Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
J Neurosci
2016 Feb 17;367:2267-82. doi: 10.1523/JNEUROSCI.2645-15.2016.
Show Gene links
Show Anatomy links
Guidance of Axons by Local Coupling of Retrograde Flow to Point Contact Adhesions.
Nichol RH, Hagen KM, Lumbard DC, Dent EW, Gómez TM.
???displayArticle.abstract???
Growth cones interact with the extracellular matrix (ECM) through integrin receptors at adhesion sites termed point contacts. Point contact adhesions link ECM proteins to the actin cytoskeleton through numerous adaptor and signaling proteins. One presumed function of growth cone point contacts is to restrain or "clutch" myosin-II-based filamentous actin (F-actin) retrograde flow (RF) to promote leading edge membrane protrusion. In motile non-neuronal cells, myosin-II binds and exerts force upon actin filaments at the leading edge, where clutching forces occur. However, in growth cones, it is unclear whether similar F-actin-clutching forces affect axon outgrowth and guidance. Here, we show in Xenopus spinal neurons that RF is reduced in rapidly migrating growth cones on laminin (LN) compared with non-integrin-binding poly-d-lysine (PDL). Moreover, acute stimulation with LN accelerates axon outgrowth over a time course that correlates with point contact formation and reduced RF. These results suggest that RF is restricted by the assembly of point contacts, which we show occurs locally by two-channel imaging of RF and paxillin. Further, using micropatterns of PDL and LN, we demonstrate that individual growth cones have differential RF rates while interacting with two distinct substrata. Opposing effects on RF rates were also observed in growth cones treated with chemoattractive and chemorepulsive axon guidance cues that influence point contact adhesions. Finally, we show that RF is significantly attenuated in vivo, suggesting that it is restrained by molecular clutching forces within the spinal cord. Together, our results suggest that local clutching of RF can control axon guidance on ECM proteins downstream of axon guidance cues. Here, we correlate point contact adhesions directly with clutching of filamentous actin retrograde flow (RF), which our findings strongly suggest guides developing axons. Acute assembly of new point contact adhesions is temporally and spatially linked to attenuation of RF at sites of forward membrane protrusion. Importantly, clutching of RF is modulated by extracellular matrix (ECM) proteins and soluble axon guidance cues, suggesting that it may regulate axon guidance in vivo. Consistent with this notion, we found that RF rates of spinal neuron growth cones were slower in vivo than what was observed in vitro. Together, our study provides the best evidence that growth cone-ECM adhesions clutch RF locally to guide axons in vivo.
Antar,
Local functions for FMRP in axon growth cone motility and activity-dependent regulation of filopodia and spine synapses.
2006, Pubmed
Antar,
Local functions for FMRP in axon growth cone motility and activity-dependent regulation of filopodia and spine synapses.
2006,
Pubmed Bamburg,
Proteins of the ADF/cofilin family: essential regulators of actin dynamics.
1999,
Pubmed Bard,
A molecular clutch between the actin flow and N-cadherin adhesions drives growth cone migration.
2008,
Pubmed
,
Xenbase Bechara,
FAK-MAPK-dependent adhesion disassembly downstream of L1 contributes to semaphorin3A-induced collapse.
2008,
Pubmed Beck,
Structure and function of laminin: anatomy of a multidomain glycoprotein.
1990,
Pubmed Bridgman,
Myosin IIB is required for growth cone motility.
2001,
Pubmed Brown,
Retrograde flow rate is increased in growth cones from myosin IIB knockout mice.
2003,
Pubmed Brown,
Disruption of the cytoskeleton during Semaphorin 3A induced growth cone collapse correlates with differences in actin organization and associated binding proteins.
2009,
Pubmed Burnette,
Myosin II activity facilitates microtubule bundling in the neuronal growth cone neck.
2008,
Pubmed Carlier,
Control of actin assembly dynamics in cell motility.
2007,
Pubmed Choi,
Actin and alpha-actinin orchestrate the assembly and maturation of nascent adhesions in a myosin II motor-independent manner.
2008,
Pubmed Chrzanowska-Wodnicka,
Rho-stimulated contractility drives the formation of stress fibers and focal adhesions.
1996,
Pubmed Ciobanasu,
Actomyosin-dependent formation of the mechanosensitive talin-vinculin complex reinforces actin anchoring.
2014,
Pubmed Cluzel,
The mechanisms and dynamics of (alpha)v(beta)3 integrin clustering in living cells.
2005,
Pubmed Craig,
Membrane tension, myosin force, and actin turnover maintain actin treadmill in the nerve growth cone.
2012,
Pubmed del Rio,
Stretching single talin rod molecules activates vinculin binding.
2009,
Pubmed Dent,
The growth cone cytoskeleton in axon outgrowth and guidance.
2011,
Pubmed Dudanova,
Integration of guidance cues: parallel signaling and crosstalk.
2013,
Pubmed Evans,
Laminin and fibronectin modulate inner ear spiral ganglion neurite outgrowth in an in vitro alternate choice assay.
2007,
Pubmed Flynn,
ADF/cofilin-mediated actin retrograde flow directs neurite formation in the developing brain.
2012,
Pubmed Forscher,
Actions of cytochalasins on the organization of actin filaments and microtubules in a neuronal growth cone.
1988,
Pubmed Gallo,
RhoA-kinase coordinates F-actin organization and myosin II activity during semaphorin-3A-induced axon retraction.
2006,
Pubmed Garcia,
Two-tiered coupling between flowing actin and immobilized N-cadherin/catenin complexes in neuronal growth cones.
2015,
Pubmed Gardel,
Traction stress in focal adhesions correlates biphasically with actin retrograde flow speed.
2008,
Pubmed Gehler,
p75 neurotrophin receptor signaling regulates growth cone filopodial dynamics through modulating RhoA activity.
2004,
Pubmed Giannone,
Multi-level molecular clutches in motile cell processes.
2009,
Pubmed Gomez,
Filopodial calcium transients promote substrate-dependent growth cone turning.
2001,
Pubmed
,
Xenbase Gomez,
Filopodia initiate choices made by sensory neuron growth cones at laminin/fibronectin borders in vitro.
1994,
Pubmed Gómez,
Working with Xenopus spinal neurons in live cell culture.
2003,
Pubmed
,
Xenbase Grashoff,
Measuring mechanical tension across vinculin reveals regulation of focal adhesion dynamics.
2010,
Pubmed Johnson,
Neuronal IP3 3-kinase is an F-actin-bundling protein: role in dendritic targeting and regulation of spine morphology.
2009,
Pubmed Jurney,
Rac1-mediated endocytosis during ephrin-A2- and semaphorin 3A-induced growth cone collapse.
2002,
Pubmed Keren,
Mechanism of shape determination in motile cells.
2008,
Pubmed Kerstein,
Mechanochemical regulation of growth cone motility.
2015,
Pubmed Knöll,
Stripe assay to examine axonal guidance and cell migration.
2007,
Pubmed Koch,
Strength in the periphery: growth cone biomechanics and substrate rigidity response in peripheral and central nervous system neurons.
2012,
Pubmed Krause,
Steering cell migration: lamellipodium dynamics and the regulation of directional persistence.
2014,
Pubmed Kubo,
Shootin1-cortactin interaction mediates signal-force transduction for axon outgrowth.
2015,
Pubmed Lin,
Myosin drives retrograde F-actin flow in neuronal growth cones.
1996,
Pubmed Lin,
Growth cone advance is inversely proportional to retrograde F-actin flow.
1995,
Pubmed Liu,
p130CAS is required for netrin signaling and commissural axon guidance.
2007,
Pubmed Lowery,
The trip of the tip: understanding the growth cone machinery.
2009,
Pubmed
,
Xenbase Mallavarapu,
Regulated actin cytoskeleton assembly at filopodium tips controls their extension and retraction.
1999,
Pubmed Marsick,
Activation of ADF/cofilin mediates attractive growth cone turning toward nerve growth factor and netrin-1.
2010,
Pubmed
,
Xenbase Medeiros,
Myosin II functions in actin-bundle turnover in neuronal growth cones.
2006,
Pubmed Mitchison,
Cytoskeletal dynamics and nerve growth.
1988,
Pubmed Mogilner,
Polymer motors: pushing out the front and pulling up the back.
2003,
Pubmed Moore,
Netrin-1 attracts axons through FAK-dependent mechanotransduction.
2012,
Pubmed Moore,
Stretchy proteins on stretchy substrates: the important elements of integrin-mediated rigidity sensing.
2010,
Pubmed Moore,
Traction on immobilized netrin-1 is sufficient to reorient axons.
2009,
Pubmed Munevar,
Distinct roles of frontal and rear cell-substrate adhesions in fibroblast migration.
2001,
Pubmed Myers,
Focal adhesion kinase modulates Cdc42 activity downstream of positive and negative axon guidance cues.
2012,
Pubmed
,
Xenbase Myers,
Focal adhesion kinase promotes integrin adhesion dynamics necessary for chemotropic turning of nerve growth cones.
2011,
Pubmed
,
Xenbase Myers,
Regulation of axonal outgrowth and pathfinding by integrin-ECM interactions.
2011,
Pubmed Nie,
Tsc2-Rheb signaling regulates EphA-mediated axon guidance.
2010,
Pubmed Petchprayoon,
Fluorescent kabiramides: new probes to quantify actin in vitro and in vivo.
2005,
Pubmed Ponti,
Two distinct actin networks drive the protrusion of migrating cells.
2004,
Pubmed Riedl,
Lifeact: a versatile marker to visualize F-actin.
2008,
Pubmed Robles,
Focal adhesion kinase signaling at sites of integrin-mediated adhesion controls axon pathfinding.
2006,
Pubmed
,
Xenbase Robles,
Src-dependent tyrosine phosphorylation at the tips of growth cone filopodia promotes extension.
2005,
Pubmed
,
Xenbase Saengsawang,
The F-BAR protein CIP4 inhibits neurite formation by producing lamellipodial protrusions.
2012,
Pubmed San Miguel-Ruiz,
The role of Arp2/3 in growth cone actin dynamics and guidance is substrate dependent.
2014,
Pubmed Santiago-Medina,
Imaging adhesion and signaling dynamics in Xenopus laevis growth cones.
2012,
Pubmed
,
Xenbase Santiago-Medina,
PAK-PIX interactions regulate adhesion dynamics and membrane protrusion to control neurite outgrowth.
2013,
Pubmed
,
Xenbase Santiago-Medina,
Regulation of ECM degradation and axon guidance by growth cone invadosomes.
2015,
Pubmed
,
Xenbase Sawada,
Force sensing by mechanical extension of the Src family kinase substrate p130Cas.
2006,
Pubmed Shimada,
Shootin1 interacts with actin retrograde flow and L1-CAM to promote axon outgrowth.
2008,
Pubmed Smilenov,
Focal adhesion motility revealed in stationary fibroblasts.
1999,
Pubmed Suter,
The Ig superfamily cell adhesion molecule, apCAM, mediates growth cone steering by substrate-cytoskeletal coupling.
1998,
Pubmed Symons,
Control of actin polymerization in live and permeabilized fibroblasts.
1991,
Pubmed Tehrani,
Src phosphorylation of cortactin enhances actin assembly.
2007,
Pubmed Thievessen,
Vinculin-actin interaction couples actin retrograde flow to focal adhesions, but is dispensable for focal adhesion growth.
2013,
Pubmed Toriyama,
Conversion of a signal into forces for axon outgrowth through Pak1-mediated shootin1 phosphorylation.
2013,
Pubmed Treloar,
Tenascin-C is an inhibitory boundary molecule in the developing olfactory bulb.
2009,
Pubmed Turney,
Laminin stimulates and guides axonal outgrowth via growth cone myosin II activity.
2005,
Pubmed Tyler,
The mechanobiology of brain function.
2012,
Pubmed Van Goor,
The role of actin turnover in retrograde actin network flow in neuronal growth cones.
2012,
Pubmed Vicente-Manzanares,
Non-muscle myosin II takes centre stage in cell adhesion and migration.
2009,
Pubmed Vitriol,
Growth cone travel in space and time: the cellular ensemble of cytoskeleton, adhesion, and membrane.
2012,
Pubmed Walter,
Avoidance of posterior tectal membranes by temporal retinal axons.
1987,
Pubmed Wang,
Focal adhesion kinase is involved in mechanosensing during fibroblast migration.
2001,
Pubmed Weitzdoerfer,
Fetal life in Down syndrome starts with normal neuronal density but impaired dendritic spines and synaptosomal structure.
2001,
Pubmed Woo,
Retinotopic mapping requires focal adhesion kinase-mediated regulation of growth cone adhesion.
2009,
Pubmed
,
Xenbase Woo,
Rac1 and RhoA promote neurite outgrowth through formation and stabilization of growth cone point contacts.
2006,
Pubmed
,
Xenbase Wu,
Local translation of RhoA regulates growth cone collapse.
2005,
Pubmed Yamashiro,
A new link between the retrograde actin flow and focal adhesions.
2014,
Pubmed Zhang,
Calcineurin-dependent cofilin activation and increased retrograde actin flow drive 5-HT-dependent neurite outgrowth in Aplysia bag cell neurons.
2012,
Pubmed
