CaMKII hypo-nitrosylatlon In age-related decline of synaptic plasticity and cognition

CaMKII 低亚硝基化在与年龄相关的突触可塑性和认知能力下降中

• 批准号: 10154827
• 负责人: Nicole Rumian
• 金额: $ 3.55万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-03-05 至 2023-03-04
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10154827
• 关键词: Affect; Age-associated memory impairment; Aging; Animals; Anxiety; Behavior; Behavioral; Behavioral Assay; Binding; Biochemical; Cognition; Cysteine; Data; Dependence; Development; Elderly; Electrophysiology (science); Excitatory Synapse; Exploratory Behavior; Frequencies; Hippocampus (Brain); Image; Impaired cognition; Impairment; Intrabody; Investigation; Knock-in Mouse; L-Type Calcium Channels; Lead; Learning; Long-Term Potentiation; Mediating; Memory; Modeling; Molecular; Monitor; Motor Activity; Movement; Mus; Mutant Strains Mice; Mutation; N-Methyl-D-Aspartate Receptors; Neurons; Output; Pharmacology; Play; Population; Problem Solving; Protein Overexpression; Proteins; Quality of life; Radial; Regulation; Role; Short-Term Memory; Synapses; Synaptic Potentials; Synaptic plasticity; Techniques; Testing; Therapeutic; Therapeutic Intervention; Wild Type Mouse; age related; aged; arm; base; calmodulin-dependent protein kinase II; cognitive function; experimental study; insight; mutant; spatial memory; water maze; young adult

PROJECT SUMMARY Synaptic plasticity, specifically long-term potentiation (LTP), is thought to underlie learning, memory, and cognition. This function is disrupted in aging populations, where cognitive decline, such as forgetfulness and decreased problem-solving capacity, is a well known impairment. LTP requires Ca2+/calmodulin (CaM)- dependent protein kinase II (CaMKII), its Ca2+-independent autonomous activity and regulated binding to N- methyl-D-aspartate receptor (NMDAR), which results in CaMKII translocation to excitatory synapses. It has recently been suggested that hypo-nitrosylation may underlie the cognitive impairment seen in aging. My preliminary live-imaging data demonstrate that application of an NO donor is sufficient to stimulate CaMKII translocation to excitatory synapses and that this effect is abolished in a genetically modified knock-in mouse line, CaMKIIΔSNO, in which CaMKII is rendered un-nitrosylable. Other preliminary data from the lab suggests that these mice demonstrate normal high frequency stimulation (HFS)-induced LTP, but have impaired theta burst stimulation (TBS)-induced LTP, which mimics what is seen in aged animals. My proposal will test the hypotheses that (i) CaMKII synaptic targeting can be regulated by nitrosylation and (ii) CaMKII hypo-nitrosylation mediates both age-related LTP and cognitive impairment. Specifically, I will use live-imaging, biochemical, electrophysiological, and behavioral techniques on wild-type and mutant mice to determine the effect of CaMKII nitrosylation on synaptic plasticity and determine if hypo-nitrosylation underlies cognitive impairment seen in aging. Notably, this project utilizes intrabodies, which allow for simultaneous imaging of multiple endogenous proteins without the normal confounds seen in protein overexpression experiments, to live-monitor endogenous CaMKII targeting and an unpublished mouse line, CaMKIIΔSNO. Further, this project will provide a comprehensive investigation of the role of CaMKII nitrosylation in synaptic plasticity and aging, as it begins at the molecular level and proceeds through to functional behavioral outputs. The results from this project will not only elucidate the cellular and molecular role of CaMKII nitrosylation in synaptic plasticity, but also provide insights into how hypo- nitrosylation may underlie the cognitive impairment seen in aging, thus leading to a better understanding of synaptic plasticity and potential therapeutic interventions for aging populations.

项目概要 突触可塑性，特别是长时程增强 (LTP)，被认为是学习、记忆和认知能力的基础。 这种功能在老龄化人群中会受到干扰，认知能力下降，例如健忘和 解决问题的能力下降是 LTP 需要 Ca2+/钙调蛋白 (CaM)- 的一种众所周知的损伤。 依赖性蛋白激酶 II (CaMKII)，其独立于 Ca2+ 的自主活性和受调节的 N- 结合 甲基-D-天冬氨酸受体 (NMDAR)，导致 CaMKII 易位至兴奋性突触。 最近有人提出，低亚硝基化可能是衰老过程中认知障碍的根源。 初步实时成像数据表明，NO 供体的应用足以刺激 CaMKII 易位到兴奋性突触，并且这种效应在基因敲除小鼠中被消除 线，CaMKIIΔSNO，其中 CaMKII 被呈现为不可亚硝基化的。实验室的其他初步数据表明： 这些小鼠表现出正常的高频刺激 (HFS) 诱导的 LTP，但 θ 爆发受损 刺激（TBS）诱导的 LTP，模仿老年动物的情况，我的建议将测试这些假设。 (i) CaMKII 突触靶向可通过亚硝基化调节，(ii) CaMKII 低亚硝基化介导 具体来说，我将使用实时成像、生化、 对野生型和突变型小鼠进行电生理学和行为技术以确定 CaMKII 的作用 亚硝基化对突触可塑性的影响，并确定低亚硝基化是否是认知障碍的基础 值得注意的是，该项目利用了体内，可以同时对多个内源性进行成像。 蛋白质没有在蛋白质过表达实验中看到的正常混杂，以实时监测内源性 CaMKII 靶向和未发表的小鼠品系 CaMKIIΔSNO 此外，该项目将提供全面的研究。 从分子水平开始研究 CaMKII 亚硝基化在突触可塑性和衰老中的作用 并继续进行功能行​​为输出。该项目的结果不仅会阐明 CaMKII 亚硝基化在突触可塑性中的细胞和分子作用，同时也提供了关于低分子机制的见解。 亚硝基化可能是衰老过程中认知障碍的基础，因此可以更好地理解 突触可塑性和针对老龄化人群的潜在治疗干预措施。