酪氨酸对固原乌鸡黑色素分布沉积及相关基因表达的影响
CSTR:
作者:
作者单位:

宁夏大学动物科技学院, 银川750021

作者简介:

高思语,E-mail:1833267671@qq.com

通讯作者:

张莉,E-mail:1457839598@qq.com

中图分类号:

S827.2

基金项目:

宁夏自然科学基金项目(2022AAC03016);宁夏引才专项(2021BEB04010);宁夏高等教育质量提升工程项目(030203042417/0304)


Effects of content of tyrosine in diet on distribution and deposition of melanin and expression of melanin-related gene in Guyuan silky fowls
Author:
Affiliation:

College of Animal Science and Technology, Ningxia University, Yinchuan 750021, China

  • 摘要
  • |
  • 图/表
  • |
  • 访问统计
  • |
  • 参考文献 [29]
  • |
  • 相似文献 [20]
  • | | |
  • 文章评论
    摘要:

    为探究固原乌鸡黑色素分布沉积特征以及在不同日粮酪氨酸水平下黑色素相关基因表达量的变化,明确酪氨酸对固原乌鸡黑色素形成的影响,选取4周龄、体质量差异不显著的健康固原乌鸡150只,随机分为5组,每组3个重复,每个重复10只。日粮酪氨酸添加水平分别为0、0.2%、0.4%、0.6%、0.8%,饲养至12周龄末,利用电镜技术观察不同组织中黑色素的分布特征,利用免疫组织化学和荧光定量PCR方法检测不同组织中黑色素相关基因表达量的变化。试验结果显示:固原乌鸡皮肤、气管与小肠中存在黑色素细胞且大都已分化成熟。TYR在肺脏组织有阳性反应,TYRP1在肌肉与肝脏组织有阳性反应,MITF、CREB1在肺脏、肝脏、小肠、胸肌组织中均有阳性结果。在不同日粮酪氨酸水平下,MITF、TYR、TYRP1基因的表达量与黑色素含量成一定的线性相关。结果表明,固原乌鸡组织黑色素沉积特征与组织中黑色素颗粒合成量相关,而黑色素合成过程中TYR、MITF可能是其主要关键调控基因,日粮中添加酪氨酸能够促进其表达。

    Abstract:

    The effects of tyrosine on the formation of melanin in Guyuan silky fowls were studied to investigate the characteristics of the distribution and deposition of melanin in Guyuan silky fowls, and the changes in the expression of melanin-related gene under different content of tyrosine in diet. 150 healthy Guyuan silky fowls at 4 weeks of age with no significant difference in body weight were randomly divided into 5 groups, with 3 replicates in each group and 10 chickens in each replicate. The content of tyrosine added in diet was 0, 0.2%, 0.4%, 0.6%, and 0.8%, respectively, and the fowls were fed until the end of 12 weeks of age. The electron microscopy was used to observe the characteristics of the distribution of melanin in different tissues of Guyuan silky fowls. The immunohistochemistry and fluorescence quantitative PCR method were used to detect changes in the expression of melanin-related genes in different tissues of Guyuan silky fowls. The results showed that there were melanocytes in the skin, trachea, and small intestine of Guyuan silky fowls, and most of them were differentiated and matured. TYR had a positive reaction in lung tissue, TYRP1 had a positive reaction in muscle and liver tissue, and MITF and CREB1 had positive results in lung, liver, small intestine, and breast muscle tissue. The expression level of MITFTYR, and TYRP1 gene was linearly correlated with the content of melanin under different content of tyrosine in diet. It is indicated that the characteristics of melanin deposition in tissues of Guyuan silky fowls are related to the amount of melanin granules synthesized in the tissues, and TYR and MITF may be the main key regulatory genes during the synthesis of melanin. Adding tyrosine to the diet can promote the expression of melanin-related genes.

    表 7 不同日粮酪氨酸水平下固原乌鸡不同组织TYRP1基因的相对表达量Table 7 The relative expression of TYRP1 gene in different tissues of Guyuan silky fowl under different dietary tyrosine levels
    表 2 荧光定量PCR引物Table 2 The primers of fluorescent quantitative PCR
    表 4 不同日粮酪氨酸水平下固原乌鸡不同组织CREB1基因的相对表达量Table 4 The relative expression of CREB1 gene in different tissues of Guyuan silky fowl under different dietary tyrosine levels
    表 6 不同日粮酪氨酸水平下固原乌鸡不同组织TYR基因的相对表达量Table 6 The relative expression of TYR gene in different tissues of Guyuan silkyfowl under different dietary tyrosine levels
    表 3 不同日粮酪氨酸水平对固原乌鸡组织中TYR、TYRP1、MITF和CREB1分布的影响Table 3 Effect of dietary tyrosine levels on the distribution of TYR,TYRP1,MITF,and CREB1 in Guyuan silky fowl
    图1 固原乌鸡皮肤黑色素细胞的超微结构Fig.1 Ultrastructure of skin melanocytes of Guyuan silky fowl
    图2 固原乌鸡气管黑色素细胞的超微结构Fig.2 Ultrastructure of trachea melanocytes of Guyuan silky fowl
    图3 固原乌鸡小肠黑色素细胞超微结构Fig.3 Ultrastructure of small intestine melanocytes of Guyuan silky fowl
    图4 固原乌鸡组织器官TYR免疫组化图(400×)Fig.4 TYR immunohistochemistry of Guyuan silky fowl
    图5 固原乌鸡组织器官TYRP1免疫组化染色图(400×)Fig.5 TYRP1 immunohistochemistry of Guyuan silky fowl
    图6 固原乌鸡组织器官MITF免疫组化染色图(400×)Fig.6 MITF immunohistochemistry of Guyuan silky fowl
    图7 固原乌鸡组织器官CREB1免疫组化染色图(400×)Fig.7 CREB1 immunohistochemistry of Guyuan silky fowl
    图8 CREB1、MITF、TYR、TYRP1基因表达量与黑色素含量间的相关性Fig.8 Correlation analysis of gene expression levels of CREB1, MITF, TYR, TYRP1 and melanin content
    表 5 不同日粮酪氨酸水平下固原乌鸡不同组织MITF基因的相对表达量Table 5 The relative expression of MITF gene in different tissues of Guyuan silky fowl under different dietary tyrosine levels
    表 1 基础饲粮组成和营养水平Table 1 The composition and nutrient levels of basic diets
    参考文献
    [1] 王晓薇,高飞涛,安克龙,等.不同群体固原鸡鸡肉主要营养成分的研究[J].黑龙江畜牧兽医,2013(9):67-70.WANG X W,GAO F T,AN K L,et al.Research on the main nutritional components of Guyuan chicken meat for different groups[J].Heilongjiang animal science and veterinary medicine,2013(9):67-70 (in Chinese).
    [2] 朱星仲,杨文清.药用珍禽:固原乌鸡[J].宁夏农林科技,2001,42(5):53.ZHU X Z,YANG W Q.Guyuan black chicken,a rare medicinal bird[J].Ningxia journal of agriculture and forestry science and technology,2001,42(5):53 (in Chinese).
    [3] 田颖刚,李慧,杨亚杰,等.乌骨鸡蛋与普通鸡蛋品质特征和氨基酸组成比较[J].南昌大学学报(理科版),2018,42(3):231-235.TIAN Y G,LI H,YANG Y J,et al.Comparison of quality characteristics and amino acid composition of eggs from black-bone silky fowl and common chicken[J].Journal of Nanchang University (natural science),2018,42(3):231-235 (in Chinese with English abstract).
    [4] MENTER J M,PATTA A M,HOLLINS T D,et al.Photoprotection of mammalian acid-soluble collagen by cuttlefish sepia melanin in vitro[J].Photochemistry and photobiology,1998,68(4):532-537.
    [5] 熊和丽,席冬梅,李国治,等.动物黑色素细胞及其相关性状形成机制的研究进展[J].中国畜牧兽医,2019,46(1):72-79.XIONG H L,XI D M,LI G Z,et al.Research advances on the formation mechanism of animal melanocytes and its related traits[J].China animal husbandry & veterinary medicine,2019,46(1):72-79 (in Chinese with English abstract).
    [6] 何洋,杜彦丽,王坤,等.乌骨鸡黑色素合成、转运及分布机制的研究进展[J].中国家禽,2020,42(9):89-95.HE Y,DU Y L,WANG K,et al.Research progress on synthesis,transport and distribute of melanin mechanisms in black-bone chicken[J].China poultry,2020,42(9):89-95(in Chinese with English abstract).
    [7] 董懿,康馨月,王春生.哺乳动物黑色素生物合成及其调控机制研究进展[J].生命科学,2022,34(8):990-1000.DONG Y,KANG X Y,WANG C S.Progress on the research of biosynthesis and regulation of mammalian melanin[J].Chinese bulletin of life sciences,2022,34(8):990-1000 (in Chinese with English abstract).
    [8] 江彦,刘青,王琤.酪氨酸对泰和乌骨鸡黑色素合成的影响[J].中国家禽,2017,39(23):47-50.JIANG Y,LIU Q,WANG Z.Effects of tyrosine on melanin synthesis in silky fowl[J].China poultry,2017,39(23):47-50(in Chinese with English abstract).
    [9] S?OMINSKI A,MOELLMANN G,KUKLINSKA E,et al.Positive regulation of melanin pigmentation by two key substrates of the melanogenic pathway,L-tyrosine and L-dopa[J].Journal of cell science,1988,89(3):287-296.
    [10] 王杰.日粮酪氨酸添加水平对淅川乌骨鸡胸肌肉质的影响及相关差异基因筛选[D].郑州:河南农业大学,2018.WANG J.Effect of dietary tyrosine addition level on breast muscle quality of Xichuan black-bone chicken and screening of related differential genes[D].Zhengzhou:Henan Agricultural University,2018 (in Chinese with English abstract).
    [11] 黎观红,游金明,瞿明仁,等.日粮酪氨酸水平对泰和乌骨鸡生产性能及组织黑色素含量的影响[J].中国兽医学报,2009,29(11):1481-1484.LI G H,YOU J M,QU M R,et al.Effects of dietary tyrosine levels on performance and melanin contents in tissues of Taihe silky fowls aged from 5 to 12 weeks of age[J].Chinese journal of veterinary science,2009,29(11):1481-1484 (in Chinese with English abstract).
    [12] 曹艳芳,张晨曦,陶艺庆,等.不同水平酪氨酸对淅川乌骨鸡组织乌度及屠宰性状的影响[J].中国畜牧杂志,2020,56(12):109-113.CAO Y F,ZHANG C X,TAO Y Q,et al.Effects of dietary tyrosine levels on tissue brightness value and slaughter traits of Xichuan black-bone chicken[J].Chinese journal of animal science,2020,56(12):109-113 (in Chinese with English abstract).
    [13] 叶宇恒,高思语,张珠明,等.日粮酪氨酸水平对固原乌鸡黑色素生成的影响[J].现代畜牧兽医,2023(8):37-42.YE Y H,GAO S Y,ZHANG Z M,et al.Effect of dietary tyrosine level on melanin production in Guyuan black-bone chicken[J].Modern journal of animal husbandry and veterinary medicine,2023(8):37-42 (in Chinese with English abstract).
    [14] 肖红.酪氨酸影响泰和乌骨鸡黑色素生成的细胞学机理[D].南昌:江西农业大学,2013.XIAO H.Cytological mechanism of tyrosine affecting melanin production in Taihe silky fowl[D].Nanchang:Jiangxi Agricultural University,2013 (in Chinese with English abstract).
    [15] MUROYA S,TANABE R,NAKAJIMA I,et al.Molecular characteristics and site specific distribution of the pigment of the silky fowl[J].The journal of veterinary medical science,2000,62(4):391-395.
    [16] PRUM R O,TORRES R.Structural colouration of avian skin:convergent evolution of coherently scattering dermal collagen arrays[J].The journal of experimental biology,2003,206(14):2409-2429.
    [17] ORTOLANI-MACHADO C,DE FREITAS P,BORGES M E,et al.Special features of dermal melanocytes in white silky chicken embryos[J].Anatomical record,2008,291(1):55-64.
    [18] 刘海霞,朱爱文,王步忠,等.黑色素形成调控机理及乌骨绵羊黑色素沉积候选基因研究进展[J].中国畜牧兽医,2021,48(12):4588-4596.LIU H X,ZHU A W,WANG B Z,et al.Research progress on mechanism of melanin deposition and related candidate genes in black-bone sheep[J].China animal husbandry & veterinary medicine,2021,48(12):4588-4596 (in Chinese with English abstract).
    [19] 谢光跃.黑素细胞分布规律及基因差异表达[D].武汉:华中农业大学,2014.XIE G Y.Distribution of melanocytes and differential expression of genes[D].Wuhan:Huazhong Agricultural University,2014 (in Chinese with English abstract).
    [20] 王良炎,田雪,庞小磊,等.黄河鲤酪氨酸酶基因的克隆、表达模式及定位分析[J].水产学报,2017,41(7):1006-1016.WANG L Y,TIAN X,PANG X L,et al.Molecular cloning,expression pattern and location of tyrosinase gene in Yellow River carp (Cyprinus carpio haematopterus)[J].Journal of fisheries of China,2017,41(7):1006-1016 (in Chinese with English abstract).
    [21] 高莉,赵英虎,刘朝亮,等.酪氨酸酶与羊驼毛色相关性的研究[J].激光生物学报,2011,20(3):319-323.GAO L,ZHAO Y H,LIU C L,et al.Relevance between TYR and different hair color of alpaca[J].Acta laser biology sinica,2011,20(3):319-323 (in Chinese with English abstract).
    [22] 成磊.毛色相关miRNAs在不同山羊品种皮肤中的表达规律及对黑色素生成的影响[D].重庆:西南大学,2021.CHENG L.Expression of coat color-related miRNAs in skin of different goat breeds and its effect on melanin production[D].Chongqing:Southwest University,2021(in Chinese with English abstract).
    [23] 祖盘玉.赤水乌骨鸡MC1R、TYR、MITF基因多态性分析及其组织表达效应研究[D].贵阳:贵州大学,2019.ZU P Y.Polymorphism analysis of MC1R,TYR and MITF genes in Chishui black-bone chicken and their tissue expression effects[D].Guiyang:Guizhou University,2019 (in Chinese with English abstract).
    [24] 张立恒.日粮酪氨酸、苯丙氨酸水平对丝毛乌骨鸡生产性能和黑色素相关基因表达的影响[D].郑州:河南农业大学,2010.ZHANG L H.Effects of dietary tyrosine and phenylalanine levels on performance and melanin-related gene expression of silky fowl[D].Zhengzhou:Henan Agricultural University,2010 (in Chinese with English abstract).
    [25] 刘薇.丝羽乌骨鸡BAC文库的构建和黑色素相关基因TYRP1和ID的研究[D].北京:中国农业大学,2004.LIU W.Construction of BAC library of silky fowl and study on melanin-related genes TYRP1 and ID[D].Beijing:China Agricultural University,2004 (in Chinese with English abstract).
    [26] 辛清武,李丽,缪中纬,等.莆田黑鸭MITF基因表达特性及其与黑色素沉积的关联分析[J].农业生物技术学报,2018,26(11):1928-1937.XIN Q W,LI L,MIAO Z W,et al.Expression of MITF gene in Putian black duck (Anas anas domesticus) and its association with melanin deposition[J].Journal of agricultural biotechnology,2018,26(11):1928-1937 (in Chinese with English abstract).
    [27] 杜晓明.边鸡MC1R和TYR基因序列及mRNA表达研究[D].太谷:山西农业大学,2013.DU X M.Sequence and mRNA expression of MC1R and TYR genes in border chickens[D].Taigu:Shanxi Agricultural University,2013 (in Chinese with English abstract).
    [28] 徐志强,李丰耘,赵净颖,等.武定乌骨鸡TYR基因多态性与黑色素沉积的关联研究[J].云南农业大学学报(自然科学),2021,36(4):608-615.XU Z Q,LI F Y,ZHAO J Y,et al.The association between TYR gene polymorphism and melanin deposition in Wuding black-boned chicken[J].Journal of Yunnan Agricultural University (natural science),2021,36(4):608-615 (in Chinese with English abstract).
    [29] 陈阳,秦立志,赵博昊,等.獭兔CREB1基因的编码区克隆、生物信息学及表达规律分析[J].江苏农业科学,2018,46(23):165-168.CHEN Y,QIN L Z,ZHAO B H,et al.Coding sequence cloning,bioinformatics and gene expression analysis of CREB1 gene in rex rabbit[J].Jiangsu agricultural sciences,2018,46(23):165-168 (in Chinese with English abstract).
    引证文献
    网友评论
    网友评论
    分享到微博
    发 布
引用本文

高思语,王晓娟,齐玮瑜,姜宇辰,张莉.酪氨酸对固原乌鸡黑色素分布沉积及相关基因表达的影响[J].华中农业大学学报,2024,43(5):193-204

复制
分享
文章指标
  • 点击次数:56
  • 下载次数: 548
  • HTML阅读次数: 26
  • 引用次数: 0
历史
  • 收稿日期:2023-11-22
  • 在线发布日期: 2024-10-08
文章二维码