The effect of alcohol withdrawal on molecular and behavioral circadian rhythms

酒精戒断对分子和行为昼夜节律的影响

• 批准号: 8061182
• 负责人: Walter Francis Wiggins
• 金额: $ 4.48万
• 依托单位: WAKE FOREST UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-30 至 2014-03-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8061182
• 关键词: Alcohol abuse; Alcohol consumption; Alcohol withdrawal syndrome; Alcoholism; Alcohols; American; Behavioral; Brain; Calcium Channel; Centers for Disease Control and Prevention (U.S.); Cessation of life; Chronic; Circadian Rhythms; Exhibits; Gene Expression; Habits; Individual; Laboratories; Molecular; Monitor; Physiological; Physiological Processes; Polymerase Chain Reaction; Postdoctoral Fellow; Process; Public Health; Regulation; Relapse; Relative (related person); Research; Research Training; Reverse Transcription; Risk; Sleep; Sleep Disorders; Sleep disturbances; T-Type Calcium Channels; Techniques; Time; Training; United States; United States National Institutes of Health; Variant; Western Blotting; Withdrawal; alcohol exposure; circadian behavioral rhythms; high risk drinking; innovation; mathematical model; mouse model; public health relevance; research study; skills training; training project; voltage

DESCRIPTION (provided by applicant): The focus of the research training project is the effect of multiple chronic alcohol exposure-withdrawal cycles on molecular and behavioral circadian rhythms. Sleep is possibly the most critical physiologic process regulated by circadian rhythms of activity in the brain. While not much is understood regarding the specific mechanisms underlying sleep and its importance, sleep disturbances can have devastating physiologic consequences, including death, if they persist over time. Problems with sleep are one of the most common complaints from individuals undergoing withdrawal from alcohol and also one of the more common reasons given for relapse into alcohol abuse. Recent research has highlighted the importance of the T -type voltage gated calcium channels in the regulation of the molecular "clock" believed to generate circadian rhythms in the brain. Our lab has identified a specific variant of the T-type calcium channels - called Ca(v)3.2 - whose expression correlates strongly with the circadian disturbances associated with chronic alcohol exposure and withdrawal. The aims of the project seek to evaluate the effect of multiple chronic alcohol exposure-withdrawal cycles on normal molecular and behavioral circadian rhythms in a mouse model in which this T -type calcium channel has been either pharmacologically inhibited or genetically silenced. The approach will provide training in both molecular and behavioral laboratory techniques, including relative quantitation of gene expression using quantitative reverse transcription polymerase chain reaction (qRT-PCR) analysis and Western blot techniques, as well as behavioral analysis and sleep scoring using continuous electroencephalographic (EEG) monitoring. Furthermore, the analysis of the results will provide training in the mathematical modeling of physiologic rhythms, as well as the statistical analysis and interpretation of subsequent findings. The guidance and training provided by the advisor, research associates, and postdoctoral fellows in the lab will assist the applicant in troubleshooting experiments, analyzing pitfalls and unexpected results, and developing innovative approaches to answering the questions that arise at each point in the process of completing the project. In summary, this project will provide the training and skills necessary to develop and carry out a rigorous study of a physiologic problem related to alcohol abuse on both a molecular and behavioral level. PUBLIC HEALTH RELEVANCE: According to the NIH and CDC, respectively, approximately 43 percent of Americans exhibit moderate-to-high risk drinking habits and 70 percent do not get enough sleep. Furthermore, 36 to 72 percent of chronic drinkers cite sleep problems as a major reason for their relapse into alcoholism from 'withdrawal and at least 30 percent of insomniacs use alcohol as a sleep-aid, putting them at increased risk of alcoholism. Hence, the relationship between alcohol and sleep disturbance is a matter of great importance to public health in the United States.

描述(申请人提供)：研究培训项目的重点是多个慢性酒精暴露-戒断周期对分子和行为昼夜节律的影响。睡眠可能是受大脑活动昼夜节律调节的最关键的生理过程。虽然人们对睡眠的具体机制及其重要性了解不多，但如果睡眠障碍持续一段时间，可能会产生毁灭性的生理后果，包括死亡。睡眠问题是戒酒患者最常见的抱怨之一，也是再次酗酒的最常见原因之一。最近的研究强调了T型电压门控钙通道在调节分子时钟中的重要性，分子时钟被认为在大脑中产生昼夜节律。我们的实验室已经发现了一种特定的T型钙通道变体--称为Ca(V)3.2--它的表达与慢性酒精暴露和戒酒相关的昼夜节律紊乱密切相关。该项目的目的是在T型钙通道被药物抑制或基因沉默的小鼠模型中，评估多个慢性酒精暴露-戒断周期对正常分子和行为昼夜节律的影响。该方法将提供分子和行为实验室技术方面的培训，包括使用定量逆转录聚合酶链式反应(qRT-PCR)分析和Western印迹技术进行基因表达的相对量化，以及使用连续脑电(EEG)监测进行行为分析和睡眠评分。此外，对结果的分析将提供生理节律的数学建模方面的培训，以及对随后发现的统计分析和解释。由导师、研究助理和博士后研究员在实验室提供的指导和培训将帮助申请人排除实验故障，分析陷阱和意外结果，并开发创新方法来回答在完成项目过程中的每一个点上出现的问题。总而言之，该项目将提供必要的培训和技能，以在分子和行为层面上开发和开展与酗酒有关的生理学问题的严格研究。 与公共健康相关：根据美国国立卫生研究院和美国疾病控制与预防中心的数据，大约43%的美国人表现出中到高风险的饮酒习惯，70%的人睡眠不足。此外，36%至72%的长期饮酒者表示，睡眠问题是他们戒酒后再次酗酒的主要原因，至少30%的失眠者将酒精作为睡眠辅助，这增加了他们患酒精中毒的风险。因此，酒精与睡眠障碍之间的关系对美国的公共健康非常重要。