Visualization Method for the Cell-Level Vesicle Transport Using Optical Flow and a Diverging Colormap

Elucidation of cell-level transport mediated by vesicles within a living cell provides key information regarding viral infection processes and also drug delivery mechanisms. Although the single-particle tracking method has enabled clear analysis of individual vesicle trajectories, information regarding the entire cell-level intracellular transport is hardly obtainable, due to the difficulty in collecting a large dataset with current methods. In this paper, we propose a visualization method of vesicle transport using optical flow, based on geometric cell center estimation and vector analysis, for measuring the trafficking directions. As a quantitative visualization method for determining the intracellular transport status, the proposed method is expected to be universally exploited in various biomedical cell image analyses.

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Intracellular transport is accelerated in early apoptotic cells

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1810017115 ◽

2018 ◽

Vol 115 (48) ◽

pp. 12118-12123 ◽

Cited By ~ 6

Author(s):

Bo Li ◽

Shuo-Xing Dou ◽

Jing-Wen Yuan ◽

Yu-Ru Liu ◽

Wei Li ◽

...

Keyword(s):

Intracellular Transport ◽

Single Particle Tracking ◽

Cellular Proteins ◽

Cell Physiology ◽

Apoptotic Cells ◽

Transport Dynamics ◽

Particle Tracking Method ◽

Endocytic Vesicles ◽

Shed Light

Intracellular transport of cellular proteins and organelles is critical for establishing and maintaining intracellular organization and cell physiology. Apoptosis is a process of programmed cell death with dramatic changes in cell morphology and organization, during which signaling molecules are transported between different organelles within the cells. However, how the intracellular transport changes in cells undergoing apoptosis remains unknown. Here, we study the dynamics of intracellular transport by using the single-particle tracking method and find that both directed and diffusive motions of endocytic vesicles are accelerated in early apoptotic cells. With careful elimination of other factors involved in the intracellular transport, the reason for the acceleration is attributed to the elevation of adenosine triphosphate (ATP) concentration. More importantly, we show that the accelerated intracellular transport is critical for apoptosis, and apoptosis is delayed when the dynamics of intracellular transport is regulated back to the normal level. Our results demonstrate the important role of transport dynamics in apoptosis and shed light on the apoptosis mechanism from a physical perspective.

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Neuronal receptors display cytoskeleton-independent directed motion on the plasma membrane

10.1101/269381 ◽

2018 ◽

Author(s):

R. D. Taylor ◽

M. Heine ◽

N. J. Emptage ◽

L. C. Andreae

Keyword(s):

Plasma Membrane ◽

Neuronal Activity ◽

Particle Tracking ◽

Hippocampal Neurons ◽

Intracellular Transport ◽

Transmembrane Proteins ◽

Single Particle Tracking ◽

Directed Motion ◽

Surface Movement ◽

Transport Vesicles

AbstractDirected transport of transmembrane proteins is generally believed to occur via intracellular transport vesicles. However, using single particle tracking in rat hippocampal neurons with a pH-sensitive quantum dot probe which specifically reports surface movement of receptors, we have identified a subpopulation of neuronal EphB2 receptors that exhibit directed motion between synapses within the plasma membrane itself. This receptor movement occurs independently of the cytoskeleton but is dependent on cholesterol and is regulated by neuronal activity.

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Subtle changes in crosslinking drive diverse anomalous transport characteristics in actin-microtubule networks

10.21203/rs.3.rs-119120/v1 ◽

2020 ◽

Author(s):

Sylas Anderson ◽

Jonathan Garamella ◽

Ryan McGorty ◽

Rae Robertson-Anderson

Keyword(s):

Intracellular Transport ◽

Single Particle Tracking ◽

Anomalous Transport ◽

Brownian Diffusion ◽

Complex Environments ◽

Materials Engineering ◽

Microtubule Networks ◽

Non Gaussian ◽

Fluid Rheology ◽

The Impact

Abstract Anomalous diffusion in crowded and complex environments is widely studied due to its importance in intracellular transport, fluid rheology and materials engineering. Specifically, diffusion through the cytoskeleton, a network comprised of semiflexible actin filaments and rigid microtubules that interact both sterically and via crosslinking, plays a principal role in viral infection, vesicle transport and targeted drug delivery. Here, we elucidate the impact of crosslinking on particle diffusion in composites of actin and microtubules with actin-actin, microtubule-microtubule and actin-microtubule crosslinking. We analyze a suite of complementary transport metrics by coupling single-particle tracking and differential dynamic microscopy. Using these orthogonal techniques, we find that particles display non-Gaussian and non-ergodic subdiffusion that is markedly enhanced by cytoskeletal crosslinking of any type, which we attribute to suppressed microtubule mobility. However, the extent to which transport deviates from normal Brownian diffusion depends strongly on the crosslinking motif – with actin-microtubule crosslinking inducing the most pronounced anomalous characteristics – due to increased actin fluctuation heterogeneity. Our results reveal that subtle changes to actin-microtubule interactions can have dramatic impacts on diffusion in the cytoskeleton, and suggest that less mobile and more locally heterogeneous networks lead to more strongly anomalous transport.

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Single particle tracking method for the analysis of intracellular movement of the silica nanoparticles

Toxicology Letters ◽

10.1016/j.toxlet.2014.06.634 ◽

2014 ◽

Vol 229 ◽

pp. S186-S187

Author(s):

Michihiko Aoyama ◽

Yasuo Yoshioka ◽

Yoshiyuki Arai ◽

Rio Ishimoto ◽

Shin-ichi Tsunoda ◽

...

Keyword(s):

Silica Nanoparticles ◽

Particle Tracking ◽

Single Particle ◽

Single Particle Tracking ◽

Tracking Method ◽

Particle Tracking Method ◽

Intracellular Movement

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Generation of an erythrocyte vesicle transport system by Plasmodium falciparum malaria parasites

Blood ◽

10.1182/blood-2003-05-1448 ◽

2003 ◽

Vol 102 (9) ◽

pp. 3420-3426 ◽

Cited By ~ 56

Author(s):

Theodore F. Taraschi ◽

Megan O'Donnell ◽

Sandra Martinez ◽

Timothy Schneider ◽

Darin Trelka ◽

...

Keyword(s):

Plasmodium Falciparum ◽

Plasma Membrane ◽

G Proteins ◽

Intracellular Transport ◽

Surface Membrane ◽

Plasmodium Falciparum Malaria ◽

Vesicle Transport ◽

Aluminum Fluoride ◽

Sensitive Factor ◽

Erythrocyte Plasma Membrane

AbstractThe asexual maturation of Plasmodium falciparum is accompanied by the transport of parasite-encoded proteins to the erythrocyte plasma membrane. Activation of G proteins by treatment with aluminum fluoride produced an accumulation within the erythrocyte cytosol of vesicles coated with Plasmodium homologues of COPII and N-ethylmaleimide-sensitive factor, proteins involved in intracellular transport between the Golgi apparatus and the endoplasmic reticulum. These vesicles contain malarial proteins that appear on the erythrocyte plasma membrane, as well as actin and myosin. It is proposed that the parasite adapted a process well established for intracellular transport to mediate the extracellular movement of its proteins through the erythrocyte cytosol to the surface membrane.

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A Quantitative Visualization Method for Wind Tunnel Experiments Based on 3D Particle Tracking Velocimetry (3D-PTV)

PAMM ◽

10.1002/1617-7061(200203)1:1<254::aid-pamm254>3.0.co;2-b ◽

2002 ◽

Vol 1 (1) ◽

pp. 254

Author(s):

M. Machacek ◽

T. Rösgen

Keyword(s):

Wind Tunnel ◽

Particle Tracking ◽

Particle Tracking Velocimetry ◽

Visualization Method ◽

Wind Tunnel Experiments ◽

Quantitative Visualization

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Single Particle Tracking Confirms That Multivalent Tat Protein Transduction Domain-induced Heparan Sulfate Proteoglycan Cross-linkage Activates Rac1 for Internalization

Journal of Biological Chemistry ◽

10.1074/jbc.m110.187450 ◽

2011 ◽

Vol 286 (12) ◽

pp. 10581-10592 ◽

Cited By ~ 26

Author(s):

Junji Imamura ◽

Yasuhiro Suzuki ◽

Kohsuke Gonda ◽

Chandra Nath Roy ◽

Hiroyuki Gatanaga ◽

...

Keyword(s):

Cell Surface ◽

Heparan Sulfate ◽

Intracellular Transport ◽

Single Particle Tracking ◽

Protein Transduction ◽

Protein Transduction Domain ◽

Tat Protein ◽

Surface Binding ◽

Future Design ◽

Tat Protein Transduction Domain

The mechanism by which HIV-1-Tat protein transduction domain (TatP) enters the cell remains unclear because of an insufficient understanding of the initial kinetics of peptide entry. Here, we report the successful visualization and tracking of TatP molecular kinetics on the cell surface with 7-nm spatial precision using quantum dots. Strong cell binding was only observed with a TatP valence of ≥8, whereas monovalent TatP binding was negligible. The requirement of the cell-surface heparan sulfate (HS) chains of HS proteoglycans (HSPGs) for TatP binding and intracellular transport was demonstrated by the enzymatic removal of HS and simultaneous observation of two individual particles. Multivalent TatP induces HSPG cross-linking, recruiting activated Rac1 to adjacent lipid rafts and thereby enhancing the recruitment of TatP/HSPG to actin-associated microdomains and its internalization by macropinocytosis. These findings clarify the initial binding mechanism of TatP to the cell surface and demonstrate the importance of TatP valence for strong surface binding and signal transduction. Our data also shed light on the ability of TatP to exploit the machinery of living cells, using HSPG signaling to activate Rac1 and alter TatP mobility and internalization. This work should guide the future design of TatP-based peptides as therapeutic nanocarriers with efficient transduction.

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Intracellular transport dynamics revealed by single-particle tracking

Biophysics Reports ◽

10.52601/bpr.2021.210035 ◽

2021 ◽

Vol 7 (5) ◽

pp. 413-427

Author(s):

Zhang Ming-Li ◽

◽

Ti Hui-Ying ◽

Wang Peng-Ye ◽

Li Hui ◽

...

Keyword(s):

Particle Tracking ◽

Single Particle ◽

Intracellular Transport ◽

Single Particle Tracking ◽

Transport Dynamics

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Resolving Cargo-motor-track Interactions in Living Cells with Bifocal Parallax Single Particle Tracking

10.1101/2020.07.06.189746 ◽

2020 ◽

Author(s):

Xiaodong Cheng ◽

Kuangcai Chen ◽

Bin Dong ◽

Seth L. Filbrun ◽

Gufeng Wang ◽

...

Keyword(s):

Particle Tracking ◽

Single Particle ◽

Molecular Motors ◽

Intracellular Transport ◽

Full Range ◽

Translational Motion ◽

Polar Angle ◽

Living Cells ◽

Single Particle Tracking ◽

Dynamic Binding

AbstractResolving coordinated biomolecular interactions in living cellular environments is vital for understanding the mechanisms of molecular nanomachines. The conventional approach relies on localizing and tracking target biomolecules and/or subcellular organelles labeled with imaging probes. However, it is challenging to gain information on rotational dynamics, which can be more indicative of the work done by molecular motors and their dynamic binding status. Herein, a bifocal parallax single particle tracking method using half-plane point spread functions has been developed to resolve the full-range azimuth angle (0-360°), polar angle, and 3D displacement in real time under complex living cell conditions. Using this method, quantitative rotational and translational motion of the cargo in a 3D cell cytoskeleton was obtained. Not only well-known active intracellular transport and free diffusion were observed but new interactions, tight attachment and tethered rotation, were discovered for better interpretation of the dynamics of cargo-motor-track interactions at various types of microtubule intersections.Significance StatementTranslation and rotational motion of cargo during pauses at the microtubule intersections in living cells were revealed by high-accuracy three-dimensional single particle rotational tracking. The current study demonstrates the potential of studying coordinated interactions in living cellular environments by resolving characteristic rotational motions.

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