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The Journal of Neuroscience, November 1, 1999, 19(21):9306-9312
Excitatory Synaptogenesis between Identified Lymnaea
Neurons Requires Extrinsic Trophic Factors and Is Mediated by Receptor
Tyrosine Kinases
Toshiro
Hamakawa1, 2,
Melanie A.
Woodin1,
Micki
C.
Bjorgum1,
Sherry D.
Painter3,
Mayumi
Takasaki2,
Ken
Lukowiak1,
Gregg T.
Nagle3, and
Naweed I.
Syed1
1 Respiratory and Neuroscience Research Groups, Faculty
of Medicine, University of Calgary, Calgary, Alberta, Canada T2N 4N1,
2 Department of Anesthesiology, Miyazaki Medical College,
Miyazaki 889-1692, Japan, and 3 Marine Biomedical Institute
and Department of Anatomy and Neurosciences, University of Texas
Medical Branch, Galveston, Texas 77555
Neurotrophic factors have well established roles in neuronal
development and adult synaptic plasticity, but their precise role in
synapse formation has yet to be determined. This paper provides the
first direct evidence that neurotrophic factors in brain conditioned
medium (CM) differentially regulate excitatory and inhibitory synapse
formation. Somata of identified presynaptic and postsynaptic neurons
were isolated from the CNS of Lymnaea and were
cultured in a soma-soma configuration in the presence (CM) or absence
[defined medium (DM)] of trophic factors. In DM, excitatory synapses
did not form. When they were paired in CM or in DM containing
Lymnaea epidermal growth factor (EGF); however, all
presynaptic neurons reestablished their specific excitatory synapses,
which had electrical properties similar to those seen in
vivo. CM-induced formation of excitatory synapses required transcription and de novo protein synthesis, as
indicated by the observations that synapse formation was blocked by the
protein synthesis inhibitor anisomycin and the protein transcription
blocker actinomycin D; the CM factor was inactivated by boiling. They were also blocked by receptor tyrosine kinase inhibitors (lavendustin A, genistein, K252a, and KT5926) but not by inactive analogs (genistin and lavendustin B), suggesting that the effect was mediated by receptor
tyrosine kinases. These results, together with our previously published
data, demonstrate that trophic factors are required for excitatory, but
not inhibitory, synapse formation and extends the role of EGF from cell
proliferation, neurite outgrowth, and survival to excitatory synapse formation.
Key words:
synapse formation; trophic factors; EGF; protein
synthesis; trk receptors; Lymnaea; soma-soma synapses
Copyright © 1999 Society for Neuroscience 0270-6474/99/19219306-07$05.00/0
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