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Determination of cytoplasmic optineurin foci sizes using mage correlation spectroscopy

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Title: Determination of cytoplasmic optineurin foci sizes using mage correlation spectroscopy
Authors: Kitamura, Akira Browse this author →KAKEN DB
Shimizu, Hiroki Browse this author
Kinjo, Masataka Browse this author →KAKEN DB
Keywords: foci
image correlation spectroscopy
inclusion body
vesicular trafficking
Issue Date: Sep-2018
Publisher: Oxford University Press
Journal Title: Journal of biochemistry
Volume: 164
Issue: 3
Start Page: 223
End Page: 229
Publisher DOI: 10.1093/jb/mvy044
Abstract: Optineurin (OPTN) plays an important role in membrane trafficking processes such as exocytosis and autophagy. The sizes and rate of formation of accumulated structures comprising OPTN, such as foci or inclusion bodies (IBs), are often disrupted by amyotrophic lateral sclerosis (ALS) and glaucoma-associated mutants of OPTN. Therefore, methods for the quantitative measurement of the size of the accumulated structure are necessary. Here, we show that, using spatial image correlation spectroscopy (ICS), the average diameter of accumulated structures of the wild-type and disease-associated mutants in living cells may he easily determined. Although OPTN was found to frequently form foci in the cytoplasm, regardless of ALS- and glaucoma-associated mutation, the diameter of OPTN foci decreased in an ALS-associated mutant and increased in a glaucoma-associated mutant. However, a portion of cells carried IBs of the AM-associated mutant that were larger than micrometre and ellipse-like shape, suggesting that this mutant accumulates non-uniformly in the IBs. The findings suggest that changes in their accumulation, determined via quantitative comparison of the OPTN foci and IBs in the cells, are involved in pathological features of ALS. In addition, this method enables rapid comparison of the average sizes of various other intracellular structures such as granules.
Type: article (author version)
Appears in Collections:生命科学院・先端生命科学研究院 (Graduate School of Life Science / Faculty of Advanced Life Science) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 北村 朗

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