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dc.contributor.authorSweeney, Carolyn G.
dc.contributor.authorKearney, Patrick
dc.contributor.authorFagan, Rita R.
dc.contributor.authorSmith, Lindsey A.
dc.contributor.authorBolden, Nicholas C.
dc.contributor.authorZhao-Shea, Rubing
dc.contributor.authorRivera, Iris V.
dc.contributor.authorKolpakova, Jenya
dc.contributor.authorXie, Jun
dc.contributor.authorGao, Guangping
dc.contributor.authorTapper, Andrew R.
dc.contributor.authorMartin, Gilles E.
dc.contributor.authorMelikian, Haley E.
dc.date2022-08-11T08:08:24.000
dc.date.accessioned2022-08-23T15:53:36Z
dc.date.available2022-08-23T15:53:36Z
dc.date.issued2019-06-03
dc.date.submitted2019-06-20
dc.identifier.citation<p>bioRxiv 658856; doi: https://doi.org/10.1101/658856. <a href="https://doi.org/10.1101/658856" target="_blank">Link to preprint on bioRxiv service.</a></p>
dc.identifier.doi10.1101/658856
dc.identifier.urihttp://hdl.handle.net/20.500.14038/29388
dc.description.abstractDopamine (DA) signaling is critical for movement, motivation, and addictive behavior. The neuronal GTPase, Rit2, is enriched in DA neurons (DANs), binds directly to the DA transporter (DAT), and is implicated in several DA-related neuropsychiatric disorders. However, it remains unknown whether Rit2 plays a role in either DAergic signaling and/or DA-dependent behaviors. Here, we leveraged the TET-OFF system to conditionally silence Rit2 in Pitx3IRES2-tTA mouse DANs. Following DAergic Rit2 knockdown (Rit2-KD), mice displayed an anxiolytic phenotype, with no change in baseline locomotion. Further, males exhibited increased acute cocaine sensitivity, whereas DAergic Rit2-KD suppressed acute cocaine sensitivity in females. DAergic Rit2-KD did not affect presynaptic TH and DAT protein levels in females, nor was TH was affected in males; however, DAT was significantly diminished in males. Paradoxically, despite decreased DAT levels in males, striatal DA uptake was enhanced, but was not due to enhanced DAT surface expression in either dorsal or ventral striatum. Finally, patch recordings in nucleus accumbens (NAcc) medium spiny neurons (MSNs) revealed reciprocal changes in spontaneous EPSP (sEPSP) frequency in male and female D1+ and D2+ MSNs following DAergic Rit2-KD. In males, sEPSP frequency was decreased in D1+, but not D2+, MSNs, whereas in females sEPSP frequency decreased in D2+, but not D1+, MSNs. Moreover, DAergic Rit2-KD abolished the ability of cocaine to reduce sEPSP frequency in D1+, but not D2+, male MSNs. Taken together, our studies are among the first to acheive AAV-mediated, conditional and inducible DAergic knockdown in vivo. Importantly, our results provide the first evidence that DAergic Rit2 expression differentially impacts striatal function and DA-dependent behaviors in males and females.
dc.language.isoen_US
dc.rightsThe copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It is made available under a CC-BY-NC-ND 4.0 International license.
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/
dc.subjectDopamine
dc.subjectaddictive behavior
dc.subjectGTPase
dc.subjectRit2
dc.subjectDA neurons
dc.subjectAmino Acids, Peptides, and Proteins
dc.subjectBehavioral Neurobiology
dc.subjectEnzymes and Coenzymes
dc.subjectNeuroscience and Neurobiology
dc.titleConditional, inducible gene silencing in dopamine neurons reveals a sex-specific role for Rit2 GTPase in acute cocaine response and striatal function [preprint]
dc.typePreprint
dc.source.journaltitlebioRxiv
dc.identifier.legacyfulltexthttps://escholarship.umassmed.edu/cgi/viewcontent.cgi?article=2624&amp;context=faculty_pubs&amp;unstamped=1
dc.identifier.legacycoverpagehttps://escholarship.umassmed.edu/faculty_pubs/1618
dc.identifier.contextkey14777788
refterms.dateFOA2022-08-23T15:53:37Z
html.description.abstract<p>Dopamine (DA) signaling is critical for movement, motivation, and addictive behavior. The neuronal GTPase, Rit2, is enriched in DA neurons (DANs), binds directly to the DA transporter (DAT), and is implicated in several DA-related neuropsychiatric disorders. However, it remains unknown whether Rit2 plays a role in either DAergic signaling and/or DA-dependent behaviors. Here, we leveraged the TET-OFF system to conditionally silence Rit2 in <em>Pitx3<sup>IRES2-tTA</sup></em> mouse DANs. Following DAergic Rit2 knockdown (Rit2-KD), mice displayed an anxiolytic phenotype, with no change in baseline locomotion. Further, males exhibited increased acute cocaine sensitivity, whereas DAergic Rit2-KD suppressed acute cocaine sensitivity in females. DAergic Rit2-KD did not affect presynaptic TH and DAT protein levels in females, nor was TH was affected in males; however, DAT was significantly diminished in males. Paradoxically, despite decreased DAT levels in males, striatal DA uptake was enhanced, but was not due to enhanced DAT surface expression in either dorsal or ventral striatum. Finally, patch recordings in nucleus accumbens (NAcc) medium spiny neurons (MSNs) revealed reciprocal changes in spontaneous EPSP (sEPSP) frequency in male and female D1+ and D2+ MSNs following DAergic Rit2-KD. In males, sEPSP frequency was decreased in D1+, but not D2+, MSNs, whereas in females sEPSP frequency decreased in D2+, but not D1+, MSNs. Moreover, DAergic Rit2-KD abolished the ability of cocaine to reduce sEPSP frequency in D1+, but not D2+, male MSNs. Taken together, our studies are among the first to acheive AAV-mediated, conditional and inducible DAergic knockdown <em>in vivo</em>. Importantly, our results provide the first evidence that DAergic Rit2 expression differentially impacts striatal function and DA-dependent behaviors in males and females.</p>
dc.identifier.submissionpathfaculty_pubs/1618
dc.contributor.departmentGraduate School of Biomedical Sciences, Neuroscience Program
dc.contributor.departmentTapper Lab
dc.contributor.departmentMartin Lab
dc.contributor.departmentMelikian Lab
dc.contributor.departmentViral Vector Core
dc.contributor.departmentGene Therapy Center
dc.contributor.departmentNeurobiology
dc.contributor.departmentBrudnick Neuropsychiatric Research Institute
dc.contributor.studentJenya Kolpakova
dc.contributor.studentCarolyn Sweeney
dc.contributor.studentRita Fagan
dc.contributor.studentPatrick Kearney
dc.contributor.studentNicholas Bolden
dc.description.thesisprogramNeuroscience
dc.description.thesisprogramMD/PhD


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The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It is made available under a CC-BY-NC-ND 4.0 International license.
Except where otherwise noted, this item's license is described as The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It is made available under a CC-BY-NC-ND 4.0 International license.