechanism of egg yolk protein on embryonic development.
The egg yolk provides nutrients such as fat and protein to the developing embryo, and can directly affect embryo development. This study was designed to explore the changes of egg yolk protein during the early embryonic development. The egg yolk protein composition at hatching for 0 and10 days was analyzed by 2-DE combined with MALDI-TOF MS/MS. A significant difference in the abundance of 57 protein spots representing 10 proteins was identified(P<0.05). Among them, eight protein spots existed only on 0 day, 24 protein spots existed only on hatching for 10 days, and another25 protein spots co-existed in hatching for 0 and 10 days, in which 9 spots exhibited up regulation and16 spots was down regulated. The vitelline membrane outer layer protein 1 was detected in the yolk at embryonic days for the first time. Two-DE gel image analysis showed that the relative molecular mass of 39 proteins were lower than that of the theoretical value. Bioinformatics enrichment analysis showed that four proteins were involved in lipid transport and lipid localization, and two proteins were involved in antigen binding. The differentially expressed proteins in egg yolk at 10 embryonic days were mostly involved in lipid transportation for embryo development. The results will help to further understand the regulation mechanism of egg yolk protein on embryonic development.
引文
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[17]Gentili C.Ovotransferrin and ovotransferrin receptor expression during chondrogenesis and endochondral bone formation in developing chick embryo[J].The Journal of Cell Biology,1994,124(4):579-588.
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[22]余燕,马艺源,吴世秀,等.肉鸡鸡胚肝脏中脂滴的发育研究[J].中国畜牧兽医,2017,44(3):692-697.
[2]Mann K,Mann M.The chicken egg yolk plasma and granule proteomes[J].Proteomics,2010,8(1):178-191.
[3]Farinazzo A,Restuccia U,Bachi A,et al.Chicken egg yolk cytoplasmic proteome,mined via combinatorial peptide ligand libraries[J].Journal of Chromatography A,2009,1216(8):1241-1252.
[4]Gao D,Qiu N,Liu Y,et al.Comparative proteome analysis of egg yolk plasma proteins during storage[J].Journal of the Science of Food and Agriculture,2017,97(8):2392-2400.
[5]Réhault-Godbert S,Mann K,Bourin M,et al.Effect of embryonic development on the chicken egg yolk plasma proteome after 12days of incubation[J].Journal of Agricultural and Food Chemistry,2014,62(12):2531-2540.
[6]吴红翔,藩东福,温庆琪,等.孵化期间鸡胚内水分、脂肪及胆固醇含量的变化[J].黑龙江畜牧兽医,2013(1):61-63.
[7]Wang S,Smith D E,Williams D L.Purification of avian vitellogeninⅢ:Comparison with vitellogenins I and II[J].Biochemistry,1983,22(26):6206-6212.
[8]Wang J,Wu J.Proteomic analysis of fertilized egg white during early incubation[J].EuPA Open Proteomics,2014,2(1):38-59.
[9]Chalghoumi R,Beckers Y,Portetelle D,et al.Hen egg yolk antibodies(IgY),production and use for passive immunization against bacterial enteric infections in chicken:A review[J].Biotechnologie Agronomie SociétéEt Environnement,2009,13(2):295-308.
[10]李彩波.鸡卵被动抗体传递吸收规律的研究[D].长春:吉林农业大学,2008.
[11]Oubre C M,Els D'Hondt,Moore R W,et al.The chicken pituitary expresses an ovoinhibitor-like protein in subpopulations of some,but not all,hormone-producing cell types[J].Domestic Animal Endocrinology,2003,25(4):389-397.
[12]Begum S,Saito A,Xianhua X,et al.Improved functional properties of the ovoinhibitor by conjugating with galactomannan[J].Journal of the Agricultural Chemical Society of Japan,2003,67(9):1897-1902.
[13]Qiu N,Ma M,Zhao L,et al.Comparative proteomic analysis of egg white proteins under various storage temperatures[J].Journal of Agricultural and Food Chemistry,2012,60(31):7746-7753.
[14]Richards M P,Huang T L.Metalloprotein analysis by capillary isoelectric focusing[J].Journal of Chromatography B,1997,690(1-2):43-54.
[15]Valenti P,Visca P,Antonini G,et al.The effect of saturation with Zn2+and other metal ions on the antibacterial activity of ovotransferrin[J].Medical Microbiology and Immunology,1987,176(3):123-130.
[16]黄蓉,徐明生,吴磊燕,等.卵转铁蛋白磷酸化修饰工艺条件优化[J].江西农业大学学报,2013,35(4):864-869.
[17]Gentili C.Ovotransferrin and ovotransferrin receptor expression during chondrogenesis and endochondral bone formation in developing chick embryo[J].The Journal of Cell Biology,1994,124(4):579-588.
[18]Lim W,Song G.Differential expression of vitelline membrane outer layer protein 1:Hormonal regulation of expression in the oviduct and in ovarian carcinomas from laying hens[J].Molecular and Cellular Endocrinology,2015,399:250-258.
[19]Guérin-Dubiard C,Pasco M,Mollé,D,et al.Proteomic analysis of hen egg white[J].Journal of Agricultural and Food Chemistry,2006,54(11):3901-3910.
[20]Quirce S,Mara?ón F,Umpiérrez A,et al.Chicken serum albumin(Gal d 5*)is a partially heat-labile inhalant and food allergen implicated in the bird-egg syndrome[J].Allergy,2015,56(8):754-762.
[21]李国喜,康相涛,韩瑞丽,等.固始鸡孵化期间蛋黄胆固醇、粗脂肪和锌含量分析[J].湖北农业科学,2006,45(4):497-499.
[22]余燕,马艺源,吴世秀,等.肉鸡鸡胚肝脏中脂滴的发育研究[J].中国畜牧兽医,2017,44(3):692-697.