Supplemental Information. C. elegans AWA Olfactory Neurons Fire. Calcium-Mediated All-or-None Action Potentials
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1 Cell, Volume 175 Supplemental Information C. elegans AWA Olfactory Neurons Fire Calcium-Mediated All-or-None Action Potentials Qiang Liu, Philip B. Kidd, May Dobosiewicz, and Cornelia I. Bargmann
2 Table S1. Strain list, related to Figures 1-7. Strain Genotype Comment Figure CX3262 kyis37[odr-10::gfp] II Fig 1-5, 7, S1- S6 CX12516 kyex3488[gcy-13::gfp 100ng + unc- RIM::GFP Fig 1, S1A, B 122::dsRed 10ng] CX13495 kyex4059[mod-1::gfp 50ng + unc- AIY::GFP Fig 1, S1A, B 122::dsRed 10ng] PY1322 oyis18[gcy-8::gfp] X AFD::GFP Fig 1, S1A, B CX17063 unc-13(e51) I; kyis37 II unc-13; Fig 2A CX4312 kyis37 II; osm-9(ky10) IV osm-9; Fig 2G, H CX16801 kyis37 II; egl-19(n582) IV egl-19; Fig 2E CX6889 kyis37 II; unc-2(e55) X unc-2; Fig 2G, S2B CX17062 kyis37 II; cca-1(ad1650) X cca-1; Fig 2G, S2B CX17189 kyis37 II; nca-2(gk5) III; nca-1(gk9) IV nca-1; nca-2; Fig 2G, S2B CX16803 (ok1581) kyis37 II ; Fig 3C-G, 3H, 4F, S5B, D, S7 CX16802 kyis37 II; slo-2(nf101) X slo-2; Fig 3H, 4H, S3A CX16955 kyis37 II; shl-1(ok1168) IV shl-1; Fig 3H, S3A CX17086 kyis37 II; slo-1(eg142) V slo-1; Fig 3H, 4I, S3A CX17216 kcnl-2(ok2818) I; kyis37 II kcnl-2; Fig S3A CX17187 kyis37 II; egl-2(rg4) V egl-2; Fig S3A CX17326 shw-1(r1159) II; kyis729[gpa- shw-1; Fig S3A 6::GCaMP6f 40ng + unc-122::dsred 15ng] CX17289 kyis37 II; egl-36(n2332sa577) X shw-2; Fig S3A (egl-36=shw-2) CX17087 kyis37 II; exp-3(n2372) V exp-3(gf); Fig S3A CX17039 (ok1581) kyis37 II; slo-2(nf101) slo-2; ; Fig 3H, 4J, S3B X CX17085 (ok1581) kyis37 II; shl- 1(ok1168) IV shl-1; ; Fig 3H, 4G, S3B, S5D CX17101 (ok1581) kyis37 II; slo-1(eg142) slo-1; ; Fig 3H, S3B V CX17217 kcnl-2(ok2818) I; (ok1581) kcnl-2; ; Fig S3B, S5B kyis37 II CX17188 (ok1581) kyis37 II; egl-2(rg4) V egl-2; ; Fig S3B CX17290 kqt-3(tm0542) (ok1581) kyis37 kqt-3; ; Fig S3B II CX17258 (ok1581) kyis37 II; slo-1(eg142) slo-1; slo-2; ; Fig 3H, S3B V; slo-2(nf101) X CX17041 kyis37 II; egl-19(ad695) IV egl-19(gf); Fig 5E-F CX17462 kyex6119[gpa-6::gcamp6f 40ng + AWA::GCaMP6f Fig 6A, B, F CX17463 unc-122::dsred 15ng] (ok1581) II; kyex6120[gpa- 6::GCaMP6f 40ng + unc-122::dsred 15ng] 1 ; AWA::GCaMP6f Fig 6C-E, G
3 Table S2. Effects of extracellular solutions and genetic mutations on spiking, related to Figure 2. Wild Type Mutants Extracellular solution Recordings with action potentials /total recordings (recorded in 2 mm [Ca 2+ ] o ) 0 mm [Ca 2+ ] o 0/3 unc-13(e51) 4/4 1 mm [Ca 2+ ] o 0/3 osm-9(ky10) 6/8 2 mm [Ca 2+ ] o 44/49 egl-19(n582lf) 0/4 5 mm [Ca 2+ ] o 5/5 unc-2(e55) 3/4 8 mm [Ca 2+ ] o 2/2 cca-1(ad1650) 4/5 8 mm [Ca 2+ ] o + Nemadipine-A 0 mm [Na + ] o 2/2 1/7 nca-1(gk9); nca-2(gk5) 2/2 Recordings with action potentials /total recordings 2
4 Table S3. AWA action potential features, related to Figures 1-5. Genotype or condition n Threshold Amplitude Width AHP Threshold Amplitude Width (ms) AHP ± ± ± ± 0.26 slo ± 0.92 **** ± 1.33 ns ± 2.43 ns 6.83 ± 1.18 **** unc ± 0.56 **** ± 1.07 ns ± 2.53 ns 3.58 ± 1.08 ns cca ± 0.44 ns ± 1.25 ns ± 2.44 ns 2.35 ± 0.69 ns + 4-AP ± 0.96 **** ± 1.65 **** ± 3.58 **** 7.29 ± 0.92 **** ± 0.64 ns ± 1.05 **** ± 8.88 **** 6.82 ± 1.22 **** shl ± 0.33 ns ± 0.76 ns ± 1.55 ns 3.61 ± 0.43 ** unc ± 1.07 ns ± 1.96 ns ± 3.75 ns 4.75 ± 1.49 ns slo ± 0.40 ns ± 1.13 ns ± 1.22 ns 3.32 ± 0.78 ns egl-19(gf) ± 0.48 ns ± 0.58 ns ± 1.24 ns 5.77 ± 1.10 **** nca-1;nca ± 0.62 *** ± 1.41 ns ± 2.17 ns 3.13 ± 0.66 ns osm ±0.35 **** ± 0.73 ns ± 1.26 ns 3.44 ± 0.44 * shw ±0.71 ** ± 0.67 ns ± 1.97 ns 3.32 ± 0.61 ns exp-3(gf) ± 0.69 * ± 1.60 ns ± 2.07 ns 3.37 ± 0.72 ns kcnl ± 1.02 **** ± 0.69 ns ± 2.02 ns 3.75 ± 0.52 * 5 mm [Ca 2+ ] o ±0.69 **** ± 1.13 **** ± 1.30 ** 7.04 ± 0.81 **** 8 mm [Ca 2+ ] o ± 0.65 **** ± 1.27 *** ± 1.03 ns 4.62 ± 0.32 ns exp ± ± ± ± 1.93 egl ± ± ± ± 1.25 shw ± ± ± ± 1.11 Threshold Amplitude Width (ms) AHP ± ± ± 8.88] 6.82 ± 1.22 slo-1; ± 0.86 ns v 1.31 ns ± 10.09] ns [1.12 ns slo-2; ± 0.75 ** ± 1.16 ns ± 13.34] ns 8.87 ± 2.18 ns kcnl-2; ± 0.64 ** ± 1.00 ns ± 15.90] ns 8.27 ± 2.18 ns slo-1;slo-2; ± 1.15 *** ± 2.02 ns ± 5.86] ns ± 1.38 ns shl-1; ± 0.63 ** ± 1.18 ns v 13.49] ns 7.47 ± 1.18 ns kqt-3; ± 0.88 ns ± 1.14 ns v 6.79] ns ± 0.94 ns egl-2; ± 1.27 ns ± 1.77 ns ± 16.14] ns ± 1.97 ns + 4-AP ± 0.96 ** ± 1.65 ns v 3.58] * 7.29 ± 0.92 ns Results are presented as mean with standard error of the mean. n, number of action potentials. Data were analyzed using Dunnett's test for multiple comparisons, and were not calculated for three genotypes with small n. ****, P<0.0001; ***, P<0.001; **P<0.01; *P<0.05; ns, not significant. Many mutations slightly affected the threshold and AHP; uniquely affected spike amplitude and width, resembling pharmacological administration of 4-AP. Single spikes (Fig 1C,D) have a narrower width and larger AHP than spikes in bursts. 3
5 Table S4. AWA Model Parameters, related to Figure 5. Symbol Description Value (/) C Membrane capacitance 1.5 pf g Ca Calcium conductance 0.1/0.13 ns v Ca Calcium reversal potential 120 mv f Calcium inactivation factor 0.4 τ c1 Calcium channel activation time scale 1.0 ms τ c2 Calcium channel inactivation time scale ms v c1 Calcium channel activation voltage mv Δv c1 Calcium channel activation slope 9.17 mv v c2 Calcium channel inactivation voltage 16.2 mv Δv c2 Calcium channel inactivation slope mv v c3 Calcium channel inactivation time voltage scale 23.0 mv Δv c3 Calcium channel inactivation time voltage slope 24.0 mv c 0 Calcium channel steady state normalization 0.55 v K Potassium reversal potential -84 mv g K1 SHK-1 conductance 1.5/0.0 ns v K1 SHK-1 activation voltage 2.0 mv Δv K1 SHK-1 activation slope 10.0 mv τ K1 SHK-1 switching time 30.0 ms g K2 First high-v potassium conductance 1.0/2.0 ns v K2 First high-v potassium channel activation voltage 13.0 mv Δv K2 First high-v potassium channel activation slope 20.0 mv τ K2 First high-v potassium channel switching time 1000 ms g K3 Second high-v potassium conductance 0.1 ns v K3 Second high-v potassium channel activation voltage mv Δv K3 Second high-v potassium channel activation slope 5.0 mv τ K3 Second high-v potassium channel switching time 1000 ms g Kr Slowly inactivating (Kr) K conductance 0.39/0.77 ns v Kr1 Slowly inactivating (Kr) K-channel activation voltage mv Δv Kr1 Slowly inactivating (Kr) K-channel activation slope 5.0 mv v Kr2 Slowly inactivating (Kr) K-channel inactivation voltage mv Δv Kr2 Slowly inactivating (Kr) K-channel inactivation slope 5.0 mv τ Kr Slowly inactivating (Kr) K-channel inactivation time 1800 ms g Ka Slowly activating (Kr a ) K-channel conductance 0.91/1.98 ns v Ka1 Slowly activating (Kr a ) K-channel activation voltage mv Δv Ka1 Slowly activating (Kr a ) K-channel activation slope 20.0 mv v Ka2 Slowly activating (Kr a ) K-channel time voltage scale mv Δv Ka2 Slowly activating (Kr a ) K-channel time voltage slope 4.0 mv τ Ka1 Slowly activating (Kr a ) K-channel slow activation time ms τ Ka2 Slowly activating (Kr a ) K-channel fast activation time 3000 ms g Kir Inward rectifying potassium channel conductance 0.8 ns I 0 Inward rectifying potassium channel saturated current 5.0 pa β Inward rectifying potassium channel smoothing factor 5.0 g L Leak current conductance 0.25 ns v L Leak current reversal potential mv 4
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