Understanding how post-translational palmitoylation influences in vivo molecular and circuit dynamics during learning

了解翻译后棕榈酰化如何影响学习过程中的体内分子和电路动力学

• 批准号: 10025185
• 负责人: Jessica C Nelson
• 金额: $ 9.84万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-25 至 2022-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10025185
• 关键词: Ablation; Acoustic Nerve; Acoustics; Acute; Affect; Age; Anatomy; Animals; Auditory; Axon; BRAIN initiative; Behavior; Behavioral; Behavioral Genetics; Biochemical; Biological; Biological Assay; Biological Process; Brain; Brain Diseases; Calcium; Cells; Cognition; Complex; Data; Decision Making; Depressed mood; Disease; Electrophysiology (science); Exhibits; Fiber; Food; Foundations; Genes; Genetic; Genetic Screening; Hair Cells; Heritability; Human; Huntington Disease; Image; Imaging Techniques; Impairment; Individual; Invertebrates; Knowledge; Laboratories; Larva; Lasers; Learning; Mauthner' s neuron; Microscopy; Modeling; Molecular; Molecular Genetics; Molecular Target; Neurobehavioral Manifestations; Neurons; Pathway interactions; Pennsylvania; Play; Population; Post-Translational Protein Processing; Potassium Channel; Process; Protein Dynamics; Proteins; Research; Role; Schizophrenia; Source; Startle Reaction; Stimulus; Synapses; Synaptic plasticity; System; Testing; Time; Training; Transgenic Organisms; Universities; Vertebrates; Zebrafish; autism spectrum disorder; awake; base; career; cognitive task; drug of abuse; experimental study; habituation; human disease; in vivo; insight; interest; knockout gene; member; mutant; neural circuit; neuronal cell body; novel; optogenetics; palmitoylation; response; screening; sound; synaptic function

Habituation is a simple form of learning in which animals reduce responsiveness to repetitive stimuli. Habituation forms a foundation for normal cognition; without the ability to filter irrelevant stimuli, animals are unable to perform more complex cognitive tasks. Indeed, habituation learning is impaired in a wide range of heritable human disorders that present with more complex cognitive symptoms, including Schizophrenia, Autism Spectrum Disorders and Huntington’s Disease. Habituation learning is also a significant component of our innate decision-making: habituation to particular foods or drugs of abuse influences our responses to these stimuli and our decisions to seek them. Beyond its relevance for human behavior and disease, habituation can provide a simple and accessible model for examining some of the most exciting mysteries that inspired the BRAIN initiative, such as how proteins are mobilized during learning to alter synapses, circuits and behavior. Despite great interest, decades of study, and relevance for human disease, there are still significant gaps in our understanding of habituation learning. This proposal is based on the candidate’s discovery that post-translational palmitoylation plays a critical role in habituation learning. Specifically, using the larval zebrafish, she has found that the palmitoyltransferase Hip14 targets the Shaker-like channel Kv1.1 to regulate learning. This novel learning pathway represents an entirely independent research niche from which the PI will establish her own laboratory. The Granato lab, although expert in behavioral genetics, has never systematically examined how post-translational modifications influence protein dynamics, synapses, and behavior in real time. The PI will receive training from world experts in palmitoylation (Dr. Eric Witze) and in vivo electrophysiology (Dr. Alberto Pereda), integrating these approaches into a well-rounded system to examine learning across genes, circuits, and behavior. The PI will be based in the laboratory of Dr. Michael Granato at the University of Pennsylvania for the entire K99 period. During this time, the PI will learn to perform electrophysiological recordings in vivo to identify how activity within individual neurons is dynamic during habituation learning, and how plasticity is disrupted in mutants lacking Hip14 or Kv1.1 (Aim 1). This approach will be combined with calcium imaging, unbiased whole brain activity mapping, and transgenic rescue experiments to identify new circuit loci for habituation learning. Simultaneously, the PI will perform biochemical and live imaging experiments to examine how protein palmitoylation changes during learning, and how palmitoylation affects target protein localization in vivo (Aim 2). Finally, the PI will conduct a candidate screen to identify additional learning-relevant targets for Hip14 palmitoylation (Aim 3). These efforts will establish a broad and independent foundation for the candidate’s independent career investigating how post-translational modifications influence synaptic plasticity within defined neural circuits as we learn.

习惯化是一种简单的学习形式，动物会降低对重复刺激的反应性。习惯化形成了正常认知的基础；如果没有过滤无关刺激的能力，动物就无法执行更复杂的认知任务。事实上，习惯性学习在广泛的可遗传人类疾病中受到损害，这些疾病呈现出更复杂的认知症状，包括精神分裂症、自闭症谱系障碍和亨廷顿病。习惯化学习也是我们与生俱来的决策的一个重要组成部分：对滥用特定食物或药物的习惯性影响我们对这些刺激的反应和我们寻求它们的决定。除了与人类行为和疾病的相关性外，习惯化还可以提供一个简单易用的模型，用来研究激发大脑倡议的一些最令人兴奋的谜团，比如在学习过程中蛋白质是如何被动员起来改变突触、回路和行为的。尽管人们对习惯性学习非常感兴趣，经过了几十年的研究，而且与人类疾病有关，但我们对习惯性学习的理解仍然存在重大差距。这一建议是基于候选人的发现，即翻译后棕榈酸化在习惯性学习中起着关键作用。具体地说，通过对斑马鱼幼虫的研究，她发现棕榈酰基转移酶Hip14以类似Shaker的通道Kv1.1为靶标来调节学习。这条新的学习途径代表了一个完全独立的研究利基，PI将在这个利基上建立自己的实验室。格拉纳托实验室虽然是行为遗传学的专家，但从未系统地研究过翻译后修饰如何实时影响蛋白质动力学、突触和行为。PI将接受棕榈酰化(Eric Witze博士)和活体电生理学(Alberto Pereda博士)方面的世界专家的培训，将这些方法整合到一个全面的系统中，以检查跨基因、电路和行为的学习。在整个K99期间，PI将在宾夕法尼亚大学Michael Granato博士的实验室工作。在此期间，PI将学习在体内进行电生理记录，以确定在习惯性学习过程中单个神经元内的活动是如何动态的，以及缺乏Hip14或Kv1.1的突变体的可塑性是如何被破坏的(目标1)。这一方法将与钙成像、无偏全脑活动图和转基因救援实验相结合，以确定新的习惯性学习回路基因座。同时，PI将进行生化和现场成像实验，以检查蛋白质棕榈酰化在学习过程中的变化，以及棕榈酸化如何影响体内目标蛋白质的定位(目标2)。最后，PI将进行候选筛选，以确定Hip14棕榈酰化的更多与学习相关的目标(目标3)。这些努力将为候选人的独立职业生涯奠定广泛和独立的基础，调查翻译后修饰如何影响定义的神经回路内的突触可塑性。