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  • Title: Synaptic properties of serotonergic growth cones in developing rat brain.
    Author: Ivgy-May N, Tamir H, Gershon MD.
    Journal: J Neurosci; 1994 Mar; 14(3 Pt 1):1011-29. PubMed ID: 7509861.
    Abstract:
    In order to gain insight into the events that take place when serotonergic growth cones are remodeled into synapses, we tested the hypothesis that neurotransmitter-related properties of presynaptic terminals are already present in these growth cones before synaptogenesis begins. The ontogeny of markers for the specific reuptake of 5-HT and for 5-HT-storing synaptic vesicles was studied in isolated growth cone (IGC) fractions from developing rat brain. High-affinity 3H-imipramine binding (a marker for the plasma membrane 5-HT transporter) was significantly enriched in IGC fractions prepared before the beginning of cortical synaptogenesis [embryonic day 15 (E15) and E20]. Radioautography with 3H-imipramine or 3H-paroxetine (another marker for the transporter) confirmed that the 5-HT transporter is present in the cerebral cortex when it contains serotonergic growth cones, but not serotonergic synapses. Specific uptake of 3H-5-HT was found in IGC fractions as early as E15; this uptake was inhibited by fluoxetine. Electron microscopic radioautography demonstrated directly that growth cones were the structures in IGC fractions that took up 3H-5-HT. The synaptic vesicle protein synaptophysin and a 45 kDa protein found specifically in serotonergic synaptic vesicles, serotonin-binding protein (SBP), were each enriched in IGC fractions from E15 to postnatal day 5; SBP immunoreactivity increased approximately 10-fold between E15 and E20. Endogenous 5-HT was detected in IGC fractions at E15 and increased in amount as development proceeded. The ratio of 5-HT to 5-hydroxyindole acetic acid suggested that 5-HT within growth cones is protected from catabolism by monoamine oxidase. Reserpine-induced depletion of 5-HT, a marker for the vesicular carrier of 5-HT, was apparent in IGC fractions at E20, but not at E15. These data suggest that properties that characterize the presynaptic components of mature serotonergic synapses develop in growth cones before synapses are formed. The early development of these properties may permit neurotransmission to be established rapidly during synaptogenesis or, alternatively, enable 5-HT to play a role in ontogeny.
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