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dc.contributor.advisorCondliffe, Stephen
dc.contributor.authorBedford, Cade Henry
dc.date.available2013-11-21T01:30:30Z
dc.date.copyright2013
dc.identifier.citationBedford, C. H. (2013). Regulation of voltage-gated calcium channels in PC12 cells by Leucine Rich Repeat Kinase 2 (Thesis, Bachelor of Biomedical Sciences with Honours). University of Otago. Retrieved from http://hdl.handle.net/10523/4478en
dc.identifier.urihttp://hdl.handle.net/10523/4478
dc.description.abstractLeucine rich repeat kinase two (LRRK2) is a widely expressed protein belonging to the Roco family of proteins, mutations in which have recently been discovered as a cause of familial Parkinson’s disease (PD). Despite an array of interacting proteins having been identified across multiple cellular systems, LRRK2’s functional role remains to be determined. Manipulation of LRRK2 expression disrupts many Ca2+ dependent cellular processes. It, therefore, may act as an upstream regulator of initial Ca2+ signalling events, which could explain LRRK2’s widespread effects. The central aim of this study was to determine whether LRRK2 alters endogenous voltage-gated Ca2+ (CaV) channel function in PC12 cells using whole cell patch clamp electrophysiology. Additionally, transiently transfected PC12 cells underwent epifluorescence imaging to identify morphological changes and identify any effects of L-type Ca2+ blockers on morphology. Peak CaV channel currents in LRRK2 transfected cells showed a significantly (p=0.0025, n≥7, one way ANOVA with Tukeys post-hoc test) higher current density across a number of holding voltages relative to untransfected and EGFP transfected controls. These results indicate that LRRK2 up regulates endogenous CaV channel function. Morphological assessment, however showed no significant effect of LRRK2 transfection on morphological parameters relative to EGFP transfected and non-transfected controls (N≥63, Kruskal-Wallis test). Furthermore, addition of the L-type Ca2+ channel blocker nifedipine had no significant effect relative to untransfected and ethanol vehicle controls (N≥43,Kruskal-Wallis test). These results suggest that LRRK2 dependent modulation of CaV channel function does not affect neurite differentiation. Overall, this study has identified a novel effect of LRRK2 on CaV channels, which may explain how LRRK2 has such widespread cellular effects and advances our understanding of LRRK2s functional role. If the effect of LRRK2 on CaV channels is responsible for pathology, CaV channel blockers currently being investigated for Parkinson’s therapy may be particularly effective in Parkinson’s patients harbouring LRRK2 mutations.
dc.language.isoen
dc.publisherUniversity of Otago
dc.rightsAll items in OUR Archive are provided for private study and research purposes and are protected by copyright with all rights reserved unless otherwise indicated.
dc.subjectLRRK2
dc.subjectVoltage
dc.subjectgated
dc.subjectcalcium
dc.subjectchannel
dc.subjectCav
dc.subjectLeucine
dc.subjectrich
dc.subjectrepeat
dc.subjectkinase
dc.subject2
dc.subjectParkinson's
dc.subjectwhole
dc.subjectcell
dc.subjectpatch
dc.subjectclamp
dc.subjectelectrophysiology
dc.subjectPC12
dc.subjectneurite
dc.subjectlength
dc.subjectmorphology
dc.titleRegulation of voltage-gated calcium channels in PC12 cells by Leucine Rich Repeat Kinase 2
dc.typeThesis
dc.date.updated2013-11-21T00:02:07Z
dc.language.rfc3066en
thesis.degree.disciplinePhysiology
thesis.degree.nameBachelor of Biomedical Sciences with Honours
thesis.degree.grantorUniversity of Otago
thesis.degree.levelHonours
otago.interloanyes
otago.openaccessAbstract Only
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