Regulation of Neurotransmitter Transporter Recyclin(RMI)

描述（由申请人提供）：通过高亲和力转运蛋白（神经转运蛋白）从突触间隙清除神经递质是调节神经元信号传导的重要阶段。调节神经转运蛋白的主要机制发生在亚细胞定位水平。神经转运蛋白从细胞表面内化到细胞内池中减少了突触间隙处可用的神经转运蛋白分子的数量。影响神经转运蛋白调节的变化在许多神经退行性疾病的进展中是重要的。在这里，我们描述了一种检测，可以发展成一个高通量筛选，以确定小分子（药物），影响神经转运蛋白的贩运。该测定测量响应于刺激物如蛋白激酶C（PKC）信号传导的荧光标记的神经转运蛋白的亚细胞定位的变化。针对该测定筛选化合物具有鉴定小分子效应物的潜力，所述小分子效应物可用作抑制神经递质清除的操纵的工具。这将有助于研究由神经转运蛋白调节改变引起的病理状况。此外，这些分子的鉴定可以产生可以开发成新的治疗药物的化合物，或提供关于潜在治疗靶点的信息。

DESCRIPTION (provided by applicant): Clearance of neurotransmitter from the synaptic cleft, by high affinity transport proteins (neurotransporters), is an important stage in the regulation of neuronal signalling. A major mechanism by which neurotransporters are regulated occurs at the level of subcellular localization. Internalization of neurotransporters away from the cell surface, into an intracellular pool reduces the number of available neurotransporter molecules at the synaptic cleft. Changes that affect the regulation of neurotransporters are important in the progression of a number of neurodegenerative diseases. Here we describe an assay that can be developed into a high throughput screen to identify small molecules (drugs) that affect neurotransporter trafficking. The assay measures changes in the subcellular localization of fluorescently tagged neurotransporters in response to stimuli such as protein kinase C (PKC) signaling. Screening chemical compounds against this assay has the potential to identify small molecule effectors that can be used as tools to pharmacologically manipulation of neurotransmitter clearance. This will aid research into pathological conditions caused by alterations of neurotransporter regulation. In addition, identification of such molecules may yield compounds that can be developed into new therapeutic drugs, or provide information about potential therapeutic targets.