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. 2017 Oct 10;4(5):ENEURO.0332-16.2017. doi: 10.1523/ENEURO.0332-16.2017

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Copyright © 2017 Allen et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license, which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.

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Figure 5. — Larger current density in cells expressing e18a-Ca_V2.2 compared to Δ18a-Ca_V2.2. A, Peak Ca_V2.2 current−voltage relationships in tsA201 cells expressing Δ18a-Ca_V2.2 or e18a-Ca_V2.2 with Ca_Vβ₃ and Ca_Vα₂δ-₁ subunits. Values are mean ± SE and N values are shown in parentheses. Data were compiled from multiple different transfections and gathered by three different experimenters. Individual current−voltage relationships were fit by Boltzmann-linear functions and Boltzmann activation midpoint (V_1/2) and slope (k) estimated. V_1/2 ± SE (mV); Δ18a-Ca_V2.2: -1.3 ± 0.9; e18a-Ca_V2.2: -4.8 ± 0.6 (Student’s unpaired t test, p = 0.0001^e). k ± SE (mV); Δ18a-Ca_V2.2: 5.2 ± 0.5; e18a-Ca_V2.2 = 4.4 ± 0.25. B, Ca_V2.2 current−voltage relationships in cells expressing Δ18a-Ca_V2.2 or e18a-Ca_V2.2 with Ca_Vβ₂ and Ca_Vα₂δ-₁. Values are average ± SE. C, Cumulative frequency plots of peak Ca_V2.2 current densities measured at 5 mV in cells expressing Δ18a-Ca_V2.2 or e18a-Ca_V2.2 with either Ca_Vβ₂ or Ca_Vβ₃. Bin size was 25 pA/pF for all plots. D, Peak Ca_V2.2 current density values are shown for individual cells (open symbols), mean (filled symbol), median (horizontal line), 25th-75th interquartile range (box), and whiskers (range) in tsA201 cells expressing Δ18a-Ca_V2.2 or e18a-Ca_V2.2 with Ca_Vβ₃ and Ca_Vβ₂. Mean (median) ± SE in pA/pF at 5 mV were for Δ18a-Ca_V2.2/Ca_Vβ₃: 95.9 (66.1) ± 17.1; e18a-Ca_V2.2/Ca_Vβ₃: 194.5 (167.7) ± 19.4; Δ18a-Ca_V2.2/Ca_Vβ₂: 42.9 (32.3) ± 9.4; e18a-Ca_V2.2/Ca_Vβ₂ 53.9 (41.6) ± 7.1 (one-way ANOVA on ranks, Bonferroni corrected p = 0.0001^f). E, Amino acid sequence of wild-type e18a exon (+18a), and two mutants. Seven serines were substituted with alanines (7S/7A) in both mutants; one mutant contains an additional lysine to alanine substitution (7S/7A, K/A). F, Peak Ca_V2.2 I-V relationships in cells expressing Δ18a-Ca_V2.2, e18a-Ca_V2.2 WT, e18a-Ca_V2.2 7S/7A, and e18a-Ca_V2.2 7S/7A, K/A. Mean and SE for each condition and N values in parentheses. Mean (median) ± SE in pA/pF at 5 mV for WTe18a-Ca_V2.2: -248.64 (232.1) ± 24.6; 7S/7A: -203.8 (250.3) ± 25.9; 7S/7A,K/A: -190.2 (170.2) ± 29.9 (one-way ANOVA on ranks, Bonferroni corrected, p_WT:7S/7A = 0.262^g and one-way ANOVA on ranks, Bonferroni corrected, p_{WT: 7S/7A,K/A} = 0.213^g).