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Role of CaMKII in memory storage

CaMKII 在记忆存储中的作用

基本信息

批准号：
10164875
负责人：
Leslie C Griffith
金额：
$ 35.55万
依托单位：
BRANDEIS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-08-01 至 2023-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10164875
关键词：
Aging Alzheimer&apos s Disease Animals Behavioral Binding Binding Proteins Brain-Derived Neurotrophic Factor Cell Death Complex Cytoplasm Development Dominant-Negative Mutation Epilepsy Fluorescence Resonance Energy Transfer Goals Healthcare Hippocampus (Brain)Hour In Vitro Learning Maintenance Memory Memory Loss Methods Modification Molecular Mutation N-Methylaspartate Nature Neurologic Dysfunctions Optical Methods Pathologic Processes Peptides Phosphotransferases Process Property Protein Synthesis Inhibitors Proteins Reaction Role Specificity Stroke Synapses Synaptic Transmission Synaptic plasticity Testing Vertebral column Viral Weight Work addiction base calmodulin-dependent protein kinase II densin-180 enhancing factor experimental study in vivo inhibitor/antagonist memory process nervous system disorder novel painful neuropathy prevent

项目摘要

Project Summary/Abstract Activity-dependent synaptic modifications (LTP/LTD) are a major candidate for the mechanism of memory. LTP involves induction, maintenance, and expression processes. This proposal seeks to elucidate the molecular basis of the maintenance process, the process that underlies the engram. The critical test of any hypothesis regarding maintenance is the “erasure test” in which an inhibitor is applied after LTP/memory is established. If this blocks LTP/memory and the effect persists after the inhibitor is removed, the inhibitor must have erased a maintenance process. We have conducted the erasure test using an inhibitor of CaMKII (CN-peptide). We found that application of this peptide after LTP induction produced erasure of saturated LTP. We now propose two experiments that test the role of CaMKIIα in memory maintenance at the behavioral level. In the first, we ask whether a dominant-negative form of CaMKIIα can erase conditioned place avoidance. We present strong preliminary evidence that it does. The second test is the “occlusion test.” It has been shown that activated kinase (CaMKIIα*) enhances synaptic transmission that occludes synaptically induced LTP. We will virally express CaMKIIα* and test two predictions: that because this maximally increases all synaptic weights it should destroy memory function, and furthermore, that learning under these conditions should not be possible. Preliminary evidence supporting these predictions is presented. Other experiments in this proposal are aimed at understanding the nature of the CaMKIIα complex that stores the engram. There are strong reasons to suspect that what maintains LTP is actually the complex of CaMKIIα with NMDAR (and perhaps also densin- 180). Studies of the binding of proteins to CaMKII have relied on in vitro work, and there has been no previous method for studying the complexes formed during actual LTP induction. Thus, crucial information regarding the complex formation and persistence is lacking. We have developed and validated a novel optical method based on FLIM-FRET. Our preliminary evidence demonstrates that LTP induction produces complex of CaMKIIα with GluN2B in spines and that the formation is synapse specific. We will determine the duration of the complex under conditions that either induce short-lasting LTP (early LTP) or produce both early and late LTP. We will also examine how the duration of the complex depends on factors that enhance (e.g., BDNF) or prevent (e.g., protein synthesis inhibitors) late LTP. This approach will be extended to study the interaction of CaMKIIα with densin-180. Having identified properties of the complex that underlies LTP, in vivo experiments will be conducted to test whether disruption of protein interactions within the complex can disrupt maintenance of the engram.

项目概要/摘要活动依赖性突触修饰（LTP/LTD）是记忆机制的主要候选者。长时程包括诱导、维持和表达过程。该提案旨在阐明分子维护过程的基础，即印迹的基础过程。任何假设的批判性检验关于维护，是“擦除测试”，在LTP/记忆建立后应用抑制剂。如果这会阻止 LTP/记忆，并且在去除抑制剂后效果仍然存在，抑制剂必须已擦除维护过程。我们使用 CaMKII（CN-肽）抑制剂进行了擦除测试。我们发现在 LTP 诱导后应用该肽会消除饱和 LTP。我们现在提议两项实验在行为层面测试 CaMKIIα 在记忆维持中的作用。首先，我们询问 CaMKIIα 的显性失活形式是否可以消除条件性场所回避。我们呈现强势初步证据表明确实如此。第二个测试是“遮挡测试”。已显示已激活激酶 (CaMKIIα*) 增强突触传递，阻断突触诱导的 LTP。我们将病毒式传播表达 CaMKIIα* 并测试两个预测：因为这最大限度地增加了所有突触权重应该会破坏记忆功能，而且，在这种情况下学习是不可能的。提出了支持这些预测的初步证据。本提案中的其他实验旨在了解存储印迹的 CaMKIIα 复合物的性质。有充分的理由怀疑维持 LTP 的实际上是 CaMKIIα 与 NMDAR（也许还有致密蛋白）的复合物 180）。蛋白质与 CaMKII 结合的研究依赖于体外工作，之前还没有研究实际 LTP 诱导过程中形成的复合物的方法。因此，关于缺乏复杂的形成和持久性。我们开发并验证了一种基于光学的新型方法在 FLIM-FRET 上。我们的初步证据表明，LTP 诱导产生 CaMKIIα 与棘中的 GluN2B 且其形成是突触特异性的。我们将确定综合体的持续时间在诱导短期 LTP（早期 LTP）或同时产生早期和晚期 LTP 的条件下。我们将还检查复合体的持续时间如何取决于增强（例如 BDNF）或预防（例如蛋白质合成抑制剂）晚期 LTP。该方法将扩展到研究 CaMKIIα 与密度素-180。确定了 LTP 基础复合物的特性后，体内实验将进行测试以测试复合物内蛋白质相互作用的破坏是否会破坏蛋白质的维持印迹。