The role of wake-associated protein kinase A transients in intrinsic plasticity and learning through aging

唤醒相关蛋白激酶 A 瞬变在内在可塑性和衰老学习中的作用

• 批准号: 10751540
• 负责人: Elizabeth Tilden
• 金额: $ 3.36万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2028-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10751540
• 关键词: Acute; Address; Adenylate Cyclase; Adult; Affect; Aging; Animal Behavior; Animals; Behavior; Behavioral; Biological Assay; Cell physiology; Cells; Cognitive deficits; Complex; Custom; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; Data; Development; Elderly; Electroencephalography; Electromyography; Electrophysiology (science); Fluorescence; Frequencies; Hippocampus; Hour; Impaired cognition; Impairment; Knowledge; Learning; Measurement; Memory; Memory impairment; Methodology; Methods; Molecular; Motivation; Mus; Nature; Neurons; Optical reporter; Optics; Pattern; Peptides; Perforation; Performance; Photometry; Physiological; Play; Population; Precision therapeutics; Process; Property; Protein Inhibition; Protein Kinase A Inhibitor; Quality of life; Role; Signal Transduction; Signaling Molecule; Sleep; Sleep Deprivation; Sleep Wake Cycle; Sleep disturbances; Slice; Synaptic plasticity; Testing; Therapeutic; Time; Work; age related; aged; aging population; cognitive change; extracellular; hippocampal pyramidal neuron; improved; memory consolidation; novel; novel therapeutics; optical sensor; patch clamp; pharmacologic; protein activation; protein function; protein kinase inhibitor peptide; temporal measurement; tool

Project Summary: Cognitive deficits including disruptions in hippocampal-dependent memory are a hallmark of aging. Predictably, aging-associated cognitive decline is exacerbated by sleep disruptions commonly seen in the aging and elderly population. However, there is a significant lack of understanding about the mechanism behind the interconnected processes of sleep, aging and learning. One significant challenge to unraveling these mechanisms has been the lack of tools to study intracellular and extracellular signals in real time with high temporal resolution. This has made it difficult to observe the modulation of these signals alongside such dynamic processes as sleep, learning and aging. To address these challenges, our lab developed a fluorescence-lifetime based optical sensor, FLIM-AKAR, which when used in combination with a custom-built fluorescence lifetime photometry (FLiP) rig has allowed us to observe the activity of cAMP-dependent protein kinase A (PKA), an important plasticity signal that has been implicated in the formation and consolidation of sleep-dependent learning and has been shown to enhance learning in aging mice. Paring 24-hour FLiP recordings in hippocampal CA1 with simultaneous electroencephalography (EEG) and electromyography (EMG) measurements revealed a synchronized, transient activation of PKA that is associated with transitions from sleep to wake. Due to its short duration, this signal has never been observed before in a behaving animal. Thus, this study aims to explore its function on both cellular and behavioral levels and elucidate how those functions may change in aging mice. Using photoactivatable adenylate cyclase (biPAC) and perforated patch clamp, I will determine whether transient PKA activation is sufficient to cause an increase in intrinsic excitability (IE), a known function of PKA and a known cellular correlate of learning. Further, by using biPAC and photoactivatable protein kinase inhibitor peptide (PA-PKI) to bidirectionally manipulate this transient PKA signal, I aim to determine whether increasing the frequency of these transients can rescue hippocampal- dependent learning deficits in aging mice or disrupt intact hippocampal-dependent learning in adult mice. Ultimately, our findings will provide a more nuanced understanding of how PKA functions at physiologic timescales and in the context of aging, sleep, and learning. This study will also stand as an example of how taking advantage of new optical tools can bolster our understanding of how the dynamics of cell signaling relate to complex behaviors.

项目摘要： 认知缺陷，包括海马依赖性记忆中的破坏是衰老的标志。可以预见， 衰老和老年人通常看到的睡眠中断会加剧与衰老相关的认知下降 人口。但是，人们对背后机制的理解有很大的了解 睡眠，衰老和学习的互连过程。揭开这些的重大挑战 机制一直缺乏实时研究细胞内和细胞外信号的工具 时间分辨率。这使得很难观察到这些信号的调制 动态过程作为睡眠，学习和衰老。为了应对这些挑战，我们的实验室开发了 基于荧光时代的光传感器Flim-Akar，当与定制的结合使用时 荧光寿命光度法（FLIP）钻机使我们能够观察CAMP依赖性蛋白的活性 激酶A（PKA），这是一个重要的可塑性信号，与形成和巩固有关 与睡眠有关的学习，已被证明可以增强老化小鼠的学习。削皮24小时翻转 同时脑电图（EEG）和肌电图的海马CA1记录 （EMG）测量结果显示PKA的同步，瞬态激活与过渡有关 从睡眠到醒来。由于其持续时间短，在表现动物中从未观察到该信号。 因此，本研究旨在探索其在细胞和行为水平上的功能，并阐明这些功能 衰老小鼠的功能可能会改变。使用光活性腺苷酸环化酶（BIPAC）和穿孔贴片 夹具，我将确定瞬态PKA激活是否足以引起内在兴奋性的增加 （即），PKA的已知功能和已知的细胞学习相关性。此外，使用BIPAC和 光活化蛋白激酶抑制剂肽（PA-PKI）双向操纵这种瞬态PKA 信号，我旨在确定增加这些瞬变频率是否可以挽救海马 - 成年小鼠衰老小鼠的依赖学习缺陷或破坏完整的海马依赖性学习。 最终，我们的发现将为PKA如何在生理学上发挥作用提供更细微的理解 时间尺度和衰老，睡眠和学习的背景。这项研究也将成为一个例子 利用新的光学工具可以增强我们对电池信号动态的理解 与复杂的行为有关。