Dendritic glutamate release produces autocrine activation of mGluR1 in cerebellar Purkinje cells -- Shin et al. 105 (2): 746 Data Supplement - HTML Page - index.htslp -- Proceedings of the National Academy of Sciences

Shin et al. 10.1073/pnas.0709407105.

Supporting Figures

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SI Figure 5
SI Figure 6
SI Figure 7
SI Figure 8

SI Figure 5

Fig. 5. DISC is present at 32°C. Single exemplar traces illustrate that DISC could be observed (a) at 32°C. Recordings at room temperature revealed that DISC was modestly attenuated with the addition of 500 nM TTX (red traces; b) and was present in Purkinje cells presoaked in an NMDA receptor antagonist, 20 mM (R)-CPP (c), or internally perfused with 135 mM L-glutamate (d). (Scale bars: 200 pA, 2 sec.) The high-pass-filtered traces are displayed using a range of -25 to 25 pA.

SI Figure 6

Fig. 6. The somatic depolarization protocol used to trigger DISC evokes a Ca²⁺ transient in the Purkinje cell dendrites and soma. Purkinje cells were filled with a morphological dye, 0.3 mM Alexa Fluor 594 hydrazide, and a Ca²⁺ indicator, 0.3 mM Fluo-5F. (a Upper) A projected image of Purkinje cell was constructed from a series of z-section images of Alexa 594 hydrazide fluorescence. (Scale bar: 20 mm.) (Lower) A 128 ´ 33 pixel ROI (corresponding to the yellow box, Upper) was set to detect changes in fluorescence from Fluo-5F in somatic, proximal, and distal dendritic areas (blue boxes). The white box represents the area where the background fluorescence was obtained. (b) The same depolarizing train used previously to induce DISC was delivered (as indicated by the black bar) and Ca²⁺ transients (measured as DF/F₀) were plotted before (black) and after (red) the application of 250 mM CdCl₂.

SI Figure 7

Fig. 7. Depolarization-evoked Ca²⁺ transients are not affected by DISC-attenuating drugs. A Ca²⁺ transient was evoked by a single 10-msec depolarizing pulse to 0 mV and measured using laser-scanning confocal microscopy. The DF/F₀ value 100 msec after the onset of Ca²⁺ transient was calculated and the average ratios of the value at t = 40 min/t = 20 min were plotted (n = 5 for each group). The same concentrations of the drugs were applied at t = 20 min (con, control; SKF, SKF96365; Bo, BoTxD; DM, DMSO; CP, CPCCOEt; JNJ, JNJ16259685; Baf, bafilomycin A1).

SI Figure 8

Fig. 8. DISC is not attenuated in EAAT3/EAAT4 DKO mice. Single exemplar records are shown. (Scale bars: 300 pA, 2 sec.) The high-pass-filtered traces are displayed using a range of -25 to 25 pA.