Frederik Ravnkilde Marlet
External, Ph.d Student, Postdoc
Biopharmaceuticals
Jagtvej 162
2100 København Ø
Molecular and Cellular Pharmacology
Jagtvej 160
2100 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.
Methods:
- 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
- Lipidomics
- Gcase activity assay
- Lysosome isolation
- qPCR
- Introduction of oxidative stress (w. MPP+ and 6-OHDA)
- MTT viability assay
Teaching
2020: SFABIL107U - Cellular and molecular biology (Bachelor in Pharmacy)
2019: SFABIL107U - Cellular and molecular biology (Bachelor in Pharmacy)
ID: 124862180
Most downloads
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140
downloads
The interplay between Glucocerebrosidase, α-synuclein and lipids in human models of Parkinson's disease
Research output: Contribution to journal › Review › Research › peer-review
Published -
59
downloads
Sphingolipid changes in Parkinson L444P GBA mutation fibroblasts promote α-synuclein aggregation
Research output: Contribution to journal › Journal article › Research › peer-review
Published -
33
downloads
Golgi fragmentation: One of the earliest organelle phenotypes in Alzheimer's disease neurons
Research output: Contribution to journal › Journal article › Research › peer-review
Published