In vitro studies
Protein stability and neuronal differentiation
We use two in-vitro assays to assess the stability and pathogenicity of the mutation. The stability of the mutated protein is tested by expressing the protein in HEK cells.
The pathogenicity of a mutation is tested by (over)expressing the protein in primary neuronal cultures. Common read-outs for this latter assay are arborization (axon and dendrite formation), soma-size and synapse number. All measurements are compared to neurons transfected with control-plasmid and to wild-type protein as well as to knock-out/knock-down of the gene of interest. This will allow us to assess if the mutations acts as a gain- or loss of function (GOF, LOF) mutation.