Article Figures & Data
Figures
- Figure 1.
Activity-dependent EPSC decrease in the absence of glutamate. A, Sampled presynaptic calcium currents (ICa), EPSCs, and presynaptic Cm induced by the first, sixth, and 10th Train1Hz applied every 30 s. The rate of Cm decay (Ratedecay) in the interval between each 20 ms depolarization during Train1Hz is also plotted. The presynaptic pipette contained 0 mm glutamate (Glu applies to A–G). B, ICa, EPSC, and Cm induced by the first 20 ms depolarization during first (black), sixth (blue), and 10th (red) Train1Hz shown in A are superimposed. C–E, The QICa, ΔCm, and QEPSC induced by each Train1Hz are plotted versus the number of Train1Hz applied every 30 s (C, n = 8), 60 s (D, n = 8), or 300 s (E, n = 5). Data were normalized to the amplitude (Amp) induced by the first Train1Hz, and expressed as the mean ± SEM (applied to all figures). F, QEPSC in C and D are plotted versus the Train1Hz number. G, The mean mEPSC amplitude recorded between Train1Hz applied every 30 s for 15 times (n = 4 synapses, no kynurenic acid in bath).
- Figure 2.
The EPSC does not decrease in the presence of glutamate. A–C, Same arrangements as in Figure 1A,B,D, respectively, except that the presynaptic pipette contained 10 mm glutamate (n = 3 synapses; Train1Hz interval: 60 s).
- Figure 3.
Block of glutamate uptake causes activity-dependent EPSC decrease. A–C, Same arrangements as in Fig. 2A–C (10 mm glutamate), respectively, except that the bath contained bafilomycin A1 (Baf, 6 μm, 20–30 min, n = 4 synapses). D, Capacitance rate of Cm decay (Ratedecay) after each 20 ms depolarization (from ∼300 to 950 ms after depolarization) during the first Train1Hz with 0 mm (left, n = 8, data analyzed from Fig. 1) or 10 mm (right, n = 3, data from Fig. 2) glutamate in the pipette. E, Fura-2 images of a calyx before (−30, −10 s) and after (+20, +40 s) whole-cell break-in with a pipette containing no fura-2. Before taking these images, the calyx was preloaded with fura-2 using another whole-cell pipette containing 150 μm fura-2, which was withdrawn ∼1 min later and the calyx resealed.
- Figure 4.
Large recycling pool. A, The accumulated QEPSC (ΣQEPSC, circles) induced by Train1Hz applied every 30 s is plotted versus the Train1Hz number in the absence of glutamate (analyzed from Fig. 1C). ΣQEPSC is normalized to the QEPSC induced by the first 20 ms depolarization of the first Train1Hz, i.e., the RRP size. Black curve is a monoexponential fit with a plateau value of 46 times the RRP size. The red curve is the predicted change assuming random mixture of recycling vesicles (see Results). B, Sampled EPSCs induced by a presynaptic AP-e train at 5 Hz for 180 s (bar) with no glutamate in the pipette (applies to B–E). C, The mEPSC amplitude (n = 8 synapses) in the first and the last 30 s during the AP-e train at 5 Hz for 180 s. Sampled mean mEPSCs at the same time period from a synapse shown in B are also plotted above the bar graph (left: averaged of 97 mEPSCs; right: 93 mEPSCs). D, The cumulative EPSC amplitude (ΣEPSC Amp) and the number of vesicles (Nvesicle) released by an AP-e train at 5 Hz (n = 8 synapses). Data are shown as the mean (solid curve) ± SEM (gray). E, The amplitude of EPSCs (bottom) and sampled EPSCs (top, from one synapse) induced by an AP-e at ∼30 s before (left) and after (right) an AP-e train at 5 Hz for 180 s (n = 8 synapses). F–H, Same arrangements as in C–E, respectively, except that the pipette contained 10 mm glutamate and the bath contained 300 μm CPPG (n = 5 synapses). mEPSC traces in F: left, averaged of 47 mEPSCs; right, 56 mEPSCs. I–K, Same arrangements as in C–E, respectively, except that the pipette contained 10 mm glutamate (no CPPG in bath, n = 5 synapses). mEPSC traces in I: left, averaged of 136 mEPSCs; right, 58 mEPSCs. L–N, Same arrangements as in C–E, respectively, except that whole-cell AP-e was replaced with an extracellular stimulation that induced axonal action potentials (see Results), and the bath contained 6 μm bafilomycin A1 (n = 4, synapses). mEPSC traces in M: left, 44 mEPSCs; right, 51 mEPSCs.
In this issue
