Striatal ∆FosB gene silencing reduces abnormal involuntary movements induced by L-DOPA in hemiparkinsonian rats Público

Fong, Kayoko (Spring 2019)

Permanent URL: https://etd.library.emory.edu/concern/etds/j098zc04s?locale=pt-BR
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Abstract

Over the course of long-term treatment of Parkinson’s Disease (PD) with L-DOPA, a condition called L-DOPA-induced dyskinesias (LID) can emerge, severely affecting the quality of life of PD patients. LID manifests as abnormal involuntary movements (AIMs) and while its pathogenesis is largely unknown, its development has been linked to transcription factors, including ∆FosB, a truncated form of FosB. Using a hemiparkinsonian 6-hydroxydopamine (6- OHDA) rat model, we investigated whether inhibiting ∆FosB expression with viral vector gene transfer would reduce and/or delay the onset of AIMs. We found evidence of the former (i.e. reduced AIMs scores), but not the latter, and these results, attributed to the role of ∆FosB, were further supported by Western blotting analysis, showing less expression of ∆FosB. Furthermore, the total and peak rotation numbers as well as other behavioral tests, i.e., cylinder and stepping, did not differ significantly between the ∆FosB and control groups, indicating similar antiparkinsonian effects of L-DOPA. These results reinforce ∆FosB’s importance in the development of AIMs and support the potential for ∆FosB gene silencing as a future therapeutic for LID.

Table of Contents

Table of Contents Introduction..................................................................................................................................... 1

Materials and Methods.................................................................................................................... 6

rAAV-Viral Vector Construction ............................................................................................ 6

6-OHDA Rat Model ................................................................................................................ 7

Preparation of 6-OHDA Solution............................................................................................ 7

Viral Injection.......................................................................................................................... 7

Apomorphine test .................................................................................................................... 8

Behavioral tests....................................................................................................................... 8

Scoring AIMs .......................................................................................................................... 8

Cylinder test............................................................................................................................. 9

Stepping test ............................................................................................................................ 9

Adjusting steps ...................................................................................................................... 10

Chronic L-DOPA treatment................................................................................................... 10

Immunohistochemistry .......................................................................................................... 11

Immunofluorescence ............................................................................................................. 12

Western blotting .................................................................................................................... 12

Statistical Analysis ................................................................................................................ 13 Results........................................................................................................................................... 13

Reduction of AIMs following ΔFosB Gene Knockdown...................................................... 13

Gene Silencing of ΔFosB did not affect antiparkinsonian effects of L-DOPA..................... 14

Efficacy of viral vector-mediated ΔFosB gene silencing ...................................................... 15 Discussion..................................................................................................................................... 27

Conclusion .................................................................................................................................... 27 Citations........................................................................................................................................ 31

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