Study of PAK3 in epilepsy-associated defects in synaptic plasticity

PAK3在癫痫相关突触可塑性缺陷中的研究

基本信息

批准号：
10046413
负责人：
Nien-Pei Tsai
金额：
$ 14.55万
依托单位：
UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-01 至 2022-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10046413
关键词：
Actin-Binding Protein Actins Address Affect Area Biological Assay Brain CDKN1A gene Chemosensitization Cognition Data Defect Development Disease Dissociation Epilepsy Foundations Genes Hippocampus (Brain)Impaired cognition Impairment In Vitro Intention Knowledge Lead Mediating Memory Memory impairment Missense Mutation Molecular Mutation Names Neurons Pathologic Patients Phosphorylation Phosphotransferases Pilot Projects Protein Isoforms Protein Kinase Proteins Proteomics Quality of life Recurrence Regulation Role Seizures Signal Transduction Synapses Synaptic plasticity Testing Ubiquitination United States Validation Work base cofilin comorbidity conditional knockout memory consolidation memory retrieval mouse model nerve stem cell novel optogenetics polymerization therapeutic effectiveness ubiquitin-protein ligase upstream kinase

项目摘要

PROJECT SUMMARY/ABSTRACT Epilepsy affects more than 3 million people in the United States. In addition to unprovoked seizures, cognitive decline and memory impairment are common comorbidities associated with epilepsy, but our knowledge in this area is limited. Our preliminary work suggests that an epilepsy associated ubiquitin E3 ligase Nedd4-2 mediates actin polymerization through promoting phosphorylation of an actin binding protein cofilin during the induction of long-term synaptic potentiation (LTP). LTP describes long‐lasting increments of synaptic efficiency and is crucial for cognition and memory formation. In an effort to identify ubiquitination substrates of Nedd4-2 responsible for cofilin phosphorylation, we identified PAK3, an IDG-eligible understudied kinase, as a potential substrate of Nedd4-2 that contributes to cofilin phosphorylation during LTP. This pilot project is formulated to test our hypothesis that induction of LTP de-represses Nedd4-2 to stabilize PAK3 and subsequently induce cofilin phosphorylation and actin polymerization. In Aim 1, we will determine how PAK3 is ubiquitinated by Nedd4-2 and how this ubiquitination is disrupted by epilepsy associated mutations in Nedd4-2. We will also determine whether induction of LTP de-represses Nedd4-2 to allow PAK3 stabilization during LTP. In Aim 2, we will employ a recently developed optogenetic approach to rapidly stabilize PAK3 in Nedd4-2 conditional knockout (cKO) neurons during the induction of LTP, with the intention to restore cofilin phosphorylation and actin polymerization, and ultimately to restore LTP. We expect our project to further our knowledge of an IDG-eligible protein PAK3 in synaptic plasticity and help explain cognitive decline in epilepsy.

项目摘要/摘要癫痫会影响联合赛季的300万人，认知能力下降和记忆障碍是与癫痫相关的常见常见常见，但是我们在这个领域的知识是有限的。连接酶NEDD4-2通过促进肌动蛋白结合蛋白的磷酸化介导肌动蛋白聚合长期突触增强（LTP）的指示期间Cofilin。突触效率，对于认知和记忆的形成至关重要。 NEDD4-2负责Cofilin磷酸化的底物，我们确定了PAK3，这是IDG符合条件的研究激酶作为NEDD4-2的潜在底物，在LTP期间有助于Cofilin磷酸化。该项目的制定是为了检验我们的假设，即LTP诱导de-bepresses nedd4-2稳定PAK3和随后诱导Cofilin磷酸化和肌动蛋白聚合。 NEDD4-2泛素化，以及这种泛素化如何受到NEDD4-2的癫痫病的纪律。我们还将确定LTP的指示是否会导致NEDD4-2允许LTP期间的PAK3稳定。在AIM 2中，我们将促进光遗传学方法来快速稳定NEDD4-2中的PAK3 有条件的敲除（CKO）神经元持续的LTP指示，并具有恢复Cofilin 磷酸化和肌动蛋白聚合，并最终恢复LTP。在突触塑料中了解IDG符合条件的蛋白质PAK3的知识，并有助于解释癫痫的下降。