Attractive axon guidance involves asymmetric membrane transport and exocytosis in the growth cone.
Asymmetric elevation of the Ca(2+) concentration in the growth cone can mediate both attractive and repulsive axon guidance. Ca(2+) signals that are accompanied by Ca(2+)-induced Ca(2+) release (CICR) trigger attraction, whereas Ca(2+) signals that are not accompanied by CICR trigger repulsion. The molecular machinery downstream of Ca(2+) signals, however, remains largely unknown. Here we report that asymmetric membrane trafficking mediates growth cone attraction. Local photolysis of caged Ca(2+), together with CICR, on one side of the growth cone of a chick dorsal root ganglion neuron facilitated the microtubule-dependent centrifugal transport of vesicles towards the leading edge and their subsequent vesicle-associated membrane-protein 2 (VAMP2)-mediated exocytosis on the side with an elevated Ca(2+) concentration. In contrast, Ca(2+) signals without CICR had no effect on the vesicle transport. Furthermore, pharmacological inhibition of VAMP2-mediated exocytosis prevented growth cone attraction, but not repulsion. These results strongly suggest that growth cone attraction and repulsion are driven by distinct mechanisms, rather than using the same molecular machinery with opposing polarities.
Tojima
Takuro
T
Laboratory for Neuronal Growth Mechanisms, Brain Science Institute, The Institute of Physical and Chemical Research (RIKEN), 2-1 Hirosawa, Wako, Saitama 351-0198, Japan.
Akiyama
Hiroki
H
Itofusa
Rurika
R
Li
Yan
Y
Katayama
Hiroyuki
H
Miyawaki
Atsushi
A
Kamiguchi
Hiroyuki
H
D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D001369:Axons / Q000187:drug effects / Q000502:physiology*; D001692:Biological Transport / Q000187:drug effects / Q000502:physiology; D002118:Calcium / Q000378:metabolism / Q000494:pharmacology; D002478:Cells, Cultured; D002614:Chelating Agents / Q000494:pharmacology; D002642:Chick Embryo; D000242:Cyclic AMP / Q000031:analogs & derivatives / Q000494:pharmacology; D004347:Drug Interactions; D004533:Egtazic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D005089:Exocytosis / Q000187:drug effects / Q000502:physiology*; D005727:Ganglia, Spinal / Q000166:cytology; D049452:Green Fluorescent Proteins / Q000378:metabolism; D020439:Growth Cones / Q000187:drug effects / Q000502:physiology*; D008666:Metalloendopeptidases / Q000494:pharmacology; D009474:Neurons / Q000166:cytology*; D015739:Nocodazole / Q000494:pharmacology; D010782:Photolysis / Q000187:drug effects; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D011726:Pyridinium Compounds; D000644:Quaternary Ammonium Compounds; D013744:Tetanus Toxin / Q000494:pharmacology; D013873:Thionucleotides / Q000494:pharmacology; D014162:Transfection / Q000379:methods; D050988:Vesicle-Associated Membrane Protein 2 / Q000378:metabolism
D000970:Antineoplastic Agents; D002614:Chelating Agents; C073804:FM1 43; D047428:Protein Kinase Inhibitors; D011726:Pyridinium Compounds; D000644:Quaternary Ammonium Compounds; D013744:Tetanus Toxin; D013873:Thionucleotides; D050988:Vesicle-Associated Membrane Protein 2; C035979:tetanospasmin; C070379:1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid acetoxymethyl ester; D049452:Green Fluorescent Proteins; C016957:adenosine-3',5'-cyclic phosphorothioate; D004533:Egtazic Acid; D000242:Cyclic AMP; D008666:Metalloendopeptidases; D015739:Nocodazole; D002118:Calcium