ZFIN ID: ZDB-PUB-021213-1
Regulation by glycogen synthase kinase-3beta of the arborization field and maturation of retinotectal projection in zebrafish
Tokuoka, H., Yoshida, T., Matsuda, N., and Mishina, M.
Date: 2002
Source: The Journal of neuroscience : the official journal of the Society for Neuroscience   22(23): 10324-10332 (Journal)
Registered Authors: Matsuda, Naoto, Mishina, Masayoshi, Yoshida, Tomoyuki
Keywords: GSK-3b; retinotectal projection; arborization field; synapse maturation; zebrafish; nicotinic acetylcholine receptor b3; vesicle-associated membrane protein 2
MeSH Terms:
  • Animals
  • Animals, Genetically Modified
  • Axons/physiology
  • Genes, Dominant
  • Genes, Reporter
  • Glycogen Synthase Kinase 3/genetics
  • Glycogen Synthase Kinase 3/metabolism*
  • Luminescent Proteins/biosynthesis
  • Luminescent Proteins/genetics
  • Membrane Proteins/metabolism
  • Molecular Sequence Data
  • Neurons/cytology
  • Neurons/metabolism
  • Presynaptic Terminals/metabolism
  • Promoter Regions, Genetic/physiology
  • R-SNARE Proteins
  • Receptors, Nicotinic/genetics
  • Retina/cytology
  • Retina/embryology
  • Retina/enzymology*
  • Retinal Ganglion Cells/cytology
  • Retinal Ganglion Cells/metabolism
  • Superior Colliculi/cytology
  • Superior Colliculi/embryology
  • Superior Colliculi/enzymology*
  • Synapses/metabolism
  • Visual Pathways/cytology
  • Visual Pathways/embryology
  • Visual Pathways/enzymology*
  • Zebrafish
PubMed: 12451132
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 beta3 (nAChRbeta3) gene promoter. Enhanced green fluorescent protein (EGFP) expression signals in living transgenic zebrafish lines carrying the nAChRbeta3 gene promoter-directed EGFP expression vector visualized the development of entire retinal ganglion cell (RGC) axon projection to the tectum. Microinjection of the nAChRbeta3 gene promoter-driven double-cassette vectors directing the expression of both dominant-negative glycogen synthase kinase-3beta (dnGSK-3beta) 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-3beta 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-3beta 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-3beta regulates the arborization field and maturation of RGC axon terminals in vivo.