scholarly journals Myosin Va binding to neurofilaments is essential for correct myosin Va distribution and transport and neurofilament density

2002 ◽  
Vol 159 (2) ◽  
pp. 279-290 ◽  
Author(s):  
Mala V. Rao ◽  
Linda J. Engle ◽  
Panaiyur S. Mohan ◽  
Aidong Yuan ◽  
Dike Qiu ◽  
...  
Keyword(s):  
Normal Distribution ◽  
Axonal Transport ◽  
Immunoelectron Microscopy ◽  
Molecular Motors ◽  
Peripheral Nerves ◽  
Molecular Motor ◽  
Neurofilament Proteins ◽  
Myosin Va ◽  
Slow Transport

The identification of molecular motors that modulate the neuronal cytoskeleton has been elusive. Here, we show that a molecular motor protein, myosin Va, is present in high proportions in the cytoskeleton of mouse CNS and peripheral nerves. Immunoelectron microscopy, coimmunoprecipitation, and blot overlay analyses demonstrate that myosin Va in axons associates with neurofilaments, and that the NF-L subunit is its major ligand. A physiological association is indicated by observations that the level of myosin Va is reduced in axons of NF-L–null mice lacking neurofilaments and increased in mice overexpressing NF-L, but unchanged in NF-H–null mice. In vivo pulse-labeled myosin Va advances along axons at slow transport rates overlapping with those of neurofilament proteins and actin, both of which coimmunoprecipitate with myosin Va. Eliminating neurofilaments from mice selectively accelerates myosin Va translocation and redistributes myosin Va to the actin-rich subaxolemma and membranous organelles. Finally, peripheral axons of dilute-lethal mice, lacking functional myosin Va, display selectively increased neurofilament number and levels of neurofilament proteins without altering axon caliber. These results identify myosin Va as a neurofilament-associated protein, and show that this association is essential to establish the normal distribution, axonal transport, and content of myosin Va, and the proper numbers of neurofilaments in axons.

scholarly journals Dense granule trafficking in Toxoplasma gondii requires a unique class 27 myosin and actin filaments

2016 ◽  
Vol 27 (13) ◽  
pp. 2080-2089 ◽  
Author(s):  
Aoife T. Heaslip ◽  
Shane R. Nelson ◽  
David M. Warshaw
Keyword(s):  
Toxoplasma Gondii ◽  
Molecular Motors ◽  
Fluorescent Protein ◽  
Molecular Motor ◽  
Structural Similarity ◽  
Dense Granule ◽  
Lytic Cycle ◽  
Secretory Vesicles ◽  
Filamentous Actin

The survival of Toxoplasma gondii within its host cell requires protein release from secretory vesicles, called dense granules, to maintain the parasite’s intracellular replicative niche. Despite the importance of DGs, nothing is known about the mechanisms underlying their transport. In higher eukaryotes, secretory vesicles are transported to the plasma membrane by molecular motors moving on their respective cytoskeletal tracks (i.e., microtubules and actin). Because the organization of these cytoskeletal structures differs substantially in T. gondii, the molecular motor dependence of DG trafficking is far from certain. By imaging the motions of green fluorescent protein–tagged DGs in intracellular parasites with high temporal and spatial resolution, we show through a combination of molecular genetics and chemical perturbations that directed DG transport is independent of microtubules and presumably their kinesin/dynein motors. However, directed DG transport is dependent on filamentous actin and a unique class 27 myosin, TgMyoF, which has structural similarity to myosin V, the prototypical cargo transporter. Actomyosin DG transport was unexpected, since filamentous parasite actin has yet to be visualized in vivo due in part to the prevailing model that parasite actin forms short, unstable filaments. Thus our data uncover new critical roles for these essential proteins in the lytic cycle of this devastating pathogen.

Modeling Bidirectional Transport of New and Used Organelles in Fast Axonal Transport in Neurons

2010 ◽  
Vol 133 (1) ◽  
Author(s):  
A. V. Kuznetsov
Keyword(s):  
Axonal Transport ◽  
Cell Body ◽  
Molecular Motors ◽  
Molecular Motor ◽  
Fast Axonal Transport ◽  
Concentration Difference ◽  
Neuron Body ◽  
Microtubule Polarity ◽  
Bidirectional Transport ◽  
The Right

This paper develops a model for simulating transport of newly synthesized material from the neuron body toward the synapse of the axon as well as transport of misfolded and aggregated proteins back to the neuron body for recycling. The model demonstrates that motor-assisted transport, much similar to diffusion, can occur due to a simple concentration difference between the cell body and the synapse; organelles heading to the synapse do not need to attach preferably to plus-end-directed molecular motors, same as organelles heading to the neuron body for recycling do not need to attach preferably to minus-end-directed molecular motors. The underlying mechanics of molecular-motor-assisted transport is such that organelles would be transported to the right place even if new and used organelles had the same probability of attachment to plus-end-directed (and minus-end-directed) motors. It is also demonstrated that the axon with organelle traps and a region with a reversed microtubule polarity would support much smaller organelle fluxes of both new and used organelles than a healthy axon. The flux of organelles is shown to decrease as the width of organelle traps increases.

A four kinetic state model of fast axonal transport: Model formulation and perturbation solution

Open Physics ◽  
2011 ◽  
Vol 9 (1) ◽  
Author(s):  
Andrey Kuznetsov
Keyword(s):  
Axonal Transport ◽  
Molecular Motors ◽  
Transport Model ◽  
Molecular Motor ◽  
Perturbation Solution ◽  
State Model ◽  
Model Formulation ◽  
Fast Axonal Transport ◽  
First Order ◽  
Proposed Model

AbstractThis paper formulates a four kinetic state model for fast axonal transport. The paper further develops the Smith-Simmons model that is based on equations governing intracellular molecular-motor-assisted transport; these equations are extended by considering four rather than three kinetic states for organelles. The model considers plus-end and minus-end-oriented organelles that can be either free (suspended in the cytosol) or attached to microtubules (MTs) (in the latter case organelles are transported by molecular motors). The paper then develops a method for uncoupling differential equations of the proposed model. A perturbation solution of this problem is obtained. The effect of transition between plus-end-oriented and minus-end oriented organelles is discussed. The accuracy of the obtained perturbation solution is evaluated by comparing the zero-order and the first-order results with a high-accuracy numerical solution.

scholarly journals De novo disease-associated mutations in KIF1A dominant negatively inhibit axonal transport of synaptic vesicle precursors

2021 ◽  
Author(s):  
Yuzu Anazawa ◽  
Tomoki Kita ◽  
Kumiko Hayashi ◽  
Shinsuke Niwa
Keyword(s):  
Synaptic Vesicle ◽  
Axonal Transport ◽  
De Novo ◽  
Molecular Motor ◽  
Model Systems ◽  
In Vitro Assays ◽  
In Vivo Model ◽  
Wild Type

KIF1A is a kinesin superfamily molecular motor that transports synaptic vesicle precursors in axons. Mutations in Kif1a lead to a group of neuronal diseases called KIF1A-associated neuronal disorder (KAND). KIF1A forms a homodimer and KAND mutations are mostly de novo and autosomal dominant; however, it is not known whether the function of wild-type KIF1A is inhibited by disease-associated KIF1A. No reliable in vivo model systems to analyze the molecular and cellular biology of KAND have been developed; therefore, here, we established Caenorhabditis elegans models for KAND using CRISPR/cas9 technology and analyzed defects in axonal transport. In the C. elegans models, heterozygotes and homozygotes exhibited reduced axonal transport phenotypes. In addition, we developed in vitro assays to analyze the motility of single heterodimers composed of wild-type KIF1A and disease-associated KIF1A. Disease-associated KIF1A significantly inhibited the motility of wild-type KIF1A when heterodimers were formed. These data indicate the molecular mechanism underlying the dominant nature of de novo KAND mutations.

scholarly journals Cargo crowding, stationary clusters and dynamical reservoirs in axonal transport

2021 ◽  
Author(s):  
Vinod Kumar ◽  
Amruta Vasudevan ◽  
Keertana Venkatesh ◽  
Reshma Maiya ◽  
Parul Sood ◽  
...  
Keyword(s):  
Axonal Transport ◽  
Molecular Motors ◽  
Experimental System ◽  
C Elegans ◽  
On Demand ◽  
Motion State ◽  
State Switching ◽  
Cargo Transport ◽  
Functional Relevance

AbstractMolecular motors drive the directed transport of presynaptic vesicles along the narrow axons of nerve cells. Stationary clusters of such vesicles are a prominent feature of axonal transport, but little is known about their physiological and functional relevance. Here, we develop a simulation model describing key features of axonal cargo transport with a view to addressing this question, benchmarking the model against our experiments in the touch neurons of C. elegans. Our simulations provide for multiple microtubule tracks and varied cargo motion states while also incorporating cargo-cargo interactions. Our model also incorporates obstacles to vesicle transport in the form of microtubule ends, stalled vesicles, and stationary mitochondria. We devise computational methodologies to simulate both axonal bleaching and axotomy, showing that our results reproduce the properties of both moving as well as stationary cargo in vivo. Increasing vesicle numbers leads to larger and more long-lived stationary clusters of vesicular cargo. Vesicle clusters are dynamically stable, explaining why they are ubiquitously seen. Modulating the rates of cargo motion-state switching allows cluster lifetimes and flux to be tuned both in simulations and experiments. We demonstrate, both in simulations and in an experimental system, that suppressing reversals leads to larger stationary vesicle clusters being formed while also reducing flux. Our simulation results support the view that the physiological significance of clusters is located in their role as dynamic reservoirs of cargo vesicles, capable of being released or sequestered on demand.

scholarly journals Brain dynein (MAP1C) localizes on both anterogradely and retrogradely transported membranous organelles in vivo.

1990 ◽  
Vol 111 (3) ◽  
pp. 1027-1037 ◽  
Author(s):  
N Hirokawa ◽  
R Sato-Yoshitake ◽  
T Yoshida ◽  
T Kawashima
Keyword(s):  
Nerve Terminal ◽  
Peripheral Nerves ◽  
Molecular Motor ◽  
Retrograde Transport ◽  
Nerve Terminals ◽  
Electron Microscopic ◽  
Electron Microscopic Immunocytochemistry ◽  
Fast Flow

Brain dynein is a microtubule-activated ATPase considered to be a candidate to function as a molecular motor to transport membranous organelles retrogradely in the axon. To determine whether brain dynein really binds to retrogradely transported organelles in vivo and how it is transported to the nerve terminals, we studied the localization of brain dynein in axons after the ligation of peripheral nerves by light and electron microscopic immunocytochemistry using affinity-purified anti-brain dynein antibodies. Different classes of organelles preferentially accumulated at the regions proximal and distal to the ligated part. Interestingly, brain dynein accumulated both at the regions proximal and distal to the ligation sites and localized not only on retrogradely transported membranous organelles but also on anterogradely transported ones. This is the first evidence to show that brain dynein associates with retrogradely transported organelles in vivo and that brain dynein is transported to the nerve terminal by fast flow. This also suggests that there may be some mechanism that activates brain dynein only for retrograde transport.

Theoretical Prediction of Run Speed Distribution for a Molecular Motor

2008 ◽  
Author(s):  
Sam Walcott ◽  
Neil M. Kad
Keyword(s):  
Molecular Motors ◽  
Molecular Motor ◽  
Realistic Model ◽  
Variable Step Size ◽  
Step Size ◽  
Run Length ◽  
Speed Distribution ◽  
Myosin Va ◽  
Cell Transforming ◽  
Run Speed

Processive molecular motors are large proteins that “walk” along filaments in a cell, transforming chemical energy into mechanical work. These microscopic motors behave, at least qualitatively, like macroscopic walking machines. The dynamics and stability of macroscopic walkers are understood by analysis of “stride functions” (Poincare´ maps from one step to the next). We show that molecular motors have linear, probabilistic stride functions. Using these functions, we derive expressions for three measurable distributions: step period, run length and average run speed. The former two distributions are well known, the latter is new. We validate our calculation with simulations of a realistic model for Myosin Va (a molecular motor). The parameters of the run speed distribution specify both the run-length and step period distributions. As step-period distributions are difficult to measure under physiologically relevant conditions, this technique provides new information. Finally, we discuss the effects of variable step size and experimental error.

scholarly journals Autoinhibition of kinesin-1 is essential to the dendrite-specific localization of Golgi outposts

2018 ◽  
Author(s):  
Michael T. Kelliher ◽  
Yang Yue ◽  
Ashley Ng ◽  
Daichi Kamiyama ◽  
Bo Huang ◽  
...  
Keyword(s):  
Structure Function ◽  
Molecular Motors ◽  
Genetic Interaction ◽  
Function Analysis ◽  
Molecular Motor ◽  
Neuronal Polarity ◽  
Growth Defects ◽  
Specific Localization ◽  
Selective Localization

AbstractNeuronal polarity relies on the selective localization of cargo to axons or dendrites. The molecular motor kinesin-1 moves cargo into axons but is also active in dendrites. This raises the question of how kinesin-1 activity is regulated to maintain the compartment-specific localization of cargo. Our in vivo structure-function analysis of endogenous Drosophila kinesin-1 reveals a novel role for autoinhibition in enabling the dendrite-specific localization of Golgi outposts. Mutations that disrupt kinesin-1 autoinhibition result in the axonal mislocalization of Golgi outposts. Autoinhibition also regulates kinesin-1 localization. Uninhibited kinesin-1 accumulates in axons and is depleted from dendrites, correlating with the change in outpost distribution and dendrite growth defects. Genetic interaction tests show that a balance of kinesin-1 inhibition and dynein activity is necessary to localize Golgi outposts to dendrites and keep them from entering axons. Our data indicate that kinesin-1 activity is precisely regulated by autoinhibition to achieve the selective localization of dendritic cargo.SummaryNeuronal polarity relies on the axon-or dendrite-specific localization of cargo by molecular motors such as kinesin-1. These studies show autoinhibition regulates both kinesin-1 activity and localization to keep dendritic cargo from entering axons.

scholarly journals Neuroglia infection by rabies virus after anterograde virus spread in peripheral neurons

2020 ◽  
Vol 8 (1) ◽  
Author(s):  
Madlin Potratz ◽  
Luca M. Zaeck ◽  
Carlotta Weigel ◽  
Antonia Klein ◽  
Conrad M. Freuling ◽  
...  
Keyword(s):  
Immune Responses ◽  
Axonal Transport ◽  
Rabies Virus ◽  
Peripheral Nerves ◽  
Innate Immune ◽  
Laser Scanning Microscopy ◽  
Innate Immune Responses ◽  
Virus Spread ◽  
Peripheral Neurons

AbstractThe highly neurotropic rabies virus (RABV) enters peripheral neurons at axon termini and requires long distance axonal transport and trans-synaptic spread between neurons for the infection of the central nervous system (CNS). Recent 3D imaging of field RABV-infected brains revealed a remarkably high proportion of infected astroglia, indicating that highly virulent field viruses are able to suppress astrocyte-mediated innate immune responses and virus elimination pathways. While fundamental for CNS invasion, in vivo field RABV spread and tropism in peripheral tissues is understudied. Here, we used three-dimensional light sheet and confocal laser scanning microscopy to investigate the in vivo distribution patterns of a field RABV clone in cleared high-volume tissue samples after infection via a natural (intramuscular; hind leg) and an artificial (intracranial) inoculation route. Immunostaining of virus and host markers provided a comprehensive overview of RABV infection in the CNS and peripheral nerves after centripetal and centrifugal virus spread. Importantly, we identified non-neuronal, axon-ensheathing neuroglia (Schwann cells, SCs) in peripheral nerves of the hind leg and facial regions as a target cell population of field RABV. This suggests that virus release from axons and infected SCs is part of the RABV in vivo cycle and may affect RABV-related demyelination of peripheral neurons and local innate immune responses. Detection of RABV in axon-surrounding myelinating SCs after i.c. infection further provided evidence for anterograde spread of RABV, highlighting that RABV axonal transport and spread of infectious virus in peripheral nerves is not exclusively retrograde. Our data support a new model in which, comparable to CNS neuroglia, SC infection in peripheral nerves suppresses glia-mediated innate immunity and delays antiviral host responses required for successful transport from the peripheral infection sites to the brain.

scholarly journals Neuroglia Infection by Rabies Virus after Anterograde Virus Spread in Peripheral Neurons

2020 ◽  
Author(s):  
Madlin Potratz ◽  
Luca M. Zaeck ◽  
Carlotta Weigel ◽  
Antonia Klein ◽  
Conrad M. Freuling ◽  
...  
Keyword(s):  
Immune Responses ◽  
Axonal Transport ◽  
Rabies Virus ◽  
Peripheral Nerves ◽  
Innate Immune ◽  
Laser Scanning Microscopy ◽  
Innate Immune Responses ◽  
Virus Spread ◽  
Peripheral Neurons

AbstractThe highly neurotropic rabies virus (RABV) enters peripheral neurons at axon termini and requires long distance axonal transport and trans-synaptic spread between neurons for the infection of the central nervous system (CNS). Whereas laboratory strains are almost exclusively detected in neurons, recent 3D imaging of field RABV-infected brains revealed an remarkably high proportion of infected astroglia, indicating that in contrast to attenuated lab strains highly virulent field viruses are able to suppress astrocyte mediated innate immune responses and virus elimination pathways. While fundamental for CNS invasion, in vivo field RABV spread and tropism in peripheral tissues is understudied. Here, we used three-dimensional light sheet and confocal laser scanning microscopy to investigate the in vivo distribution patterns of a field RABV clone in cleared high-volume tissue samples after infection via a natural (intramuscular; hind leg) and an artificial (intracranial) inoculation route. Immunostaining of virus and host markers provided a comprehensive overview of RABV infection in the CNS and peripheral nerves after centripetal and centrifugal virus spread. Importantly, we identified non-neuronal,axon-ensheathing neuroglia (Schwann cells, SCs) in peripheral nerves of the hind leg and facial regions as a target cell population of field RABV. This suggest that virus release from axons and infected SCs is part of the RABV in vivo cycle and may affect RABV-related demyelination of peripheral neurons and local innate immune responses. Detection of RABV in axon surrounding myelinating SCs after i.c. infection further provided evidence for anterograde spread of RABV, highlighting that RABV axonal transport and spread of infectious virus in peripheral nerves is not exclusively retrograde. Our data support a new model in which, comparable to CNS neuroglia, SC infection in peripheral nerves suppresses glia-mediated innate immunity and delays antiviral host responses required for successful transport from the peripheral infection sites to the brain.

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