Developmental expression of NMDA receptor subunits and the emergence of glutamate neurotoxicity in primary cultures of murine cerebral cortical neurons (original) (raw)
Abstract.
Using primary cultures of murine cerebral cortices, we investigated the developmental expression of _N_-methyl-D-aspartate (NMDA) receptor subunits in relation to the appearance of NMDA receptor-mediated glutamate neurotoxicity. The cultures were not affected by glutamate exposure on culture days 7 – 9, but became sensitive to glutamate neurotoxicity on day 11. The expression of NMDA receptor subunit messenger RNAs (mRNAs) was investigated by means of reverse transcription polymerase chain reaction (RT-PCR). The _ε_3-NR2C and _ε4-_NR2D transcripts could not be detected in the culture. The _ε_2-NR2B and _ζ_1-NR1 subunit mRNAs, on the other hand, could be detected clearly and continuously from the culture initiation, and the _ε1-_NR2A subunit m-RNA became clearly detectable on culture day 4. The expression of these three subunits' proteins in the glutamate-insensitive stage (culture day 8) and the sensitive stage (day 11) were studied by means of Western blotting. The _ε_2-NR2B and _ζ_1-NR1 subunit proteins were clearly expressed on culture days 8 and 11, but the _ε1-_NR2A subunit protein could hardly be detected on either day 8 or day 11. These results suggest that the glutamate neurotoxicity in the primary culture was mediated mainly by _ε_2/_ζ_1 NMDA receptors. The time lag between the protein expression of the _ε_2-NR2B and _ζ_1-NR1 subunits and the emergence of glutamate neurotoxicity may be necessary for the maturation of functional NMDA receptor systems, including heteromeric receptor formation, increase in receptor density and maturation of the postreceptor signal transduction system.
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Authors and Affiliations
- Department of Cell Biology, Institute for Virus Research, Kyoto University, Sakyo-ku, Kyoto 606 (Japan), , , , , , JP
I. Mizuta, M. Katayama & K. Ishii - Department of Anatomy, Hokkaido University School of Medicine, Sapporo 060 (Japan), , , , , , JP
M. Watanabe - Department of Molecular Neurobiology and Pharmacology, School of Medicine, University of Tokyo, Tokyo 113 (Japan), , , , , , JP
M. Mishina
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- I. Mizuta
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Received 7 April 1998; accepted 30 April 1998
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Mizuta, I., Katayama, M., Watanabe, M. et al. Developmental expression of NMDA receptor subunits and the emergence of glutamate neurotoxicity in primary cultures of murine cerebral cortical neurons.CMLS, Cell. Mol. Life Sci. 54, 721–725 (1998). https://doi.org/10.1007/s000180050199
- Issue Date: July 1998
- DOI: https://doi.org/10.1007/s000180050199