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[人类胎儿和成人脑皮质的蛋白质组学分析]

[Proteomic analysis of human fetal and adult brain cortex].

作者信息

Fan Wei-jia, Huang Hui-ling, Mo Li-dong, Wang Chen, Wu Yan-hong, Cai Ying

机构信息

Department of Graduate School, Tianjin Medical University, Tianjin, 300070 P. R. China.

出版信息

Zhonghua Yi Xue Yi Chuan Xue Za Zhi. 2011 Jun;28(3):323-7. doi: 10.3760/cma.j.issn.1003-9406.2011.03.019.

Abstract

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.

摘要

目的

采用蛋白质组学技术研究人类胎儿和成人脑皮质中蛋白质表达水平的差异,为脑发育过程中蛋白质的变化提供初步数据。

方法

分别提取胎儿(3个月和5个月)及成人(30岁)脑颞叶中的蛋白质,经双向凝胶电泳(2DE)分离,然后用考马斯亮蓝胶体染色以生成蛋白质组的高分辨率图谱。用PDQuest 7.0软件分析差异蛋白质点,选取在脑发育过程中逐渐减少或逐渐增加且差异超过两倍的8个蛋白质点,通过基质辅助激光解吸电离飞行时间质谱(MALDI-TOF/TOF)进行鉴定。

结果

(1)成人、3个月胎儿和5个月胎儿脑颞叶平均分别可获得642、511和527个蛋白质点,图像匹配率为87%。成人脑中的碱性蛋白质明显多于胎儿;(2)成人、3个月胎儿和5个月胎儿脑颞叶分别有172、171和152个独特蛋白质点;(3)与成人相比,3个月和5个月胎儿分别有131和115个不同蛋白质点。差异超过2倍的蛋白质点分别有60和40个,其中24和17个下调,36和23个上调;(4)3组中血清白蛋白、磷酸丙糖异构酶等蛋白质存在差异表达。脂肪酸结合蛋白7和未命名蛋白质仅在3个月龄脑中高表达;核酮糖-1,5-二磷酸羧化酶/加氧酶大亚基和转导素β-1亚基在成人脑中上调。血清白蛋白随脑发育逐渐降低。然而,ATP合酶、线粒体F0复合物和磷酸丙糖异构酶随脑发育逐渐增加。

结论

人类脑皮质蛋白质从胚胎期到成人期有明显变化。差异显示的蛋白质可能有助于进一步了解人类脑的发育。

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