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. 2021 May 26;19(5):e3001279. doi: 10.1371/journal.pbio.3001279

Fig 8. Neurons in the contralateral homotopic cortical area in RHEBp.P37L mice show increased excitability that can be reversed by blocking vesicles release.

Fig 8

(A) Schematic representation of the timeline and experimental conditions of the IUE and ex vivo whole-cell patch clamp recordings showing the targeted cells patched in the targeted S1, L2/3, and nontargeted L2/3 cells in the ipsilateral and contralateral sides. (B) Analysis of the passive membrane properties (Cm, Rm, and Vm) of pyramidal cells in L2/3 (targeted and nontargeted) of control EV mice and targeted and nontargeted pyramidal cells in L2/3 of RHEBp.P37L mice; numbers in the legend indicate number of targeted mice (N = 3, N = 6) and number of cells (n = 37, n = 20, n = 22, n = 21) analyzed; data are presented as mean ± SEM and single data points indicate the values of each cell; for statistics, see S4 Table. (C) Example traces and number of action potentials in response to increasing depolarizing currents; number of mice and cells is as indicated in (B); data are presented as mean ± SEM and the red dashed line represents the pooled mean value ± SEM of targeted and nontargeted cells in empty vector control mice (N = 3) shown separately in S5A Fig, for comparison; for statistics, see S4 Table. (D) Schematic representation of the timeline and experimental conditions IUE, tamoxifen injections, and ex vivo whole-cell patch clamp recordings in L2/3 of ipsilateral and contralateral S1 cortex. (E) Number of action potentials in response to increasing depolarizing currents of cells expressing both RHEBp.P37L and LSL-TeTxLC in L2/3 ipsilateral S1 and nontargeted cells in L2/3 contralateral S1; data are presented as mean ± SEM and dashed lines represent the mean values ± SEM of the pooled control cells from EV mice shown in S5A Fig and of the RHEBp.P37L mice from Fig 8C, for comparison; N = number of mice, n = number of cells analyzed; for statistics see S5 Table. (F) Analysis of passive membrane properties (Cm, Rm, and Vm) of pyramidal cells in L2/3 of mice targeted with RHEBp.P37L and LSL-TeTxLC in ipsilateral S1 and nontargeted cells on the contralateral side; data are presented as mean ± SEM and the dashed lines indicate the mean values of Cm, Rm, and Vm of RHEBp.P37L targeted cells in L2/3 and contralateral cells shown in Fig 8B, for comparison; for statistics see S5 Table. The data underlying this figure can be found in S8 Data. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Cm, capacitance; ctx, cortex; EV, empty vector; GE, ganglion eminence; IUE, in utero electroporation; L2/3, layer 2/3; LSL-TeTxLC, LSL-tetanus toxin light chain; Rm, membrane resistance; Vm, resting membrane potential.