title: A new method for quantifying mitochondrial axonal transport.
abstract: Axonal transport of mitochondria is critical for neuronal survival and function. Automatically quantifying and analyzing mitochondrial movement in a large quantity remain challenging. Here, we report an efficient method for imaging and quantifying axonal mitochondrial transport using microfluidic-chamber-cultured neurons together with a newly developed analysis package named "MitoQuant". This tool-kit consists of an automated program for tracking mitochondrial movement inside live neuronal axons and a transient-velocity analysis program for analyzing dynamic movement patterns of mitochondria. Using this method, we examined axonal mitochondrial movement both in cultured mammalian neurons and in motor neuron axons of Drosophila in vivo. In 3 different paradigms (temperature changes, drug treatment and genetic manipulation) that affect mitochondria, we have shown that this new method is highly efficient and sensitive for detecting changes in mitochondrial movement. The method significantly enhanced our ability to quantitatively analyze axonal mitochondrial movement and allowed us to detect dynamic changes in axonal mitochondrial transport that were not detected by traditional kymographic analyses.
authors.affiliation: University of Chinese Academy of Sciences, Beijing, 100049, China.|Department of Neurology, Center for Genetic Medicine, Lurie Cancer Center, Northwestern University Feinberg School of Medicine, Chicago, IL, 60611, USA.
authors.forename: Mengmeng
authors.initials: M
authors.lastname: Chen
authors.affiliation: School of Electronic Science & Engineering, Nanjing University, Nanjing, 210093, China. yogo@nju.edu.cn.|Department of Neurology, Center for Genetic Medicine, Lurie Cancer Center, Northwestern University Feinberg School of Medicine, Chicago, IL, 60611, USA. yogo@nju.edu.cn.
authors.forename: Yang
authors.initials: Y
authors.lastname: Li
authors.affiliation: University of Chinese Academy of Sciences, Beijing, 100049, China.|Department of Neurology, Center for Genetic Medicine, Lurie Cancer Center, Northwestern University Feinberg School of Medicine, Chicago, IL, 60611, USA.
authors.forename: Mengxue
authors.initials: M
authors.lastname: Yang
authors.affiliation: Department of Neurology, Center for Genetic Medicine, Lurie Cancer Center, Northwestern University Feinberg School of Medicine, Chicago, IL, 60611, USA.
authors.forename: Xiaoping
authors.initials: X
authors.lastname: Chen
authors.affiliation: School of Electronic Science & Engineering, Nanjing University, Nanjing, 210093, China.
authors.forename: Yemeng
authors.initials: Y
authors.lastname: Chen
authors.affiliation: School of Electronic Science & Engineering, Nanjing University, Nanjing, 210093, China.
authors.forename: Fan
authors.initials: F
authors.lastname: Yang
authors.affiliation: School of Electronic Science & Engineering, Nanjing University, Nanjing, 210093, China.
authors.forename: Sheng
authors.initials: S
authors.lastname: Lu
authors.affiliation: Department of Neurology, Center for Genetic Medicine, Lurie Cancer Center, Northwestern University Feinberg School of Medicine, Chicago, IL, 60611, USA.
authors.forename: Shengyu
authors.initials: S
authors.lastname: Yao
authors.affiliation: Department of Neurology, Center for Genetic Medicine, Lurie Cancer Center, Northwestern University Feinberg School of Medicine, Chicago, IL, 60611, USA.
authors.forename: Timothy
authors.initials: T
authors.lastname: Zhou
authors.affiliation: State Key Laboratory for Brain & Cognitive Science, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101, China.
authors.forename: Jianghong
authors.initials: J
authors.lastname: Liu
authors.affiliation: State Key Laboratory for Brain & Cognitive Science, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101, China.
authors.forename: Li
authors.initials: L
authors.lastname: Zhu
authors.affiliation: School of Electronic Science & Engineering, Nanjing University, Nanjing, 210093, China.
authors.forename: Sidan
authors.initials: S
authors.lastname: Du
authors.affiliation: State Key Laboratory for Brain & Cognitive Science, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101, China. jane-wu@northwestern.edu.|Department of Neurology, Center for Genetic Medicine, Lurie Cancer Center, Northwestern University Feinberg School of Medicine, Chicago, IL, 60611, USA. jane-wu@northwestern.edu.
authors.forename: Jane Y
authors.initials: JY
authors.lastname: Wu
mesh_terms: D000818:Animals; D001370:Axonal Transport / Q000502:physiology*; D002540:Cerebral Cortex / Q000166:cytology / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000166:cytology / Q000378:metabolism; D004622:Embryo, Mammalian; D015870:Gene Expression; D056656:Lab-On-A-Chip Devices; D018613:Microscopy, Confocal; D008928:Mitochondria / Q000378:metabolism* / Q000648:ultrastructure; D009046:Motor Neurons / Q000378:metabolism* / Q000648:ultrastructure; D009068:Movement; D009154:Mutation; D061251:Primary Cell Culture; D034702:RNA-Binding Protein FUS / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D012984:Software
chemical_list: D034702:RNA-Binding Protein FUS
keywords: FUS proteinopathy and mitochondrial transport defect; image processing and analysis; mitochondrial transport