Ray W J, Gottlieb D I
Department of Anatomy and Neurobiology, Washington University School of Medicine, St. Louis, Missouri 63110.
Biochem Biophys Res Commun. 1993 Dec 30;197(3):1475-82. doi: 10.1006/bbrc.1993.2643.
P19 embryonal carcinoma cells can be induced to differentiate into neuron-like cells by retinoic acid. P19 neurons were recently shown to express both NMDA and non-NMDA type glutamate receptor-mediated currents and be susceptible to glutamate excitotoxicity. In this study, we used RT-PCR to survey differentiated P19 cultures for glutamate receptor transcript expression. The following transcripts were detected: at least one member of the GluR1-4 family, GluR5, GluR6, GluR7, KA1, KA2, NMDAR1, and NMDAR2B. Nuclease protection assays revealed a large quantitative induction of GluR6 transcripts following retinoic acid treatment. Inotropic glutamate receptors are a fundamental and major feature of CNS neurons which are not expressed by the cell lines commonly used as experimental models for mammalian neurons. The present results show that P19 cells express multiple genes involved in glutamate receptor biology. Since the stem cells can be manipulated genetically, the system has the basic requirements for analyzing mechanisms involved in glutamate receptor gene expression.
维甲酸可诱导P19胚胎癌细胞分化为神经元样细胞。最近研究表明，P19神经元可同时表达NMDA型和非NMDA型谷氨酸受体介导的电流，并对谷氨酸兴奋性毒性敏感。在本研究中，我们用逆转录-聚合酶链反应（RT-PCR）检测分化的P19培养物中谷氨酸受体转录本的表达。检测到以下转录本：GluR1-4家族的至少一个成员、GluR5、GluR6、GluR7、KA1、KA2、NMDAR1和NMDAR2B。核酸酶保护分析显示，维甲酸处理后GluR6转录本大量增加。促离子型谷氨酸受体是中枢神经系统神经元的一个基本且主要的特征，常用作哺乳动物神经元实验模型的细胞系并不表达该受体。目前的结果表明，P19细胞表达多个与谷氨酸受体生物学相关的基因。由于干细胞可进行基因操作，该系统具备分析谷氨酸受体基因表达相关机制的基本条件。
