Cargos Rotate at Microtubule Intersections during Intracellular Trafficking

Gao, Yuan; Anthony, Stephen M.; Yu, Yanqi; Yi, Yi

doi:10.1016/j.bpj.2018.05.010

Cited by 25 publications

(25 citation statements)

References 51 publications

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“…In their study, the circumventing motion of a vesicle was detected when the vesicle encountered an obstacle, such as an intersection of microtubules. Furthermore, the rotation of a cargo at the microtubule intersection was also recently reported using fluorescently anisotropic particle and two-dimension image analysis [42]. Therefore, one of the possible explanations for the rotational motion of the vesicle reported in the present study might be the result of the interaction with such obstacles including the microtubule intersection in the cytoplasmic area.…”

Section: Discussionsupporting

confidence: 81%

3D rotational motion of an endocytic vesicle on a complex microtubule network in a living cell

Lee

Higuchi

2019

Biomed. Opt. Express

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The transport dynamics of endocytic vesicles in a living cell contains essential biomedical information. Although the movement mechanism of a vesicle by motor proteins has been revealed, understanding the precise movement of vesicles on the cytoskeleton in a living cell has been considered challenging, due to the complex 3D network of cytoskeletons. Here, we specify the shape of the 3D interaction between the vesicle and microtubule, based on the theoretically estimated location of the microtubule and the vesicle trajectory data acquired at high spatial and temporal precision. We detected that vesicles showed more frequent direction changes with either in very acute or in obtuse angles than right angles, on similar time scales in a microtubule network. Interestingly, when a vesicle interacted with a relatively longer (> 400 nm) microtubule filament, rotational movement along the axis of the microtubule was frequently observed. Our results are expected to give in-depth insight into understanding the actual 3D interactions between the intracellular molecule and complex cytoskeletal network.

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Section: Discussionsupporting

confidence: 81%

3D rotational motion of an endocytic vesicle on a complex microtubule network in a living cell

Lee

Higuchi

2019

Biomed. Opt. Express

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“…When motors engage in a tug-of-war, they apply forces on each other (Fig. 6B) (5,50,51). An understanding of how a tugof-war between teams of motors can result in stationary liposomes can be derived from single MyoVa studies in which opposing loads are applied.…”

Section: The Molecular Basis For Modes Of Cargo Motion: Directed Motionmentioning

confidence: 99%

Myosin Va transport of liposomes in three-dimensional actin networks is modulated by actin filament density, position, and polarity

Lombardo

Nelson

Kennedy

et al. 2019

Proc. Natl. Acad. Sci. U.S.A.

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The cell’s dense 3D actin filament network presents numerous challenges to vesicular transport by teams of myosin Va (MyoVa) molecular motors. These teams must navigate their cargo through diverse actin structures ranging from Arp2/3-branched lamellipodial networks to the dense, unbranched cortical networks. To define how actin filament network organization affects MyoVa cargo transport, we created two different 3D actin networks in vitro. One network was comprised of randomly oriented, unbranched actin filaments; the other was comprised of Arp2/3-branched actin filaments, which effectively polarized the network by aligning the actin filament plus-ends. Within both networks, we defined each actin filament’s 3D spatial position using superresolution stochastic optical reconstruction microscopy (STORM) and its polarity by observing the movement of single fluorescent reporter MyoVa. We then characterized the 3D trajectories of fluorescent, 350-nm fluid-like liposomes transported by MyoVa teams (∼10 motors) moving within each of the two networks. Compared with the unbranched network, we observed more liposomes with directed and fewer with stationary motion on the Arp2/3-branched network. This suggests that the modes of liposome transport by MyoVa motors are influenced by changes in the local actin filament polarity alignment within the network. This mechanism was supported by an in silico 3D model that provides a broader platform to understand how cellular regulation of the actin cytoskeletal architecture may fine tune MyoVa-based intracellular cargo transport.

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“…Paclitaxel (also known as taxol), a drug that induces tubulin assembly (Caplow et al, 1994), is one such example. It can be used as a constantly fluorescent derivative (Abal et al, 2001; Lillo et al, 2002; Barasoain et al, 2010) or as a modified reagent that only attains fluorescence inside the cell – known as Tubulin Tracker TM (Thermo Fisher Scientific), available with green and deep-red fluorophores (Grau et al, 2013; Zarrouk et al, 2015; Gao et al, 2018; Wang et al, 2018). These derivatives are membrane-permeable and can label microtubules in live cells when simply added to the cellular medium, but cannot be used in fixed samples or for long-term imaging as they are not retained well inside the cells.…”

Section: Methodsmentioning

confidence: 99%

Interrogating Synaptic Architecture: Approaches for Labeling Organelles and Cytoskeleton Components

Reshetniak

Rizzoli

2019

Front. Synaptic Neurosci.

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Synaptic transmission has been studied for decades, as a fundamental step in brain function. The structure of the synapse, and its changes during activity, turned out to be key aspects not only in the transfer of information between neurons, but also in cognitive processes such as learning and memory. The overall synaptic morphology has traditionally been studied by electron microscopy, which enables the visualization of synaptic structure in great detail. The changes in the organization of easily identified structures, such as the presynaptic active zone, or the postsynaptic density, are optimally studied via electron microscopy. However, few reliable methods are available for labeling individual organelles or protein complexes in electron microscopy. For such targets one typically relies either on combination of electron and fluorescence microscopy, or on super-resolution fluorescence microscopy. This review focuses on approaches and techniques used to specifically reveal synaptic organelles and protein complexes, such as cytoskeletal assemblies. We place the strongest emphasis on methods detecting the targets of interest by affinity binding, and we discuss the advantages and limitations of each method.

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Cargos Rotate at Microtubule Intersections during Intracellular Trafficking

Cited by 25 publications

References 51 publications

3D rotational motion of an endocytic vesicle on a complex microtubule network in a living cell

3D rotational motion of an endocytic vesicle on a complex microtubule network in a living cell

Myosin Va transport of liposomes in three-dimensional actin networks is modulated by actin filament density, position, and polarity

Interrogating Synaptic Architecture: Approaches for Labeling Organelles and Cytoskeleton Components

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