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dc.contributor.authorNimitvilai, Sudarat
dc.contributor.authorArora, Devinder S
dc.contributor.authorMcElvain, Maureen A
dc.contributor.authorBrodie, Mark S
dc.date.accessioned2017-05-03T16:12:36Z
dc.date.available2017-05-03T16:12:36Z
dc.date.issued2013
dc.date.modified2013-12-19T22:42:36Z
dc.identifier.issn1662-5102
dc.identifier.doi10.3389/fncel.2013.00007
dc.identifier.urihttp://hdl.handle.net/10072/55216
dc.description.abstractSelective suppression of protein function in the brain can be achieved using specific silencing RNAs administered in vivo. A viral delivery system is often employed to transfect neurons with small hairpin RNA (shRNA) directed against specific proteins, and intervals of several days are allowed between microinjection of the shRNA-containing virus into the brain and experiments to assess suppression of gene function. Here we report studies using extracellular recording of dopaminergic neurons of the ventral tegmental area (DA VTA neurons) recorded in brain slices in which lentivirus containing shRNA directed against Gq was included in the recording pipette, and suppression of Gq-related function was observed within the time frame of the recording. The action of neurotensin (NT) is associated with activation of Gq, and the firing rate of DA VTA neurons is increased by NT. With shRNA directed against Gq in the pipette, there was a significant reduction of NT excitation within 2?h. Likewise, time-dependent dopamine desensitization, which we have hypothesized to be Gq-dependent, was not observed when shRNA directed against Gq was present in the pipette and dopamine was tested 2?h after initiation of recording. As the time interval (2?h) is relatively short, we tested whether blockade of protein synthesis with cycloheximide delivered via the recording pipette would alter Gq-linked responses similarly. Both NT-induced excitation and dopamine desensitization were inhibited in the presence of cycloheximide. Inclusion of shRNA in the recording pipette may be an efficient and selective way to dampen responses linked to Gq, and, more generally, the use of lentiviral-packaged shRNA in the recording pipette is a means to produce selective inhibition of the function of specific proteins in experiments.
dc.description.peerreviewedYes
dc.description.publicationstatusYes
dc.format.extent1665476 bytes
dc.format.mimetypeapplication/pdf
dc.languageEnglish
dc.language.isoeng
dc.publisherFrontiers Research Foundation
dc.publisher.placeSwitzerland
dc.relation.ispartofstudentpublicationN
dc.relation.ispartofpagefrom7.1
dc.relation.ispartofpageto7.6
dc.relation.ispartofjournalFrontiers in Cellular Neuroscience
dc.relation.ispartofvolume7
dc.rights.retentionY
dc.subject.fieldofresearchCentral Nervous System
dc.subject.fieldofresearchBiochemistry and Cell Biology
dc.subject.fieldofresearchNeurosciences
dc.subject.fieldofresearchcode110903
dc.subject.fieldofresearchcode0601
dc.subject.fieldofresearchcode1109
dc.titleSuppression of Gq Function Using Intra-Pipette Delivery of shRNA during Extracellular Recording in the Ventral Tegmental Area
dc.typeJournal article
dc.type.descriptionC1 - Articles
dc.type.codeC - Journal Articles
gro.rights.copyright© The Author(s) 2013. The attached file is reproduced here in accordance with the copyright policy of the publisher. For information about this journal please refer to the journal’s website or contact the author[s].
gro.date.issued2013
gro.hasfulltextFull Text
gro.griffith.authorArora, Devinder S.


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