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Expression of mRNAs encoding subunits of the N-methyl-D-aspartate receptor in cultured cortical neurons.
Mol Pharmacol. 1994 May; 45(5):846-53.MP

Abstract

The expression of mRNAs encoding subunits of the N-methyl-D-aspartate (NMDA) receptor was examined in cortical neurons maintained in primary culture. Cultures were prepared from embryonic day 17 rat neocortex. At this developmental age, levels of NR1, NR2A, NR2B, and NR2C mRNA were low or undetectable. Expression of NR1 mRNA increased progressively between days 1 and 21 in vitro. The amount of NR2A mRNA did not change between days 1 and 7 but increased between days 7 and 21. In contrast, levels of NR2B mRNA increased between days 1 and 7, with little further change after day 7. The level of NR2B mRNA was approximately 4-fold higher than that of NR2A mRNA in 21-day cultures. Using ligand binding assays, the proportion of NMDA receptors having a low affinity for ifenprodil was also found to increase over time in culture. The increase in the expression of receptors having a low affinity for ifenprodil and the increase in NR1 and NR2A mRNAs were reduced or prevented by maintaining cells in medium with a low concentration of serum. The results are consistent with the hypothesis that inclusion of the NR2A subunit in native NMDA receptors is responsible for their low affinity for ifenprodil. Splice variants of NR1 lacking the 5' (amino-terminal) insert were found to be the predominant forms of NR1 in cultured neurons. Variants containing the 5' insert represented only a small (< or = 5%) fraction of total NR1 mRNA, and their proportion was not altered as a function of time in culture. Time-dependent changes in the properties of NMDA receptors and in the expression of subunit mRNA occurring in cultured neurons are similar to changes observed in developing rat brain. Thus, the developmental sequence of NMDA receptor expression that occurs in vivo is partially retained in neurons maintained in vitro.

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Authors+Show Affiliations

Zhong J
Department of Pharmacology, University of Pennsylvania School of Medicine, Philadelphia 19104-6084.
Russell SL
No affiliation info available
Pritchett DB
No affiliation info available
Molinoff PB
No affiliation info available
Williams K
No affiliation info available

MeSH

AnimalsCells, CulturedCerebral CortexCulture Media, Serum-FreeGene Expression RegulationNeuronsRNA SplicingRNA, MessengerRatsReceptors, N-Methyl-D-AspartateTime Factors

Pub Type(s)

Journal Article
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

8190101

Citation

Zhong, J, et al. "Expression of mRNAs Encoding Subunits of the N-methyl-D-aspartate Receptor in Cultured Cortical Neurons." Molecular Pharmacology, vol. 45, no. 5, 1994, pp. 846-53.
Zhong J, Russell SL, Pritchett DB, et al. Expression of mRNAs encoding subunits of the N-methyl-D-aspartate receptor in cultured cortical neurons. Mol Pharmacol. 1994;45(5):846-53.
Zhong, J., Russell, S. L., Pritchett, D. B., Molinoff, P. B., & Williams, K. (1994). Expression of mRNAs encoding subunits of the N-methyl-D-aspartate receptor in cultured cortical neurons. Molecular Pharmacology, 45(5), 846-53.
Zhong J, et al. Expression of mRNAs Encoding Subunits of the N-methyl-D-aspartate Receptor in Cultured Cortical Neurons. Mol Pharmacol. 1994;45(5):846-53. PubMed PMID: 8190101.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Expression of mRNAs encoding subunits of the N-methyl-D-aspartate receptor in cultured cortical neurons. AU - Zhong,J, AU - Russell,S L, AU - Pritchett,D B, AU - Molinoff,P B, AU - Williams,K, PY - 1994/5/1/pubmed PY - 1994/5/1/medline PY - 1994/5/1/entrez SP - 846 EP - 53 JF - Molecular pharmacology JO - Mol Pharmacol VL - 45 IS - 5 N2 - The expression of mRNAs encoding subunits of the N-methyl-D-aspartate (NMDA) receptor was examined in cortical neurons maintained in primary culture. Cultures were prepared from embryonic day 17 rat neocortex. At this developmental age, levels of NR1, NR2A, NR2B, and NR2C mRNA were low or undetectable. Expression of NR1 mRNA increased progressively between days 1 and 21 in vitro. The amount of NR2A mRNA did not change between days 1 and 7 but increased between days 7 and 21. In contrast, levels of NR2B mRNA increased between days 1 and 7, with little further change after day 7. The level of NR2B mRNA was approximately 4-fold higher than that of NR2A mRNA in 21-day cultures. Using ligand binding assays, the proportion of NMDA receptors having a low affinity for ifenprodil was also found to increase over time in culture. The increase in the expression of receptors having a low affinity for ifenprodil and the increase in NR1 and NR2A mRNAs were reduced or prevented by maintaining cells in medium with a low concentration of serum. The results are consistent with the hypothesis that inclusion of the NR2A subunit in native NMDA receptors is responsible for their low affinity for ifenprodil. Splice variants of NR1 lacking the 5' (amino-terminal) insert were found to be the predominant forms of NR1 in cultured neurons. Variants containing the 5' insert represented only a small (< or = 5%) fraction of total NR1 mRNA, and their proportion was not altered as a function of time in culture. Time-dependent changes in the properties of NMDA receptors and in the expression of subunit mRNA occurring in cultured neurons are similar to changes observed in developing rat brain. Thus, the developmental sequence of NMDA receptor expression that occurs in vivo is partially retained in neurons maintained in vitro. SN - 0026-895X UR - https://www.unboundmedicine.com/medline/citation/8190101/Expression_of_mRNAs_encoding_subunits_of_the_N_methyl_D_aspartate_receptor_in_cultured_cortical_neurons_ L2 - http://molpharm.aspetjournals.org/cgi/pmidlookup?view=long&amp;pmid=8190101 DB - PRIME DP - Unbound Medicine ER -