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The Journal of Neuroscience, December 1, 2002, 22(23):10324-10332
Regulation by Glycogen Synthase Kinase-3 of the Arborization
Field and Maturation of Retinotectal Projection in Zebrafish
Hirofumi
Tokuoka,
Tomoyuki
Yoshida,
Naoto
Matsuda, and
Masayoshi
Mishina
Department of Molecular Neurobiology and Pharmacology, Graduate
School of Medicine, University of Tokyo, and Solution-Oriented
Research for Science and Technology, Japan Science and
Technology Corporation, Tokyo 113-0033, Japan
The retinotectal projection is one of the best systems to study the
molecular basis of synapse formation in the CNS because of the
well characterized topographic connections and activity-dependent refinement. Here, we developed a presynaptic neuron-specific gene manipulation system in the zebrafish retinotectal projection in vivo using the nicotinic acetylcholine receptor  3
(nAChR3) gene promoter. Enhanced green fluorescent protein (EGFP)
expression signals in living transgenic zebrafish lines carrying the
nAChR3 gene promoter-directed EGFP expression vector
visualized the development of entire retinal ganglion cell (RGC) axon
projection to the tectum. Microinjection of the
nAChR3 gene promoter-driven double-cassette vectors
directing the expression of both dominant-negative glycogen synthase
kinase-3 (dnGSK-3) and EGFP enabled us to follow the development of individual RGCs and to examine the effect of the molecule on the axonal arborization and maturation of the same neurons
in living zebrafish. We found that the expression of the dominant-negative form of zebrafish GSK-3 suppressed the
arborization field of RGC axon terminals in the tectum as estimated by
the reduction of arbor branch length and arbor areas. Furthermore, the
suppression of GSK-3 activity increased the size of
vesicle-associated membrane protein 2-EGFP puncta in RGC axon
terminals at the early stage of innervation to the tectum. These
results suggest that GSK-3 regulates the arborization field and
maturation of RGC axon terminals in vivo.
Key words:
GSK-3; retinotectal projection; arborization field; synapse maturation; zebrafish; nicotinic acetylcholine receptor 3; vesicle-associated membrane protein 2
Copyright © 2002 Society for Neuroscience 0270-6474/02/222310324-09$05.00/0
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