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Protein Cell. 2016 Nov;7(11):804-819. doi: 10.1007/s13238-016-0268-3. Epub 2016 May 25.

A new method for quantifying mitochondrial axonal transport.

Chen M1,2, Li Y3,4, Yang M1,2, Chen X2, Chen Y5, Yang F5, Lu S5, Yao S2, Zhou T2, Liu J6, Zhu L6, Du S5, Wu JY7,8.

Author information

1
University of Chinese Academy of Sciences, Beijing, 100049, China.
2
Department of Neurology, Center for Genetic Medicine, Lurie Cancer Center, Northwestern University Feinberg School of Medicine, Chicago, IL, 60611, USA.
3
School of Electronic Science & Engineering, Nanjing University, Nanjing, 210093, China. yogo@nju.edu.cn.
4
Department of Neurology, Center for Genetic Medicine, Lurie Cancer Center, Northwestern University Feinberg School of Medicine, Chicago, IL, 60611, USA. yogo@nju.edu.cn.
5
School of Electronic Science & Engineering, Nanjing University, Nanjing, 210093, China.
6
State Key Laboratory for Brain & Cognitive Science, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101, China.
7
State Key Laboratory for Brain & Cognitive Science, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101, China. jane-wu@northwestern.edu.
8
Department of Neurology, Center for Genetic Medicine, Lurie Cancer Center, Northwestern University Feinberg School of Medicine, Chicago, IL, 60611, USA. jane-wu@northwestern.edu.

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.

KEYWORDS:

FUS proteinopathy and mitochondrial transport defect; image processing and analysis; mitochondrial transport

PMID:
27225265
PMCID:
PMC5084152
DOI:
10.1007/s13238-016-0268-3
[Indexed for MEDLINE]
Free PMC Article

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