Imaging LTP of presynaptic release of FM1‐43 from the rapidly recycling vesicle pool of Schaffer collateral–CA1 synapses in rat hippocampal slices

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Abstract
Recent studies using the styryl dye FM1‐43 and two‐photon microscopy to directly visualize transmitter release at CA3–CA1 excitatory synapses in the hippocampus have demonstrated that activity‐dependent long‐term potentiation (LTP) and long‐term depression are associated with alterations in vesicular release. It is not known whether particular vesicle pools preferentially express these alterations or what second messenger cascades are involved. To address these questions, we selectively loaded FM1‐43 into the rapidly recycling pool (RRP) of vesicles by use of a brief hypertonic shock to release and load the RRP. We demonstrate here that the induction of LTP can lead to a selective long‐lasting enhancement in presynaptic release from the RRP, while reserve pool kinetics remain unchanged. LTP of RRP release was N‐methyl‐d‐aspartate receptor‐dependent and also required production of the intercellular messenger NO and activation of receptor tyrosine kinase. Measurement of FM1‐43 stimulus‐evoked uptake rates following induction of LTP confirmed that LTP produces more rapid recycling of vesicles released by electrical stimulation, consistent with an enhanced release probability from the RRP.