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1 Department of Molecular and Cellular Neurobiology, Free University, Amsterdam, Holland, The Netherlands
2 Department of Cell Biology and Anatomy, University of Calgary, Calgary, Alberta, Canada
* To whom correspondence should be addressed. E-mail: nisyed{at}ucalgary.ca.
The involvement of neuronal protein synthetic machinery and extrinsic trophic factors during synapse formation is poorly understood. Here we determine the roles of these processes by reconstructing synapses between the axons severed from identified Lymnaea neurons in cell culture, either in the presence or absence of trophic factors. We demonstrate that, although synapses are maintained between isolated pre- and postsynaptic axons for several days, the presynaptic, but not the postsynaptic cell body, however, is required for new synapse formation between soma-axon pairs. The formation of cholinergic synapses between presynaptic soma and postsynaptic axon requires gene transcription and protein synthesis solely in the presynaptic neuron. We show that this synaptogenesis is contingent upon extrinsic trophic factors present in brain conditioned medium (CM). The CM-induced excitatory synapse formation is mediated through receptor tyrosine kinases. We further demonstrate that, although the postsynaptic axon does not require new protein synthesis for synapse formation, its contact with the presynaptic cell in CM, but not in defined medium (DM-no trophic factors), differentially alters its responsiveness to exogenously applied acetylcholine at synaptic compared to extra-synaptic sites. Together, these data suggest a synergetic action of cell-cell signaling and trophic factors to bring about specific changes in both pre- and postsynaptic neurons during synapse formation.
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