NDLI: NMDA receptor characterization and subunit expression in rat cultured mesencephalic neurones

Content Provider	PubMed Central
Author	Allgaier, C. Scheibler, P. Müller, D. Feuerstein, T. J. Illes, P.
Copyright Year	1999
Abstract	NMDA-induced changes in free intracellular Ca2+ concentration ([Ca2+]i) were determined in individual cultured rat mesencephalic neurones by the fura-2 method. mRNA expression encoding NMDA receptor subunits (NR1, NR2A-D) was examined by RT–PCR. NMDA (1–100 μM, plus 10 μM glycine) induced a concentration-dependent increase in [Ca2+]i (EC50=5.7 μM). The effect of NMDA was virtually insensitive to tetrodotoxin (0.3 μM) and nitrendipine (1 μM), but dependent on extracellular Ca2+. 5,7-Dichlorokynurenic acid (10 μM), a specific antagonist at the glycine binding site on the NMDA receptor, abolished the NMDA response. Memantine, an open-channel blocker, and ifenprodil, a preferential non-competitive NR1/NR2B receptor antagonist diminished the NMDA effect with an IC50 value of 0.17 and 1 μM, respectively. Ethanol at 50 and 100 mM caused about 25 and 45%-inhibition, respectively. Agarose gel analysis of the PCR products followed by ethidium bromide fluorescence or CSPD chemiluminescence detection revealed an almost exclusive expression of the NR1 splice variants lacking exon (E) 5 and E22. The 3′ splice form without both E21 and E22 exceeded that containing E21 by approximately 4 fold. The relative amounts of NR2A, NR2B, NR2C corresponded to approximately 1:2:1. NR2D mRNA was also detectable. In conclusion, mesencephalic neurones bear ethanol-sensitive NMDA receptors which might be involved in the development of ethanol dependence and withdrawal. The high affinity of NMDA to this receptor, its sensitivity to ifenprodil and memantine may suggest that the mesencephalic NMDA receptor comprises the NR1 splice variant lacking E5, NR2B, and NR2C, respectively.
Related Links	http://dx.doi.org/10.1038/sj.bjp.0702284
Ending Page	130
Page Count	10
Starting Page	121
File Format	PDF
ISSN	00071188
Journal	British Journal of Pharmacology
Issue Number	1
Volume Number	126
Language	English
Publisher Date	1999-01-01
Access Restriction	Open
Subject Keyword	Pharmacology Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Pharmacology

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