VAChT Expression Across Sleep-Wakefulness

VAChT 在睡眠-觉醒状态下的表达

• 批准号: 7023715
• 负责人: MARY ANN GRECO
• 金额: $ 4.55万
• 依托单位: SRI INTERNATIONAL
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-06-15 至 2007-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7023715
• 关键词: REM sleep; acetylcholine; cerebral cortex; choline acetyltransferase; electroencephalography; electromyography; fos protein; gene expression; hippocampus; immunocytochemistry; laboratory rat; messenger RNA; neurotransmitter biosynthesis; neurotransmitter transport; prosencephalon; protein quantitation /detection; sleep; sleep deprivation; synaptic vesicles; transport proteins; wakefulness; western blottings

DESCRIPTION (provided by applicant): Over 70 million people in the U.S. are estimated to suffer from alterations in sleep-wakefulness. Existing therapies are of limited utility in treating these disorders. The difficulties in devising more effective treatments stem, in large part, from the fact that there is no one sleep center in the brain. As a result, the biological function(s) of sleep have not been elucidated. The neuronal circuitry underlying sleep-wakefulness is beginning to be understood. The intracellular events associated with the activity of these neurons across sleep-wakefulness, however, are not well defined. Understanding cellular mechanisms that occur during sleep is key to the identification of biological function(s). Acetylcholine (ACh) is associated with sleep-wakefulness. Preliminary results examining the ACh synthesis enzyme, choline acetyltransferase (ChAT) are consistent with the hypothesis that sleep may play a role in maintaining neurotransmitter levels that are depleted during waking via changes in mRNA during sleep. The vesicular acetylcholine transporter (VAChT) performs the essential function of packaging nascent acetylcholine into vesicles. The gene encoding yacht lies within the gene encoding chat in all species examined, an extremely rare mammalian genomic organization. Preliminary studies suggest that chat and vacht mRNA levels are differentially expressed during sleep. In these studies, we focus first on delineating VAChT expression. In the first specific aim, we test the hypothesis that vacht mRNA and protein vary across individual sleep-wake bouts (i.e., waking, SWS and REM sleep) occurring over a 24h period. Preliminary studies of vacht mRNA suggest that it is higher during waking than during sleep. The second specific aim tests the hypothesis that mRNA and protein are associated with wakefulness. Initial results show vacht mRNA did not vary following six hours of sleep deprivation compared to controls. Specific aims 3 and 4 focus on comparisons of vacht and chat mRNAs, given the genomic organization of the genes and preliminary results that show that vacht mRNA contrasts with chat mRNA expression. Specific aim 3 tests the hypothesis that chat and vacht mRNAs are differentially expressed during sleep, while specific aim 4 tests the hypothesis that vacht mRNA subtypes are differentially expressed in response to sleep-wakefulness. Identification of these patterns of expression are key to understanding cellular mechanisms underlying sleep and wakefulness.

描述（由申请人提供）：据估计，在美国，超过7000万人会遭受睡眠消耗的改变。现有的疗法在治疗这些疾病方面的效用有限。设计更有效的治疗方法的困难在很大程度上是由于大脑中没有一个睡眠中心。结果，尚未阐明睡眠的生物学功能。开始吸引睡眠的神经元电路开始被理解。然而，与这些神经元在睡眠消除性范围内的活性相关的细胞内事件尚未很好地定义。了解睡眠期间发生的细胞机制是鉴定生物学功能的关键。乙酰胆碱（ACH）与睡眠效果有关。初步的结果检查了ACH合成酶，胆碱乙酰转移酶（CHAT）与假设可能在维持神经递质水平中发挥作用的假设是一致的，而神经递质水平在醒来期间通过睡眠过程中的mRNA醒来而耗尽的神经递质水平。囊泡乙酰胆碱转运蛋白（VACHT）在包装新生乙酰胆碱中的基本功能。编码游艇的基因在于所检查的所有物种中编码聊天的基因，这是一个极为罕见的哺乳动物基因组组织。初步研究表明，在睡眠期间，聊天和VACHT mRNA水平差异表达。在这些研究中，我们首先关注描述VACHT表达。在第一个特定目的中，我们检验了以下假设：在24小时内发生的vacht mRNA和蛋白质在单个睡眠唤醒爆发中有所不同（即醒来，SWS和REM睡眠）。 VACHT mRNA的初步研究表明，在醒来期间，它比睡眠时高。第二个特定目的检验了mRNA和蛋白质与清醒相关的假设。最初的结果表明，与对照组相比，六个小时的睡眠剥夺后，Vacht mRNA没有变化。鉴于基因的基因组组织和初步结果表明，Vacht mRNA与CHAT mRNA表达形成鲜明对比。特定的目标3检验了聊天和VACHT mRNA在睡眠期间差异表达的假设，而特定的目标4检验了以下假设：VACHT mRNA亚型响应于睡眠效果而差异表达。这些表达模式的识别是理解睡眠和清醒潜在的细胞机制的关键。