Fig. 1: BTSP in single dendritic spines with glutamate uncaging.
From: Dendritic, delayed, stochastic CaMKII activation in behavioural time scale plasticity
a, Top, schematic of the experimental set up. Bottom, BTSP and cBTSP protocols: 5 uncaging pulses were given at a spine before (orange) or after (purple) a 600 pA current injection (300 ms width) with a 750 ms delay. b, Representative image of a CA1 dendrite in an organotypic hippocampal slice (top left), uncaging-evoked EPSPs (ten recordings and average) before and after BTSP from the spine (top right) and electrical trace during BTSP protocol (bottom). c, Same as b, but with the cBTSP protocol. d,e, Averaged time course (d) and summary of magnitude of potentiation (25–30 min) (e) of uncaging-evoked normalized (Norm.) EPSP amplitudes (0–5 min) at stimulated (Stim) (n = 26) and adjacent (Adj) spines (n = 17) in response to BTSP (arrow). Two-tailed unpaired t-test, (t40 = 4.293, P = 0.0001). f,g, Same as d and e, but with the cBTSP protocol (stimulated spines, n = 15; adjacent spines, n = 14). Two-tailed unpaired t-test, (t27 = 2.366, P = 0.025). h, Left, representative Alexa-594 filled image of a CA1 neuron and dendritic shaft in acute hippocampal slices, in which BTSP was induced. Right, uncaging-evoked EPSP traces of stimulated spine (ten recordings and average) before and after BTSP induction. i,j, Same as d and e, but with the BTSP protocol in acute hippocampal slices (stimulated spines, n = 12; adjacent spines, n = 8). Two-tailed unpaired t-test (t18 = 2.276, P = 0.0353). k,l, Same as d and e, but with the BTSP protocol under various conditions (basal, n = 10; distal, n = 8; TTX stimulated, n = 14, adjacent, n = 11; APV, n = 12; QX314, n = 12; MK801, n = 12; T286A stimulated, n = 15; adjacent, n = 14). Two-way analysis of variance (ANOVA) (F17,176 = 4.395, P = 0.00000016) with Dunnett’s multiple comparison test (all P values are shown in the source data file). The data in d–g and i–l are presented as the mean ± s.e.m. The plus symbol in images depicts a BTSP-stimulated spine. *P < 0.05, ***P < 0.001. Scale bars, 2.5 μm (b,c), 5 μm (h, right) or 50 μm (h, left).
