The Journal of Neuroscience, February 22, 2006, ():
Characterization of AMPA Receptors Targeted by the Climbing Fiber Transmitter Mediating Presynaptic Inhibition of GABAergic Transmission at Cerebellar Interneuron-Purkinje Cell Synapses
J. Neurosci. Satake et al.
26: 2278
Supplemental data
Files in this Data Supplement:
- supplemental material
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Supplementary Figure 1. Electrophysiological and pharmacological profiles of AMPARs in BG and BCs. A, C, Effects of PhTX (10 µM) on the AMPA-induced currents recorded from a BG (A) and a BC (C). AMPA was iontophoretically applied with constant current pulses (40–70 mA for 10–30 ms) to each cell in normal ACSF (upper two sets) and a Na+ free, 10 mM Ca2+ ACSF (lower two sets) before (control) and after treatment with PhTX (10 µM) while the holding potential was changed in each set of records from –80 to +60 mV in 20 mV steps. Cyclothiazide (10 µM) was added to ACSF throughout all recordings. B, D, I-V relations of AMPA-induced currents in BG (B) and BCs (D) in the absence (open circles) or presence (closed circles) of PhTX (10 µM). The I-V curves were obtained in normal ACSF (upper) and Na+ free, 10 mM Ca2+ ACSF (lower) in (B) and (D). Each point represents the mean ± SEM (n = 5–7).
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Supplementary Figure 2. Immunohistochemical staining of AMPARs and GAD65 in the cerebellar cortex. A, B, Confocal micrographs of double immunofluorescence detected by antibodies for AMPARs (GluR2/3 subunits, green) and GAD65 (red). Sections in (A) and (B) were derived from 8- and 2-week-old rats, respectively. C–E, Higher magnification images of a rectangular area indicated in (A): GluR2/3 (C), GAD65 (D) and merged (E). Note co-localized products (yellowish dots) that surround the PC soma. F–H, Enlarged from the micrograph (B) as in (C–E): GluR2/3 (F), GAD65 (G) and merged (H). Arrows indicate the pinceau, and arrowheads represent examples for co-localized immunoreaction products. No enzymatic treatments were applied to these preparations. Scale bars (µm): 20 for (A) and (B), and 10 for (C–H).
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Supplementary Figure 3. Co-localization of immunolabeling of GluR2/3 subunits in GFP-tagged terminals derived in the cerebellar cortex from GAD67-GFP knock-in mice. A–C, Fluorescence channel of GFP-tagged GABAergic cells; somata of most interneurons in the molecular layer appear bright; dotted profiles indicate bodies of two PCs (also shown in B and C). Scale bar, 30 µm (applies to A–C). B, Fluorescence channel of GluR2-Rhodamine in the same tissue fragment; dotted circles
