The Journal of Neuroscience, February 21, 2007, ():
Number and Density of AMPA Receptors in Individual Synapses in the Rat Cerebellum as Revealed by SDS-Digested Freeze-Fracture Replica Labeling
J. Neurosci. Masugi-Tokita et al.
27: 2135
Supplemental Data
Files in this Data Supplement:
- supplemental material
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Supplemental Figure 1. AMPAR labeling at PF-PC synapses of GluR2 and GluR3 double knockout mice
Immunolabeling for AMPAR (5 nm gold particles) was mostly abolished in PF-PC synapses identified by labeling for GluRδ2 (15 nm). Cross-fracture face of the parallel fiber terminal is clearly seen in (A). One PF terminal makes synapses with two PC spines in (B). Unidirectional shadowing was used. Scale bars: 200 nm.
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Supplemental Figure 2. Comparison of PF-PC synapses in replicas prepared with different concentrations of fixative
(A-C) AMPAR labeling at PF-PC synapses of 2% paraformaldehyde-fixed mouse. (A) In 2% paraformaldehyde-fixed tissue, most synapses were fractured at the neck of the spine (arrows). Occasionally incomplete areas of the synaptic face were visible (enclosed area is enlarged in B). (C) Density of immunolabeling for AMPAR (5 nm) at PF-PC synapses identified by GluRδ2 labeling (15 nm) is highly variable. (D, E) AMPAR labeling at PF-PC synapses of 0.5% paraformaldehyde-fixed mouse. Many synapses (arrows) were flattened and complete areas of the synaptic face were visible. Enclosed area in (D) is enlarged in (E). Scale bars: 500 nm.
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Supplemental Figure 3. Homogeneous density of AMPAR labeling in CF-PC and PF-interneuron synapses revealed with different antibody concentrations
Number and density of gold particles for AMPARs in PF-PC (A-D), CF-PC (E-H) and PF-interneuron synapses (I-L) are shown. Similar patterns of AMPAR labeling with two different antibody concentrations (3 μg/ml in A, C, E, G, I and K, 0.3 μg/ml in B, D, F, H, J, and L) indicate that homogeneous density of AMPAR labeling in CF-PC and PF-interneuron synapses is not caused by saturation.
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Supplemental Figure 4. Density of extrasynaptic AMPAR labeling
No correlation was detected between the density of extrasynaptic particles and distance from the center of PF-PC synapses.
