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

基本信息

批准号：
9892327
负责人：
Jessica C Nelson
金额：
$ 9.9万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-09-25 至 2021-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9892327
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&apos 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

项目摘要

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，其中动物减少对重复刺激的反应，形成了正常认知的基础；如果没有过滤不相关刺激的能力，动物就无法执行更复杂的认知任务。一系列具有更复杂认知症状的遗传性人类疾病，包括精神分裂症、自闭症谱系障碍和亨廷顿舞蹈病，习惯学习也是我们先天决策的重要组成部分：对特定食物的习惯。药物或滥用会影响我们对这些刺激的反应以及我们寻求这些刺激的决定，除了与人类行为和疾病的相关性之外，习惯化还可以提供一个简单且易于理解的模型来研究激发 BRAIN 计划的一些最令人兴奋的谜团，例如如何进行这些刺激。尽管经过数十年的研究和人类疾病的相关性，人们对学习改变突触、回路和行为的过程中的蛋白质进行了动员，但该提议是基于候选人的发现，即后-翻译的具体而言，她利用斑马鱼幼虫发现棕榈酰转移酶 Hip14 以 Shaker 样通道 Kv1.1 为靶点来调节学习，这种新颖的学习途径代表了 PI 的一个完全独立的研究领域。 Granato 实验室虽然是行为遗传学方面的专家，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。接受棕榈酰化（Eric Witze 博士）和体内电生理学（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 博士的实验室进行电生理记录，以识别在习惯化学习过程中单个神经元内的活动如何动态变化。缺乏 Hip14 或 Kv1.1 的突变体的可塑性被破坏（目标 1）。这种方法将与钙成像、无偏全脑活动图谱和转基因救援实验相结合，以识别新的回路基因座以进行习惯化学习。通过生化和实时成像实验来检查蛋白质棕榈酰化在学习过程中如何变化，以及棕榈酰化如何影响体内目标蛋白质定位（目标 2）。确定 Hip14 棕榈酰化的其他与学习相关的目标（目标 3），这些努力将为候选人的独立职业生涯奠定广泛且独立的基础，研究翻译后修饰如何影响我们学习的定义神经回路内的突触可塑性。