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. 2021 Feb 22;118(9):e1916337118. doi: 10.1073/pnas.1916337118

Fig. 2.

Fig. 2.

Electrophysiological effects of rAb-IP2 on murine hippocampal CA1 pyramidal neurons. (A) Exemplary traces showing sIPSCs recorded from rOCB-MS3-s1– (control; Left) and rAb-IP2–incubated (Right) pyramidal neurons in the hippocampal region CA1. (B) Scatter plot showing that incubation with rAb-IP2 leads to a significant decrease of the number of sIPSCs recorded in an overall period of 10 min in comparison to control (rAb-IP2: 371.8 ± 65.8, n = 6; rOCB-MS3-s1: 1,316.0 ± 171.8, n = 5; unpaired Student’s t test: t = 5.511, degrees of freedom [df] = 9, P = 0.0004). (C) Bar graphs show a tendency to reduced amplitudes of sIPSCs in rAb-IP2–incubated pyramidal neurons in comparison to controls (rAb-IP2: 19.19 ± 0.60 pA, n = 3; rOCB-MS3-s1: 23.9 ± 5.6 pA, n = 4; Mann–Whitney U test: P = 0.4). (D) Scatter plot showing the RMP of rAb-IP2–incubated neurons and controls (rAbIP2: −67.20 ± 1.56, n = 10; rOCB-MS3-s1: −64.5 ± 1.83 mV, n = 8; unpaired Student’s t test: t = 1.128, df = 16, P = 0.2764). (E) Scatter plot showing the Rin of rAb-IP2–incubated neurons and controls (rAb-IP2: 145.6 ± 10.3 MΩ, n = 9; rOCB-MS3-s1: 104.3 ± 20.2 MΩ, n = 7, respectively; Mann–Whitney U test: P = 0.11). (F) Scatter plot showing the whole-cell capacitance of rAb-IP2–incubated neurons and controls (rAb-IP2: 16.12 ± 1.44 pF, n = 10; rOCB-MS3-s1: 14.07 ± 1.82 pF, n = 8, respectively; unpaired Student’s t test: t = 0.8988, df = 16, P = 0.382). (G) Bar graphs showing quantification of APs generated in response to depolarizing current steps of increasing intensity (from +20 to +240 pA, 20-pA increments, duration of 2.5 ms) in rAb-IP2–incubated pyramidal neurons and controls [mixed design two-way ANOVA, main effect of rAb-IP2- vs. rOCB-MS3-s1-incubation: F(1,165) = 6.538, P = 0.02, Sidak’s multiple comparisons test for rAb-IP2- vs. rOCB-MS3-s1: at 120 pA, P = 0.0459; at 140 pA, P = 0.0127; at 160 pA, P = 0.0115; at 180 pA, P = 0.0055; at 200 pA, P = 0.0076 and at 220 pA, P = 0.0257]. Of note, for both groups the input/output relation reflected by the proportional increase of the number of APs in response to the increasing current intensity did not differ, indicating a lower threshold for AP firing in rAb-IP2–incubated cells rather than a defective AP generation in rOCB-MS3-s1–incubated cells (main effect of input/output relation: F(11, 165) = 24,90, P < 0,0001, Sidak’s multiple comparisons test: no significance between groups). (H) Exemplary traces showing APs generated in response to a +140-pA current step at resting membrane potential in rAb-IP2–incubated (red) and rOCB-MS3-s1–incubated (control; black) pyramidal neurons.