© The Rockefeller University Press,
0021-9525/2000//411 $5.00
The Journal of Cell Biology, Volume 149, Number 2,
, 2000 411-422
Semaphorin3a Enhances Endocytosis at Sites of Receptor–F-Actin Colocalization during Growth Cone Collapse
Alyson E. Fourniera,b,
Fumio Nakamuraa,b,
Susumu Kawamotod,
Yoshio Goshimae,
Robert G. Kalba,c, and
Stephen M. Strittmattera,b
a Department of Neurology, Yale University School of Medicine, New Haven, Connecticut 06520
b Department of Neurobiology, Yale University School of Medicine, New Haven, Connecticut 06520
c Department of Pharmacology, Yale University School of Medicine, New Haven, Connecticut 06520
d Department of Bacteriology, Yokohama City University School of Medicine, Yokohama 236, Japan
e Department of Pharmacology, Yokohama City University School of Medicine, Yokohama 236, Japan
Department of Neurology, Yale University School of Medicine, 333 Cedar Street, P.O. Box 208018, New Haven, CT 06520.(203) 785-5098(203) 785-4878
stephen.strittmatter{at}yale.edu
Axonal growth cone collapse is accompanied by a reduction in filopodial F-actin. We demonstrate here that semaphorin 3A (Sema3A) induces a coordinated rearrangement of Sema3A receptors and F-actin during growth cone collapse. Differential interference contrast microscopy reveals that some sites of Sema3A-induced F-actin reorganization correlate with discrete vacuoles, structures involved in endocytosis. Endocytosis of FITC-dextran by the growth cone is enhanced during Sema3A treatment, and sites of dextran accumulation colocalize with actin-rich vacuoles and ridges of membrane. Furthermore, the Sema3A receptor proteins, neuropilin-1 and plexin, and the Sema3A signaling molecule, rac1, also reorganize to vacuoles and membrane ridges after Sema3A treatment. These data support a model whereby Sema3A stimulates endocytosis by focal and coordinated rearrangement of receptor and cytoskeletal elements. Dextran accumulation is also increased in retinal ganglion cell (RGC) growth cones, in response to ephrin A5, and in RGC and DRG growth cones, in response to myelin and phorbol-ester. Therefore, enhanced endocytosis may be a general principle of physiologic growth cone collapse. We suggest that growth cone collapse is mediated by both actin filament rearrangements and alterations in membrane dynamics.
Key Words: membrane dynamics ephrins dextran uptake axon guidance axon repulsion
© 2000 The Rockefeller University Press
Abbreviations used in this paper: DIC, differential interference contrast; DRG, dorsal root ganglia; NP, neuropilin; RGC, retinal ganglion cell; Sema3A, semaphorin 3A; Sema3C, semaphorin 3C; TPA, phorbol-12 myristate 13-acetate.

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