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A more recent version of this article appeared on March 1, 2006
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Submitted on July 18, 2005
Revised on December 5, 2005
Accepted on December 20, 2005
*Membrane Traffic in Neuronal and Epithelial Morphogenesis, INSERM Avenir Team, 75005 Paris, France; Institut Jacque Monod, UMR 7592 CNRS, Universities Paris 6 and 7, 75005 Paris, France; INSERM U536, Institut du Fer-à-Moulin, 75005 Paris, France; ||Rho GTPases, Adhesion and Skeletal Muscle, CRBM-CNRS FRE 2593, 34293 Montpellier, France
Monitoring Editor: Jennifer Lippincott-Schwartz
Tetanus neurotoxin Insensitive-Vesicle Associated Membrane Protein (TI-VAMP)-mediated fusion of intracellular vesicles with the plasma membrane is crucial for neurite outgrowth, a pathway not requiring synaptobrevin-dependent exocytosis. Yet, it is not known how the TI-VAMP membrane trafficking pathway is regulated or how it is coordinated with cytoskeletal dynamics within the growth cone that guide neurite outgrowth. Here, we demonstrate that TI-VAMP, but not synaptobrevin 2, concentrates in the peripheral, F-actin-rich region of the growth cones of hippocampal neurons in primary culture. Its accumulation correlates with and depends upon the presence of F-actin. Moreover, acute stimulation of actin remodeling by homophilic activation of the adhesion molecule L1 induces a site directed, actin dependent recruitment of the TI-VAMP compartment. Expression of a dominant-positive mutant of Cdc42, a key regulator of cell polarity, stimulates formation of F-actin- and TI-VAMP-rich filopodia outside the growth cone Furthermore, we report that Cdc42 activates exocytosis of pHLuorin tagged TI-VAMP in an actin-dependent manner. Collectively, our data suggest that Cdc42 and regulated assembly of the F-actin network control the accumulation and exocytosis of TI-VAMP-containing membrane vesicles in growth cones to coordinate membrane trafficking and actin remodeling during neurite outgrowth.
Present address: Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06520.
Address correspondence to:
Thierry Galli (thierry{at}tgalli.net)
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