CAJ | 학술논문

目的应用蛋白质组技术研究成年和胚胎脑皮质蛋白质组表达差异,初步探索在胎脑与成年人中脑蛋白的变化情况.方法收集成人无病变区正常脑皮质1份和水囊引产胎脑组织脑皮质2份(3月胎脑组织和5月胎脑组织),提取总蛋白,通过双相电泳分离蛋白,每组样品均重复电泳,使用PDQuest7.0图像分析软件分析电泳图谱,得到具有表达差异的蛋白质,分别选取在大脑发育过程中逐步降低和逐步升高的差异点及在胎脑成脑蛋白表达中差异两倍以上的蛋白点,应用胶内酶切和基质辅助激光解析电离飞行时间质谱(matrix-assisted laser desorption/ionization-time of flight-time of flight,MALDI-TOF-TOF)进行鉴定,得到肽指纹图谱,并查阅蛋白质数据库得到蛋白质的相关信息.结果 (1)成人脑组织、3月胎脑组织和5月胎脑组织的脑组织蛋白图谱分别测得642、511和527个点,图象匹配率达87%.成人脑蛋白图谱中碱性蛋白点明显多于胎脑的;(2)成人脑组织、3月胎脑组织和5月胎脑组织的脑组织单独特有蛋白分别为:172、171和152个;(3)与成人脑组织比较,3月胎脑组织和5月胎脑组织的差异蛋白质分别为131和115个;差异在2倍以上的蛋白点为60和40个,其中低表达的蛋白点分别有24和17个,高表达蛋白点为36和23个;(4)白蛋白、磷酸丙糖异构酶等8种蛋白表达均有不同的变化.脂肪酸结合蛋白7和未命名蛋白只在3月胎脑高表达,而5月胎脑和成人组均无表达;核酮糖1,5-二磷酸羧化酶/加氧酶和转导素只在成人脑中高表达,胎脑无表达.血清白蛋白却随着大脑发育逐渐降低.ATP合成酶、线粒体F0复合体亚基、磷酸丙糖异构酶随着大脑发育而逐渐升高.结论人类从胚胎期到成人发育过程中脑部蛋白质可能会发生明显改变,这些差异蛋白的发现为研究脑发育机理提供了极其有益的线索.
목적 응용단백질조기술연구성년화배태뇌피질단백질조표체차이,초보탐색재태뇌여성년인중뇌단백적변화정황.방법 수집성인무병변구정상뇌피질1빈화수낭인산태뇌조직뇌피질2빈(3월태뇌조직화5월태뇌조직),제취총단백,통과쌍상전영분리단백,매조양품균중복전영,사용PDQuest7.0도상분석연건분석전영도보,득도구유표체차이적단백질,분별선취재대뇌발육과정중축보강저화축보승고적차이점급재태뇌성뇌단백표체중차이량배이상적단백점,응용효내매절화기질보조격광해석전리비행시간질보(matrix-assisted laser desorption/ionization-time of flight-time of flight,MALDI-TOF-TOF)진행감정,득도태지문도보,병사열단백질수거고득도단백질적상관신식.결과 (1)성인뇌조직、3월태뇌조직화5월태뇌조직적뇌조직단백도보분별측득642、511화527개점,도상필배솔체87%.성인뇌단백도보중감성단백점명현다우태뇌적;(2)성인뇌조직、3월태뇌조직화5월태뇌조직적뇌조직단독특유단백분별위:172、171화152개;(3)여성인뇌조직비교,3월태뇌조직화5월태뇌조직적차이단백질분별위131화115개;차이재2배이상적단백점위60화40개,기중저표체적단백점분별유24화17개,고표체단백점위36화23개;(4)백단백、린산병당이구매등8충단백표체균유불동적변화.지방산결합단백7화미명명단백지재3월태뇌고표체,이5월태뇌화성인조균무표체;핵동당1,5-이린산최화매/가양매화전도소지재성인뇌중고표체,태뇌무표체.혈청백단백각수착대뇌발육축점강저.ATP합성매、선립체F0복합체아기、린산병당이구매수착대뇌발육이축점승고.결론 인류종배태기도성인발육과정중뇌부단백질가능회발생명현개변,저사차이단백적발현위연구뇌발육궤리제공료겁기유익적선색.
Objective To study the differences of protein expression levels in the brain cortex of human fetus and adult with proteomics technique, and provide preliminary data on the change of proteins during brain development. Methods Proteins extracted from human temporal lobes in fetal (3 month and 5 month respectively) and adult (30 years old) brain were separated by two-dimensional gel electrophoresis (2DE).The proteins were then stained with colloidal Coomassie blue to produce a high-resolution map of the proteiome.The differential protein spots were analyzed by PDQuest 7.0 software and 8 spots,which were gradually reduced or gradually increased in brain development process and the protein spots of difference over two-fold in the brain,were identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF/TOF).Results (1) On average, 642, 511 and 527 protein spots could be obtained in the temporal lobes of adult, 3 month and 5 month fetus. The matching rate of images was 87%. The basic proteins in adult brain were obviously much more than that in the fetus; (2) There were 172, 171 and 152 singular protein spots in temporal lobes of adult, 3 month and 5 month fetus respectively. (3) Compared with adult, there were 131 and 115 different protein spots in the 3 month and 5 month fetus respectively. There were 60 and 40 protein spots with more than 2 fold difference, among which 24 and 17 were down-regulated, and 36 and 23 were up-regulated respectively. (4) There was different expression in proteins such as serum albumin, triosephosphate isomerase, etc. in the 3 groups. Fatty acid binding protein 7 and unnamed proteins were only highly expressed in the 3 month brain;ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit and transducin beta-1 subunit were up-regulated in adult brain. Serum albumin decreases gradually with brain development. However, ATP synthase, mitochondrial F0 complex, and triosephosphate isomerase increase gradually with brain development. Conclusion The proteins of human brain cortex were obviously changed from embryonic stage to adult. The differentially displayed proteins may provide further insight into the understanding of development of human brain.