title: LRRK2 kinase activity regulates synaptic vesicle trafficking and neurotransmitter release through modulation of LRRK2 macro-molecular complex.
abstract: Mutations in Leucine-rich repeat kinase 2 gene (LRRK2) are associated with familial and sporadic Parkinson's disease (PD). LRRK2 is a complex protein that consists of multiple domains executing several functions, including GTP hydrolysis, kinase activity, and protein binding. Robust evidence suggests that LRRK2 acts at the synaptic site as a molecular hub connecting synaptic vesicles to cytoskeletal elements via a complex panel of protein-protein interactions. Here we investigated the impact of pharmacological inhibition of LRRK2 kinase activity on synaptic function. Acute treatment with LRRK2 inhibitors reduced the frequency of spontaneous currents, the rate of synaptic vesicle trafficking and the release of neurotransmitter from isolated synaptosomes. The investigation of complementary models lacking LRRK2 expression allowed us to exclude potential off-side effects of kinase inhibitors on synaptic functions. Next we studied whether kinase inhibition affects LRRK2 heterologous interactions. We found that the binding among LRRK2, presynaptic proteins and synaptic vesicles is affected by kinase inhibition. Our results suggest that LRRK2 kinase activity influences synaptic vesicle release via modulation of LRRK2 macro-molecular complex.
authors.affiliation: Division of Neuroscience, San Raffaele Scientific Institute and Vita-Salute University Milan, Italy ; Department of Molecular and Cellular Pharmacology, National Research Council, Neuroscience Institute Milan, Italy.
authors.forename: Maria D
authors.initials: MD
authors.lastname: Cirnaru
authors.affiliation: Department of Experimental Medicine, University of Genova Genova, Italy.
authors.forename: Antonella
authors.initials: A
authors.lastname: Marte
authors.affiliation: Department of Biology, University of Padova Padova, Italy.
authors.forename: Elisa
authors.initials: E
authors.lastname: Belluzzi
authors.affiliation: Department of Biology, University of Padova Padova, Italy.
authors.forename: Isabella
authors.initials: I
authors.lastname: Russo
authors.affiliation: Department of Molecular and Cellular Pharmacology, National Research Council, Neuroscience Institute Milan, Italy ; Department of Medical Biotechnology and Translational Medicine, University of Milan Milan, Italy.
authors.forename: Martina
authors.initials: M
authors.lastname: Gabrielli
authors.affiliation: Department of Medical Science and National Institute of Neuroscience, University of Ferrara Ferrara, Italy.
authors.forename: Francesco
authors.initials: F
authors.lastname: Longo
authors.affiliation: Department of Medical Science and National Institute of Neuroscience, University of Ferrara Ferrara, Italy.
authors.forename: Ludovico
authors.initials: L
authors.lastname: Arcuri
authors.affiliation: Department of Molecular and Cellular Pharmacology, National Research Council, Neuroscience Institute Milan, Italy.
authors.forename: Luca
authors.initials: L
authors.lastname: Murru
authors.affiliation: Department of Biology, University of Padova Padova, Italy.
authors.forename: Luigi
authors.initials: L
authors.lastname: Bubacco
authors.affiliation: Department of Medical Biotechnology and Translational Medicine, University of Milan Milan, Italy ; Humanitas Clinical and Research Center, Pharmacology and Brain Pathology Rozzano, Italy.
authors.forename: Michela
authors.initials: M
authors.lastname: Matteoli
authors.affiliation: Department of Pharmacy, University of Genoa Genoa, Italy.
authors.forename: Ernesto
authors.initials: E
authors.lastname: Fedele
authors.affiliation: Department of Molecular and Cellular Pharmacology, National Research Council, Neuroscience Institute Milan, Italy ; Department of Medical Biotechnology and Translational Medicine, University of Milan Milan, Italy.
authors.forename: Carlo
authors.initials: C
authors.lastname: Sala
authors.affiliation: Department of Molecular and Cellular Pharmacology, National Research Council, Neuroscience Institute Milan, Italy.
authors.forename: Maria
authors.initials: M
authors.lastname: Passafaro
authors.affiliation: Department of Medical Science and National Institute of Neuroscience, University of Ferrara Ferrara, Italy.
authors.forename: Michele
authors.initials: M
authors.lastname: Morari
authors.affiliation: Department of Biology, University of Padova Padova, Italy.
authors.forename: Elisa
authors.initials: E
authors.lastname: Greggio
authors.affiliation: Department of Experimental Medicine, University of Genova Genova, Italy.
authors.forename: Franco
authors.initials: F
authors.lastname: Onofri
authors.affiliation: Division of Neuroscience, San Raffaele Scientific Institute and Vita-Salute University Milan, Italy ; Department of Molecular and Cellular Pharmacology, National Research Council, Neuroscience Institute Milan, Italy.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Piccoli
keywords: LRRK2; kinase; presynaptic vesicle; protein interaction; synaptic activity