AMPA Receptor Regulation at the mRNA and Protein Level in Rat Primary Cortical Cultures 英文参考文献.docVIP
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AMPA Receptor Regulation at the mRNA and Protein Level in Rat Primary Cortical Cultures 英文参考文献
AMPAReceptorRegulationatthemRNAandProtein
LevelinRatPrimaryCorticalCultures
CesareOrlandi,LucaLaVia,DanielaBonini,CristinaMora,IsabellaRusso,AlessandroBarbon,Sergio
Barlati*
DivisionofBiologyandGenetics,DepartmentofBiomedicalSciencesandBiotechnologiesandNationalInstituteofNeuroscience,UniversityofBrescia,Brescia,Italy
Abstract
Ionotropicglutamatea-amino-3-hydroxy-5-methyl-4-isoxazolepropionicacid(AMPA)receptorsarethemajormediatorsof
fastsynapticneurotransmission.Inthiswork,weusedprimarycorticalculturesfromratsasamodelsystemtostudyAMPA
receptor regulation duringin vitrocell maturation andaftersynaptic activitymodifications. The levelsofAMPAreceptor
mRNAandprotein,alongwiththealternativesplicing andRNAeditingoftheAMPAreceptorsubunit(GluR1-4)mRNAs,
wereanalyzedinimmature(DIV5)andmature(DIV26)ratneuronalcultures.Weobservedanincreaseintheexpressionofall
four AMPA receptor subunits during in vitro neuronal maturation. This finding might be due to the formation of new
synapses between neurons during the development of a complex neuronal network. We also analyzed the effects of
stimulation(KClandglutamate)andinhibition(APV/TTX)onratmatureneuronalcultures(DIV26):stimulationwithKClled
to an overall down-regulation of GluR1 and GluR3 AMPA receptor subunits and an up-regulation of the GluR2 subunit.
Similarly,glutamatetreatmentinducedasignificantdown-regulationofGluR1togetherwithanup-regulationofGluR2.In
contrast, thechronic blockadeof neuronalactivitythat resultedfromAPV/TTX treatmentup-regulatedGluR1andGluR3
withaparalleldown-regulationofGluR2andGluR4.RNAeditingattheR/Gsiteincreasedduringneuronalcellmaturation
forallAMPAreceptors(from8–39%atDIV5to28–67%atDIV26).Unexpectedly,allthetreatmentstestedinducedamarked
reduction(rangingfrom29%to252%)ofR/Geditinglevelsinmatureneurons,primarilyforthemRNAflipvariant.
In
summary, we showed that cultured rat cortical neurons are able to vary the stoichiometric ratios of the AMPA receptor
subunits and to control post-transcriptional
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