Fig. 3: Characterization of CKAR3 in neurons from brain slices.
From: A high-performance genetically encoded sensor for cellular imaging of PKC activity in vivo
Representative images (a), average traces (b), and quantification (c) of CKAR sensors responding to acetylcholine (Ach, 20 μM) in CA1 neurons of cultured slices. n (cells) = 7, 9,9 for CKAR, CKAR2, CKAR3, respectively. One-way ANOVA [F(2, 22) = 14.11, p = 0.0001] with Bonferroni post hoc test. (p = 2 × 10–4 and 1 × 10–3, respectively). Response traces of CKAR3 in CA1 neurons to acetylcholine compared to pre-treatment with scopolamine (scop., 1 μM) (d), mutCKAR3 (e), and their quantifications (f). In panel (f) from left to right, n (cells) = 9, 5, and 3. One-way ANOVA [F(2, 14) = 6.73, p = 0.009] with Bonferroni post hoc test [p = 0.02 and 0.04, respectively]. Representative images (g), average traces (h), and collective quantifications (i) of CKAR3 in M1 neurons in response to acetylcholine (responders are threshold 2x baseline s.d.) or with scopolamine pretreatments (blue), or mutCKAR3 responding to acetylcholine. In panel (i) from left to right, n (cells/mice) = 5/3, 15/3, 13/2, 14/2, and 15/3. One-way ANOVA [F(4, 57) = 70.74, p = 1.9 × 10–21] with Bonferroni post hoc test. p = 0 for all comparisons. j mEPSC amplitudes from paired adjacent L5 M1 pyramidal neurons with and without CKAR3 expression. n (pairs/mice) = 5/4, two-sided paired t test [t(4) = 0.7, p = 0.52]. k Representative recordings (left) at holding potential of –70 (red) and +55 (black) mV and quantified AMPA/NMDA receptor current ratios (right) from paired adjacent L5 M1 pyramidal neurons with and without CKAR3 expression. n (pairs/mice) = 5/2, [W = 9, p = 0.79], two-sided Wilcoxon signed-rank test. l Normalized amplitudes of slow afterhyperpolarizations (sAHPs) in response to a moderate concentration of muscarine (100 nM), with the majority followed by scopolamine (scop.) administrations, in CA1 neurons with and without CKAR3 expression. From left to right, n (cells/slices) = 8/8, 8/8, 6/6, 7/7, 7/7, and 5/5. Kruskal-Wallis test. n.s.: not significant [H(1) = 0.48, p = 0.51]. Mean (dark lines and bars) and s.e.m. (shaded area and error bars) are shown. n.s.: not significant, *: p < 0.05, **: p < 0.01, ***: p < 0.01. Source data are provided as a Source Data file.
