title: No dopamine cell loss or changes in cytoskeleton function in transgenic mice expressing physiological levels of wild type or G2019S mutant LRRK2 and in human fibroblasts.
abstract: Mutations within the LRRK2 gene have been identified in Parkinson's disease (PD) patients and have been implicated in the dysfunction of several cellular pathways. Here, we explore how pathogenic mutations and the inhibition of LRRK2 kinase activity affect cytoskeleton dynamics in mouse and human cell systems. We generated and characterized a novel transgenic mouse model expressing physiological levels of human wild type and G2019S-mutant LRRK2. No neuronal loss or neurodegeneration was detected in midbrain dopamine neurons at the age of 12 months. Postnatal hippocampal neurons derived from transgenic mice showed no alterations in the seven parameters examined concerning neurite outgrowth sampled automatically on several hundred neurons using high content imaging. Treatment with the kinase inhibitor LRRK2-IN-1 resulted in no significant changes in the neurite outgrowth. In human fibroblasts we analyzed whether pathogenic LRRK2 mutations change cytoskeleton functions such as cell adhesion. To this end we compared the adhesion characteristics of human skin fibroblasts derived from six PD patients carrying one of three different pathogenic LRRK2 mutations and from four age-matched control individuals. The mutant LRRK2 variants as well as the inhibition of LRRK2 kinase activity did not reveal any significant cell adhesion differences in cultured fibroblasts. In summary, our results in both human and mouse cell systems suggest that neither the expression of wild type or mutant LRRK2, nor the inhibition of LRRK2 kinase activity affect neurite complexity and cellular adhesion.
authors.affiliation: Department of Neurodegeneration, Hertie-Institute for Clinical Brain Research and DZNE, German Center for Neurodegenerative Diseases, 72076 Tuebingen, Germany.
authors.forename: Marta
authors.initials: M
authors.lastname: Garcia-Miralles
authors.affiliation: Department of Cellular Neurology, Hertie-Institute for Clinical Brain Research and DZNE, German Center for Neurodegenerative Diseases, 72076 Tuebingen, Germany.
authors.forename: Janaky
authors.initials: J
authors.lastname: Coomaraswamy
authors.affiliation: Department of Medical Genetics and Applied Genomics, Institute of Human Genetics, University of Tuebingen, 72076 Tuebingen, Germany.
authors.forename: Karina
authors.initials: K
authors.lastname: Häbig
authors.affiliation: Department of Cellular Neurology, Hertie-Institute for Clinical Brain Research and DZNE, German Center for Neurodegenerative Diseases, 72076 Tuebingen, Germany.
authors.forename: Martin C
authors.initials: MC
authors.lastname: Herzig
authors.affiliation: Department of Neurodegeneration, Hertie-Institute for Clinical Brain Research and DZNE, German Center for Neurodegenerative Diseases, 72076 Tuebingen, Germany.
authors.forename: Natalja
authors.initials: N
authors.lastname: Funk
authors.affiliation: Boehringer Ingelheim Pharma GmbH & Co. KG, CNS Research, 88397 Biberach an der Riss, Germany.
authors.forename: Frank
authors.initials: F
authors.lastname: Gillardon
authors.affiliation: Department of Neurodegeneration, Hertie-Institute for Clinical Brain Research and DZNE, German Center for Neurodegenerative Diseases, 72076 Tuebingen, Germany.
authors.forename: Martina
authors.initials: M
authors.lastname: Maisel
authors.affiliation: Department of Cellular Neurology, Hertie-Institute for Clinical Brain Research and DZNE, German Center for Neurodegenerative Diseases, 72076 Tuebingen, Germany.
authors.forename: Mathias
authors.initials: M
authors.lastname: Jucker
authors.affiliation: Department of Neurodegeneration, Hertie-Institute for Clinical Brain Research and DZNE, German Center for Neurodegenerative Diseases, 72076 Tuebingen, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Gasser
authors.affiliation: Department of Neuroscience, Karolinska Institutet, 17177 Stockholm, Sweden.
authors.forename: Dagmar
authors.initials: D
authors.lastname: Galter
authors.affiliation: Department of Neurodegeneration, Hertie-Institute for Clinical Brain Research and DZNE, German Center for Neurodegenerative Diseases, 72076 Tuebingen, Germany.
authors.forename: Saskia
authors.initials: S
authors.lastname: Biskup
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D003599:Cytoskeleton / Q000187:drug effects / Q000378:metabolism*; D004298:Dopamine / Q000378:metabolism*; D005260:Female; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D008875:Middle Aged; D009154:Mutation; D016501:Neurites / Q000187:drug effects / Q000378:metabolism; D010300:Parkinson Disease / Q000201:enzymology / Q000235:genetics; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000235:genetics*
chemical_list: D047428:Protein Kinase Inhibitors; C495280:LRRK2 protein, human; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2; D017346:Protein Serine-Threonine Kinases; D004298:Dopamine
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