Are oligomers as toxic as we think ?

INTRODUCTION

Parkinson`s disease (PD) is one of the most common neurodegenerative diseases. Dopaminergic neurons in substantia nigra and noradrenergic neurons in locus coeruleus develop characteristic filamentous inclusions in Parkinson’s disease known as Lewy bodies and neuritis.

Progression of PD is associated with cell death of DA neurons by permeabilization of oligomeric αS. Monomeric alpha-synuclein may interfere with the protofibril-specific permeabilization process.

ASSIGNMENT

You will deal with amyloid pore hypothesis which suggests that α-synuclein (αS) oligomers permeabilize cellular membranes and interfere with Ca homeostasis and cellular pathways. As a model system for membrane-αS oligomer interactions you will use giant unilamellar vesicles (GUVs). Binding of oligomeric and monomeric αS to GUVs will be monitored using confocal microscopy. You will study oligomer and monomer membrane permeabilization and binding using a dye release assay.

Your assignment will be:

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Implementing various analytical techniques (size exclusion chromatography, confocal microscopy, gel and gradient gel electrophoresis, fluorescence spectroscopy, dynamic light scattering)

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Making oligomeric and monomeric α-synuclein

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Learning to make vesicles (GUVs and LUVs) as a cell membrane model system

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Doing membrane binding experiments with oligomeric and monomeric α-synuclein (various mutants and wild type) and physiologically relevant lipid compositions

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Observing protein and lipid interactions using leakage experiments and fluorescence spectroscopy.

Image7C:\Documents and Settings\stefanovica\Mijn documenten\Mijn afbeeldingen\Picture3.tif

Confocal images of DOPE-Rhod labeled GUVs and αS wt-140C-fluorescein monomers-binding experiments (scale bar corresponds to 10 µm). Lipids (left) and monomers (right).

MORE INFORMATION

If you are interested in this Master assigment, you can contact:

Anja Stefanović, MSc in medical biochemistry,

PhD student

Nanobiophysiscs group

MESA+ Institute for Nanotechnology

Faculty of Science and Technology

Zuidhorst ZH 167

Drienerlolaan 5

7522 NB Enschede

The Netherlands

phone: +31 53 489 4612

phone 2: +31 53 489 3157

e-mail: a.stefanovic@utwente.nl