scholarly journals Dynein activator Hook1 is required for trafficking of BDNF-signaling endosomes in neurons

2018 ◽  
Vol 218 (1) ◽  
pp. 220-233 ◽  
Author(s):  
Mara A. Olenick ◽  
Roberto Dominguez ◽  
Erika L.F. Holzbaur
Keyword(s):  
Axonal Transport ◽  
Hippocampal Neurons ◽  
Neuronal Development ◽  
Molecular Motor ◽  
Cytoplasmic Dynein ◽  
Differential Association ◽  
Primary Hippocampal Neurons ◽  
Bdnf Signaling ◽  
Microfluidic Chambers

Axonal transport is required for neuronal development and survival. Transport from the axon to the soma is driven by the molecular motor cytoplasmic dynein, yet it remains unclear how dynein is spatially and temporally regulated. We find that the dynein effector Hook1 mediates transport of TrkB–BDNF-signaling endosomes in primary hippocampal neurons. Hook1 comigrates with a subpopulation of Rab5 endosomes positive for TrkB and BDNF, which exhibit processive retrograde motility with faster velocities than the overall Rab5 population. Knockdown of Hook1 significantly reduced the motility of BDNF-signaling endosomes without affecting the motility of other organelles. In microfluidic chambers, Hook1 depletion resulted in a significant decrease in the flux and processivity of BDNF-Qdots along the mid-axon, an effect specific for Hook1 but not Hook3. Hook1 depletion inhibited BDNF trafficking to the soma and blocked downstream BDNF- and TrkB-dependent signaling to the nucleus. Together, these studies support a model in which differential association with cargo-specific effectors efficiently regulates dynein in neurons.

scholarly journals Dynein efficiently navigates the dendritic cytoskeleton to drive the retrograde trafficking of BDNF/TrkB signaling endosomes

2017 ◽  
Vol 28 (19) ◽  
pp. 2543-2554 ◽  
Author(s):  
Swathi Ayloo ◽  
Pedro Guedes-Dias ◽  
Amy E. Ghiretti ◽  
Erika L. F. Holzbaur
Keyword(s):  
Real Time ◽  
Axonal Transport ◽  
Hippocampal Neurons ◽  
Cytoplasmic Dynein ◽  
Microtubule Dynamics ◽  
Neuronal Function ◽  
Retrograde Trafficking ◽  
Basic Understanding ◽  
Dependent Transport ◽  
Dynamic Microtubule

The efficient transport of cargoes within axons and dendrites is critical for neuronal function. Although we have a basic understanding of axonal transport, much less is known about transport in dendrites. We used an optogenetic approach to recruit motor proteins to cargo in real time within axons or dendrites in hippocampal neurons. Kinesin-1, a robust axonal motor, moves cargo less efficiently in dendrites. In contrast, cytoplasmic dynein efficiently navigates both axons and dendrites; in both compartments, dynamic microtubule plus ends enhance dynein-dependent transport. To test the predictions of the optogenetic assay, we examined the contribution of dynein to the motility of an endogenous dendritic cargo and found that dynein inhibition eliminates the retrograde bias of BDNF/TrkB trafficking. However, inhibition of microtubule dynamics has no effect on BDNF/TrkB motility, suggesting that dendritic kinesin motors may cooperate with dynein to drive the transport of signaling endosomes into the soma. Collectively our data highlight compartment-specific differences in kinesin activity that likely reflect specialized tuning for localized cytoskeletal determinants, whereas dynein activity is less compartment specific but is more responsive to changes in microtubule dynamics.

scholarly journals Roles of Lycopene During Neurite Development of Primary Hippocampal Neurons

2021 ◽  
Vol 5 (Supplement_2) ◽  
pp. 923-923
Author(s):  
Madison Scott ◽  
Joseph Jansen ◽  
Mary Margaret Hayden ◽  
Katheryn Broman ◽  
Han-A Park
Keyword(s):  
Hippocampal Neurons ◽  
Developmental Disorders ◽  
Neuronal Development ◽  
Stage Iv ◽  
Neurite Growth ◽  
Redox Balance ◽  
University Of Alabama ◽  
Mitochondrial Superoxide ◽  
Primary Hippocampal Neurons ◽  
Neurite Development

Abstract Objectives Neurite outgrowth and branching is critical during neuronal development. Failure to achieve proper neurite complexity is highly associated with developmental disorders. We have previously shown that oxidative stress contributes to alteration of neurite morphology. Therefore, nutrients capable of regulating neuronal redox balance may help maintain normal neurite development. We have recently found that lycopene, a nutritional antioxidant, protects mitochondria by preventing generation of mitochondrial superoxide. In this study, we hypothesize that treatment with lycopene improves development of primary hippocampal neurons under physiological conditions. Methods Primary hippocampal neurons were grown in neurobasal media with or without 0.1 μM lycopene for 1, 3, 5, 7, and 14 days, and imaged using a Zeiss Axio Vert.A1 microscope. Neurons were classified into 4 different categories: stage I, circular and non-polar neurons; stage II, presence of minor neurites without polarity; stage III, neurons with polarity; and stage IV, neurons with dendritic branches. We also performed Sholl analysis to quantify intersections of neurites evaluating neurite complexity. Results Primary hippocampal neurons grown in media containing lycopene showed advanced neuronal development. In particular, lycopene treatment significantly advanced polarity and dendritic arborization in immature neurons. In addition, neurons grown in lycopene showed improved neurite branching at their maturity. Conclusions This study shows that lycopene supports neurite growth and branching during development. Therefore, we suggest novel role of lycopene during physiological development of the brain, or potential therapeutic effect against developmental disorders. Funding Sources RGC Program (University of Alabama) Crenshaw Research Fund (University of Alabama).

scholarly journals Tissue-based metabolic labeling of polysialic acids in living primary hippocampal neurons

2015 ◽  
Vol 112 (3) ◽  
pp. E241-E248 ◽  
Author(s):  
Kyungtae Kang ◽  
Sunghoon Joo ◽  
Ji Yu Choi ◽  
Sujeong Geum ◽  
Seok-Pyo Hong ◽  
...  
Keyword(s):  
Sialic Acid ◽  
Hippocampal Neurons ◽  
Neuronal Development ◽  
Polysialic Acid ◽  
Metabolic Labeling ◽  
Primary Hippocampal Neurons ◽  
Polysialic Acids ◽  
Conventional Cell ◽  
Vitro Analysis

The posttranslational modification of neural cell-adhesion molecule (NCAM) with polysialic acid (PSA) and the spatiotemporal distribution of PSA-NCAM play an important role in the neuronal development. In this work, we developed a tissue-based strategy for metabolically incorporating an unnatural monosaccharide, peracetylated N-azidoacetyl-d-mannosamine, in the sialic acid biochemical pathway to present N-azidoacetyl sialic acid to PSA-NCAM. Although significant neurotoxicity was observed in the conventional metabolic labeling that used the dissociated neuron cells, neurotoxicity disappeared in this modified strategy, allowing for investigation of the temporal and spatial distributions of PSA in the primary hippocampal neurons. PSA-NCAM was synthesized and recycled continuously during neuronal development, and the two-color labeling showed that newly synthesized PSA-NCAMs were transported and inserted mainly to the growing neurites and not significantly to the cell body. This report suggests a reliable and cytocompatible method for in vitro analysis of glycans complementary to the conventional cell-based metabolic labeling for chemical glycobiology.

scholarly journals Role of Dynein and Dynactin (DCTN-1) in Neurodegenerative Diseases

2019 ◽  
pp. 53-58
Author(s):  
Rajib Dutta ◽  
Swatilekha Roy Sarkar
Keyword(s):  
Neurodegenerative Diseases ◽  
Axonal Transport ◽  
Molecular Motor ◽  
Neuronal Cell ◽  
Cytoplasmic Dynein ◽  
Retrograde Axonal Transport ◽  
Sporadic Amyotrophic Lateral Sclerosis ◽  
Motor Neuropathy ◽  
Excitatory Neurotransmitter ◽  
Charcot Marie Tooth Disease

The pathophysiology and concept of degeneration in central nervous system is very complex and overwhelming at times. There is a complex mechanism which exists among different molecules in the cytoplasm of cell bodies of neurons, antegrade and retrograde axonal transport of cargoes and accumulation of certain substances and proteins which can influence the excitatory neurotransmitter like glutamate initiating the process of neurodegeneration. Neurons have extensive processes and communication between those processes and the cell body is crucial to neuronal function, viability and survival over time with progression of age. Researchers believe neurons are uniquely dependent on microtubule-based cargo transport. There is enough evidence to support that deficits in retrograde axonal transport contribute to pathogenesis in multiple neurodegenerative diseases. Cytoplasmic dynein and its regulation by Dynactin (DCTN1) is the major molecular motor cargo involved in autophagy, mitosis and neuronal cell survival. Mutation in dynactin gene located in 2p13.1,is indeed studied very extensively and is considered to be involved directly or indirectly to various conditions like Perry syndrome, familial and sporadic Amyotrophic lateral sclerosis, Hereditary spastic paraplegia, Spinocerebellar Ataxia (SCA-5), Huntingtons disease, Alzheimers disease, Charcot marie tooth disease, Hereditary motor neuropathy 7B, prion disease, parkinsons disease, malformation of cortical development, polymicrogyria to name a few with exception of Multiple Sclerosis (MS).

Primary hippocampal neurons as suitable model to evaluate the influence of neurotoxicants on neuronal development

2017 ◽  
Vol 280 ◽  
pp. S278
Author(s):  
Natalia Marchetti ◽  
Laura Gerosa ◽  
Mariaserena Boraso ◽  
Barbara Viviani
Keyword(s):  
Hippocampal Neurons ◽  
Neuronal Development ◽  
Suitable Model ◽  
Primary Hippocampal Neurons

scholarly journals Dexmedetomidine attenuates the neurotoxicity of propofol toward primary hippocampal neurons in vitro via Erk1/2/CREB/BDNF signaling pathways

2019 ◽  
Vol Volume 13 ◽  
pp. 695-706 ◽  
Author(s):  
Youbing Tu ◽  
Yubing Liang ◽  
Yong Xiao ◽  
Jing Lv ◽  
Ruicong Guan ◽  
...  
Keyword(s):  
Signaling Pathways ◽  
Hippocampal Neurons ◽  
Primary Hippocampal Neurons ◽  
Bdnf Signaling

scholarly journals Molecular motor function in axonal transport in vivo probed by genetic and computational analysis in Drosophila

2012 ◽  
Vol 23 (9) ◽  
pp. 1700-1714 ◽  
Author(s):  
Gerald F. Reis ◽  
Ge Yang ◽  
Lukasz Szpankowski ◽  
Carole Weaver ◽  
Sameer B. Shah ◽  
...  
Keyword(s):  
Axonal Transport ◽  
Imaging System ◽  
Molecular Motor ◽  
Cytoplasmic Dynein ◽  
Movement Velocity ◽  
Neuronal Viability ◽  
And Function ◽  
Vesicle Movement

Bidirectional axonal transport driven by kinesin and dynein along microtubules is critical to neuronal viability and function. To evaluate axonal transport mechanisms, we developed a high-resolution imaging system to track the movement of amyloid precursor protein (APP) vesicles in Drosophila segmental nerve axons. Computational analyses of a large number of moving vesicles in defined genetic backgrounds with partial reduction or overexpression of motor proteins enabled us to test with high precision existing and new models of motor activity and coordination in vivo. We discovered several previously unknown features of vesicle movement, including a surprising dependence of anterograde APP vesicle movement velocity on the amount of kinesin-1. This finding is largely incompatible with the biophysical properties of kinesin-1 derived from in vitro analyses. Our data also suggest kinesin-1 and cytoplasmic dynein motors assemble in stable mixtures on APP vesicles and their direction and velocity are controlled at least in part by dynein intermediate chain.

ATRA-induced increase in intracellular Ca2+concentration in primary hippocampal neurons

2010 ◽  
Vol 34 (8) ◽  
pp. S74-S74
Author(s):  
Tingyu Li ◽  
Xiaojuan Zhang ◽  
Xuan Zhang ◽  
Jian Hea ◽  
Yang Bi Youxue Liu ◽  
...  
Keyword(s):  
Hippocampal Neurons ◽  
Primary Hippocampal Neurons

Spastin Interacts with CRMP2 to Regulate Neurite Outgrowth by Controlling Microtubule Dynamics through Phosphorylation Modifications

2020 ◽  
Vol 19 ◽  
Author(s):  
Sumei Li ◽  
Jifeng Zhang ◽  
Jiaqi Zhang ◽  
Jiong Li ◽  
Longfei Cheng ◽  
...  
Keyword(s):  
Neurite Outgrowth ◽  
Hippocampal Neurons ◽  
Neuronal Development ◽  
Phosphorylation Site ◽  
Microtubule Dynamics ◽  
Microtubule Assembly ◽  
C Terminus ◽  
Mediator Protein ◽  
Branch Formation

Aims: Our work aims to revealing the underlying microtubule mechanism of neurites outgrowth during neuronal development, and also proposes a feasible intervention pathway for reconstructing neural network connections after nerve injury. Background: Microtubule polymerization and severing are the basis for the neurite outgrowth and branch formation. Collapsin response mediator protein 2 (CRMP2) regulates axonal growth and branching as a binding partner of the tubulin heterodimer to promote microtubule assembly. And spastin participates in the growth and regeneration of neurites by severing microtubules into small segments. However, how CRMP2 and spastin cooperate to regulate neurite outgrowth by controlling the microtubule dynamics needs to be elucidated. Objective: To explore whether neurite outgrowth was mediated by coordination of CRMP2 and spastin. Method: Hippocampal neurons were cultured in vitro in 24-well culture plates for 4 days before being used to perform the transfection. Calcium phosphate was used to transfect the CRMP2 and spastin constructs and their control into the neurons. An interaction between CRMP2 and spastin was examined by using pull down, CoIP and immunofluorescence colocalization assays. And immunostaining was also performed to determine the morphology of neurites. Result: We first demonstrated that CRMP2 interacted with spastin to promote the neurite outgrowth and branch formation. Furthermore, our results identified that phosphorylation modification failed to alter the binding affinities of CRMP2 for spastin, but inhibited their binding to microtubules. CRMP2 interacted with the MTBD domain of spastin via its C-terminus, and blocking the binding sites of them inhibited the outgrowth and branch formation of neurites. In addition, we confirmed one phosphorylation site S210 at spastin in hippocampal neurons and phosphorylation spastin at site S210 promoted the neurite outgrowth but not branch formation by remodeling microtubules. Conclusion: Taken together, our data demonstrated that the interaction of CRMP2 and spastin is required for neurite outgrowth and branch formation and their interaction is not regulated by their phosphorylation.

Schisandrin Restores the Amyloid β-Induced Impairments on Mitochondrial Function, Energy Metabolism, Biogenesis, and Dynamics in Rat Primary Hippocampal Neurons

Pharmacology ◽  
2021 ◽  
pp. 1-11
Author(s):  
Zhongyuan Piao ◽  
Lin Song ◽  
Lifen Yao ◽  
Limei Zhang ◽  
Yichan Lu
Keyword(s):  
Energy Metabolism ◽  
Cytochrome C ◽  
Mitochondrial Biogenesis ◽  
Mitochondrial Function ◽  
Hippocampal Neurons ◽  
Citrate Synthase ◽  
Mitochondrial Permeability Transition ◽  
Amyloid Β ◽  
Mitochondrial Functions ◽  
Primary Hippocampal Neurons

Introduction: Schisandrin which is derived from Schisandra chinensis has shown multiple pharmacological effects on various diseases including Alzheimer’s disease (AD). It is demonstrated that mitochondrial dysfunction plays an essential role in the pathogenesis of neurodegenerative disorders. Objective: Our study aims to investigate the effects of schisandrin on mitochondrial functions and metabolisms in primary hippocampal neurons. Methods: In our study, rat primary hippocampal neurons were isolated and treated with indicated dose of amyloid β1–42 (Aβ1–42) oligomer to establish a cell model of AD in vitro. Schisandrin (2 μg/mL) was further subjected to test its effects on mitochondrial function, energy metabolism, mitochondrial biogenesis, and dynamics in the Aβ1–42 oligomer-treated neurons. Results and Conclusions: Our findings indicated that schisandrin significantly alleviated the Aβ1–42 oligomer-induced loss of mitochondrial membrane potential and impaired cytochrome c oxidase activity. Additionally, the opening of mitochondrial permeability transition pore and release of cytochrome c were highly restricted with schisandrin treatment. Alterations in cell viability, ATP production, citrate synthase activity, and the expressions of glycolysis-related enzymes demonstrated the relief of defective energy metabolism in Aβ-treated neurons after the treatment of schisandrin. For mitochondrial biogenesis, elevated expression of peroxisome proliferator-activated receptor γ coactivator along with promoted mitochondrial mass was found in schisandrin-treated cells. The imbalance in the cycle of fusion and fission was also remarkably restored by schisandrin. In summary, this study provides novel mechanisms for the protective effect of schisandrin on mitochondria-related functions.

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