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PKC Delta in Ethanol Regulation of GABA-A Receptors and Behavior

GABA-A 受体和行为的乙醇调节中的 PKC Delta

基本信息

批准号：
8494465
负责人：
ROBERT O. MESSING
金额：
$ 29.13万
依托单位：
UNIVERSITY OF TEXAS AT AUSTIN
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-20 至 2015-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8494465
关键词：
Adult Alanine Alcohol consumption Alcoholic Intoxication Amygdaloid structure Ataxia Behavior Behavior Control Behavioral Biological Assay Blood - brain barrier anatomy Brain Brain region Cells Chemosensitization Cultured Cells Development Dose Ethanol Fibroblasts Financial compensation GABA-A Receptor Genes Goals Health Hippocampus (Brain)Human In Vitro Injection of therapeutic agent Intoxication Knock-in Mouse Knock-out Knockout Mice Life Mediating Molecular Mus Neurons Phenotype Phosphorylation Phosphorylation Site Phosphotransferases Play Property Protein Kinase C Proteins Regulation Research Personnel Resistance Rodent Role Self Administration Signal Pathway Signal Transduction Site Slice Specificity Testing Thalamic structure Tissues alcohol behavior alcohol effect alcohol response alcohol sensitivity alcohol use disorder analog base chemical genetics dentate gyrus drinking drinking water gamma-Aminobutyric Acid hypnotic interest kinase inhibitor mature animal mouse model mutant novel novel therapeutics preference protein kinase C-delta receptor social tool

项目摘要

DESCRIPTION (provided by applicant): Studies with protein kinase C delta knockout (PKC4-/-) mice indicate that PKC4 regulates ethanol intoxication and self-administration. These effects may be related to PKC4 actions at extrasynaptic GABAA receptors, since thalamic and hippocampal neurons from PKC4-/- mice lack ethanol enhancement of tonic inhibitory GABA currents. The main hypothesis of this project is that PKC4 regulates ethanol intoxication and self-administration by phosphorylating proteins that alter the function of extrasynaptic GABAA receptors in brain regions controlling these behaviors. This hypothesis will be tested using a novel mouse model, a knock-in mouse that expresses an ATP analog-sensitive mutant of PKC4 (AS-PKC4). AS-PKC4 can be selectively and potently inhibited by analogs of the general kinase inhibitor PP1 that cross the blood brain barrier and can be administered orally or by i.p. injection. Studies will determine if inhibition of AS-PKC4 mice increases signs of ethanol intoxication and enhances ethanol self-administration in adult mice. Electrophysiological studies in brain slices will investigate whether inhibiting AS-PKC4 blocks ethanol potentiation of tonic GABA currents in hippocampal, thalamic, and amygdala neurons. In vitro kinase assays will determine if 22, 23, and 4 subunits are possible substrates of PKC4. Novel ATP analogs will be used with tissues from AS-PKC4 mice to identify PKC4 phosphorylation sites on GABAA receptor-associated proteins. Studies in cells that heterologously express 1422/34 receptors and receptor-associated proteins with alanine substitutions at PKC4 phosphorylation sites, will determine if these sites regulate the ethanol sensitivity of 1422/34 receptors. The overall goal of this project is to identify a novel PKC4 signaling pathway that regulates the ethanol sensitivity of extrasynaptic GABAA receptors and may contain targets for the development of new therapeutics to treat alcohol use disorders.

描述（由申请人提供）：对蛋白激酶 C δ 敲除（PKC4-/-）小鼠的研究表明 PKC4 调节乙醇中毒和自我给药。这些效应可能与 PKC4 对突触外 GABAA 受体的作用有关，因为 PKC4-/- 小鼠的丘脑和海马神经元缺乏乙醇增强强直抑制性 GABA 电流的作用。该项目的主要假设是 PKC4 通过磷酸化蛋白质来调节乙醇中毒和自我给药，这些蛋白质改变控制这些行为的大脑区域中突触外 GABAA 受体的功能。这一假设将使用一种新型小鼠模型进行测试，该模型是一种表达 PKC4 ATP 类似物敏感突变体 (AS-PKC4) 的敲入小鼠。 AS-PKC4 可以被一般激酶抑制剂 PP1 的类似物选择性地、有效地抑制，PP1 的类似物可以穿过血脑屏障，可以口服或腹腔注射给药。注射。研究将确定抑制 AS-PKC4 小鼠是否会增加成年小鼠的乙醇中毒症状并增强乙醇的自我给药。脑切片的电生理学研究将探讨抑制 AS-PKC4 是否会阻断海马、丘脑和杏仁核神经元中强直 GABA 电流的乙醇增强作用。体外激酶测定将确定 22、23 和 4 个亚基是否是 PKC4 的可能底物。新型 ATP 类似物将与 AS-PKC4 小鼠的组织一起使用，以鉴定 GABAA 受体相关蛋白上的 PKC4 磷酸化位点。对异源表达 1422/34 受体和 PKC4 磷酸化位点处丙氨酸取代的受体相关蛋白的细胞进行研究，将确定这些位点是否调节 1422/34 受体的乙醇敏感性。该项目的总体目标是确定一种新的 PKC4 信号通路，该通路可调节突触外 GABAA 受体的乙醇敏感性，并可能包含开发治疗酒精使用障碍的新疗法的靶点。