DOPAMINE & SLEEP HOMEOSTASIS--MOLECULAR GENETIC APPROACH

代偿性睡眠反应（CSR），以睡眠增加为特征 时间和深度），在小鼠完全睡眠一段时间后观察到 剥夺（TSD）表明睡眠是一个稳态过程。 的 这种自我平衡驱动的机制尚不清楚，但很可能是 与某些睡眠障碍的病因学有关。 我们发现 类似于TSD，甲基苯丙胺，一种细胞膜抑制剂 多巴胺（DA）转运蛋白（DAT）和细胞内DA转运蛋白 （VMAT2）在小鼠中产生睡眠丧失，随后是CSR。 相比之下 DAT特异性抑制剂GBRI2909产生睡眠丧失，而没有随后的 CSR。 根据这些观察，我们推测细胞内DA 在睡眠的稳态中，储存起着关键的作用。 这项建议 解决了与这个理论相关的三个问题。 守灵是不是 GBR 12909仅通过抑制DAT9？ 是 甲基苯丙胺的促醒作用和随后的CSR 通过抑制DAT和细胞内DA储存介导？ 做 细胞膜（DAT）和囊泡（VMAT2）DA的遗传改变 转运体将CSR改变为TSD？ 我们提出了一种靶向分子遗传学 解决这些问题的方法。为了确定分子位置 的作用的GBRI2909和甲基苯丙胺，我们将研究的影响 这些药物对DAT和VMAT2基因改变的小鼠的睡眠影响 表情 为了确定DAT和VMAT2在生理学上的作用， 睡眠在没有药理学操作的情况下，我们还将研究 将CSR转化为TSD。

The Compensatory Sleep Response (CSR; characterized by increased sleep time and depth) that is seen in mice after a period of total sleep deprivation (TSD) indicates that sleep is a homeostatic process. The mechanism for this homeostatic drive is unknown, but is likely to be involved in the etiology of some sleep disorders. We have found that similar to TSD, methamphetamine, an inhibitor of the cell membrane dopamine (DA) transporter (DAT) and of the intracellular DA transporter (VMAT2), produces sleep loss followed by a CSR in mice. In contrast the DAT-specific inhibitor GBRI2909 produces sleep loss without a subsequent CSR. In light of these observations we theorize that intracellular DA stores play a critical role in sleep homeostasis. This proposal addresses three issues relevant to this theory. Is the wake promoting effect GBR12909 mediated exclusively through inhibition of DAT9? Are the wake promoting effect of methamphetamine and the subsequent CSR mediated through inhibition of DAT and of intracellular DA stores? Do genetic alterations of cell membrane (DAT) and vesicular (VMAT2) DA transporters alter CSR to TSD? We propose a targeted molecular genetic approach to address these questions. To determine the molecular sites of action of GBRI2909 and methamphetamine, we will study the effects of these drugs on sleep in mice with genetic alterations of DAT and VMAT2 expression. To determine the role of DAT and VMAT2 in physiological sleep in the absence of pharmacological manipulation, we will also study the CSR to TSD in these genetically engineered mice.