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The Journal of Neuroscience, July 7, 2004, 24(27):6161-6170; doi:10.1523/JNEUROSCI.1476-04.2004

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Development/Plasticity/Repair
Correlation between Semaphorin3A-Induced Facilitation of Axonal Transport and Local Activation of a Translation Initiation Factor Eukaryotic Translation Initiation Factor 4E

Chanxia Li,¹ Yukio Sasaki,¹ Kohtaro Takei,^1,4 Hiroshi Yamamoto,¹ Masayuki Shouji,¹ Yoshinobu Sugiyama,¹ Tadashi Kawakami,² Fumio Nakamura,¹ Takeshi Yagi,³ Toshio Ohshima,⁴ and Yoshio Goshima^1,5

¹Department of Molecular Pharmacology and Neurobiology, Yokohama City University Graduate School of Medicine, Yokohama 236-0004, Japan, ²Department of Physiology, Kitasato University School of Medicine, Sagamihara 228-8555, Japan, ³Division of Molecular Genetics, Section of Gene Networks and Gene Functions, Institute for Molecular and Cellular Biology, Osaka University, Suita 565-0871, Japan, ⁴Laboratory for Developmental Neurobiology, Brain Science Institute, The Institute of Physical and Chemical Research, Wako 351-0198, Japan, and ⁵Core Research for Evolutional Science and Technology, Japan Science and Technology Corporation, Kawaguchi 332-0012, Japan

An impressive body of evidence has been accumulated indicating that local protein synthesis is implicated in navigation of neurite extension induced by guidance cues, such as semaphorin3A (Sema3A). We found previously that a Src type tyrosine kinase Fyn and cyclin-dependent kinase 5 (Cdk5) mediate Sema3A-signaling. We also showed that Sema3A elicits axonal transport through neuropilin-1, a receptor for Sema3A, located at the growth cones. Here, we investigate the relationship between Sema3A-induced local signaling, protein synthesis, and axonal transport. Lavendustin A, a tyrosine kinase inhibitor, and olomoucine, a cyclin-dependent kinase inhibitor, suppressed Sema3A-induced facilitation of anterograde and retrograde axonal transport in dorsal root ganglion (DRG) neuron with and without the cell body. Sema3A-induced facilitation of axonal transport was attenuated in DRG neurons of fyn- (fyn^-/-) and a Cdk5 activator, p35 (p35^-/-)-deficient mice when compared with those of wild-type or heterozygous mice. Inhibition of protein synthesis suppressed Sema3A-induced facilitation of axonal transport in the DRG neuron with and without the cell body. Sema3A enhanced the level of immunoreactivity of phosphorylated eukaryotic translation initiation factor 4E (eIF-4E) within 5 min in growth cones in a time course similar to that of the facilitated axonal transport. This enhanced signal for phospho-eIF4E was blocked by lavendustin A or olomoucine and was not detected in the fyn^-/- and p35^-/- neurons. These results provide evidence for a mutual regulatory mechanism between local protein synthesis and axonal transport.

Key words: axon guidance; Sema3A; Fyn; Cdk5; axonal transport; growth cones; local protein synthesis

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Received Dec 4, 2003; revised May 17, 2004; accepted May 17, 2004.

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