Mechanisms in genetic models of Parkinson Disease

Description of projects available to graduate students:
Loss of PINK1 function causes early onset familial PD. Using stable knockdown of endogenous PINK1 and transfection with PINK1 mutants, we have defined a set of mitochondrial, neuritic and autophagic phenotypes. We have also discovered that dominant mutations in LRRK2, which cause late onset familial and sporadic PD, increase vulnerability to excitotoxic dendritic remodeling through a mechanism involving perturbed mitochondrial calcium homeostasis.

We use a combination of 2D-gel analysis, proteomics, molecular cell biology and microscopy to study mechanisms by which PINK1 and LRRK2 regulate mitochondria and neurites.

Current questions include:
1) Proteomic and mutagenesis in functional characterization of PINK1 interacting targets.
2) Employ knock-in culture models to study the role of PINK1 mutants on mitochondrial function, biogenesis, trafficking and degradation
3) study mechanisms underlying altered mitochondrial function in mutant LRRK2 transgenic mouse brains.

Techniques graduate student will learn:
primary neuron culture and differentiated neuronal lines

RNAi, mutagenesis and transfection

multi-label immunofluorescence, western blot, electron microscopy, computer assisted quantitative image analysis

Live cell imaging and Seahorse metabolic profiling of living neurons and cells

2D-gel analysis, immunoprecipitation, mass spectrometry