Integrins and proteoglycans in LTP

LTP 中的整合素和蛋白聚糖

基本信息

批准号：
6359021
负责人：
RICHARD G LEBARON
金额：
$ 19.07万
依托单位：
UNIVERSITY OF TEXAS SAN ANTONIO
依托单位国家：
美国
项目类别：
财政年份：
2000
资助国家：
美国
起止时间：
2000-09-01 至 2001-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6359021
关键词：
brain mapping cell adhesion molecules cooperative study electrophysiology extracellular matrix proteins hippocampus integrins laboratory rabbit laboratory rat long term potentiation neural plasticity neutralizing antibody proteoglycan synapses

项目摘要

The proposed study will provide a better understanding of experience- dependent structural plasticity in area CA1 of the hippocampus. The results will provide insights into cellular mechanisms that might contribute to neuropathology such as Alzheirner's Disease as it relates to Long-term potentiation (LTP). LTP is believed to be an important component of some forms of learning and memory. A current focus of neuroscience is to understand the molecular mechanisms of LTP. Recent findings demonstrated an integrin-binding peptide and a heparan sulfate proteoglycan (HSPG)-binding peptide antagonized maintenance of LTP in area CA1 of rat hippocampus. The general goal of the collaborative research proposed is to understand whether integrins and HSPGs play a role in maintenance of LTP and whether they may associate to form adhesive and signaling structures in synapses that are analogous to focal adhesions. Studies are proposed to test the working hypothesis that integrins and HSPGs play a role in LTP and share a signaling pathway that is necessary for maintenance of LTP. Application of peptide on hippocampus slice will reveal whether the effects on LTP are time- dependent. The precise integrins and HSP(3s in area CA1 that affect LTP will be tested for by cross-linking experiments and mass spectrometry mass map analysis of protein cross-linked protein. Immunohistochemistry using antibodies to integrins and HSPGs will verify their expression in area CA1. Patch-clamp introduction of antibody in neurons in area CA1 and whole-cell recording will confirm integrin and HSPG functions in LTP. The expression and co-localization of integrins, HSPGs and components of focal adhesions will be tested for by immunofluorescence and transmission electron microscopy. The proposed project will be conducted as an integrated and coordinated collaboration between Dr. Richard G. LeBaron and Dr. Joe L. Martinez Jr. at The University of Texas at San Antonio (UTSA). Dr. LeBaron will focus on the component of the project that cross-links integrins and HSPGs that may function in synapses and for immunohistochemical analyses. Dr. Martinez will focus on 'the component of the project that tests physiological effects of peptide and antibody on LTP. Dr. LeBaron is an expert in the field of cell adhesion and extracellular matrix interactions and Dr. Martinez is an expert in the field of LTP. Because the proposed research focuses on the functional role of cell-surface adhesion proteins in synapses active in LTP, the experimental design proin6tes a strong and complementary application of the expertise of Drs. LeBaron and Martinez. The proposed collaboration will also provide opportunities for students with diverse backgrounds and fellows at The UTSA to obtain training in the neuroscience techniques that will be implemented to conduct the proposed research.

拟议的研究将有助于更好地理解海马 CA1 区的经验依赖性结构可塑性。这些结果将提供对可能导致神经病理学（例如阿尔茨海纳氏病）的细胞机制的见解，因为它与长时程增强（LTP）有关。 LTP 被认为是某些形式的学习和记忆的重要组成部分。当前神经科学的一个焦点是了解 LTP 的分子机制。最近的研究结果表明，整合素结合肽和硫酸乙酰肝素蛋白聚糖 (HSPG) 结合肽可拮抗大鼠海马 CA1 区 LTP 的维持。所提出的合作研究的总体目标是了解整合素和 HSPG 是否在 LTP 的维持中发挥作用，以及它们是否可以在突触中形成类似于粘着斑的粘附和信号结构。提出的研究旨在检验整合素和 HSPG 在 LTP 中发挥作用并共享维持 LTP 所需的信号通路的工作假设。将肽应用于海马切片将揭示对 LTP 的影响是否具有时间依赖性。将通过交联实验和蛋白质交联蛋白的质谱质量图分析来测试CA1区中影响LTP的精确整合素和HSP(3s)。使用整合素和HSPG的抗体的免疫组织化学将验证它们在CA1区中的表达。在 CA1 区神经元中膜片钳引入抗体并进行全细胞记录将证实整合素和 HSPG 在 LTP 中的功能整合素的表达和共定位。 HSPG 和粘着斑成分将通过免疫荧光和透射电子显微镜进行测试。拟议的项目将由德克萨斯大学 Richard G. LeBaron 博士和 Joe L. Martinez Jr. 博士进行综合协调合作进行。 LeBaron 博士将重点研究可能在突触中发挥作用的整合素和 HSPG 的交联部分，以及 Martinez 博士将进行的免疫组织化学分析。重点关注“测试肽和抗体对 LTP 生理效应的项目组成部分”。 LeBaron 博士是细胞粘附和细胞外基质相互作用领域的专家，Martinez 博士是 LTP 领域的专家。由于拟议的研究重点是细胞表面粘附蛋白在 LTP 中活跃的突触中的功能作用，因此实验设计表明了 Drs. 的专业知识的强大和补充应用。勒巴伦和马丁内斯。拟议的合作还将为 UTSA 具有不同背景的学生和研究员提供获得神经科学技术培训的机会，这些技术将用于开展拟议的研究。