41 Neuroscience Application RevC Draft_No Tau data

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1、ApplicationNoteNeuroscienceVisualizegeneexpressionandsplicejunctionsatthesingle-celllevelusingtheRNAscope®andBaseScope™insituhybridizationassaysSummaryInsitudetectionofG-proteinFigures1-4providebothfluorescentandcoupledreceptors(GPCRs)inthechromogenicimagesforfourtypesof

2、GPCRsThenervoussystemconsists-readilydetectedasmRNAtargetsbytheofnumerousspecializedcelladultmousebrainusingtheRNAscope®assay-inhippocampalandstriataltypesthatremaintobefullyRNAscope®ISHAssaycharacterizedatthemolecularnormalmousebrainareas:Dopaminergiclevel.Duetothehighd

3、egreeRaisingantibodiestoG-proteincoupledreceptorsReceptorsD1andD2(Drd1andDrd2),Cannabinoidofstructuralandfunctional(GPCRs)canbechallengingduetodifficultiesReceptor1(Cnr1),OpioidReceptorμ(OPRM1),δheterogeneityandtheintricateinobtainingsuitableantigenaccessibilitysince(OPR

4、D1),andκ(OPRK1),andCholinergicReceptorspatialorganizationoftheseGPCRsareoftenexpressedatlowlevelsandareMuscarinic3(Chrm3).cells,itisofspecialimportancetoveryunstablewhenpurified(Hutchingsetal.2010analyzegeneexpressionwithinthe(1)).Hereweshowexamplesforthedetectionof:morp

5、hologicalandspatialtissuecontext.TheRNAscope®insitu-DopaminergicReceptorshybridization(ISH)assayenables-CannabinoidReceptorhighlysensitiveandspecificgene-CholinergicReceptorexpressionvisualizationwithinthe-OpioidReceptorsnervoussystem.BaseScope™isaISHassaythatallowsvisua

6、lizationofsplicejunctionsbetweenadjacentexonsand/orretainedintronsinordertocharacterizealternativesplicingeventsincellsandtissues.ThisapplicationnotewillhighlightcasestudiesforbothISHassaysinthenervoussystem:•InsitudetectionofG-protein-FIGURE1.DetectionofDopamineReceptor

7、D1(Drd1,Red)andDopamineFIGURE 2.DetectionofCannabinoidreceptor1(Cnr1,Red)andcoupledreceptors(GPCRs)inReceptorD2(Drd2,Green)inmousebrainstriatumusingtheRNAscope®DopaminergicreceptorD1(Drd1,Green)innormalmousebrainstriatumtheadultmousebrainusingtheMultiplexFluorescentassay

8、onfreshfrozentissuesamples.DelineatedusingtheRNAscope®2.5HDDuplexChromogenicassayonFFPEtissueRNAscope®I