Cytoskeletal Alterations and Biomechanical Properties of parkin-Mutant Human Primary Fibroblasts.
Parkinson's disease (PD) is one of the most common neurodegenerative diseases. Genes which have been implicated in autosomal-recessive PD include PARK2 which codes for parkin, an E3 ubiquitin ligase that participates in a variety of cellular activities. In this study, we compared parkin-mutant primary fibroblasts, from a patient with parkin compound heterozygous mutations, to healthy control cells. Western blot analysis of proteins obtained from patient's fibroblasts showed quantitative differences of many proteins involved in the cytoskeleton organization with respect to control cells. These molecular alterations are accompanied by changes in the organization of actin stress fibers and biomechanical properties, as revealed by confocal laser scanning microscopy and atomic force microscopy. In particular, parkin deficiency is associated with a significant increase of Young's modulus of null-cells in comparison to normal fibroblasts. The current study proposes that parkin influences the spatial organization of actin filaments, the shape of human fibroblasts, and their elastic response to an external applied force.
Department of Biological and Environmental Sciences and Technologies, University of Salento, Lecce, Italy.|Laboratory of Clinical Proteomic, ''Giovanni Paolo II'' Hospital, ASL-Lecce, Italy.
Daniele
D
Vergara
Department of Biological and Environmental Sciences and Technologies, University of Salento, Lecce, Italy.|Institute of Nanoscience-NNL, CNR, Via Arnesano, 16, Lecce, 73100, Italy.
Marzia M
MM
Ferraro
Institute of Nanoscience-NNL, CNR, Via Arnesano, 16, Lecce, 73100, Italy.|Department of Mathematics and Physics "Ennio De Giorgi", University of Salento, Lecce, Italy.
Mariafrancesca
M
Cascione
Institute of Nanoscience-NNL, CNR, Via Arnesano, 16, Lecce, 73100, Italy.
Loretta L
LL
del Mercato
Institute of Nanoscience-NNL, CNR, Via Arnesano, 16, Lecce, 73100, Italy.
Stefano
S
Leporatti
Department of Basic Medical Sciences, Neurosciences and Organs of Senses, University of Bari 'A. Moro', Bari, Italy.
Anna
A
Ferretta
Department of Basic Medical Sciences, Neurosciences and Organs of Senses, University of Bari 'A. Moro', Bari, Italy.
Paola
P
Tanzarella
Department of Pharmacology, Faculty of Medicine, Universitè de Montreal, 2900 Boulevard Edouard-Montpetit, Montreal, QC, H3T1J4, Canada.
Consiglia
C
Pacelli
Institute of Science of Food Production, CNR, Lecce, Italy.
Angelo
A
Santino
Department of Biological and Environmental Sciences and Technologies, University of Salento, Lecce, Italy.|Laboratory of Clinical Proteomic, ''Giovanni Paolo II'' Hospital, ASL-Lecce, Italy.
Michele
M
Maffia
Department of Basic Medical Sciences, Neurosciences and Organs of Senses, University of Bari 'A. Moro', Bari, Italy.
Tiziana
T
Cocco
Institute of Nanoscience-NNL, CNR, Via Arnesano, 16, Lecce, 73100, Italy.|Department of Mathematics and Physics "Ennio De Giorgi", University of Salento, Lecce, Italy.
Ross
R
Rinaldi
Institute of Nanoscience-NNL, CNR, Via Arnesano, 16, Lecce, 73100, Italy. antonio.gaballo@nano.cnr.it.
Antonio
A
Gaballo
D001696:Biomechanical Phenomena; D048430:Cell Shape; D003598:Cytoskeletal Proteins / Q000378:metabolism; D003599:Cytoskeleton / Q000378:metabolism*; D004548:Elasticity; D005347:Fibroblasts / Q000473:pathology*; D006801:Humans; D055595:Mechanical Phenomena; D009154:Mutation; D010300:Parkinson Disease / Q000235:genetics / Q000473:pathology; D044767:Ubiquitin-Protein Ligases / Q000235:genetics*
D003598:Cytoskeletal Proteins; D044767:Ubiquitin-Protein Ligases; C111567:parkin protein
Atomic force microscopy; Cofilin; Cytoskeleton; Fibers anisotropy; Fibroblasts; Parkin