scholarly journals Ser/Thr kinase Trc controls neurite outgrowth in Drosophila by modulating microtubule-microtubule sliding

2019 ◽  
Author(s):  
Rosalind Norkett ◽  
Urko del Castillo ◽  
Wen Lu ◽  
Vladimir I. Gelfand
Keyword(s):  
Neurite Outgrowth ◽  
Neuronal Development ◽  
Live Imaging ◽  
Biochemical Methods ◽  
Interphase Cells ◽  
Ndr Kinase

AbstractCorrect neuronal development requires tailored neurite outgrowth. Neurite outgrowth is driven by microtubule sliding – the transport of microtubules along each other. We have recently demonstrated that a “mitotic” kinesin-6 (Pavarotti in Drosophila) effectively inhibits microtubule-sliding and neurite outgrowth. However, mechanisms of Pavarotti regulation in interphase cells and specifically in neurite outgrowth are unknown. Here, we use a combination of live imaging and biochemical methods to show that the inhibition of microtubule sliding by Pavarotti is controlled by phosphorylation. We identify the Ser/Thr NDR kinase Tricornered (Trc) as a Pavarotti-dependent regulator of microtubule sliding in neurons. Further, we show that Trc-mediated phosphorylation of Pavarotti promotes its interaction with 14-3-3 proteins. 14-3-3 binding is necessary for Pavarotti to interact with microtubules and inhibit sliding. Thus, we propose a pathway by which microtubule sliding can be up or down regulated in neurons to control neurite outgrowth, and establish parallels between microtubule sliding in mitosis and post-mitotic neurons.

scholarly journals Ser/Thr kinase Trc controls neurite outgrowth in Drosophila by modulating microtubule-microtubule sliding

eLife ◽  
2020 ◽  
Vol 9 ◽  
Author(s):  
Rosalind Norkett ◽  
Urko del Castillo ◽  
Wen Lu ◽  
Vladimir I Gelfand
Keyword(s):  
Neurite Outgrowth ◽  
Neuronal Development ◽  
Live Imaging ◽  
Biochemical Methods ◽  
Interphase Cells ◽  
Ndr Kinase

Correct neuronal development requires tailored neurite outgrowth. Neurite outgrowth is driven in part by microtubule-sliding – the transport of microtubules along each other. We have recently demonstrated that a ‘mitotic’ kinesin-6 (Pavarotti in Drosophila) effectively inhibits microtubule-sliding and neurite outgrowth. However, mechanisms regulating Pavarotti itself in interphase cells and specifically in neurite outgrowth are unknown. Here, we use a combination of live imaging and biochemical methods to show that the inhibition of microtubule-sliding by Pavarotti is controlled by phosphorylation. We identify the Ser/Thr NDR kinase Tricornered (Trc) as a Pavarotti-dependent regulator of microtubule sliding in neurons. Further, we show that Trc-mediated phosphorylation of Pavarotti promotes its interaction with 14-3-3 proteins. Loss of 14-3-3 prevents Pavarotti from associating with microtubules. Thus, we propose a pathway by which microtubule-sliding can be up- or downregulated in neurons to control neurite outgrowth, and establish parallels between microtubule-sliding in mitosis and post-mitotic 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.

scholarly journals Golgi-resident TRIO regulates membrane trafficking during neurite outgrowth

2019 ◽  
Vol 294 (28) ◽  
pp. 10954-10968 ◽  
Author(s):  
Tao Tao ◽  
Jie Sun ◽  
Yajing Peng ◽  
Yeqiong Li ◽  
Pei Wang ◽  
...  
Keyword(s):  
Neurite Outgrowth ◽  
Membrane Trafficking ◽  
Rho Gtpases ◽  
Neuronal Development ◽  
Membrane Vesicles ◽  
Cerebellar Granule Neurons ◽  
Granule Neurons ◽  
Nucleotide Exchange ◽  
Cerebellar Granule ◽  
Cytoskeletal Dynamics

Neurite outgrowth requires coordinated cytoskeletal rearrangements in the growth cone and directional membrane delivery from the neuronal soma. As an essential Rho guanine nucleotide exchange factor (GEF), TRIO is necessary for cytoskeletal dynamics during neurite outgrowth, but its participation in the membrane delivery is unclear. Using co-localization studies, live-cell imaging, and fluorescence recovery after photobleaching analysis, along with neurite outgrowth assay and various biochemical approaches, we here report that in mouse cerebellar granule neurons, TRIO protein pools at the Golgi and regulates membrane trafficking by controlling the directional maintenance of both RAB8 (member RAS oncogene family 8)– and RAB10-positive membrane vesicles. We found that the spectrin repeats in Golgi-resident TRIO confer RAB8 and RAB10 activation by interacting with and activating the RAB GEF RABIN8. Constitutively active RAB8 or RAB10 could partially restore the neurite outgrowth of TRIO-deficient cerebellar granule neurons, suggesting that TRIO-regulated membrane trafficking has an important functional role in neurite outgrowth. Our results also suggest cross-talk between Rho GEF and Rab GEF in controlling both cytoskeletal dynamics and membrane trafficking during neuronal development. They further highlight how protein pools localized to specific organelles regulate crucial cellular activities and functions. In conclusion, our findings indicate that TRIO regulates membrane trafficking during neurite outgrowth in coordination with its GEF-dependent function in controlling cytoskeletal dynamics via Rho GTPases.

scholarly journals c-Jun Amino-Terminal Kinase is Involved in Valproic Acid-Mediated Neuronal Differentiation of Mouse Embryonic NSCs and Neurite Outgrowth of NSC-Derived Neurons

2017 ◽  
Vol 42 (4) ◽  
pp. 1254-1266 ◽  
Author(s):  
Lu Lu ◽  
Hengxing Zhou ◽  
Bin Pan ◽  
Xueying Li ◽  
Zheng Fu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Central Nervous System ◽  
Valproic Acid ◽  
Nervous System ◽  
Neurite Outgrowth ◽  
Neuronal Differentiation ◽  
Neuronal Development ◽  
Neuroprotective Effect ◽  
Amino Terminal ◽  
Jnk Activation

Abstract Valproic acid (VPA), an anticonvulsant and mood-stabilizing drug, can induce neuronal differentiation, promote neurite extension and exert a neuroprotective effect in central nervous system (CNS) injuries; however, comparatively little is known regarding its action on mouse embryonic neural stem cells (NSCs) and the underlying molecular mechanism. Recent studies suggested that c-Jun N-terminal kinase (JNK) is required for neurite outgrowth and neuronal differentiation during neuronal development. In the present study, we cultured mouse embryonic NSCs and treated the cells with 1 mM VPA for up to 7 days. The results indicate that VPA promotes the neuronal differentiation of mouse embryonic NSCs and neurite outgrowth of NSC-derived neurons; moreover, VPA induces the phosphorylation of c-Jun by JNK. In contrast, the specific JNK inhibitor SP600125 decreased the VPA-stimulated increase in neuronal differentiation of mouse embryonic NSCs and neurite outgrowth of NSC-derived neurons. Taken together, these results suggest that VPA promotes neuronal differentiation of mouse embryonic NSCs and neurite outgrowth of NSC-derived neurons. Moreover, JNK activation is involved in the effects of VPA stimulation.

scholarly journals Opposing functions of F-BAR proteins in neuronal membrane protrusion, tubule formation, and neurite outgrowth

2019 ◽  
Vol 2 (3) ◽  
pp. e201800288 ◽  
Author(s):  
Kendra L Taylor ◽  
Russell J Taylor ◽  
Karl E Richters ◽  
Brandon Huynh ◽  
Justin Carrington ◽  
...  
Keyword(s):  
Neurite Outgrowth ◽  
Cortical Neurons ◽  
Neuronal Development ◽  
Actin Dynamics ◽  
Neuronal Membrane ◽  
Membrane Protrusion ◽  
Tubule Formation ◽  
Primary Cortical Neurons ◽  
Splice Isoforms ◽  
Neurite Formation

The F-BAR family of proteins play important roles in many cellular processes by regulating both membrane and actin dynamics. The CIP4 family of F-BAR proteins is widely recognized to function in endocytosis by elongating endocytosing vesicles. However, in primary cortical neurons, CIP4 concentrates at the tips of extending lamellipodia and filopodia and inhibits neurite outgrowth. Here, we report that the highly homologous CIP4 family member, FBP17, induces tubular structures in primary cortical neurons and results in precocious neurite formation. Through domain swapping and deletion experiments, we demonstrate that a novel polybasic region between the F-BAR and HR1 domains is required for membrane bending. Moreover, the presence of a poly-PxxP region in longer splice isoforms of CIP4 and FBP17 largely reverses the localization and function of these proteins. Thus, CIP4 and FBP17 function as an antagonistic pair to fine-tune membrane protrusion, endocytosis, and neurite formation during early neuronal development.

scholarly journals Versican V1 Isoform Induces Neuronal Differentiation and Promotes Neurite Outgrowth

2004 ◽  
Vol 15 (5) ◽  
pp. 2093-2104 ◽  
Author(s):  
Yaojiong Wu ◽  
Wang Sheng ◽  
Liwen Chen ◽  
Haiheng Dong ◽  
Vivian Lee ◽  
...  
Keyword(s):  
Epidermal Growth Factor Receptor ◽  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Neurite Outgrowth ◽  
Neuronal Differentiation ◽  
Hippocampal Neurons ◽  
Neuronal Development ◽  
Growth Factor Receptor ◽  
Adult Brain ◽  
Epidermal Growth

The chondroitin sulfate proteoglycan versican is one of the major extracellular components in the developing and adult brain. Here, we show that isoforms of versican play different roles in neuronal differentiation and neurite outgrowth. Expression of versican V1 isoform in PC12 cells induced complete differentiation, whereas expression of V2 induced an aborted differentiation accompanied by apoptosis. V1 promoted neurite outgrowth of hippocampal neurons, but V2 failed to do so. V1 transfection enhanced expression of epidermal growth factor receptor and integrins, and facilitated sustained extracellular signal-regulated kinase/MAPK phosphorylation. Blockade of the epidermal growth factor receptor, β1 integrin, or Src significantly inhibited neuronal differentiation. Finally, we demonstrated that versican V1 isoform also promoted differentiation of neural stem cells into neurons. Our results have implications for understanding how versican regulates neuronal development, function, and repair.

Adhesion Molecules and Axon Pathfinding

The Neuron ◽  
2015 ◽  
pp. 387-414
Author(s):  
Irwin B. Levitan ◽  
Leonard K. Kaczmarek
Keyword(s):  
Neurite Outgrowth ◽  
Adhesion Molecules ◽  
Growth Cone ◽  
Neuronal Development ◽  
Axon Pathfinding ◽  
Nervous Systems ◽  
Development And Differentiation ◽  
Adult Neuron ◽  
Developing Neurons ◽  
Extracellular Environment

Developing neurons extend neurites, which become the axons and dendrites of the adult neuron. These neurites follow specific paths and branch in characteristic ways. The leading tip of the neurite, the growth cone, appears to sample the extracellular environment and contribute to decisions about the direction of extension. Various molecules are essential for appropriate pathfinding by growing neurites. For example, neurites grow selectively toward or away from guidance molecules such as semaphorins, netrins, slits, and ephrins. In addition, adhesion molecules such as fibronectin and laminin mediate specific adhesion of the neurite to the substrate over which it is growing, while CAMs and cadherins promote the adhesion of neurites of different cells to each other in specific patterns. Some molecules and mechanisms that regulate neuronal development and differentiation may also regulate neurite outgrowth in adult nervous systems, either during recovery from injury or in response to novel stimuli from the environment.

scholarly journals RalA, PLD and mTORC1 Are Required for Kinase-Independent Pathways in DGKβ-Induced Neurite Outgrowth

Biomolecules ◽  
2021 ◽  
Vol 11 (12) ◽  
pp. 1814
Author(s):  
Takuya Kano ◽  
Ryosuke Tsumagari ◽  
Akio Nakashima ◽  
Ushio Kikkawa ◽  
Shuji Ueda ◽  
...  
Keyword(s):  
Neurite Outgrowth ◽  
Neuronal Development ◽  
Mammalian Target Of Rapamycin ◽  
Small Gtpase ◽  
Wild Type ◽  
Common Pathway ◽  
Human Neuroblastoma ◽  
Brain Functions ◽  
Emotion And Memory ◽  
Higher Brain Functions

Diacylglycerol kinase β (DGKβ) is an enzyme that converts diacylglycerol to phosphatidic acid and is mainly expressed in the cerebral cortex, hippocampus and striatum. We previously reported that DGKβ induces neurite outgrowth and spinogenesis, contributing to higher brain functions, including emotion and memory. To elucidate the mechanisms involved in neuronal development by DGKβ, we investigated the importance of DGKβ activity in the induction of neurite outgrowth using human neuroblastoma SH-SY5Y cells. Interestingly, both wild-type DGKβ and the kinase-negative (KN) mutant partially induced neurite outgrowth, and these functions shared a common pathway via the activation of mammalian target of rapamycin complex 1 (mTORC1). In addition, we found that DGKβ interacted with the small GTPase RalA and that siRNA against RalA and phospholipase D (PLD) inhibitor treatments abolished DGKβKN-induced neurite outgrowth. These results indicate that binding of RalA and activation of PLD and mTORC1 are involved in DGKβKN-induced neurite outgrowth. Taken together with our previous reports, mTORC1 is a key molecule in both kinase-dependent and kinase-independent pathways of DGKβ-mediated neurite outgrowth, which is important for higher brain functions.

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