REGULATION OF THE HUMAN NOREPINEPHRINE TRANSPORTER

描述(摘自申请人的摘要)： 人去甲肾上腺素转运体(Hnet)是一种高亲和力结合蛋白 许多精神治疗化合物的网站，包括那些 抗抑郁药效(例如三环类抗抑郁药)。吸收了 由Net产生的去甲肾上腺素(NE)是 去甲肾上腺素的作用终止于去甲肾上腺素能突触。规管 因此，质膜中的净活性代表着一种重要的 去甲肾上腺素能调节的候选机制 可能会发生重传。尽管许多具有精神活性的化合物 与网络结合，这些配体诱导的网络功能的调节 人们对此知之甚少。我们的初步数据表明，某些 Net的抑制剂(抗抑郁药)下调Net 功能。事实上，接触某些净值抑制剂会降低净值。 在他没有抑制剂的一段时间内作用更大 比最初的曝光期要长。这一数据的含义是 职业诱导的网络功能下调可能有助于 与网结合的药物的治疗/药理作用。这个 这项提议的目标是检查Net配体的能力 在体外和体内制剂中诱导Net下调，并 阐明配体诱导网络形成的分子机制 下调监管。在连续曝光完整的网络表达之后 细胞到净配体，研究将检查：(1)净摄取能力 (功能)在体外摄取试验中，(2)有可能 网从质膜表面快速重新分布，(3) 蛋白激酶在配体诱导的网络调节中的作用，以及(4) 净蛋白周转率和净信使RNA水平。生物学的 Net配体诱导的Net体外调节的相关性将是 通过研究治疗后脑片的网络功能而建立的 具有净配体的大鼠。重点放在体外和体内研究上 被放在监管和复苏的时间方面，因为 从抑制剂诱导的下调中缓慢恢复可能意味着 抗抑郁化合物不需要按顺序出现在网中 以抑制摄取。这些信息可能会对 网络抑制剂治疗方案在精神疾病治疗中的应用 疾病。总的来说，建议的研究将揭示以下基本原则 毒品暴露与净管制和意愿之间的关系 提供了与药理作用相关的重要线索 绑定到网中的精神活性物质。

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.