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Visualization of changes in presynaptic function during long-term synaptic plasticity

Abstract

Controversy exists regarding the site of modification of synaptic transmission during long-term plasticity in the mammalian hippocampus. Here we used a fluorescent marker of presynaptic activity, FM 1-43, to directly image changes in presynaptic function during both short-term and long-term forms of plasticity at presynaptic boutons of CA3–CA1 excitatory synapses in acute hippocampal slices. We demonstrated enhanced presynaptic function during long-term potentiation (LTP) induced either chemically (with tetraethylammonium), or by high-frequency (200-Hz) electrical stimulation. Both of these forms of LTP required activation of L-type voltage-gated calcium channels and NMDA receptors in the postsynaptic CA1 neuron. These results thus implied that a long-lasting increase in the efficacy of synaptic transmission is likely to depend, at least in part, on enhanced transmitter release from the presynaptic neuron.

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Figure 1: FM 1-43 visualization of synaptic boutons in acute hippocampal slices.
Figure 2: The rate of FM 1-43 unloading was proportional to the probability of neurotransmitter release.
Figure 3: Chemically induced LTP increased the rate of FM 1-43 unloading.
Figure 4: Induction of LTP with 50- or 100-Hz tetanic stimulation did not change the rate of FM 1-43 unloading.
Figure 5: Induction of LTP with a strong, 200-Hz tetanic stimulation protocol accelerated the rate of FM 1-43 unloading.
Figure 6: Comparison of FM 1-43 unloading kinetics in individual slices before and after induction of 200 Hz LTP revealed an increase in the rate of puncta unloading without an increase in the number or size of presynaptic boutons.

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Acknowledgements

We thank R. Hawkins, E. Kandel, M. Nolan, S. Patterson and V. Unni for comments on the manuscript, and E. Odell for help with artwork. This work was partially supported by a grant from the NIH.

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Correspondence to Steven A. Siegelbaum.

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Zakharenko, S., Zablow, L. & Siegelbaum, S. Visualization of changes in presynaptic function during long-term synaptic plasticity. Nat Neurosci 4, 711–717 (2001). https://doi.org/10.1038/89498

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