Frederik Ravnkilde Marlet
Peptides and Proteins
Jagtvej 162, København Ø
Identification of lipid changes associated with Parkinson’s Disease and alpha-synuclein pathology in dopaminergic neuronal cells.
My current focuses are:
1. Development of novel PD model by using the human derived neuroblastoma cell line, SH-SY5Y. The aim is to investigate the lipid composition in both the healthy and diseased model. PD-like symptoms are established in the SH-SY5Y cells by GBA knockout and oxidative stress repectively.
2. Investigate the affect of lipid vesicles on a-synuclein aggregation. The vesicles are prepared from various sphingolipids differing in size and side chains bound to the sphingosine backbone. These lipids are proven to be the most abundant lipids in the brain and the levels of these indivdual lipids are perturbed in PD.
3. Identification of fibril characteristics by electron microscopy. Fibrils formed by aggregation of a-synuclein in presence of lipid vesicles prepared from different sphingolipids, are used.
4. Investigation if Ambroxol can restore the effects on the whole cell lipid profile generated from expression of mutated GCase. Patient-derived fibroblasts expressing mutated GCase will be used as model.
- Protein expression and purification
- Tht aggregation experiment
- Protein-lipid binding assay (Circular dichroism)
- Preparation of lipid vesicles
- Fibril formation
- Electron microscopy
- Cell culturing
- Lipid extraction
- Gcase activity assay
- Lysosome isolation
- Introduction of oxidative stress (w. MPP+ and 6-OHDA)
- MTT viability assay
2020: SFABIL107U - Cellular and molecular biology (Bachelor in Pharmacy)
2019: SFABIL107U - Cellular and molecular biology (Bachelor in Pharmacy)