A Bioluminescence Reporter Assay for Retinoic Acid Control of Translation of the GluR1 Subunit of the AMPA Glutamate Receptor.
A Bioluminescence Reporter Assay for Retinoic Acid Control of Translation of the GluR1 Subunit of the AMPA Glutamate Receptor.
复制标题
视黄酸控制 AMPA 谷氨酸受体 GluR1 亚基翻译的生物发光报告基因测定。
DOI：
10.1007/978-1-0716-2453-1_15
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发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Khatib T
中科院分区：
文献类型：
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作者：
Khatib T
视黄酸（RA）通过视黄酸受体（RAR）调节基因转录来调节中枢神经系统功能的许多方面。然而，RA具有独立于基因转录（非基因组作用）的重要作用，并且在大脑中，稳态可塑性的关键调节因子是谷氨酸受体亚基1（GluR 1）翻译的RAR控制。定量RAR调节GluR 1翻译的测定将有利于研究调节该系统的分子组分，并筛选影响大脑中学习和记忆这一关键机制的药物。开发了一种生物发光报告基因检测方法，该方法在RAR结合的GluR 1 5′非翻译区的控制下表达萤火虫荧光素酶。将该试验引入SH-SY 5 Y细胞中，并用于证明RARα在RA调节GluR 1翻译中的作用。合成RAR和RXR配体的筛选表明，只有这些配体的一个子集激活GluR 1翻译。结果证明了该试验用于探索RARα对该途径的贡献的实用性，并且RAR配体激活翻译的能力是有限数量化合物的质量限制，其RAR选择性和潜在的药物使用特异性具有意义。
Retinoic acid (RA) regulates numerous aspects of central nervous system function through modulation of gene transcription via retinoic acid receptors (RARs). However, RA has important roles independent of gene transcription (non-genomic actions) and in the brain a crucial regulator of homeostatic plasticity is RAR control of glutamate receptor subunit 1 (GluR1) translation. An assay to quantify RAR regulation of GluR1 translation would be beneficial both to study the molecular components regulating this system and screen drugs that influence this critical mechanism for learning and memory in the brain. A bioluminescence reporter assay was developed that expresses firefly luciferase under the control of the GluR1 5′ untranslated region bound by RAR. This assay was introduced into SH-SY5Y cells and used to demonstrate the role of RARα in RA regulation of GluR1 translation. A screen of synthetic RAR and RXR ligands indicated that only a subset of these ligands activated GluR1 translation. The results demonstrate the practicality of this assay to explore the contribution of RARα to this pathway and that the capacity of RAR ligands to activate translation is a quality restricted to a limited number of compounds, with implications for their RAR selectivity and potentially their specificity in drug use.