Ahmed Md Mahiuddin, Dhanasekaran A Ranjitha, Block Aaron, Tong Suhong, Costa Alberto C S, Gardiner Katheleen J
Linda Crnic Institute for Down Syndrome, Department of Pediatrics;
Mol Cell Proteomics. 2014 Apr;13(4):919-37. doi: 10.1074/mcp.M113.035568. Epub 2014 Jan 27.
Analysis of the molecular basis of learning and memory has revealed details of the roles played by many genes and the proteins they encode. Because most individual studies focus on a small number of proteins, many complexities of the relationships among proteins and their dynamic responses to stimulation are not known. We have used the technique of reverse phase protein arrays (RPPA) to assess the levels of more than 80 proteins/protein modifications in subcellular fractions from hippocampus and cortex of mice trained in Context Fear Conditioning (CFC). Proteins include components of signaling pathways, several encoded by immediate early genes or involved in apoptosis and inflammation, and subunits of glutamate receptors. At one hour after training, levels of more than half the proteins had changed in one or more fractions, among them multiple components of the Mitogen-activated protein kinase, MAPK, and Mechanistic Target of Rapamycin, MTOR, pathways, subunits of glutamate receptors, and the NOTCH pathway modulator, NUMB homolog (Drosophila). Levels of 37 proteins changed in the nuclear fraction of hippocampus alone. Abnormalities in levels of thirteen proteins analyzed have been reported in brains of patients with Alzheimer's Disease. We therefore further investigated the protein profiles of mice treated with memantine, a drug approved for treatment of AD. In hippocampus, memantine alone induced many changes similar to those seen after CFC and altered the levels of seven proteins associated with Alzheimer's Disease abnormalities. Lastly, to further explore the relevance of these datasets, we superimposed responses to CFC and memantine onto components of the long term potentiation pathway, a process subserving learning and memory formation. Fourteen components of the long term potentiation pathway and 26 proteins interacting with components responded to CFC and/or memantine. Together, these datasets provide a novel view of the diversity and complexity in protein responses and interactions following normal learning.
对学习和记忆分子基础的分析揭示了许多基因及其编码蛋白质所起作用的细节。由于大多数个体研究仅关注少数蛋白质,因此蛋白质之间关系的许多复杂性及其对刺激的动态反应尚不为人所知。我们使用反相蛋白质阵列(RPPA)技术评估了在情境恐惧条件反射(CFC)训练的小鼠海马体和皮质亚细胞组分中80多种蛋白质/蛋白质修饰的水平。这些蛋白质包括信号通路的组成部分,其中一些由即刻早期基因编码或参与细胞凋亡和炎症反应,以及谷氨酸受体的亚基。训练后一小时,超过一半的蛋白质在一个或多个组分中的水平发生了变化,其中包括丝裂原活化蛋白激酶(MAPK)和雷帕霉素作用靶点(MTOR)通路的多个组成部分、谷氨酸受体亚基以及NOTCH通路调节剂NUMB同源物(果蝇)。仅海马体细胞核组分中就有37种蛋白质的水平发生了变化。在阿尔茨海默病患者的大脑中,已报道了所分析的13种蛋白质水平异常。因此,我们进一步研究了美金刚治疗的小鼠的蛋白质谱,美金刚是一种已被批准用于治疗阿尔茨海默病的药物。在海马体中,单独使用美金刚诱导了许多与CFC后相似的变化,并改变了与阿尔茨海默病异常相关的7种蛋白质的水平。最后,为了进一步探索这些数据集的相关性,我们将对CFC和美金刚的反应叠加到长时程增强通路的组成部分上,长时程增强是一个支持学习和记忆形成的过程。长时程增强通路的14个组成部分以及与这些组成部分相互作用的26种蛋白质对CFC和/或美金刚有反应。这些数据集共同提供了一个关于正常学习后蛋白质反应和相互作用的多样性和复杂性的新视角。