Medizinische Universität Graz - Research portal
Selected Publication:
Ramadani, J.
Development of novel fluorescent protein based biosensors to dynamically visualize mitochondria associated membranes (MAMs) on the single cell level.
[ Diplomarbeit/Master Thesis (UNI) ] University of Graz; 2017.
FullText
- Authors Med Uni Graz:
- Advisor:
- Malli Roland
- Altmetrics:
- Abstract:
- Despite the availability of sophisticated methods and approaches for the investigation of mitochondria associated membranes (MAMs), little is known, if any, about the dynamic regulation and formation/degradation of these organelle contact sides in a real-time manner. An excellent approach to visualize cellular events on the level of individual cells offer genetically encoded fluorescent sensors (GES). Hence, in this diploma thesis we attempted to develop a novel approach to visualize MAMs in live. For this purpose, we designed and generated a palette of various novel putative MAMs sensors that were differentially targeted to the ER surface and outer mitochondrial membrane. First application of these probes in HeLa cells combined with co-labeling unveiled that only a small fraction of these two organelles are in interaction to form MAMs under physiological conditions. In order to probe these novel sensors, we utilized a cell model of mouse embryonic fibroblasts (MEFs) and a respective Mfn2-/- variant that is supposed to display an altered MAMs tethering. This approach builds the basis to further develop and refine fluorescent protein- (FP) based techniques to visualize MAMs with high spatial and temporal resolution.