Selective transgene expression in cerebellar Purkinje cells and granule cells using adeno-associated viruses together with specific promoters

Title
Selective transgene expression in cerebellar Purkinje cells and granule cells using adeno-associated viruses together with specific promoters
Authors
김윤희김태곤이준규이동원게이코 야마모토유키오 야마모토
Keywords
Adeno-associatedvirus; Cerebellum; Purkinje cell; Granule cell; Stereotaxic injection; Gene delivery
Issue Date
2015-09
Publisher
Brain research
Citation
VOL 1620, 1-16
Abstract
Adeno-associatedvirus(AAV)isapowerfultoolforgenedeliveryintothebrainand has been usedfortransgeneexpressioninthecerebellarcortex.Althoughtheefficacies of different AAVserotypestotransducecerebellarPurkinjecellswereexamined,ithasbeen difficult toachievecell-typespecific transgeneexpression.HereweusedAAVserotype1 with twospecific promoters,namely,Ca2þ/calmodulin-dependentproteinkinaseII α (CaMKIIα) andtheminimumregionoftheGABAA receptor α6 subunit(GABRα6) promoters, and comparedtheirexpressionpatternsinthecerebellarcortexwiththeexpression patterns ofubiquitouspromotersthatareoftenusedforAAV-mediatedexpression. Whereas AAVwithubiquitouspromoters,thecytomegalovirusearlyenhancer/chicken β-actin promoter,andasmallfragmentofthesynapsin-1genepromotercausedubiquitous expressioninallcerebellarneuronstested,AAVwiththeCaMKIIα promoter injectedinto 10-day-old miceenabledselectiveexpressioninPurkinjecells.Furthermore,wedeveloped AAV withtheGABRα6 promoter,andsucceededforthe first timetoexpressthetransgene exclusively ingranulecells.FreshcerebellarslicesofmiceinjectedwiththeseAAVswere applicable forphysiologicalexperiments,suchaspatchclamprecording,optogenetic imaging, andstimulation.Thus,theseAAVvectorsareusefultoolstowardsunderstanding the basicpropertiesofcerebellarneuronsormechanismsofcerebellarfunctions. Particularly,selectiveexpressioninPurkinjeorgranulecellsisusefulforanalysesusing genetically-modified animals,suchasknockoutmice.
URI
http://pubs.kist.re.kr/handle/201004/49964
ISSN
00068993
Appears in Collections:
KIST Publication > Article
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