Investigations of non-NMDA receptor-induced toxicity in serum-free antioxidant-rich primary cultures of murine cerebellar granule cells

• Published in: Neurochemistry international 1998-07, Vol.33 (1), p.23-28
• Main Authors: Carroll, F.Y, Cheung, N.S, Beart, P.M
• Format: Article
• Language: English
• Subjects: alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid - toxicity; AMPA; Animals; Antioxidants - metabolism; Biological and medical sciences; Cells, Cultured; Central nervous system; Central neurotransmission. Neuromudulation. Pathways and receptors; Cerebellar granule cells; Cerebellum - cytology; Cerebellum - drug effects; Culture Media, Serum-Free; Cytoplasmic Granules - drug effects; Excitatory Amino Acid Agonists - toxicity; Excitotoxicity; Fundamental and applied biological sciences. Psychology; Kainate; Kainic Acid - toxicity; L-glutamate; Mice; Receptors, AMPA - agonists; Receptors, AMPA - physiology; Receptors, Kainic Acid - agonists; Receptors, Kainic Acid - physiology; Receptors, N-Methyl-D-Aspartate - physiology; Serum-free; Vertebrates: nervous system and sense organs
• ISSN: 0197-0186; 1872-9754
• DOI: 10.1016/S0197-0186(05)80004-X

A culture system was developed whereby murine cerebellar granule cells were grown under serum-free conditions in chemically defined B27-supplemented neurobasal medium TM plus depolarizing K+ levels, to allow the investigation of the role of agonists at the kainate and α-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) receptors in glutamate-mediated neurotoxicity. Neurones were killed in a concentration-dependent manner by L-glutamate, kainate and its analogues, domoate and 4-(2-methoxyphenyl)-2-carboxy-3-pyrrolidineacetic acid, but not by (S)-AMPA or (S)-5-~uorowillardiine. Kainate (60% maximal cell death at 1mM) was markedly more toxic than NMDA (40% maximal cell death at 1mM) and was shown to be the predominant cause of excitatory amino acid-induced toxicity in these cells as the neuronal death induced by KA was attenuated by the non-NMDA antagonist CNQX, but not the AMPA antagonist LY293558. This study suggests that serum-free cultures of cerebellar granule cells in B27-supplemented neurobasal medium provide a valuable model system for investigations of the role of the kainate receptor in excitatory amino acid-induced neurodegeneration.