Neuron
Volume 68, Issue 4, 18 November 2010, Pages 639-653
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Neurotechnique
Single-Synapse Analysis of a Diverse Synapse Population: Proteomic Imaging Methods and Markers

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Summary

A lack of methods for measuring the protein compositions of individual synapses in situ has so far hindered the exploration and exploitation of synapse molecular diversity. Here, we describe the use of array tomography, a new high-resolution proteomic imaging method, to determine the composition of glutamate and GABA synapses in somatosensory cortex of Line-H-YFP Thy-1 transgenic mice. We find that virtually all synapses are recognized by antibodies to the presynaptic phosphoprotein synapsin I, while antibodies to 16 other synaptic proteins discriminate among 4 subtypes of glutamatergic synapses and GABAergic synapses. Cell-specific YFP expression in the YFP-H mouse line allows synapses to be assigned to specific presynaptic and postsynaptic partners and reveals that a subpopulation of spines on layer 5 pyramidal cells receives both VGluT1-subtype glutamatergic and GABAergic synaptic inputs. These results establish a means for the high-throughput acquisition of proteomic data from individual cortical synapses in situ.

Highlights

► Resolution of individual synapses in situ by high-throughput array tomography ► Measurement of multiple (e.g., 17) molecular markers at individual synapses ► Discrimination of defined subtypes of glutamate and GABA synapses ► Detection of both VGluT1-type glutamate and GABA synapses on single spines

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