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

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

About this Honors Thesis

Rights statement

• Permission granted by the author to include this thesis or dissertation in this repository. All rights reserved by the author. Please contact the author for information regarding the reproduction and use of this thesis or dissertation.

School	Emory College
Department	Human Health
Degree	B.A.
Submission	Honors Thesis
Language	English
Research Field	Biology, Neuroscience
Palavra-chave	L-DOPA-induced dyskinesias (LID) ∆FosB Parkinson's Disease
Committee Chair / Thesis Advisor	Dr. Stella Papa, Emory University
Committee Members	Dr. Ellen Hess, Emory University Dr. Michelle Lampl, Emory University

Última modificação

Primary PDF

Thumbnail	Title	Date Uploaded	Actions
	Striatal ∆FosB gene silencing reduces abnormal involuntary movements induced by L-DOPA in hemiparkinsonian rats ()	2019-04-18 19:28:14 -0400	Press to Select an action Download

Supplemental Files

Thumbnail	Title	Date Uploaded	Actions