Expression of mRNAs encoding subunits of the N-methyl-D-aspartate receptor in cultured cortical neurons
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 undetectab...
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| Veröffentlicht in: | Molecular pharmacology 1994-05, Vol.45 (5), p.846-853 |
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| creator | JIE ZHONG RUSSELL, S. L PRITCHETT, D. B MOLINOFF, P. B WILLIAMS, K |
| description | 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. |
| format | Article |
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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.</description><identifier>ISSN: 0026-895X</identifier><identifier>EISSN: 1521-0111</identifier><identifier>PMID: 8190101</identifier><identifier>CODEN: MOPMA3</identifier><language>eng</language><publisher>Bethesda, MD: American Society for Pharmacology and Experimental Therapeutics</publisher><subject>Animals ; Biological and medical sciences ; Cells, Cultured ; Central nervous system ; Central neurotransmission. Neuromudulation. Pathways and receptors ; Cerebral Cortex - cytology ; Cerebral Cortex - embryology ; Cerebral Cortex - metabolism ; Culture Media, Serum-Free - pharmacology ; Fundamental and applied biological sciences. Psychology ; Gene Expression Regulation ; Neurons - metabolism ; Rats ; Receptors, N-Methyl-D-Aspartate - genetics ; RNA Splicing ; RNA, Messenger - genetics ; RNA, Messenger - metabolism ; Time Factors ; Vertebrates: nervous system and sense organs</subject><ispartof>Molecular pharmacology, 1994-05, Vol.45 (5), p.846-853</ispartof><rights>1994 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=4159724$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/8190101$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>JIE ZHONG</creatorcontrib><creatorcontrib>RUSSELL, S. L</creatorcontrib><creatorcontrib>PRITCHETT, D. B</creatorcontrib><creatorcontrib>MOLINOFF, P. B</creatorcontrib><creatorcontrib>WILLIAMS, K</creatorcontrib><title>Expression of mRNAs encoding subunits of the N-methyl-D-aspartate receptor in cultured cortical neurons</title><title>Molecular pharmacology</title><addtitle>Mol Pharmacol</addtitle><description>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.</description><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>Cells, Cultured</subject><subject>Central nervous system</subject><subject>Central neurotransmission. Neuromudulation. Pathways and receptors</subject><subject>Cerebral Cortex - cytology</subject><subject>Cerebral Cortex - embryology</subject><subject>Cerebral Cortex - metabolism</subject><subject>Culture Media, Serum-Free - pharmacology</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene Expression Regulation</subject><subject>Neurons - metabolism</subject><subject>Rats</subject><subject>Receptors, N-Methyl-D-Aspartate - genetics</subject><subject>RNA Splicing</subject><subject>RNA, Messenger - genetics</subject><subject>RNA, Messenger - metabolism</subject><subject>Time Factors</subject><subject>Vertebrates: nervous system and sense organs</subject><issn>0026-895X</issn><issn>1521-0111</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1994</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqF0E1LxDAQBuAiyrqu_gQhB_VWyCRt2h4Xv0FWEAVvJZtOtpE2qUmK7r-3sotXT3N4H15m5iCZQ84gpQBwmMwpZSItq_z9ODkJ4YNSyPKSzpJZCRUFCvNkc_s9eAzBOEucJv3LahkIWuUaYzckjOvRmhh-o9giWaU9xnbbpTepDIP0UUYkHhUO0XliLFFjF0ePDVHOR6NkRyyO3tlwmhxp2QU8289F8nZ3-3r9kD493z9eL5_SlgHEtBAouUYmBJQyUyipgFyyhnJRas5BaY6YaSiBNVpQXJeCV-sKs0JDThnli-Rq1zt49zliiHVvgsKukxbdGOpCZJXg4n8IYtqhYGyC53s4rnts6sGbXvptvX_hlF_scxmmg7WXVpnwxzLIq4JlE7vcsdZs2i_jsR5a6XupXOc2E8vrvC4zwX8AFcqIRA</recordid><startdate>199405</startdate><enddate>199405</enddate><creator>JIE ZHONG</creator><creator>RUSSELL, S. L</creator><creator>PRITCHETT, D. B</creator><creator>MOLINOFF, P. B</creator><creator>WILLIAMS, K</creator><general>American Society for Pharmacology and Experimental Therapeutics</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>7TK</scope><scope>7X8</scope></search><sort><creationdate>199405</creationdate><title>Expression of mRNAs encoding subunits of the N-methyl-D-aspartate receptor in cultured cortical neurons</title><author>JIE ZHONG ; RUSSELL, S. L ; PRITCHETT, D. B ; MOLINOFF, P. B ; WILLIAMS, K</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-h211t-76ea3fe26618a4cea0615a2d0368f331cf3ee4f1812df60eb8639b9e47f150203</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1994</creationdate><topic>Animals</topic><topic>Biological and medical sciences</topic><topic>Cells, Cultured</topic><topic>Central nervous system</topic><topic>Central neurotransmission. Neuromudulation. Pathways and receptors</topic><topic>Cerebral Cortex - cytology</topic><topic>Cerebral Cortex - embryology</topic><topic>Cerebral Cortex - metabolism</topic><topic>Culture Media, Serum-Free - pharmacology</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gene Expression Regulation</topic><topic>Neurons - metabolism</topic><topic>Rats</topic><topic>Receptors, N-Methyl-D-Aspartate - genetics</topic><topic>RNA Splicing</topic><topic>RNA, Messenger - genetics</topic><topic>RNA, Messenger - metabolism</topic><topic>Time Factors</topic><topic>Vertebrates: nervous system and sense organs</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>JIE ZHONG</creatorcontrib><creatorcontrib>RUSSELL, S. L</creatorcontrib><creatorcontrib>PRITCHETT, D. B</creatorcontrib><creatorcontrib>MOLINOFF, P. B</creatorcontrib><creatorcontrib>WILLIAMS, K</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>Neurosciences Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Molecular pharmacology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>JIE ZHONG</au><au>RUSSELL, S. L</au><au>PRITCHETT, D. B</au><au>MOLINOFF, P. B</au><au>WILLIAMS, K</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Expression of mRNAs encoding subunits of the N-methyl-D-aspartate receptor in cultured cortical neurons</atitle><jtitle>Molecular pharmacology</jtitle><addtitle>Mol Pharmacol</addtitle><date>1994-05</date><risdate>1994</risdate><volume>45</volume><issue>5</issue><spage>846</spage><epage>853</epage><pages>846-853</pages><issn>0026-895X</issn><eissn>1521-0111</eissn><coden>MOPMA3</coden><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.</abstract><cop>Bethesda, MD</cop><pub>American Society for Pharmacology and Experimental Therapeutics</pub><pmid>8190101</pmid><tpages>8</tpages></addata></record> |
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| source | MEDLINE; EZB-FREE-00999 freely available EZB journals |
| subjects | Animals Biological and medical sciences Cells, Cultured Central nervous system Central neurotransmission. Neuromudulation. Pathways and receptors Cerebral Cortex - cytology Cerebral Cortex - embryology Cerebral Cortex - metabolism Culture Media, Serum-Free - pharmacology Fundamental and applied biological sciences. Psychology Gene Expression Regulation Neurons - metabolism Rats Receptors, N-Methyl-D-Aspartate - genetics RNA Splicing RNA, Messenger - genetics RNA, Messenger - metabolism Time Factors Vertebrates: nervous system and sense organs |
| title | Expression of mRNAs encoding subunits of the N-methyl-D-aspartate receptor in cultured cortical neurons |
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