Regulation of ChAT Expression During Sleep-Wakefulness

睡眠-觉醒期间 ChAT 表达的调节

• 批准号: 6538116
• 负责人: MARY ANN GRECO
• 金额: $ 36.13万
• 依托单位: SRI INTERNATIONAL
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-07-01 至 2005-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6538116
• 关键词: REM sleep; acetylcholine; brain metabolism; choline acetyltransferase; enzyme activity; gene expression; laboratory rat; messenger RNA; neurochemistry; neurotransmitter biosynthesis; protein metabolism; sleep; sleep deprivation; wakefulness

The neuronal circuitry underlying sleep-wakefulness is beginning to be understood. Changes in the release of neurotransmitters across sleep-wakefulness are well documented. The intracellular events associated with the activity of these neurons across sleep-wakefulness, however, are not clear. The available evidence suggests that the transcriptional activation of genes occurs during sleep, indicating that changes in protein and mRNA expression are likely to occur during sleep as well as waking states. The function of sleep is not known. One hypothesis is that sleep serves to replenish biomolecules depleted during waking. If sleep serves such a function, The intracellular environment of brain centers involved in sleep- wakefulness will support biosynthesis. The main hypothesis of this proposal is that changes in the expression of key enzymes associated with neurotransmitter synthesis occur sleep- wakefulness to maintain neurotransmitter stores. To test this hypothesis, the expression of choline acetyltransferase (ChAT), the enzyme that synthesizes acetylcholine will be examined. Specific aim 1 will test the hypothesis that chat mRNA is increased during sleep. In these experiments, mRNA levels will be examined during waking, non-REM and REM sleep states that occur across a 24-hour period. Specific aim 2 tests the hypothesis that protein activity and/or protein levels vary across individual sleep bouts over a 24-hour period. Specific aims 3 and 4 will test the hypothesis that sleep loss effects mRNA expression (Specific aim 3) and protein expression (Specific aim 4). In these experiments, the reversibility of the effects of sleep loss will also be monitored during the recovery sleep period. Sleep disorders affect approximately 70 million Americans with associated costs estimated at billions of dollars in lost revenue per year. By identifying how sleep might replenish key enzymes in neurotransmitter synthesis, we will advance our knowledge of sleep. The experiments proposed are the first steps in the identification of cellular mechanisms effecting acetylcholine synthesis across sleep-wakefulness.

开始吸引睡眠的神经元电路开始被理解。 有充分的文献记录了神经递质遍布睡眠的变化。然而，与这些神经元在睡眠消除性遍及睡眠中的活性相关的细胞内事件尚不清楚。 现有证据表明，基因的转录激活发生在睡眠期间，表明蛋白质和mRNA表达的变化可能在睡眠期间以及醒来状态发生。 睡眠功能尚不清楚。 一个假设是睡眠可以补充醒来过程中耗尽的生物分子。 如果睡眠具有这样的功能，则与睡眠觉醒有关的大脑中心的细胞内环境将支持生物合成。 该提议的主要假设是，与神经递质合成相关的关键酶的表达变化发生睡眠清醒以维持神经递质储存。 为了检验该假设，将检查胆碱乙酰转移酶（CHAT）的表达，将检查合成乙酰胆碱的酶。特定的目标1将测试在睡眠期间增加聊天mRNA的假设。 在这些实验中，将在24小时内发生在醒来，非REM和REM睡眠状态期间检查mRNA水平。 具体目标2检验了以下假设：蛋白质活性和/或蛋白质水平在24小时内各个睡眠回合中有所不同。 具体目的3和4将检验以下假设：睡眠损失影响mRNA表达（特定目标3）和蛋白质表达（特定目标4）。 在这些实验中，还将在恢复睡眠期间监测睡眠损失影响的可逆性。 睡眠障碍影响约7000万美国人，相关费用估计每年收入损失数十亿美元。 通过确定睡眠如何补充神经递质合成中的关键酶，我们将提高对睡眠的了解。 提出的实验是鉴定在跨睡眠吸引力的乙酰胆碱合成的细胞机制的第一步。