Reference : The use of primary human fibroblasts for monitoring mitochondrial phenotypes in the f...
Scientific journals : Article
Life sciences : Genetics & genetic processes
http://hdl.handle.net/10993/18775
The use of primary human fibroblasts for monitoring mitochondrial phenotypes in the field of Parkinson's disease.
English
Burbulla, Lena F. [> >]
Krüger, Rejko mailto [University of Luxembourg > Faculty of Science, Technology and Communication (FSTC) > Life Science Research Unit]
2012
Journal of visualized experiments : JoVE
68
Yes
International
1940-087X
1940-087X
United States
[en] Biopsy ; Fibroblasts/pathology ; Flow Cytometry/methods ; Humans ; Membrane Potential, Mitochondrial ; Microscopy, Fluorescence/methods ; Mitochondria/pathology ; Parkinson Disease/pathology ; Phenotype ; Skin/pathology
[en] Parkinson's disease (PD) is the second most common movement disorder and affects 1% of people over the age of 60 (1). Because ageing is the most important risk factor, cases of PD will increase during the next decades (2). Next to pathological protein folding and impaired protein degradation pathways, alterations of mitochondrial function and morphology were pointed out as further hallmark of neurodegeneration in PD (3-11). After years of research in murine and human cancer cells as in vitro models to dissect molecular pathways of Parkinsonism, the use of human fibroblasts from patients and appropriate controls as ex vivo models has become a valuable research tool, if potential caveats are considered. Other than immortalized, rather artificial cell models, primary fibroblasts from patients carrying disease-associated mutations apparently reflect important pathological features of the human disease. Here we delineate the procedure of taking skin biopsies, culturing human fibroblasts and using detailed protocols for essential microscopic techniques to define mitochondrial phenotypes. These were used to investigate different features associated with PD that are relevant to mitochondrial function and dynamics. Ex vivo, mitochondria can be analyzed in terms of their function, morphology, colocalization with lysosomes (the organelles degrading dysfunctional mitochondria) and degradation via the lysosomal pathway. These phenotypes are highly relevant for the identification of early signs of PD and may precede clinical motor symptoms in human disease-gene carriers. Hence, the assays presented here can be utilized as valuable tools to identify pathological features of neurodegeneration and help to define new therapeutic strategies in PD.
Luxembourg Centre for Systems Biomedicine (LCSB): Clinical & Experimental Neuroscience (Krüger Group)
http://hdl.handle.net/10993/18775

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