Fig. 5.
Two-photon imaging of the effect of CfICR and of T-588 on [Ca2+]i dynamics in PCs. (A) Diagram of caffeine increase and T-588 block of CfICR. (B) Effect of caffeine on response to PC depolarization. (a) Low-power image of PC filled with calcium-sensitive dyes (calcium green 1 and 2). (b and c) Magnified views of the region outlined by the box in a during PC depolarization before (b) and after (c) addition of 10 mM caffeine to the bathing solution. (C) Effect of T-588. (a) Low-power image of dye-filled PC. (b and c) Magnified views of the region outlined by the box in a during PC depolarization in the presence of T-588 before (b) and after (c) addition of caffeine (10 mM). (D) The mean ratio of stimuli-induced calcium changes in a PC dendrite before (first green column) and after (blue column) caffeine infusion (10 mM). Mean ratio in the presence of T-588 before (second green column), and after (red column) addition of 10 mM caffeine. Note that T-588 reduced the increase of [Ca+]i by caffeine (mean ± SEM; *, P < 0.05, paired t test). (E) Time course of calcium changes immediately after depolarizing pulse in a PC dendrite under control conditions before (Left green symbols) and after (blue symbols) caffeine infusion (10 mM). Same protocol but in the presence of T-588 (10 μM) before (Right green symbols) and after (red symbols) addition of caffeine (10 mM) (mean ± SEM). A significant difference was seen between control before and after caffeine infusion (P < 0.05, Scheffé's post hoc test). (Scale bars, 20 μm in Ba and Ca; 2 μm in Bb, Bc, Cb, and Cc.)