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Development of a method to examine primary cell cultures and their response to dopamine and glutamate using a High-Content Imaging system
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Pharmaceutical Biosciences.
2016 (English)Independent thesis Advanced level (degree of Master (One Year)), 20 credits / 30 HE creditsStudent thesis
Abstract [en]

HSPB5, a small heat shock protein, has been found to be notably up-regulated in brain tissue from parkinsonian rats, as well as post mortem Parkinson’s disease (PD) patients. Recent studies indicate that HSPB5 might have a crucial role in neurological diseases, acting as a protective chaperone or under certain conditions, as a pathogenic factor. It is believed to play a role in the development of levodopa-induced dyskinesia (LID), a severe adverse effect from levodopa (L-DOPA) treatment of PD. One hypothesis is that oligodendrocytes are linked to the progression of LID, a process that might be facilitated by exosome release and excretion of HSPB5 under dopamine and glutamate stimulation.

The overall aim of this project was to investigate HSPB5s role in the development of L-DOPA-induced dyskinesia. A new method was developed to examine how dopamine and glutamate affects glial cells and exosome release. Primary cortical cell cultures were labeled with fluorescent markers and live cells were analyzed with a High-Content Imaging System after dopamine and glutamate stimulation.

The results indicate that dopamine and glutamate stimulate glial cells to release exosomes, although the new method shows promising potential it needs to be further optimized in order to enable statistical analysis. 

Immunocytochemistry on the primary cell cultures revealed large amounts of astrocytes, among other glial cells, meaning that no conclusions can be made whether observed cell effects are carried through oligodendrocytes, or any other specific type of glial cell. 

Place, publisher, year, edition, pages
2016.
National Category
Medical and Health Sciences
Identifiers
URN: urn:nbn:se:uu:diva-274225OAI: oai:DiVA.org:uu-274225DiVA: diva2:896092
Subject / course
Toxicology
Educational program
Master of Science Programme in Pharmacy
Supervisors
Examiners
Available from: 2016-10-06 Created: 2016-01-20 Last updated: 2016-10-06Bibliographically approved

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