Research paperDevelopmental maturation of synaptic vesicle cycling as a distinctive feature of central glutamatergic synapses
Section snippets
Cell culture
Microisland cultures of neocortical neurons were prepared as described previously Segal 1991, Lessmann and Heumann 1997. In brief, the occipital neocortex was removed from Wistar rat pups at P0-P2, and the tissue was dissociated after trypsin (0.1%) treatment. Dissociated neurons were seeded on cultured glial cells at a density of 2.5×105 cells/dish. For attachment to glial islands, dissociated cells were incubated for 4–5 h in Dulbecco’s Minimal Essential Medium containing fetal calf serum
Developmental increase in the size of the cycling synaptic vesicle pool in cultured neocortical neurons
To characterize the developmental maturation of the presynaptic vesicle population, we visualized cycling synaptic vesicles with the styryl dye FM1-43 Betz and Bewick 1992, Cochilla et al 1999, Murthy 1999. Saturating stainings (incubation times >2 min) were performed using a depolarizing (50-mM K+) extracellular solution to stimulate synaptic vesicle cycling in the presence of FM1-43. This procedure led to a punctate labeling of functional presynaptic terminals (Fig. 1). To confirm that
Discussion
The characterization of the cellular and molecular mechanisms involved in formation and maturation of synapses is of crucial importance for the understanding of mammalian CNS development. In particular, differentiation of presynaptic terminals plays a major role in the establishment of mature functional properties at central synapses. Ultrastructural studies have well documented that synaptic vesicles are progressively accumulated inside developing presynaptic terminals (Blue and Parnavelas,
Acknowledgements
We want to thank Dr. P. Caroll for providing BDNF knock-out mice. We further thank Dr. M. Hartmann for help with FM1-43 imaging and H. Bartel for excellent technical assistance. This work was supported by the Deutsche Forschungsgemeinschaft.
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2017, Journal of EthnopharmacologyCitation Excerpt :At the presynaptic terminal level, a modification of the size of the pool of synaptic vesicles available for release is a sign of functional synaptic plasticity (Malenka and Siegelbaum, 2001). To assess potential modulatory effects of RPE and puerarin on functional presynaptic plasticity in cultured hippocampal neurons, we visualized cycling synaptic vesicles using the styryl dye FM1-43 (Bhuiyan et al., 2015a; Mohibbullah et al., 2016; Mohrmann et al., 2003). To stimulate exocytosis and eventually visualize cycling synaptic vesicles, hippocampal cultures were incubated for 3 min in a depolarizing extracellular solution containing FM1-43 (10 μM), which caused the saturated staining of cycling synaptic vesicles.
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2008, NeuroscienceCitation Excerpt :In contrast to these postsynaptic mechanisms in input specificity, the sensitivity of the thalamocortical input to higher stimulation frequencies and the selective expression of paired-pulse depression at this synapse most likely reflect the depletion of the readily releasable pool of vesicles (for review Thomson, 2000). A slow time course for this process is generally observed in young synapses (Mohrmann et al., 2003). Our data argue for a slower vesicle recycling process at thalamocortical synapses as compared with intra-subplate synapses.
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Present address: Department of Physiology and Pathophysiology, Johannes-Gutenberg-University Mainz, Duesbergweg 6, D-55099 Mainz, Germany.