MAP1B coordinates microtubule and actin filament remodeling in adult mouse Schwann cell tips and DRG neuron growth cones

2007 ◽  
Vol 36 (2) ◽  
pp. 235-247 ◽  
Author(s):  
Céline Bouquet ◽  
Michèle Ravaille-Veron ◽  
Friedrich Propst ◽  
Fatiha Nothias
Development ◽  
2000 ◽  
Vol 127 (10) ◽  
pp. 2099-2111 ◽  
Author(s):  
J. Zhang ◽  
M. Granato

En route to their targets, motor axons encounter choice points at which they select their future path. Experimental studies predict that at each choice point specialized cells provide local guidance to pathfinding motor axons, however, the identity of these cells and their signals is unknown. Here, we identify the zebrafish unplugged gene as a key component for choice point navigation of pioneering motor axons. We show that in unplugged mutant embryos, motor neuron growth cones reach the choice point but make inappropriate pathway decisions. Analysis of chimeric embryos demonstrates that unplugged activity is produced by a selective group of mesodermal cells located adjacent to the choice point. As the first motor growth cones approach the choice point, these mesodermal cells migrate away, suggesting that unplugged activity influences growth cones by a contact-independent mechanism. These data suggest that unplugged defines a somite-derived signal that elicits differential guidance decisions in motor growth cones.


2005 ◽  
Vol 25 (22) ◽  
pp. 9973-9984 ◽  
Author(s):  
Nariko Arimura ◽  
Céline Ménager ◽  
Yoji Kawano ◽  
Takeshi Yoshimura ◽  
Saeko Kawabata ◽  
...  

ABSTRACT Collapsin response mediator protein 2 (CRMP-2) enhances the advance of growth cones by regulating microtubule assembly and Numb-mediated endocytosis. We previously showed that Rho kinase phosphorylates CRMP-2 during growth cone collapse; however, the roles of phosphorylated CRMP-2 in growth cone collapse remain to be clarified. Here, we report that CRMP-2 phosphorylation by Rho kinase cancels the binding activity to the tubulin dimer, microtubules, or Numb. CRMP-2 binds to actin, but its binding is not affected by phosphorylation. Electron microscopy revealed that CRMP-2 localizes on microtubules, clathrin-coated pits, and actin filaments in dorsal root ganglion neuron growth cones, while phosphorylated CRMP-2 localizes only on actin filaments. The phosphomimic mutant of CRMP-2 has a weakened ability to enhance neurite elongation. Furthermore, ephrin-A5 induces phosphorylation of CRMP-2 via Rho kinase during growth cone collapse. Taken together, these results suggest that Rho kinase phosphorylates CRMP-2, and inactivates the ability of CRMP-2 to promote microtubule assembly and Numb-mediated endocytosis, during growth cone collapse.


1992 ◽  
Vol 119 (5) ◽  
pp. 1219-1243 ◽  
Author(s):  
A K Lewis ◽  
P C Bridgman

The organization and polarity of actin filaments in neuronal growth cones was studied with negative stain and freeze-etch EM using a permeabilization protocol that caused little detectable change in morphology when cultured nerve growth cones were observed by video-enhanced differential interference contrast microscopy. The lamellipodial actin cytoskeleton was composed of two distinct subpopulations: a population of 40-100-nm-wide filament bundles radiated from the leading edge, and a second population of branching short filaments filled the volume between the dorsal and ventral membrane surfaces. Together, the two populations formed the three-dimensional structural network seen within expanding lamellipodia. Interaction of the actin filaments with the ventral membrane surface occurred along the length of the filaments via membrane associated proteins. The long bundled filament population was primarily involved in these interactions. The filament tips of either population appeared to interact with the membrane only at the leading edge; this interaction was mediated by a globular Triton-insoluble material. Actin filament polarity was determined by decoration with myosin S1 or heavy meromyosin. Previous reports have suggested that the polarity of the actin filaments in motile cells is uniform, with the barbed ends toward the leading edge. We observed that the actin filament polarity within growth cone lamellipodia is not uniform; although the predominant orientation was with the barbed end toward the leading edge (47-56%), 22-25% of the filaments had the opposite orientation with their pointed ends toward the leading edge, and 19-31% ran parallel to the leading edge. The two actin filament populations display distinct polarity profiles: the longer filaments appear to be oriented predominantly with their barbed ends toward the leading edge, whereas the short filaments appear to be randomly oriented. The different length, organization and polarity of the two filament populations suggest that they differ in stability and function. The population of bundled long filaments, which appeared to be more ventrally located and in contact with membrane proteins, may be more stable than the population of short branched filaments. The location, organization, and polarity of the long bundled filaments suggest that they may be necessary for the expansion of lamellipodia and for the production of tension mediated by receptors to substrate adhesion molecules.


PLoS ONE ◽  
2016 ◽  
Vol 11 (8) ◽  
pp. e0159405 ◽  
Author(s):  
Benjamin Rappaz ◽  
Karen Lai Wing Sun ◽  
James P. Correia ◽  
Paul W. Wiseman ◽  
Timothy E. Kennedy

2012 ◽  
Vol 12 (05) ◽  
pp. 1250032 ◽  
Author(s):  
CHENG-TAO CHANG ◽  
CHOU-CHING K. LIN ◽  
MING SHAUNG JU

A cost-effective method that integrates high-resolution morphology using an atomic force microscope and immunofluorescence imaging for measuring the local mechanical properties of a cell was developed. By considering the normal indentation conditions and the distribution of the underlying cytoskeleton, a criterion for selecting indentation sites was proposed. PC-12 cells cultivated under normal and high D-glucose medium are employed to demonstrate the applicability of the proposed method. The apparent Young's modulus for each indentation site was estimated by fitting the data with a pyramidal punch contact mechanics model. The results showed that the cell bodies cultivated in the high D-glucose medium were higher but their growth cones were shorter than those cultivated in a normal medium. The Young's moduli of the growth cones were positively correlated with the density of the actin filament in the cytoskeleton. The Young's moduli at the growth cone and the nucleus region of cells cultivated in the high D-glucose medium were higher and lower, respectively, than those of the control group. The results demonstrated the integrated method could correlate local mechanical properties and distribution of actin filament of the growth cone of PC-12 cell.


1991 ◽  
Vol 115 (2) ◽  
pp. 381-395 ◽  
Author(s):  
J H Sabry ◽  
T P O'Connor ◽  
L Evans ◽  
A Toroian-Raymond ◽  
M Kirschner ◽  
...  

The growth of an axon toward its target results from the reorganization of the cytoskeleton in response to environmental guidance cues. Recently developed imaging technology makes it possible to address the effect of such cues on the neural cytoskeleton directly. Although high resolution studies can be carried out on neurons in vitro, these circumstances do not recreate the complexity of the natural environment. We report here on the arrangement and dynamics of microtubules in live neurons pathfinding in response to natural guidance cues in situ using the embryonic grasshopper limb fillet preparation. A rich microtubule network was present within the body of the growth cone and normally extended into the distal growth cone margin. Complex microtubule loops often formed transiently within the growth cone. Branches both with and without microtubules were regularly observed. Microtubules did not extend into filopodia. During growth cone steering events in response to identified guidance cues, microtubule behaviour could be monitored. In turns towards guidepost cells, microtubules selectively invaded branches derived from filopodia that had contacted the guidepost cell. At limb segment boundaries, microtubules displayed a variety of behaviors, including selective branch invasion, and also invasion of multiple branches followed by selective retention in branches oriented in the correct direction. Microtubule invasion of multiple branches also was seen in growth cones migrating on intrasegmental epithelium. Both selective invasion and selective retention generate asymmetrical microtubule arrangements within the growth cone, and may play a key role in growth cone steering events.


1973 ◽  
Vol 56 (3) ◽  
pp. 713-735 ◽  
Author(s):  
Mary Bartlett Bunge

The leading tips of elongating nerve fibers are enlarged into "growth cones" which are seen in tissue culture to continually undergo changes in conformation and to foster numerous transitory slender extensions (filopodia) and/or a veillike ruffling sheet. After explantation of 1-day-old rat superior cervical ganglia (as pieces or as individual neurons), nerve fibers and tips were photographed during growth and through the initial stages of aldehyde fixation and then relocated after embedding in plastic. Electron microscopy of serially sectioned tips revealed the following. The moving parts of the cone, the peripheral flange and filopodia, contained a distinctive apparently filamentous feltwork from which all organelles except membranous structures were excluded; microtubules were notably absent from these areas. The cone interior contained varied forms of agranular endoplasmic reticulum, vacuoles, vesicles, coated vesicles, mitochondria, microtubules, and occasional neurofilaments and polysomes. Dense-cored vesicles and lysosomal structures were also present and appeared to be formed locally, at least in part from reticulum. The possible roles of the various forms of agranular membranous components are discussed and it is suggested that structures involved in both the assembly and degradation of membrane are present in the cone. The content of these growing tips resembles that in sensory neuron growth cones studied by others.


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