雄激素刺激AP发育前后差异蛋白筛选及DIGE分析  

DIGE Analysis and Screening of Differential Proteins Before and After AP Develpoment Induced by Androgen

作　　者：刘慧[1] 刘振[1] 王权威 董振[1] 杨雪雯[1] 王桂武[1] 杨福合[1] 李春义[1] 

机构地区：[1]中国农业科学院特产研究所/特种动物分子生物学国家重点实验室,长春130112

出　　处：《农业生物技术学报》2017年第7期1119-1129,共11页Journal of Agricultural Biotechnology

基　　金：中国梅花鹿基因组计划(No.20140309016YY);中国农业科学院中央级公益性科研院所基本科研业务费专项(No.Y2016PT46和No.1610342016001);吉林省科技发展计划项目(No.20170307007YY)

摘　　要：鹿茸是一特殊的骨性器官,内部布满血管和神经。鹿茸发生是指雄性仔鹿进入青春期后角柄和初角茸的形成过程。研究已经证实鹿额外脊上的生茸区骨膜(antlerogenic periosteum,AP)是鹿茸发生的组织基础。AP承担了鹿茸器官发育的蓝图。研究表明,雄激素在鹿茸发生中起到重要作用。为了研究AP发育前后蛋白质组学变化,深入探讨雄激素诱导鹿茸发生的分子机制。本研究利用雄激素刺激雌性梅花鹿(Cervus nippon)生茸区骨膜发育,应用荧光差异双向凝胶电泳(two-dimensional fluorescence difference gelelectrophoresis,2D-DIGE)技术,分别提取雄激素刺激前生茸骨膜对照组和刺激后实验组的组织总蛋白,经2D-DIGE技术分析获得蛋白点的差异表达信息,运用基质辅助激光解吸电离飞行时间质谱法(matrixassisted laser desorption/ionization time of flight mass spectrometry,MALDI-TOF-MS)鉴定差异蛋白质,并根据数据库检索分析鉴定出的差异蛋白。实验数据分析表明,共筛选出318个差异蛋白点,其中163个蛋白点上调,155个蛋白下调。筛选其中93个蛋白点采用质谱鉴定技术,成功鉴定17个蛋白,分属5种结构和功能各异的蛋白,包括结合相关蛋白,酶调活性相关蛋白,结构分子活性相关蛋白,催化活性相关蛋白和抗氧化活性相关蛋白。本研究为鹿茸生长调控机制提供理论依据,进而为器官发生机制研究提供参考数据。Antler is a special osseous organs coverd with blood vessel, nerve inside. Antler development refers to the forming process of male fawn pedicle and first antler after puberty. It has confirmed that the piece of periosteum overlying the lateral crest of prepubertal deer frontal bone is a foundation of antler development, which is known as antlerogenic periosteum （AP）. AP holds the patterning information for antler formation. Studies suggest that androgens play an important role in antler development. The aim of the study is to investigate the changes of proteomics before and after AP development, which will help to reveal the underlying molecular mechanisms of androgen induced antler development. The study used androgen stimulated female deer until antlerogenic periosteum developed and extracted total proteins from the treated and control groups AP and applied two-dimensional fluorescence difference gelelectrophoresis （2D-DIGE） to obtain differential expression information of proteins. After that, differentially expressed proteins were identified by MALDI-TOF/MS and analysed the proteins based on the database retrieval. The results showed that there were 318 differentially expressed proteins searched and identified by database selected. Among them, 163 proteins were up-regulated and 155 were down-regulated significantly. MALDI-TOF/MS was used to identify 93 of 318 proteins and there were 17 proteins identified successfully. The 17 identified proteins were classified into 5 categories with different structures and functions and these proteins were associated with binding, enzyme activity, catalytic activity and antioxidant activity. Furthermore, the study had classified the mechanism of antler morphogenesis and offers the experience for further studies. What＇s more, among the 17 successfully identified proteins, the proteins were classified into 5 categories with different structures and functionsand these proteins were associated with binding, enzyme activity, catalytic activity and antioxidant activity

关 键 词：生茸区骨膜 发生 蛋白质组学 荧光差异双向凝胶电泳(2D-DIGE) 

分 类 号：S825[农业科学—畜牧学]