The Michael J. Fox Foundation for Parkinson's Research

2 ARTICLES PUBLISHED IN JoVE

Biochemistry

Human Peripheral Blood Neutrophil Isolation for Interrogating the Parkinson's Associated LRRK2 Kinase Pathway by Assessing Rab10 Phosphorylation

Ying Fan^*1, Francesca Tonelli^*1, Shalini Padmanabhan², Marco A.S. Baptista², Lindsey Riley², Danielle Smith³, Connie Marras⁴, Andrew Howden⁵, Dario R. Alessi¹, Esther Sammler^1,6

¹MRC Protein Phosphorylation and Ubiquitylation Unit, School of Life Sciences, University of Dundee, ²The Michael J. Fox Foundation for Parkinson's Research, ³Department of Medical and Molecular Genetics, Indiana University School of Medicine, ⁴Morton and Gloria Shulman Movement Disorders Centre and the Edmond J. Safra Program in Parkinson's Disease, Toronto Western Hospital, University of Toronto, ⁵Division of Cell Signalling and Immunology, School of Life Sciences, University of Dundee, ⁶Division of Molecular and Clinical Medicine, School of Medicine, Ninewells Hospital and Medical School, University of Dundee

Mutations in the leucine rich repeat kinase 2 gene (LRRK2) cause hereditary Parkinson’s disease. We have developed an easy and robust method for assessing LRRK2-controlled phosphorylation of Rab10 in human peripheral blood neutrophils. This may help identify individuals with increased LRRK2 kinase pathway activity.

Neuroscience

Generation of Alpha-Synuclein Preformed Fibrils from Monomers and Use In Vivo

Joseph R. Patterson¹, Nicole K. Polinski², Megan F. Duffy^1,3, Christopher J. Kemp¹, Kelvin C. Luk⁴, Laura A. Volpicelli-Daley⁵, Nicholas M. Kanaan¹, Caryl E. Sortwell^1,3,6

¹Department of Translational Science and Molecular Medicine, Michigan State University, ²The Michael J. Fox Foundation for Parkinson's Research, ³Neuroscience Program, Michigan State University, ⁴Center for Neurodegenerative Disease Research, Department of Pathology and Laboratory Medicine, University of Pennsylvania Perelman School of Medicine, ⁵Center for Neurodegeneration and Experimental Therapeutics, University of Alabama at Birmingham, ⁶Mercy Health Hauenstein Neuroscience Medical Center

The goal of this article is to outline the steps required for the generation of fibrils from monomeric alpha-synuclein, subsequent quality control, and use of the preformed fibrils in vivo.