Alterations in mitochondrial dynamics, including their intracellular trafficking, are common early manifestations of neuronal degeneration. However, current methodologies used to study mitochondrial trafficking events rely on parameters that are primarily altered in later stages of neurodegeneration. Our objective was to establish a reliable applied statistical analysis to detect early alterations in neuronal mitochondrial trafficking. We propose a novel quantitative analysis of mitochondria trajectories based on innovative movement descriptors, including straightness, efficiency, anisotropy, and kurtosis. We evaluated time- and dose-dependent alterations in trajectory descriptors using biological data from differentiated SH-SY5Y cells treated with the mitochondrial toxicants 6-hydroxydopamine and rotenone. MitoTracker Red CMXRos-labelled mitochondria movement was analyzed by total internal reflection fluorescence microscopy followed by computational modelling to describe the process. Based on the aforementioned trajectory descriptors, this innovative analysis of mitochondria trajectories provides insights into mitochondrial movement characteristics and can be a consistent and sensitive method to detect alterations in mitochondrial trafficking occurring in the earliest time points of neurodegeneration.

Centre for Bioinformatics Faculty of Mathematics and Natural Sciences University of Oslo Oslo Norway

CFisUC Department of Physics University of Coimbra Coimbra Portugal

CISUC Department of Informatics Engineering University of Coimbra Coimbra Portugal

CNC Center for Neuroscience and Cell Biology UC Biotech Cantanhede Portugal

Coimbra Polytechnic ISEC Coimbra Portugal

Institute of Biotechnology Czech Academy of Sciences Prague West Czech Republic

OCBE Faculty of Medicine University of Oslo Oslo Norway

School of Medical Science Griffith University Southport Queensland Australia

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