Asymmetric N-Cadherin Expression Results in Synapse Dysfunction, Synapse Elimination, and Axon Retraction in Cultured Mouse Neurons.docVIP

AsymmetricN-CadherinExpressionResultsinSynapse Dysfunction,SynapseElimination,andAxonRetraction inCulturedMouseNeurons KimN.Pielarski1,BerndvanStegen1,AksanaAndreyeva1,KatjaNieweg1,KayJu¨ngling1¤, ChristophRedies2,KurtGottmann1* 1Institute of Neuro- and Sensory Physiology, Medical Faculty, Heinrich-Heine-University Du¨sseldorf, Du¨sseldorf, Germany, 2Institute of Anatomy I, University of Jena SchoolofMedicine,JenaUniversityHospital,Jena,Germany Abstract Synapseeliminationandpruningofaxoncollateralsarecrucialdevelopmentaleventsintherefinementofneuronalcircuits. Whileacontrolofsynapseformationbyadhesionmoleculesiswellestablished,theinvolvementofadhesionmoleculesin developmental synapse loss is poorly characterized. To investigate the consequences of mis-match expression of ahomophilicsynapticadhesionmolecule,weanalysedanasymmetric,exclusivelypostsynapticexpressionofN-cadherin. ThiswasinducedbytransfectingindividualneuronsinculturesofN-cadherinknockoutmouseneuronswithaN-cadherin expression vector. 2 days after transfection, patch-clamp analysis of AMPA receptor-mediated miniature postsynaptic currentsrevealedanimpairedsynapticfunctionwithoutareductioninthenumberofpresynapticvesicleclusters.Long- term asymmetric expression of N-cadherin for 8 days subsequently led to synapse elimination as indicated by a loss of colocalizationofpresynapticvesiclesandpostsynapticPSD95protein.Wefurtherstudiedlong-termasymmetricN-cadherin expressionbyconditional,Cre-inducedknockoutofN-cadherininindividualneuronsinculturesofN-cadherinexpressing corticalmouseneurons.ThisresultedinastrongretractionofaxonalprocessesinindividualneuronsthatlackedN-cadherin protein.Moreover,aninvivoasymmetricexpressionofN-cadherininthedevelopmentallytransientcortico-tectalprojection wasindicatedbyin-situhybridizationwithlayerVneuronslackingN-cadherinexpression.Thus,mis-matchexpressionofN- cadherinmightcontributetoselectivesynapticconnectivity. Citation:PielarskiKN,vanStegenB,AndreyevaA,NiewegK,Ju¨nglingK,etal.(