REGULATION OF THE HUMAN NOREPINEPHRINE TRANSPORTER

描述（来自申请人摘要）： 人去甲肾上腺素转运蛋白（hNET）是一种高亲和力结合蛋白， 网站的许多精神治疗化合物，包括那些与 抗抑郁功效（例如三环类抗抑郁药）。的摄取 去甲肾上腺素（NE）的NET是主要的机制， NE的作用终止于去甲肾上腺素能突触。调控 因此，细胞膜中的NET活性代表了一种重要的 调节去甲肾上腺素能神经递质的 可以发生传输。尽管许多精神活性化合物 与NET结合，这些配体诱导的NET功能调节 是很难理解的。我们的初步数据显示， NET抑制剂（抗抑郁药）下调NET 功能事实上，接触某些NET抑制剂会降低NET 在没有抑制剂的情况下， 比最初的暴露期。这些数据的含义是， 职业引起的NET功能下调可能有助于 与NET结合的药物的治疗/药理作用。的 该提案的目标是检查NET配体的能力， 在体外和体内制剂中诱导NET下调，以及 阐明配体诱导NET的分子机制 下调。在连续暴露于完整的NET表达的 细胞NET配体，研究将检查：（1）NET摄取能力 （功能）在体外摄取试验中，（2）存在 NET从质膜表面迅速重新分布，（3） 蛋白激酶在配体诱导的NET调节中的作用，以及（4） NET蛋白的周转和NET信使RNA的水平。生物 NET配体诱导的NET体外调节的相关性将是 通过研究治疗后脑切片中的NET功能建立 NET配体的大鼠。体外和体内研究的重点将 应放在监管和恢复的时间方面，因为 缓慢恢复从走廊诱导下调可能意味着， 抗抑郁药化合物不需要存在于NET中， 以抑制摄取。这些信息可能会对 NET抑制剂治疗精神病的方案 疾病总的来说，拟议的研究将揭示基本原则 药物暴露与NET监管之间的关系， 提供了重要的线索有关的药理作用， 与网结合的精神活性剂

DESCRIPTION (from applicant's abstract): The human norepinephrine transporter (hNET) is a high affinity binding site for many psychotherapeutic compounds, including those with antidepressant efficacy (e.g. tricyclic antidepressants). The uptake of norepinephrine (NE) by the NET is the principal mechanism by which the action of NE is terminated at the noradrenergic synapse. Regulation of NET activity in the plasma membrane, therefore, represents an important candidate mechanism through which modulation of noradrenergic ransmission can occur. Despite the fact that many psychoactive compounds bind to the NET, the regulation of NET function induced by these ligands is poorly understood. Our preliminary data demonstrate that certain inhibitors (those which are antidepressants) of NET down-regulate NET function. In fact, exposure to certain NET inhibitors reduces NET function in he absence of the inhibitor for a period of time greater than the initial exposure period. The implication of this data is that occupation-induced down-regulation of NET function may contribute to the therapeutic/ pharmacological action of drugs that bind to the NET. The goals of this proposal are to examine the ability of NET ligands to induce NET down-regulation in in vitro and in vivo preparations, and to elucidate the molecular mechanisms responsible for ligand-induced NET down-regulation. Following continuous exposure of intact NET-expressing cells to NET ligands, studies will examine: (1) NET uptake capacity (function) in in vitro uptake assays, (2) the possibility that there is a rapid redistribution of NET from the plasma membrane surface,(3) the role of protein kinases in ligand-induced NET regulation, and (4) the turnover of NET protein and levels of NET messenger RNA. The biological relevance of NET ligand-induced regulation of NET in vitro will be established by studying NET function in brain slices following treatment of rats with NET ligands. Emphasis in in vitro and in vivo studies will be placed on the temporal aspects of regulation and recovery, because slow recovery from inhibitorinduced down-regulation may imply that antidepressant compounds do not need to be present at the NET in order to inhibit uptake. This information may have a significant impact on treatment regimens of NET inhibitors in the management of psychiatric diseases. Overall, the proposed studies will reveal the basic principles of the relationship between drug exposure and NET regulation and will provide important clues relevant to the pharmacological actions of psychoactive agents that bind to the net.