Paired pre–postsynaptic recordings can be integrated with biocytin labeling and morphological analysis, allowing correlative investigation of synapse structure and function. Single presynaptic hippocampal mossy fiber terminals are stimulated minimally invasively in the bouton-attached configuration, in which the cytoplasmic content remains unperturbed, or in the whole-bouton configuration, in which the cytoplasmic composition can be precisely controlled. Slice preparation takes ~2 h, recording ~3 h and post hoc morphological analysis 2 d. Here, we describe a protocol for improved subcellular patch-clamp recording in rat and mouse brain slices, with the synapse in a largely intact environment. Cortical presynaptic terminals have been largely inaccessible to direct presynaptic recording, due to their small size. However, this has been achieved at only a limited number of model synapses, including the squid giant synapse and the mammalian calyx of Held. Rigorous investigation of synaptic transmission requires analysis of unitary synaptic events by simultaneous recording from presynaptic terminals and postsynaptic target neurons.
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