title: Striatal neurons but not nigral dopaminergic neurons in neonatal primary cell culture express endogenous functional N-methyl-D-aspartate receptors.
abstract: Developmental expression of N-methyl-D-aspartate (NMDA) receptor subunits were determined and compared in striatal and nigral neurons in neonatal primary cell cultures. In striatal neurons, NR1, NR2A and NR2B mRNAs and immunoreactivity, and NR2D mRNA were found and the maximal levels of NR1 mRNA and immunoreactivity expression were found at 6 day-in-vitro (DIV). NMDA receptors found at this stage in striatal neurons are likely to contain NR1 plus NR2A, NR2B and NR2D subunits. In nigral neurons, NR1 and NR2B mRNAs and immunoreactivity, and NR2D mRNA were found and the maximal level of NR1 immunoreactivity expression was found at 10 DIV. Unlike striatal neurons, NMDA receptors found in nigral neurons are likely to contain NR1 plus NR2B and NR2D subunits only. NMDA-induced toxicity assays showed that striatal neurons were most susceptible to cell death at around 10 DIV but nigral neurons were not susceptible to NMDA-induced cell death at all stages. In addition, patch clamp analysis revealed that functional NMDA receptors could only be found in striatal neurons but not in nigral dopaminergic neurons in vitro. The present results indicate that striatal and nigral neurons are programmed to express distinct NMDA receptor subunits during their endogenous development in cell cultures. Despite dopaminergic neurons in culture display NMDA receptor subunits, functional NMDA receptors are not assembled. The present findings have demonstrated that dopaminergic neurons in vitro may behave very differently to their counterparts in vivo in terms of NMDA receptor-mediated responses. Our results also have implications in transplantations using dopaminergic neurons in vitro in treatments of Parkinson's disease.
authors.affiliation: Department of Biology, Hong Kong Baptist University, Kowloon Tong, Hong Kong, PR China.
authors.forename: P W
authors.initials: PW
authors.lastname: Lui
authors.forename: K C
authors.initials: KC
authors.lastname: Suen
authors.forename: Y S
authors.initials: YS
authors.lastname: Chan
authors.forename: W H
authors.initials: WH
authors.lastname: Yung
authors.forename: K K L
authors.initials: KK
authors.lastname: Yung
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D015214:Antigens, Differentiation, Myelomonocytic / Q000494:pharmacology; D002452:Cell Count; D002478:Cells, Cultured; D004298:Dopamine / Q000378:metabolism*; D005455:Fluorescent Antibody Technique / Q000379:methods; D005968:Glutamate Decarboxylase / Q000378:metabolism; D007527:Isoenzymes / Q000378:metabolism; D008564:Membrane Potentials / Q000187:drug effects; D017072:Neostriatum / Q000166:cytology* / Q000378:metabolism; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*; D018408:Patch-Clamp Techniques; D021122:Protein Subunits / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D016194:Receptors, N-Methyl-D-Aspartate / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D013378:Substantia Nigra / Q000166:cytology* / Q000378:metabolism; D013997:Time Factors; D014157:Transcription Factors / Q000494:pharmacology; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism
chemical_list: D015214:Antigens, Differentiation, Myelomonocytic; D007527:Isoenzymes; C075373:MNDA protein, human; D021122:Protein Subunits; D012333:RNA, Messenger; D016194:Receptors, N-Methyl-D-Aspartate; D014157:Transcription Factors; D014446:Tyrosine 3-Monooxygenase; D005968:Glutamate Decarboxylase; C402565:glutamate decarboxylase 1; D004298:Dopamine