Polarization of hippocampal neurons with competitive surface stimuli: contact guidance cues are preferred over chemical ligands

Natalia Gomez; Shaochen Chen; Christine E Schmidt

doi:10.1098/rsif.2006.0171

Polarization of hippocampal neurons with competitive surface stimuli: contact guidance cues are preferred over chemical ligands

Journal of The Royal Society Interface ◽

10.1098/rsif.2006.0171 ◽

2006 ◽

Vol 4 (13) ◽

pp. 223-233 ◽

Cited By ~ 82

Author(s):

Natalia Gomez ◽

Shaochen Chen ◽

Christine E Schmidt

Keyword(s):

Hippocampal Neurons ◽

Artificial Substrates ◽

Contact Guidance ◽

Neural Engineering ◽

Environmental Signals ◽

Guidance Cues ◽

Unique System ◽

Extracellular Stimuli ◽

Regeneration Processes ◽

Microfabrication Techniques

Neuronal behaviour is profoundly influenced by extracellular stimuli in many developmental and regeneration processes. Understanding neuron responses and integration of environmental signals could impact the design of successful therapies for neurodegenerative diseases and nerve injuries. Here, we have investigated the influence of localized extracellular cues on polarization (i.e. axon formation) of hippocampal neurons. Electron-beam lithography, microfabrication techniques and protein immobilization were used to create a unique system that provided simultaneous and independent chemical and physical cues to individual neurons. In particular, we analysed competitive responses between simultaneous stimulation with chemical ligands, including immobilized nerve growth factor and laminin, and contact guidance cues mediated by surface topography (i.e. microchannels). Contact guidance cues were preferred 70% of the time over chemical ligands by neurons extending axons, which suggests a stronger stimulation mechanism triggered by topography. This investigation contributes to the understanding of neuronal behaviour on artificial substrates, which is applicable to the creation of artificial environments for neural engineering applications.

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Impaired Neurite Contact Guidance in Ubiquitin Ligase E3a (Ube3a)-Deficient Hippocampal Neurons on Nanostructured Substrates

Advanced Healthcare Materials ◽

10.1002/adhm.201500815 ◽

2016 ◽

Vol 5 (7) ◽

pp. 850-862 ◽

Cited By ~ 13

Author(s):

I. Tonazzini ◽

S. Meucci ◽

G. M. Van Woerden ◽

Y. Elgersma ◽

M. Cecchini

Keyword(s):

Ubiquitin Ligase ◽

Hippocampal Neurons ◽

Contact Guidance ◽

Nanostructured Substrates

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Anomalous ipsilateral optic fibre projection in Xenopus induced by larval tectal ablation

Development ◽

10.1242/dev.50.1.111 ◽

1979 ◽

Vol 50 (1) ◽

pp. 111-122

Author(s):

Charles Straznicky ◽

John Glastonbury

Keyword(s):

Contact Guidance ◽

Retinal Projection ◽

Guidance Cues ◽

Optic Fibre ◽

The Right

Unilateral left tectal ablation was carried out in Xenopus between stage 48 and 1 month after metamorphosis. Six to 12 weeks after metamorphosis the retinal projection from the right eye was examined with the use of [3H]proline autoradiography. The autoradiographs indicated that optic fibres whose tectal target was destroyed recrossed to the ipsilateral tectum and basal optic nucleus via the posterior and pretectal commissures. No anomalous recrossing occurred if the tectal ablation was carried out at stage 58 or later. The aberrant optic fibres were restricted to the rostrolateral and central parts of the ipsilateral tectum and they terminated in a discontinuous manner. It is concluded that available surfaces serving as contact guidance cues are needed to direct aberrant optic fibres to the ipsilateral tectum.

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Transfer of assembled collagen fibrils to flexible substrates for mechanically tunable contact guidance cues

Integrative Biology ◽

10.1039/c8ib00127h ◽

2018 ◽

Vol 10 (11) ◽

pp. 705-718 ◽

Cited By ~ 2

Author(s):

Juan Wang ◽

Joseph Koelbl ◽

Anuraag Boddupalli ◽

Zhiqi Yao ◽

Kaitlin M. Bratlie ◽

...

Keyword(s):

Wound Healing ◽

Cancer Invasion ◽

Collagen Fibrils ◽

Contact Guidance ◽

Flexible Substrates ◽

Guidance Cues ◽

Aligned Fibers

Contact guidance or bidirectional migration along aligned fibers modulates many physiological and pathological processes such as wound healing and cancer invasion.

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Contact guidance of CNS neurites on grooved quartz: influence of groove dimensions, neuronal age and cell type

Journal of Cell Science ◽

10.1242/jcs.110.23.2905 ◽

1997 ◽

Vol 110 (23) ◽

pp. 2905-2913 ◽

Cited By ~ 2

Author(s):

A. Rajnicek ◽

S. Britland ◽

C. McCaig

Keyword(s):

Hippocampal Neurons ◽

Growth Cones ◽

Companion Paper ◽

Contact Guidance ◽

Neuronal Polarity ◽

Cell Type ◽

Spinal Cord Neurons ◽

In Vitro System ◽

Guidance Information

We used an in vitro system that eliminates competing guidance cues found in embryos to determine whether substratum topography alone provides important neurite guidance information. Dissociated embryonic Xenopus spinal cord neurons and rat hippocampal neurons were grown on quartz etched with a series of parallel grooves. Xenopus neurites grew parallel to grooves as shallow as 14 nm and as narrow as 1 microm. Hippocampal neurites grew parallel to deep, wide grooves but perpendicular to shallow, narrow ones. Grooved substrata determined the sites at which neurites emerged from somas: Xenopus neurites sprouted from regions parallel to grooves but presumptive axons on rat hippocampal neurons emerged perpendicular to grooves and presumptive dendrites emerged parallel to them. Neurites grew faster in the favored direction of orientation and turned through large angles to align on grooves. The frequency of perpendicular alignment of hippocampal neurites depended on the age of the embryos from which neurons were isolated, suggesting that contact guidance is regulated in development. Collectively, the data indicate that substratum topography is a potent morphogenetic factor for developing CNS neurons and suggest that in addition to a role in pathfinding the geometry of the embryo assists in establishing neuronal polarity. In the companion paper (A. M. Rajnicek and C. D. McCaig (1997) J. Cell Sci. 110, 2915–2924) we explore the cellular mechanism for contact guidance of growth cones.

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Electric fields, contact guidance and the direction of nerve growth

Development ◽

10.1242/dev.94.1.245 ◽

1986 ◽

Vol 94 (1) ◽

pp. 245-255

Author(s):

Colin D. McCaig

Keyword(s):

Electric Field ◽

Electric Fields ◽

Applied Electric Field ◽

Free Choice ◽

Contact Guidance ◽

Orthogonal Direction ◽

Nerve Growth ◽

Guidance Cues

Nerve orientation in response to electrical guidance cues in one direction and contact guidance cues in an orthogonal direction has been studied. Where neurites had a free choice between following contact guidance cues or electrical cues, the direction of nerve growth was determined predominantly by the vector of the applied electric field.

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Noncanonical scaffolding of Gαi and β-arrestin by G protein-coupled receptors

10.1101/629576 ◽

2019 ◽

Author(s):

Jeffrey S. Smith ◽

Thomas F. Pack ◽

Asuka Inoue ◽

Claudia Lee ◽

Xinyu Xiong ◽

...

Keyword(s):

G Protein ◽

G Protein Coupled Receptors ◽

Environmental Signals ◽

Extracellular Signal Regulated Kinase ◽

Intracellular Signals ◽

Extracellular Signal ◽

Extracellular Stimuli ◽

Protein Isoform ◽

Sense And Respond ◽

G Protein Coupled

SummaryG-protein-coupled receptors (GPCRs) enable cells to sense and respond appropriately to hormonal and environmental signals, and are a target of ~30% of all FDA-approved medications. Canonically, each GPCR couples to distinct Gα proteins, such as Gαs, Gαi, Gαq or Gα12/13, as well as β-arrestins. These transducer proteins translate and integrate extracellular stimuli sensed by GPCRs into intracellular signals through what are broadly considered separable signalling pathways. However, the ability of Gα proteins to directly interact with β-arrestins to integrate signalling has not previously been appreciated. Here we show a novel interaction between Gαi protein family members and β-arrestin. Gαi:β-arrestin complexes were formed by all GPCRs tested, regardless of their canonical G protein isoform coupling, and could bind both GPCRs as well as the extracellular signal-regulated kinase (ERK). This novel paradigm of Gαi:β-arrestin scaffolds enhances our understanding of GPCR signalling.

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Requirement of Pax6 for the integration of guidance cues in cell migration

Royal Society Open Science ◽

10.1098/rsos.170625 ◽

2017 ◽

Vol 4 (10) ◽

pp. 170625 ◽

Cited By ~ 2

Author(s):

Miguel Arocena ◽

Ann M. Rajnicek ◽

Jon Martin Collinson

Keyword(s):

Cell Migration ◽

Electric Fields ◽

Neural Development ◽

Mouse Embryos ◽

Contact Guidance ◽

Wild Type ◽

General Direction ◽

Vast Array ◽

Guidance Cues ◽

Flat Substrate

The intricate patterns of cell migration that are found throughout development are generated through a vast array of guidance cues. Responding integratively to distinct, often conflicting, migratory signals is probably crucial for cells to reach their correct destination. Pax6 is a master transcription factor with key roles in neural development that include the control of cell migration. In this study, we have investigated the ability of cells derived from cortical neurospheres from wild-type (WT) and Pax6 −/− mouse embryos to integrate diverging guidance cues. We used two different cues, either separately or in combination: substratum nanogrooves to induce contact guidance, and electric fields (EFs) to induce electrotaxis. In the absence of an EF, both WT and Pax6 −/− cells aligned and migrated parallel to grooves, and on a flat substrate both showed marked electrotaxis towards the cathode. When an EF was applied in a perpendicular orientation to grooves, WT cells responded significantly to both cues, migrating in highly oblique trajectories in the general direction of the cathode. However, Pax6 −/− cells had an impaired response to both cues simultaneously. Our results demonstrate that these neurosphere derived cells have the capacity to integrate diverging guidance cues, which requires Pax6 function.

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The role of ubiquitin ligase E3A in polarized contact guidance and rescue strategies in UBE3A-deficient hippocampal neurons

Molecular Autism ◽

10.1186/s13229-019-0293-1 ◽

2019 ◽

Vol 10 (1) ◽

Cited By ~ 5

Author(s):

Ilaria Tonazzini ◽

Geeske M. Van Woerden ◽

Cecilia Masciullo ◽

Edwin J. Mientjes ◽

Ype Elgersma ◽

...

Keyword(s):

Focal Adhesion ◽

Hippocampal Neurons ◽

Treatment Strategies ◽

Contact Guidance ◽

Cell Contractility ◽

Actin Fiber ◽

Mediated Contact ◽

Focal Adhesion Pathway

Abstract Background Although neuronal extracellular sensing is emerging as crucial for brain wiring and therefore plasticity, little is known about these processes in neurodevelopmental disorders. Ubiquitin protein ligase E3A (UBE3A) plays a key role in neurodevelopment. Lack of UBE3A leads to Angelman syndrome (AS), while its increase is among the most prevalent genetic causes of autism (e.g., Dup15q syndrome). By using microstructured substrates that can induce specific directional stimuli in cells, we previously found deficient topographical contact guidance in AS neurons, which was linked to a dysregulated activation of the focal adhesion pathway. Methods Here, we study axon and dendrite contact guidance and neuronal morphological features of wild-type, AS, and UBE3A-overexpressing neurons (Dup15q autism model) on micrograting substrates, with the aim to clarify the role of UBE3A in neuronal guidance. Results We found that loss of axonal contact guidance is specific for AS neurons while UBE3A overexpression does not affect neuronal directional polarization along microgratings. Deficits at the level of axonal branching, growth cone orientation and actin fiber content, focal adhesion (FA) effectors, and actin fiber–binding proteins were observed in AS neurons. We tested different rescue strategies for restoring correct topographical guidance in AS neurons on microgratings, by either UBE3A protein re-expression or by pharmacological treatments acting on cytoskeleton contractility. Nocodazole, a drug that depolymerizes microtubules and increases cell contractility, rescued AS axonal alignment to the gratings by partially restoring focal adhesion pathway activation. Surprisingly, UBE3A re-expression only resulted in partial rescue of the phenotype. Conclusions We identified a specific in vitro deficit in axonal topographical guidance due selectively to the loss of UBE3A, and we further demonstrate that this defective guidance can be rescued to a certain extent by pharmacological or genetic treatment strategies. Overall, cytoskeleton dynamics emerge as important partners in UBE3A-mediated contact guidance responses. These results support the view that UBE3A-related deficits in early neuronal morphogenesis may lead to defective neuronal connectivity and plasticity.

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