Gu Feng, Shi Jianting, Wen Ying, Fan Hui, Hu Jinfeng, Hu Yinghe, Zhao Zheng
Key Laboratory of Brain Functional Genomics, MOE and STCSM, Shanghai Institute of Brain Functional Genomics, East China Normal University, Shanghai 200062, China.
Brain Res. 2009 Jan 23;1250:1-13. doi: 10.1016/j.brainres.2008.10.039. Epub 2008 Oct 30.
The N-methyl-D-aspartate (NMDA) receptor 2B subunit (NR2B) is important for long-term potentiation (LTP) and synaptic plasticity. The NR2B transgenic mice exhibited larger LTP in the hippocampal CA1 region and enhanced behavioral performance in several learning and memory tasks. In this study, we applied two-dimensional liquid chromatography-based proteomic approach to examine the expression levels of cerebral cortical proteins from 6-month NR2B transgenic (Tg) and their wild-type (WT) mice that were maintained on the same genetic background. Proteins were separated according to the pI in the first dimension using a chromatofocusing column and the hydrophobicity in the second dimension using a nonporous reversed-phase silica column. The DeltaVue software was applied to examine the differential expression of protein samples. Twenty six differentially expressed proteins were identified by matrix assisted laser desorption/ionization time-of-flight (MALDI-TOF/TOF) mass spectrometry, including glutamine synthetase (GS), guanine nucleotide-releasing factor 1, carbonic anhydrase, clathrin light chain B (Lcb), enolase 1, ATP synthase, cytochrome c, THO complex 4, and M-phase phosphoprotein 1. The findings were further corroborated in an independent group of NR2B Tg and WT mice by Western blot analysis of two selected proteins. The results revealed a unique profile of cortical proteins in the NR2B transgenic mice. A close association of functional activation of NR2B with the excitatory neurotransmission and neuroplasticity has been discussed.
N-甲基-D-天冬氨酸（NMDA）受体2B亚基（NR2B）对长时程增强（LTP）和突触可塑性至关重要。NR2B转基因小鼠在海马CA1区表现出更大的LTP，并在多项学习和记忆任务中行为表现增强。在本研究中，我们应用基于二维液相色谱的蛋白质组学方法，检测了在相同遗传背景下饲养的6个月大的NR2B转基因（Tg）小鼠及其野生型（WT）小鼠大脑皮质蛋白的表达水平。蛋白质在第一维根据pI使用聚焦色谱柱进行分离，在第二维根据疏水性使用无孔反相硅胶柱进行分离。应用DeltaVue软件检测蛋白质样品的差异表达。通过基质辅助激光解吸/电离飞行时间（MALDI-TOF/TOF）质谱鉴定出26种差异表达蛋白，包括谷氨酰胺合成酶（GS）、鸟嘌呤核苷酸释放因子1、碳酸酐酶、网格蛋白轻链B（Lcb）、烯醇化酶1、ATP合酶、细胞色素c、THO复合物4和M期磷蛋白1。通过对两种选定蛋白的蛋白质印迹分析，在另一组独立的NR2B Tg和WT小鼠中进一步证实了这些发现。结果揭示了NR2B转基因小鼠皮质蛋白的独特图谱。讨论了NR2B的功能激活与兴奋性神经传递和神经可塑性的密切关联。
