The type III intermediate filament vimentin regulates organelle distribution and modulates autophagy

Biskou, Olga; Casanova, Víctor; Hooper, Kirsty M.; Kemp, Sadie; Wright, G. Payling; Satsangi, Jack; Barlow, Peter G.; Stevens, Craig

doi:10.1371/journal.pone.0209665

Cited by 31 publications

(30 citation statements)

References 52 publications

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“…Vimentin is involved in vesicular membrane traffic and vimentin filaments are required for endo-lysosomal vesicle transport and positioning of endosomes and lysosomes [ 26 ]. Recently, some evidence has emerged that vimentin is involved in autophagy progression [ 27 ]. In HEK293 cells, vimentin aggregation by whitaferin-A increases LC3-II levels and causes the juxtanuclear clustering of autophagosomes and lysosomes with only limited colocalization [ 27 ].…”

Section: Resultsmentioning

confidence: 99%

“…Recently, some evidence has emerged that vimentin is involved in autophagy progression [ 27 ]. In HEK293 cells, vimentin aggregation by whitaferin-A increases LC3-II levels and causes the juxtanuclear clustering of autophagosomes and lysosomes with only limited colocalization [ 27 ]. While the latter finding would suggest that vimentin aggregation interferes with the fusion of autophagosomes with lysosomes, vimentin depletion by gene silencing does not seem to affect basal autophagy or autophagosome distribution [ 27 ].…”

Section: Resultsmentioning

confidence: 99%

“…In HEK293 cells, vimentin aggregation by whitaferin-A increases LC3-II levels and causes the juxtanuclear clustering of autophagosomes and lysosomes with only limited colocalization [ 27 ]. While the latter finding would suggest that vimentin aggregation interferes with the fusion of autophagosomes with lysosomes, vimentin depletion by gene silencing does not seem to affect basal autophagy or autophagosome distribution [ 27 ]. We investigated only the levels of the protein and no cell imaging study was performed that could inform us whether d-limonene and CQ affect the intracellular distribution of vimentin, which indeed deserves further investigation.…”

Section: Resultsmentioning

confidence: 99%

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Effects of the autophagy modulators d-limonene and chloroquine on vimentin levels in SH-SY5Y cells

Gentile

Berliocchi

Russo

et al. 2020

Biochemical and Biophysical Research Communications

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The molecular target and mechanism by which d-limonene induces LC3 lipidation and autophagosome formation remain elusive. Here, we report that this monoterpene rapidly enhances Ca 2+ levels in SH-SY5Y cells; yet this effect does not lead to calpain- or caspase-mediated proteolysis of α-spectrin, nor calpain activity is required for the established enhancement of LC3-II levels by d-limonene. However, d-limonene rapidly reduced vimentin levels, an unexpected effect also induced by the autophagy inhibitor chloroquine (CQ). The magnitude of vimentin reduction parallels accumulation of LC3-II caused by a brief incubation with d-limonene or CQ. For longer exposure (48 h), d-limonene does not reduce vimentin, nor it increases LC3-II levels; conversely, a clear reduction of vimentin along with a massive accumulation of LC3-II is evident in cells treated with CQ. Vimentin participates in organelle positioning and in other cellular processes that have linked this intermediate filament protein to various diseases, including cancer, inflammatory and autoimmune disorders, and to virus replication and internalization. Our findings suggest an inverse relationship between vimentin reduction and LC3-II accumulation, whose causal link needs to be examined. Further experiments are needed to dissect the role of vimentin reduction in the mechanisms through which CQ impairs fusion of autophagosome with lysosomes as well as in other effects of this drug.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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Effects of the autophagy modulators d-limonene and chloroquine on vimentin levels in SH-SY5Y cells

Gentile

Berliocchi

Russo

et al. 2020

Biochemical and Biophysical Research Communications

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“…This opens up the potential to study lysosome transport dynamics and their interaction with the cytoskeleton 24 in cells under controlled mechanical load. Recent reports show that the vimentin cytoskeleton plays a central role in regulating lysosome trafficking in autophagy 25 . To illustrate the ability of the system to visualize fine detail while scanning at a high frame rate, we fixed the mEmerald vimentin cells and stained them with AlexaFluor 594 phalloidin for F-actin.…”

Section: Supplementary Video 1 and Supplementarymentioning

confidence: 99%

Combined Atomic Force Microscope and Volumetric Light Sheet System for Correlative Force and Fluorescence Mechanobiology Studies

Nelsen

Hobson

Kern

et al. 2020

Sci Rep

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the central goals of mechanobiology are to understand how cells generate force and how they respond to environmental mechanical stimuli. A full picture of these processes requires high-resolution, volumetric imaging with time-correlated force measurements. Here we present an instrument that combines an open-top, single-objective light sheet fluorescence microscope with an atomic force microscope (AfM), providing simultaneous volumetric imaging with high spatiotemporal resolution and high dynamic range force capability (10 pN-100 nN). With this system we have captured lysosome trafficking, vimentin nuclear caging, and actin dynamics on the order of one second per single-cell volume. to showcase the unique advantages of combining Line Bessel light sheet imaging with AfM, we measured the forces exerted by a macrophage during fcɣR-mediated phagocytosis while performing both sequential two-color, fixed plane and volumetric imaging of F-actin. This unique instrument allows for a myriad of novel studies investigating the coupling of cellular dynamics and mechanical forces. Cells interact mechanically with their environment by generating and responding to forces. A focus on the mechanical dynamics of cell phenomena such as motility, division and phagocytosis is essential for understanding stem cell fate 1 , cancer progression 2 and innate immunity 3. These mechanical processes are inherently three-dimensional (3D) and are regulated both by very local (nm) interactions as well as whole-cell scale (μm) biochemical and mechanical signaling. Obtaining a more complete picture of a cell's mechanical interaction with its environment requires monitoring local and global structures in 3D while simultaneously measuring associated forces. These goals necessitate integrating high spatial and temporal resolution volumetric imaging methods with high resolution force acquisition. Light sheet florescence microscopy (LSFM) enables acquisition of volumetric, multicolor time series at a high resolution and frame rate with low background fluorescence and low phototoxicity 4-6. Among single cell force methods 7 , Atomic Force Microscopy (AFM) 8 is unique in combining a large force range (10-11-10-6 N) that enables molecular-scale to tissue-level mechanics measurements, with high bandwidth temporal resolution (μs) and sub-nanometer spatial control. LSFM provides sufficient spatiotemporal resolution for studying cell-wide protein specific dynamics that AFM imaging cannot. Therefore, by using the AFM for force spectroscopy and LSFM for imaging we have exploited the strengths of each technique resulting in a system that is greater than its parts. Combining LSFM with AFM (AFM-LS) imposes significant geometrical constraints to the optical system design, and demands low vibration operation to accommodate sensitive force measurements. To address this challenge, we used a single-objective selective plane microscopy (soSPIM) technique integrated with, and time-synchronized to, an AFM. As described in a report on our first generation system 9 , we gene...

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Section: Two-color Volumetric Imagingmentioning

confidence: 99%

Combined Atomic Force Microscope and Volumetric Light Sheet System for Mechanobiology

Nelsen

Hobson

Kern

et al. 2019

Preprint

View full text Add to dashboard Cite

The central goals of mechanobiology are to understand how cells generate force and how they respond to environmental mechanical stimuli. A full picture of these processes requires high-resolution, volumetric imaging with time-correlated force measurements. Here we present an instrument that combines an open-top, single-objective light sheet fluorescence microscope with an atomic force microscope (AFM), providing simultaneous volumetric imaging with high spatiotemporal resolution and high dynamic range force capability (10 pN – 100 nN). With this system we have captured lysosome trafficking, vimentin nuclear caging, and actin dynamics on the order of one second per volume. To showcase the unique advantages of combining Line Bessel light sheet imaging with AFM, we measured the forces exerted by a macrophage during FcɣR-mediated phagocytosis while performing both sequential two-color, fixed plane and volumetric imaging of F-actin. This unique instrument allows for a myriad of novel studies investigating the coupling of cellular dynamics and mechanical forces.

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The type III intermediate filament vimentin regulates organelle distribution and modulates autophagy

Cited by 31 publications

References 52 publications

Effects of the autophagy modulators d-limonene and chloroquine on vimentin levels in SH-SY5Y cells

Effects of the autophagy modulators d-limonene and chloroquine on vimentin levels in SH-SY5Y cells

Combined Atomic Force Microscope and Volumetric Light Sheet System for Correlative Force and Fluorescence Mechanobiology Studies

Combined Atomic Force Microscope and Volumetric Light Sheet System for Mechanobiology

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