Molecular Mechanisms of Ethanol Sensitivity in ILS and ISS Mice

ILS 和 ISS 小鼠乙醇敏感性的分子机制

• 批准号: 6967079
• 负责人: MICHAEL D BROWNING
• 金额: $ 34.05万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-08-01 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6967079
• 关键词: GABA receptor; NMDA receptors; ethanol; genetic strain; inbreeding; laboratory mouse; neuropharmacology; pharmacogenetics; pharmacokinetics; phosphorylation; receptor expression; receptor sensitivity; sleep

DESCRIPTION (provided by applicant): We have previously shown that GABAARs in ILS mice exhibit ethanol potentiation while ethanol had no effect on GABAARs in hippocampi of ISS mice. Similarly, preliminary data presented in this proposal show that ethanol potentiates the GABAAR in cortex of ILS but not ISS mice and inhibits the NMDAR in hippocampus and cortex of ILS but not ISS mice. These dramatic differences in ILS and ISS mice in terms of the ability of ethanol to affect the GABAAR and NMDAR in hippocampus and cortex will be the principal focus of our proposal. We are proposing to study the molecular mechanisms that underlie the differences in ethanol sensitivity of the GABAAR and NMDAR in ILS and ISS mice. We anticipate that these two strains of mice will thus provide us with a unique opportunity to test hypotheses about these mechanisms. This will be the principal goal of the proposed studies. In pursuit of this goal our proposal will have three Specific Aims. Aim #1. Test the hypothesis that differences in the phosphorylation of the NMDAR and/or GABAAR in ILS and ISS mice may underlie the differences in ethanol effects on these receptors in the two strains of mice. We and others have recently shown that phosphorylation may play an important role in the ethanol sensitivity of the NMDAR and the GABAAR. And, we have shown that GABAAR and NMDAR of ILS mice are sensitive to ethanol and those of ISS mice are not. Therefore, in this aim we will compare the basal and ethanol-stimulated phosphorylation of specific subunits and/or phosphosites of the NMDAR and GABAAR in ILS and ISS mice. Aim #2. Test the hypothesis that differences in the ethanol sensitivity of the trafficking mechanisms for the NMDAR and/or GABAAR in ILS and ISS mice may underlie the differences in ethanol effects on these receptors in the two strains of mice. We and others have shown that NMDAR and GABAAR membrane trafficking may play important roles in the function of these receptors. Moreover, we have recently obtained preliminary data that ethanol reduces membrane surface expression of the NMDAR in ILS but not in ISS mice. Therefore, in this aim we will confirm and extend this observation by comparing the basal and ethanol- stimulated surface expression of the NMDAR and GABAAR in ILS and ISS mice. Aim #3 Test the hypothesis that interventions that modulate phosphorylation and/or surface expression of the NMDAR and GABAAR will reduce the effect of ethanol on GABAA R and NMDAR responses in ILS mice and and "rescue" ethanol effects on these receptors in ISS mice. Based on results from Aims 1 and 2 above, we will intervene to pharmacologically modulate phosphorylation and/or surface expression. We will then test these interventions to determine whether the biochemical differences seen in Aims 1 and 2 or either necessary or sufficient to explain the differences in ethanol effects on the evoked responses of these receptors in ILS and ISS mice.

描述（由申请人提供）：我们先前表明，ILS小鼠中的Gabaars表现出乙醇增强，而乙醇对ISS小鼠海马的Gabaars没有影响。同样，本提案中提出的初步数据表明，乙醇在ILS的皮层中增强了Gabaar，但不抑制IS的NMDAR，并抑制了ILS的海马和皮质的NMDAR，但不抑制ISS小鼠的NMDAR。在ILS和ISS小鼠中，这些巨大的差异就乙醇影响海马和Cortex的Gabaar和NMDAR的能力而言将是我们建议的主要重点。我们建议研究ILS和ISS小鼠中GABAAR和NMDAR乙醇敏感性差异的基础的分子机制。我们预计这两种小鼠将为我们提供一个独特的机会来检验有关这些机制的假设。这将是拟议研究的主要目标。为了实现这一目标，我们的建议将具有三个具体目标。目标＃1。检验以下假设：IL和ISS小鼠中NMDAR和/或GABAAR磷酸化的差异可能是小鼠两种菌株中乙醇对这些受体效应的差异的基础。我们和其他人最近表明，磷酸化可能在NMDAR和GABAAR的乙醇敏感性中起重要作用。而且，我们已经表明，ILS小鼠的Gabaar和NMDAR对乙醇很敏感，而ISS小鼠的小鼠也不是。因此，在此目标中，我们将比较ILS和ISS小鼠中NMDAR和GABAAR的特定亚基和/或磷脂的基底和乙醇刺激的磷酸化。目标＃2。检验以下假设：ILS和ISS小鼠在IL和ISS小鼠中对NMDAR和/或GABAAR运输机制的乙醇敏感性差异可能是两种小鼠菌株中乙醇对这些受体的差异的基础。我们和其他人表明，NMDAR和GABAAR膜运输可能在这些受体的功能中起重要作用。此外，我们最近获得了乙醇的初步数据，这些数据可降低IL中NMDAR的膜表面表达，但在ISS小鼠中却没有。因此，在此目标中，我们将通过比较IL和ISS小鼠中NMDAR和GABAAR的基底和乙醇刺激的表面表达来确认和扩展这一观察结果。 AIM＃3检验了以下假设：NMDAR和GABAAR调节磷酸化和/或表面表达的干预措施将减少乙醇对ILS小鼠中GABAA R和NMDAR反应的影响，以及“ Rescue”乙醇对ISS小鼠中这些受体的影响。根据上面的目标1和2的结果，我们将干预药理调节磷酸化和/或表面表达。然后，我们将测试这些干预措施，以确定目标1和目标2中看到的生化差异是否需要或足以解释乙醇对ILS和ISS小鼠中这些受体诱发反应的影响的差异。