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First published online 30 June 2004
doi: 10.1242/dev.01228

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PlexinA4 is necessary as a downstream target of Islet2 to mediate Slit signaling for promotion of sensory axon branching

Toshio Miyashita^1,*, Sang-Yeob Yeo^1,, Yoshikazu Hirate¹, Hiroshi Segawa¹, Hironori Wada¹, Melissa H. Little², Toshiya Yamada^2,, Naoki Takahashi⁴ and Hitoshi Okamoto^1,3,

¹ Laboratory for Developmental Gene Regulation, RIKEN Brain Science Institute, 2-1, Hirosawa, Wako, Saitama 351-0198, Japan
² Center for Molecular and Cellular Biology, The University of Queensland, Queensland 4072, Australia
³ Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Corporation (JST), 3-4-5 Nihonbashi, Chuo-ku, Tokyo 103-0027, Japan
⁴ Graduate School of Biological Sciences, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara 630-0101, Japan

Author for correspondence (e-mail: hitoshi{at}brain.riken.go.jp)

Accepted 2 April 2004

Slit is a secreted protein known to repulse the growth cones of commissural neurons. By contrast, Slit also promotes elongation and branching of axons of sensory neurons. The reason why different neurons respond to Slit in different ways is largely unknown. Islet2 is a LIM/homeodomain-type transcription factor that specifically regulates elongation and branching of the peripheral axons of the primary sensory neurons in zebrafish embryos. We found that PlexinA4, a transmembrane protein known to be a co-receptor for class III semaphorins, acts downstream of Islet2 to promote branching of the peripheral axons of the primary sensory neurons. Intriguingly, repression of PlexinA4 function by injection of the antisense morpholino oligonucleotide specific to PlexinA4 or by overexpression of the dominant-negative variant of PlexinA4 counteracted the effects of overexpression of Slit2 to induce branching of the peripheral axons of the primary sensory neurons in zebrafish embryos, suggesting involvement of PlexinA4 in the Slit signaling cascades for promotion of axonal branching of the sensory neurons. Colocalized expression of Robo, a receptor for Slit2, and PlexinA4 is observed not only in the primary sensory neurons of zebrafish embryos but also in the dendrites of the pyramidal neurons of the cortex of the mammals, and may be important for promoting the branching of either axons or dendrites in response to Slit, as opposed to the growth cone collapse.

Key words: Zebrafish, PlexinA4, Slit, Islet2, LIM/homeodomain protein, Axon branching, Zebrafish

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