Regulation of Neurotransmitter Transporter Recyclin(RMI)

描述（由申请人提供）：高亲和力转运蛋白（神经转运蛋白）清除突触间隙中的神经递质是神经元信号传导调节的重要阶段。神经递质调控的主要机制发生在亚细胞定位水平。神经递质从细胞表面内化，进入细胞内池，减少了突触间隙中可用神经递质分子的数量。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，我们描述了一种可以发展成高通量筛选来识别影响神经转运蛋白运输的小分子（药物）的测定方法。该实验测量了在刺激如蛋白激酶C （PKC）信号的刺激下，荧光标记的神经转运蛋白亚细胞定位的变化。筛选化学化合物对抗该试验有潜力识别小分子效应物，可以用作药理学操纵神经递质清除的工具。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，对这些分子的鉴定可能产生可以开发成新的治疗药物的化合物，或提供有关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.