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Paul Morley Paul MacPherson James F. Whitfield †Eric W. Harris Michael W. McBurney 《Journal of neurochemistry》1995,65(3):1093-1099
Abstract: Retinoic acid-treated murine P19 embryonal carcinoma cells differentiate into cells with neuronal morphology that display typical neuronal markers. In this study, the presence of glutamate receptors linked to Ca2+ -signaling mechanisms on these neurons was demonstrated by testing the effects of glutamate agonists and antagonists on the intracellular calcium ion concentration ([Ca2+ ]i ). Glutamate (1 m M ) induced either sustained or transient increases in [Ca2+ ]i . The sustained glutamate-induced increase in [Ca2+ ]i was mimicked by NMDA (40 µ M ). The NMDA-triggered [Ca2+ ]i response was abolished by incubating the cells in Ca2+ -free medium or by pretreating them with Mg2+ (2 m M ) or MK-801 (0.1 µ M ). These responses were unaffected by the non-NMDA antagonist CNQX (10 µ M ), but they required glycine (3–30 µ M ). Kainate (40 µ M ) and AMPA (40 µ M ) did not affect [Ca2+ ]i . Without external Ca2+ , glutamate triggered transient, sometimes oscillating, increases in [Ca2+ ]i . These responses were mimicked by the metabotropic agonist trans -(1 S ,3 R )-1-amino-1,3-cyclopentanedicarboxylic acid (300 µ M ). These results suggest that neurons derived from P19 embryonal carcinoma cells have NMDA and metabotropic, but not AMPA/kainate receptors, which are linked to Ca2+ -signaling mechanisms. These cells could provide a consistent and reproducible model with which to study neuronal differentiation, neurotoxicity, and glutamate receptor-signaling mechanisms. 相似文献
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