Journal: The Journal of Neuroscience
Article Title: Number and Density of AMPA Receptors in Individual Synapses in the Rat Cerebellum as Revealed by SDS-Digested Freeze-Fracture Replica Labeling
Figure Lengend Snippet: AMPAR distribution in adult PF–PC, CF–PC, and PF–interneuron synapses. A , Immunolabeling for GluRδ2 (10 nm particles) is observed in IMP clusters found on the E-face of PC spines, showing very homogeneous labeling within individual synapses and also in different synapses. B , Immunolabeling for AMPAR (5 nm) with high (left) or low (right) densities of immunogold is observed in PF–PC synapses identified by labeling for GluRδ2 (15 nm). C , Some synapses are fractured with partial synaptic areas (open arrow), whereas others are flattened and fractured completely, showing whole areas of synapses (arrows). D , E , Localization of AMPAR labeling (5 nm) in the CF–PC synapses identified by labeling for VGluT2 (15 nm) on opposing P-face of presynaptic membrane in the molecular layer. Note that the central area of E was not shadowed with platinum. This makes the 5 nm gold easily discerned on IMPs (above bottom center), but they are more difficult to discern when the IMPs were shadowed with platinum (right side). F , Immunolabeling for Kv4.3 (15 nm) is diffusely observed on the P-face of interneurons. Smaller particles are for mGluR1α, which is not related to this report. G , Immunolabeling for AMPARs (5 nm) is dense and homogeneous in IMP clusters (arrows) in E-face of interneurons identified by matching the complementary P-face shown in F , which is labeled for Kv4.3. Inset shows one of the synapses (open arrow). Rotary shadowing was used for this figure. Scale bars, A–C , F , G , 250 nm; D , E , 100 nm.
Article Snippet: The specificity of pan-AMPA , GluR2 , VGluT1 and VGluT2 , and Kv4.3 ( ) (Alomone Labs) antibodies has been extensively characterized previously.
Techniques: Immunolabeling, Labeling