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五个大鼠脑区中对尼古丁产生差异表达的蛋白质:基于二维电泳/质谱的蛋白质组学方法鉴定

Proteins differentially expressed in response to nicotine in five rat brain regions: identification using a 2-DE/MS-based proteomics approach.

作者信息

Hwang Yoon Y, Li Ming D

机构信息

Department of Psychiatric Medicine, University of Virginia, 1670 Discovery Drive, Charlottesville, VA 22911, USA.

出版信息

Proteomics. 2006 May;6(10):3138-53. doi: 10.1002/pmic.200500745.

Abstract

To determine protein expression patterns within the central nervous system (CNS) in response to nicotine, 2-DE/MS was performed on samples from five brain regions of rats that had received nicotine bitartrate by osmotic minipump infusion at a dose of 3.15 mg/kg/day for 7 days. After spot matching and statistical analysis, 41 spots in the amygdala, 49 in the nucleus accumbens (NA), 46 in the prefrontal cortex (PFC), 36 in the striatum, and 28 in the ventral tegmental area (VTA) showed significant differences in the nicotine-treated compared with control samples. Using MALDI-TOF MS peptide fingerprinting, 14 proteins in the amygdala, 11 in the NA, 19 in the PFC, 13 in the striatum, and 19 in the VTA were identified. Several proteins (e.g. dynamin 1, laminin receptors, aldolase A, enolase 1 alpha, Hsc70-ps1, and N-ethylmaleimide-sensitive fusion protein) were differentially expressed in multiple brain regions. By gene ontology analysis, these differentially expressed proteins were grouped into biological process categories, such as energy metabolism, synaptic function, and oxidative stress metabolism. These data, in combination with microarray analysis of mRNA transcripts, have the potential to identify the CNS gene products that show coordinated changes in expression at both the RNA and protein levels in response to nicotine.

摘要

为了确定中枢神经系统(CNS)内对尼古丁的蛋白质表达模式,对通过渗透微型泵以3.15mg/kg/天的剂量输注酒石酸尼古丁7天的大鼠的五个脑区样本进行了双向电泳/质谱分析。经过斑点匹配和统计分析,与对照样本相比,杏仁核中有41个斑点、伏隔核(NA)中有49个斑点、前额叶皮层(PFC)中有46个斑点、纹状体中有36个斑点以及腹侧被盖区(VTA)中有28个斑点在尼古丁处理组中显示出显著差异。使用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)肽指纹图谱,鉴定出杏仁核中有14种蛋白质、NA中有11种、PFC中有19种、纹状体中有13种以及VTA中有19种蛋白质。几种蛋白质(如发动蛋白1、层粘连蛋白受体、醛缩酶A、烯醇化酶1α、热休克蛋白70-ps1和N-乙基马来酰亚胺敏感融合蛋白)在多个脑区中差异表达。通过基因本体分析,这些差异表达的蛋白质被归类到生物过程类别中,如能量代谢、突触功能和氧化应激代谢。这些数据与mRNA转录本的微阵列分析相结合,有可能识别出在RNA和蛋白质水平上对尼古丁反应时表达呈现协同变化的中枢神经系统基因产物。

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