Molecular basis of low-and high-pHi hippocampal neurons

低pHi和高pHi海马神经元的分子基础

• 批准号: 6998615
• 负责人: Michelle L KELLY
• 金额: $ 4.83万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-01 至 2007-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6998615
• 关键词: acid base balance; acidity /alkalinity; bicarbonates; gene expression; hippocampus; hypoxia; immunocytochemistry; laboratory rat; membrane transport proteins; neurons; neuroregulation; phenotype; polymerase chain reaction; postdoctoral investigator; tissue /cell culture

DESCRIPTION (provided by applicant): Hippocampal (HC) neurons exist in two populations based on initial intracellular pH (pHi); one consisting of neurons with low initial pHi values and the other with high initial pHi values. High-pHi neurons recover faster from acid loads than do low-pHi neurons. There are two specific aims: (1) to explore the molecular basis of low- vs. high-pHi neurons and determine whether the difference relates to neurotransmitter content and/or properties of specific HCO3 transporters expressed by a neuron. The approach will be to monitor pHi in HC neurons grown on gridded coverslips and assess activities of HCO3 transporters in identified cells. Immunocytochemistry and single-cell PCR will be used to assess molecular phenotype in the same cells. A knockdown technique will show how the expression of specific HCO3 transporters affects the low/high pHi state. (2) To use chronic hypoxia, and the return to normoxia, as tools to shift the low- vs. high-pHi status of HC neurons. The approach will be to monitor individual neurons. The results from these studies are significant because the pHi differences between low- and high-pHi neurons are large enough to have a major impact on neuronal activity. In other words, pHi shifts could be a form of neuromodulation.

描述（由申请人提供）：基于初始细胞内pH（PHI），海马（HC）神经元存在于两个种群中；一个由初始PHI值低的神经元组成，另一个由具有较高初始PHI值的神经元组成。与低phi神经元相比，高pHI神经元从酸负荷中恢复的速度更快。有两个具体的目的：（1）探索低和高phi神经元的分子基础，并确定差异是否与神经元表达的特定HCO3转运蛋白的神经递质含量和/或特性有关。该方法将是监测在网格盖玻片上生长的HC神经元中的PHI，并评估识别细胞中HCO3转运蛋白的活性。免疫细胞化学和单细胞PCR将用于评估同一细胞中的分子表型。敲低技术将显示特定HCO3转运蛋白的表达如何影响低/高PHI状态。 （2）使用慢性缺氧以及回到常规氧的恢复，作为移动HC神经元低和高phi状态的工具。该方法将是监测单个神经元。这些研究的结果很重要，因为低phi神经元之间的PHI差异足够大，可以对神经元活性产生重大影响。换句话说，PHI转移可能是神经调节的一种形式。