Article Figures & Data
Figures
- Figure 1.
Retrieval of appetitive and novel taste increases excitability in LIV–VI aIC-BLA projection neurons. A, Diagrammatic representation of experimental procedures. Following surgery and stereotaxic delivery of ssAAV_retro2-hSyn1-chi-mCherry-WPRE-SV40p(A) into the BLA, mice were allowed four weeks of recovery. Animals were subsequently assigned to treatment groups and trained to drink from pipettes (see Materials and Methods). We compared the intrinsic properties of LIV–VI aIC-BLA neurons among the Water (n = 6 animals, 23 cells), Saccharin 1x (n = 5 animals, 20 cells), Saccharin 1x (4 h) (n = 4 animals, 17 cells), Saccharin 5x (n = 6 animals, 18 cells), and Quinine 1x groups (n = 4 animals, 19 cells), as well as a Cage Control group (n = 4 animals, 19 cells) that underwent surgery and stereotaxic delivery of ssAAV_retro2-hSyn1-chi-mCherry-WPRE-SV40p(A) at the BLA without water restriction. B, Graph showing the water consumption before treatment (mean ± SD). There was no significant difference between water intakes between the groups before the treatment. One-way ANOVA, p = 0.9766. C, Representative traces of LIV–VI aIC-BLA projecting neurons from the six treatment groups. Scale bars: 20 mV vertical and 50 ms horizontal from 300-pA step. D, The dependence of firing rate on current step magnitude in LIV–VI aIC-BLA neurons was significantly different among the treatment groups. Excitability in the Saccharin 1x was increased compared with all other groups. Two-way repeated measures ANOVA, Current × Treatment: p < 0.0001; Cage Control versus Saccharin 1x: **p < 0.01, ***p < 0.001; Saccharin 1x versus Saccharin 1x (4 h): #p < 0.05, ##p < 0.01, ####p < 0.0001; Water versus Saccharin 1x: ̂p < 0.05, ̂̂p < 0.01, ̂̂̂p < 0.001; Saccharin 1x versus Quinine 1x: $p < 0.05, $$p < 0.01; Saccharin 1x versus Saccharin 5x: -p < 0.05; Saccharin 1x (4 h) versus Saccharin 5x: +p < 0.05. E, Representative of all fAHP measurements in response to 500-ms step current injections. Scale bars: 20 mV vertical and 50 ms horizontal. F, Representative of all action potential properties were taken. Scale bars: 20 mV vertical and 5 ms horizontal. G, Measurements for all input resistance, sag ratio, and membrane time constants were analyzed in response to 1-s, −150-pA step current injection. P, peak voltage; S, steady state voltage. Scale bars: 5 mV vertical and 100 ms horizontal. H, Significant differences were observed among the treatment groups in terms of fAHP. Cage Control (9.191 ± 1.449 mV), Water (8.150 ± 0.8288 mV), Saccharin 1x (3.016 ± 0.9423 mV), Quinine 1x (13.58 ± 1.562 mV), Saccharin 5x (8.158 ± 1.356 mV), Saccharin 1x (4 h) (5.989 ± 1.074 mV), one-way ANOVA, p < 0.0001. I, Action potential half-width in the Saccharin 1x group (0.6005 ± 0.03260 ms) was significantly decreased compared with Saccharin 1x (4 h) (0.7765 ± 0.03641 ms), one-way ANOVA, p = 0.0065. J, The membrane time constant was significantly different between the Cage Control (15.03 ± 1.376 ms) and Saccharin 1x (4 h) (26.21 ± 2.421 ms), Water (19.24 ± 1.620 ms) and Saccharin 1x (4 h) (26.21 ± 2.421 ms), Saccharin 1x (14.82 ± 1.485 ms) and Saccharin 1x (4 h) (26.21 ± 2.421 ms), and Saccharin 5x (17.30 ± 1.660 ms) and Saccharin 1x (4 h) (26.21 ± 2.421 ms) groups. One-way ANOVA, p < 0.0001. For panels D, H–J: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. All data are shown as mean ± SEM. Histologic verification of viral delivery at the IC and BLA, as well as locations of whole-cell patch-clamp recording electrode (see Extended Data Fig. 1-1). Individual IC neurons were classified as burst-spiking and regular-spiking by post hoc analysis of responses to rheobase current injections (see Extended Data Fig. 1-2). The intrinsic properties of burst spiking LIV–VI aIC-BLA projecting neurons are differentially modulated by taste valence in the context of novelty (see Extended Data Fig. 1-3).
- Figure 2.
Learned aversive taste memory retrieval decreases the excitability of LIV–VI aIC-BLA projecting neurons. A, Experimental design of behavioral procedures conducted to compare the intrinsic properties of LIV–VI aIC-BLA neurons following learned aversive taste memory retrieval (CTA Retrieval, n = 8 animals, 27 cells), appetitive retrieval for the same tastant (Saccharin 2x, n = 5 animals, 20 cells). B, Mice showed a significantly reduced saccharin consumption following learned aversive memory retrieval (n = 8) compared with appetitive retrieval mice (n = 5) group. p = 0.0085, Mann–Whitney test. C, Representative traces of LIV–VI aIC-BLA projecting neurons from the two treatment groups. Scale bars: 20 mV vertical and 50 ms horizontal from 300-pA step. D, The excitability of LIV–VI aIC-BLA in the Saccharin 2x group was significantly enhanced compared with CTA Retrieval. Two-way repeated measures ANOVA, Current × Treatment: p < 0.0001. E, Representative traces showing action potential measurements for both groups. Scale bar: 20 mV vertical and 2 ms horizontal. F, Representative traces showing the input resistance and sag ratio measurements. Scale bar: 10 mV vertical and 100 ms horizontal. G, Action potential amplitude in the CTA Retrieval (56.21 ± 0.9978 mV) group was increased compared with Saccharin 2x (49.14 ± 1.568 mV), p = 0.0005, Mann–Whitney test. H, Input resistance in the CTA Retrieval group (136.4 ± 9.064 MΩ) was significantly decreased compared with Saccharin 2x (181.1 ± 11.7 MΩ). p = 0.0036, Unpaired t test. I, SAG ratio following CTA Retrieval (13.41 ± 1.31) was significantly enhanced compared with Saccharin 2x (7.815 ± 1.176). p = 0.0037, Unpaired t test. Data are shown as mean ± SEM *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Learned aversive taste memory retrieval decreases the excitability of burst spiking LIV–VI aIC-BLA neurons (see Extended Data Fig. 2-1).
- Figure 3.
Extinction of CTA enhances, whereas reinstatement decreases, the excitability of LIV–VI aIC-BLA projecting neurons. A, Experimental design of behavioral procedures conducted to compare the intrinsic properties of LIV–VI aIC-BLA neurons following CTA Extinction (n = 5, animals, 14 cells) and Reinstatement (n = 3 animals, 15 cells). B, The graph showing the reduced aversion following the successful extinction in both treatment groups. C, Data showing the saccharin consumption on the test day following successful extinction and Reinstatement of CTA. CTA reinstated mice showed significantly reduced saccharin consumption compared with extinguished mice. p = 0.0179, Mann–Whitney test. D, Representative traces of LIV–VI aIC-BLA projection neurons firing from the two treatment groups. Scale bars: 20 mV and 50 ms horizontal from 300-pA step. E, Excitability in LIV–VI aIC-BLA neurons was significantly different among the treatment groups. Two-way repeated measures ANOVA, Current × Treatment: p < 0.0001. F, Action potential threshold in the Reinstatement group (−29.43 ± 1.731 mV) was enhanced compared with Extinction (−36.06 ± 1.481 mV). p = 0.0076, Unpaired t test. G, The membrane time constant following Reinstatement (25.48 ± 1.58 ms) was significantly enhanced compared with Extinction (17.55 ± 2.684 ms, p = 0.047). p = 0.0153, Unpaired t test. For panels D–F: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. All data are shown as mean ± SEM. Extinction of CTA enhances the excitability of burst spiking LIV–VI aIC-BLA projecting neurons compared with CTA reinstatement (see Extended Data Fig. 3-1).
- Figure 4.
Innately aversive taste is correlated with high fAHP, and prolonged conflicting experiences is correlated with an increased AP half-width in LIV–VI aIC-BLA projecting neurons. We compared the intrinsic properties of LIV–VI aIC-BLA neurons among the Saccharin 1x (n = 5 animals, 20 cells), Quinine 1x (n = 4 animals, 19 cells), Saccharin 2x (n = 5 animals, 20 cells), CTA Retrieval (n = 8, 27 cells), Extinction (n = 5 animals, 14 cells), and Reinstatement (n = 3 animals, 15 cells) groups. A, Groups associated with positive taste valence (Saccharin 1x, Saccharin 2x, Extinction), exhibited significantly increased excitability compared with innate or learned negative taste valence groups (Quinine 1x, CTA Retrieval, and Reinstatement). Two-way repeated measures ANOVA, Current × Treatment: p < 0.0001; Saccharin 2x versus CTA Retrieval: *p; Saccharin 2x versus Reinstatement: #p: Saccharin 2x versus Quinine 1x: p$; Saccharin 1x versus CTA Retrieval: p̂; Saccharin 1x versus Quinine 1x: p%; Saccharin 1x versus reinstatement: p+; Extinction versus CTA Retrieval: p@; Extinction versus Reinstatement: p&; Extinction versus Quinine 1x: p-. B, fAHP was significantly enhanced in response to Quinine 1x (13.56 ± 1.562 mV) compared with all other groups. Significant differences were also observed between Saccharin 1x (3.016 ± 0.9423 mV), Saccharin 2x (5.223 ± 0.8217 mV), and CTA Retrieval (7.97 ± 1.018 mV, p = 0.0036). Extinction (4.731 ± 1.021 mV) and Reinstatement (5.932 ± 1.292 mV). One-way ANOVA, p < 0.0001. C, Input resistance was significantly different between Saccharin 2x (181.1 ± 11.7 MΩ) and CTA Retrieval (136.4 ± 9.064 MΩ), p = 0.0352. Conversely, input resistance in Saccharin 1x (145.8 ± 12.56), Quinine 1x (146 ± 9.094), Extinction (151.1 ± 15.63), and Reinstatement groups was similar. One-way ANOVA, p = 0.0213. D, SAG ratio was significantly different between Saccharin 2x (7.815 ± 1.176) and CTA Retrieval (13.41 ± 1.31), p = 0.0209. Conversely, SAG ratio in Saccharin 1x (10.89 ± 1.621), Quinine 1x (12.13 ± 1.23), Extinction (12.37 ± 1.471), and Reinstatement (9.245 ± 0.884) groups was similar one-way ANOVA, p = 0.0286. E, Action potential amplitude in the Quinine 1x group (57.11 ± 1.376 mV), and CTA Retrieval (56.21 ± 0.9978 mV), was significantly increased compared with Saccharin 2x (49.14 ± 1.568 mV, p = 0.0175 and 0.0229, respectively). Conversely, action potential attitude in the Saccharin 1x (52.03 ± 1.308 mV), Extinction (55.09 ± 2.122 mV), and Reinstatement (53.1 ± 2.906 mV) groups was similar. One-way ANOVA, p = 0.0061. F, Action potential half-width following Extinction (0.7386 ± 0.03145 ms) and Reinstatement (0.8187 ± 0.06929 ms) was elevated compared with Saccharin 1x (0.6005 ± 0.03260 ms), Saccharin 2x (0.5780 ± 0.02994 ms) as well as CTA Retrieval (0.5959 ± 0.02080 ms, but no with Quinine 1x (0.6300 ± 0.03555 ms). One-way ANOVA, p = 0.0002. G, The membrane time constant in the Saccharin 1x (14.82 ± 1.485 ms) group was significantly decreased compared with Reinstatement (25.48 ± 1.58 ms, p = 0.0043) groups was. Differences between CTA Retrieval (20.96 ± 1.724 ms, p = 0.0189), Quinine 1x (21.55 ± 1.638 ms), Saccharin 2x (19.28 ± 1.837 ms), and Extinction (17.55 ± 2.684 ms) groups failed to reach significance. One-way ANOVA, p = 0.0047.
- Figure 5.
The intrinsic properties of burst spiking LIV–VI aIC-BLA projecting neurons represent taste experience and the probability for further learning. A, Data across all intrinsic properties from BS LIV–VI aIC-BLA neurons of the Saccharin 1x, Saccharin 2x, and Extinction groups were combined and assigned to the Low predictive following memory group (32 BS cells). Conversely, the intrinsic properties of BS LIV–VI aIC-BLA neurons from animals having undergone CTA Retrieval, 5× Saccharin, and Reinstatement were combined and assigned to the High predictive following memory group (31 BS cells). The resultant three-dimensional scatter representation of the two groups encompassed Excitability at 350 pA; AP amplitude, AP halfwidth, AP threshold; fAHP, mAHP, sAHP; IR, Rheobase, RMP, SAG ratio and τ in BS LIV–VI aIC-BLA neurons. See Extended Data Figure 5-1. B, Three-dimensional representation of the contribution of individual parameters (loadings matrix) to the principal components segregating the two groups of treatments (scores matrix). PCA variable contributions and component loadings of BS and RS LIV–VI aIC-BLA projecting neurons in Extended Data Figure 5-2.
Tables
Groups RMP (mV) mAHP (mV) Input resistance
(MΩ)Sag ratio (%) Time constant
(ms)AP thresh (mV) AP Amp (mV) AP half-width (ms) Rheobase (pA) LI–III aIC-BLA
Water−70.84 ± 1.09 (14) −2.411 ± 0.6767 (14) 136.4 ± 16.85 (14) 3.172 ± 1.082 (14) 11.17 ± 1.169 (14) −31.35 ± 1.547 (14) 56 ± 1.719 (14) 0.6557 ± 0.03222 (14) 140.7 ± 31.83 (14) LI–III aIC-BLA
Saccharin 1x−72.27 ± 1.212 (15) −1.752 ± 0.4953 (15) 131.6 ± 13.14 (15) 3.997 ± 0.9166 (15) 11.56 ± 1.672 (15) −31.44 ± 1.621 (15) 56.33 ± 1.323 (15) 0.684 ± 0.02767 (15) 136.1 ± 26.44 (15) LI–III aIC-BLA
Saccharin 2x−73.44 ± 1.295 (15) −2.165 ± 0.684 (15) 142.5 ± 13.17 (15) 5.793 ± 1.449 (15) 18.63 ± 1.9 (15) −31.47 ± 2.511 (15) 49.76 ± 2.065 (15) 0.564 ± 0.03708 (15) 104.7 ± 25.03 (15) LI–III aIC-BLA
CTA Retrieval−72 ± 1.117 (17) −3.087 ± 2.914 (17) 171.1 ± 22.28 (17) 6.307 ± 1.368 (17) 18.94 ± 1.667 (17) −34.24 ± 1.445 (17) 48.58 ± 1.472 (17) 0.5518 ± 0.02698 (17) 114.3 ± 31.57 (17) Values are expressed as mean ± SEM. The number of cells is in parentheses. Statistical analysis was performed by Student’s t test. RMP, resting membrane potential; mAHP, medium after hyperpolarization potentials; AP Thresh, action potential threshold; AP Amp, action potential amplitude; AP half-width, action potential half-width.
Groups RMP
(mV)fAHP
(mV)mAHP
(mV)sAHP
(mV)Input
resistance
(MΩ)Sag
ratio
(%)Time
constant
(ms)AP
threshold
(mV)AP
amplitude
(mV)AP
half-width
(ms)Rheobase
(pA)LIV–VI
aIC-BLA
Cage Control−68.28 ± 0.8506
(19)9.191 ± 1.449
(19)**##−3.73 ± 0.4241
(19)−1.881 ± 0.3376
(19)120.7 ± 7.686
(19)12.41 ± 1.938
(19)15.03 ± 1.376
(19)−30.76 ± 2.139
(19)56.28 ± 0.8818
(19)̂̂0.6774 ± 0.03816
(19)87.47 ± 9.127
(19)LIV–VI
aIC-BLA
Water−69.3 ± 0.9051
(23)8.15 ± 0.8288
(23)**−5.535 ± 0.6754
(23)−3.277 ± 0.4603
(23)139.1 ± 9.021
(23)8.909 ± 1.306
(23)19.24 ± 1.62
(23)−31.05 ± 1.30
(23)52.43 ± 1.034
(23)0.6243 ± 0.02021
(23)85 ± 11.76
(23)LIV–VI
aIC-BLA
Saccharin 1x−69.21 ± 0.94
(20)3.016 ± 0.9423
(20)####−4.17 ± 0.4542
(20)−2.521 ± 0.2735
(20)145.8 ± 12.56
(20)10.89 ± 1.621
(20)14.82 ± 1.485
(20)#−30.9 ± 2.141
(20)52.03 ± 1.308
(20)0.6005 ± 0.0326
(20)74.25 ± 11.39
(20)LIV–VI
aIC-BLA
Quinine 1x−67.32 ± 1.092
(19)13.56 ± 1.562
(19)$$$$∼−5.858 ± 0.5613
(18)−3.634 ± 0.3632
(18)146 ± 9.094
(19)12.13 ± 1.23
(19)21.55 ± 1.638
(19)−29.74 ± 1.989
(19)57.11 ± 1.376
(19)*̂̂̂0.6 ± 0.03555
(19)69.89 ± 8.932
(19)LIV–VI
aIC-BLA
Saccharin 5x−66.53 ± 1.358
(18)8.158 ± 1.356
(18)**###−3.999 ± 0.653
(18)−2.695 ± 0.5083
(18)144.6 ± 14.68
(18)9.392 ± 2.127
(18)17.3 ± 1.66
(18)−32.66 ± 1.783
(18)56.48 ± 1.337
(18)0.6539 ± 0.04814
(18)78.61 ± 10.75
(18)LIV–VI
aIC-BLA
Saccharin 1x
(4 h)−69.47 ± 0.7569
(17)5.989 ± 1.074
(17)**−4.411 ± 0.8962
(17)−2.631 ± 0.6949
(17)153.9 ± 11.10
(17)10.97 ± 1.475
(17)26.21 ± 2.421
(17)***$$$%&&−31.03 ± 1.511
(17)52.24 ± 2.311
(17)0.7765 ± 0.03641
(17)**78.88 ± 9.274
(17)LIV–VI
aIC-BLA
Saccharin 2x−70.79 ± 1.242
(20)5.223 ± 0.8217
(20)###−5.301 ± 0.7863
(20)−3.351 ± 0.3798
(20)181.1 ± 11.7
(20)7.815 ± 1.176
(20)19.28 ± 1.837
(20)−32.85 ± 1.447
(20)49.14 ± 1.568
(20)###0.578 ± 0.02994
(20)69.6 ± 10.71
(20)LIV–VI
aIC-BLA
CTA Retrieval−68.78 ± 0.8419
(27)7.97 ± 1.018
(27)##−5.213 ± 0.4544
(27)−2.69 ± 0.3064
(27)136.4 ± 9.064
(27)̂̂13.41 ± 1.31
(27)̂̂20.96 ± 1.724
(27)*−31.61 ± 2.68
(27)56.21 ± 0.9978
(27)̂̂̂0.5959 ± 0.0208
(27)90.44 ± 17.56
(27)LIV–VI
aIC-BLA
Extinction−65.98 ± 1.457
(14)4.731 ± 1.021
(14)−5.076 ± 0.6981
(14)−2.895 ± 0.6547
(14)151.1 ± 15.63
(14)12.37 ± 1.471
(14)17.55 ± 2.684
(14)∼−36.06 ± 1.481
(14)∼55.09 ± 2.122
(14)̂0.7386 ± 0.03145
(14)69.21 ± 7.454
(14)LIV–VI
aIC-BLA
Reinstatement−68.57 ± 0.936
(15)5.932 ± 1.292
(15)−5.673 ± 0.4288
(15)−3.612 ± 0.3033
(15)178.7 ± 12.1
(15)9.245 ± 0.884
(15)25.48 ± 1.58
(15)**−29.43 ± 1.731
(15)53.1 ± 2.906
(15)0.8187 ± 0.06929
(15)67.6 ± 8.753
(15)Values are expressed in mean ± SEM. The number of cells is in parentheses. Statistical analysis was performed by one-way ANOVA post hoc Tukey’s and Dunn’s multiple comparisons. Student’s t test was performed for the comparison between two groups. RMP, resting membrane potential; fAHP, mAHP and sAHP, fast, medium, and slow after hyperpolarization potentials, respectively; AP Thresh, action potential threshold; AP Amp, action potential amplitude; AP half-width, action potential half-width. Data are shown as mean ± SEM *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, with respect to the corresponding symbols.
$Vs. Cage Control *Vs. Saccharin 1x ̂Vs. Saccharin 2x ∼Vs. Reinstatement.
%Vs. Water #Vs. Quinine 1x &Vs. Saccharin 5x.
Groups RMP
(mV)fAHP
(mV)mAHP
(mV)sAHP
(mV)Input resistance
(MΩ)Sag ratio
(%)Time constant
(ms)AP thresh
(mV)AP Amp
(mV)AP half-width
(ms)Rheobase
(pA)BS LIV–VI
aIC-BLA
Cage Control−68.28 ± 0.9705
(13)9.192 ± 2.061
(13)−4.194 ± 0.5072
(13)−2.075 ± 0.4500
(13)118.4 ± 9.771
(13)14.91 ± 2.195
(13)14.71 ± 1.944
(13)−31.83 ± 2.971
(13)56.27 ± 1.147
(13)0.6692 ± 0.05460
(13)74.54 ± 8.471
(13)BS LIV–VI
aIC-BLA
Water−69.00 ± 1.639
(11)7.800 ± 1.607(11) −4.870 ± 0.8838
(11)−2.826 ± 0.6069
(11)136.5 ± 14.40
(11)8.751 ± 2.021
(11)18.03 ± 2.309
(11)−29.27 ± 2.060
(11)54.21 ± 1.572
(11)0.6736 ± 0.03111
(11)85.91 ± 13.44
(11)BS LIV–VI
aIC-BLA
Saccharin 1x−68.80 ± 1.065
(17)2.870 ± 1.044
(17)−4.339 ± 0.5083
(17)−2.564 ± 0.3032
(17)146.6 ± 14.22
(17)11.67 ± 1.790
(17)14.27 ± 1.666
(17)−30.73 ± 2.385
(17)51.64 ± 1.473
(17)0.5976 ± 0.03555
(17)63.65 ± 7.679
(17)BS LIV–VI
aIC-BLA
Quinine 1x−67.37 ± 1.682
(9)13.67 ± 2.681(9) ## −6.131 ± 0.6514
(8)−3.58 ± 0.5788
(8)139.2 ± 16.86
(9)14.15 ± 2.159
(9)23.21 ± 2.717
(9)−29.35 ± 3.071
(9)58.86 ± 2.003
(9)0.6378 ± 0.0491
(9)59.56 ± 12.28
(9)BS LIV–VI
aIC-BLA
Saccharin 5x−67.20 ± 1.624
(10)11.30 ± 1.727
(10) ##−5.174 ± 0.8427
(10)−3.609 ± 0.7205
(10)156.1 ± 22.85
(10)11.92 ± 3.395
(10)17.11 ± 2.296
(10)−30.38 ± 2.493
(10)58.40 ± 1.812
(10)0.7140 ± 0.07349
(10)76.60 ± 15.32
(10)BS LIV–VI
aIC-BLA
Saccharin 1x
(4 h)−68.20 ± 1.293
(6)3.433 ± 0.9245(6) −6.693 ± 1.442
(6)−3.130 ± 1.637
(6)154.9 ± 22.41(6) 14.99 ± 2.770
(6)26.09 ± 5.331
(6)−34.61 ± 2.174
(6)46.79 ± 4.359
(6)0.8850 ± 0.05943
(6) #60.83 ± 11.36
(6)BS LIV–VI
aIC-BLA
Saccharin 2x−71.33 ± 1.641
(13)4.169 ± 0.9225
(13)−5.368 ± 0.9616
(13)−3.226 ± 0.4899
(13)180.3 ± 15.15
(13) **7.017 ± 1.317
(13) ***19.77 ± 2.447
(13)−34.20 ± 1.987
(13)46.18 ± 1.666
(13) ***0.5331 ± 0.03522
(13)77.54 ± 15.69
(13)BS LIV–VI
aIC-BLA
CTA Retrieval−67.37 ± 1.21
(12)5.473 ± 1.464
(12)−4.633 ± 0.5831
(12)−1.932 ± 0.4462
(12)110.9 ± 12.98
(12)16.8 ± 1.869
(12)17.06 ± 2.608
(12)−34.28 ± 1.771
(12)57.87 ± 1.678
(12)0.6367 ± 0.03961
(12)88.75 ± 9.847
(12)BS LIV–VI
aIC-BLA
Extinction−67.36 ± 1.43
(11)3.943 ± 1.111
(11)−4.816 ± 0.8447
(11)−2.104 ± 0.4466
(11) ∼131.1 ± 13.93
(11)13.69 ± 1.541
(11)14.52 ± 2.714
(11) ∼ ∼−37.41 ± 1.636
(11) ∼ ∼57.3 ± 2.023
(11)0.7155 ± 0.03674
(11)81 ± 6.932
(11)BS LIV–VI
aIC-BLA
Reinstatement−68.88 ± 1.163
(10)6.432 ± 1.737
(10)−5.243 ± 0.5853
(10)−3.804 ± 1.339
(10)157.4 ± 10.56
(10)9.124 ± 1.03
(10)26.93 ± 1.893
(10)−27.5 ± 2.195
(10)57.36 ± 3.001
(10)0.769 ± 0.03494
(10)73.9 ± 12.45
(10)Values are expressed in mean ± SEM. The number of cells is in parentheses. Statistical analysis was performed by one-way ANOVA post hoc Tukey’s and Dunn’s multiple comparisons. Student’s t test was performed for the comparison between two groups. RMP, resting membrane potential; fAHP, mAHP, and sAHP, fast, medium, and slow after hyperpolarization potentials, respectively; AP Thresh, action potential threshold; AP Amp, action potential amplitude; AP half-width, action potential half-width.
*BS LV/VI aIC-BLA Saccharin 2x vs. CTA Retrieval, *p < 0.05, **p < 0.01, ***p < 0.001.
∼BS LV/VI aIC-BLA Extinction vs. Reinstatement, ∼p < 0.05, ∼∼p < 0.01, ∼∼∼p < 0.001.
#BS LV/VI aIC-BLA Saccharin 1x vs. Quinine 1x, Saccharin 5x and Saccharin 1x (4hr), #p < 0.05, ##p < 0.01.
Groups RMP
(mV)fAHP
(mV)mAHP
(mV)sAHP
(mV)Input resistance
(MΩ)Sag ratio
(%)Time constant
(ms)AP thresh
(mV)AP Amp
(mV)AP half-width
(ms)Rheobase
(pA)RS LIV–VI
aIC-BLA
Cage Control−68.29 ±1.830
(6)9.188 ± 1.351
(6)−2.725 ± 0.6462
(6)−1.460 ± 0.4409
(6)125.7 ± 13.03
(6)6.993 ± 3.030
(6)15.73 ± 1.333
(6)−28.44 ± 2.166
(6)56.30 ± 1.422
(6)0.6950 ± 0.03170
(6)115.5 ± 18.62
(6)RS LIV–VI
aIC-BLA
Water−69.57 ± 0.9422
(12)8.472 ± 0.6792
(12)−6.144 ± 1.013
(12)−3.690 ± 0.6876
(12)141.5 ±11.75
(12)9.735 ± 1.400
(12)20.35 ± 2.321
(12)−32.67 ± 1.561
(12)50.79 ± 1.238
(12)0.5792 ± 0.01928
(12)84.17 ± 19.48
(12)RS LIV–VI
aIC-BLA
Saccharin 1x−71.57 ± 1.120
(3)3.840 ± 2.530
(3)−3.210 ± 0.9005
(3)−2.277 ± 0.7297
(3)141.3 ±28.45
(3)6.455 ± 3.099
(3)17.95 ± 2.856
(3)−31.86 ± 5.650
(3)54.23 ± 2.660
(3)0.6167 ± 0.09939
(3)134.3 ± 58.52
(3)RS LIV–VI
aIC-BLA
Quinine 1x−67.28 ±1.505
(10)11.63 ± 1.616
(10)−5.639 ± 0.8918
(10)−3.678 ± 0.4895
(10)151.4 ± 9.055
(10)10.31 ± 1.115
(10)23.6 ± 1.77(10) −30.1 ± 2.732
(10)55.53 ± 1.845
(10)0.6230 ± 0.0535
(10)79.20 ± 12.73
(10)RS LIV–VI
aIC-BLA
Saccharin 5x−65.70 ± 2.378
(8)4.235 ± 1.141
(8)−2.530 ± 0.7960
(8)−1.553 ±0.4915
(8)130.1 ± 16.85
(8)6.237 ± 1.910
(8)17.54 ± 2.561
(8)−35.50 ± 2.302
(8)54.09 ± 1.734
(8)0.5788 ± 0.05034
(8)81.13 ±15.88
(8)RS LIV–VI
aIC-BLA
Saccharin 1x
(4 h)−70.17 ± 0.9075
(11)7.383 ± 1.439
(11)−3.165 ± 0.9897
(11)−2.359 ± 0.6647
(11)153.3 ± 12.96
(11)8.782 ± 1.389
(11)26.28 ± 2.596
(11)−28.43 ± 1.652
(11)51.66 ± 3.053
(11)0.7773 ± 0.04702
(11)88.73 ± 12.25
(11)RS LIV–VI
aIC-BLA
Saccharin 2x−69.79 ± 1.921
(7)7.180 ± 1.402
(7)−5.016 ± 1.460
(7)−3.581 ± 0.6324
(7)182.6 ± 19.62
(7)9.297 ± 2.347
(7)18.36 ± 2.842
(7)−30.36 ± 1.638
(7)54.62 ± 2.058
(7)0.6614 ± 0.04149
(7)54.86 ± 8.207
(7)RS LIV–VI
aIC-BLA
CTA Retrieval−69.9 ± 1.116
(15)9.967 ± 1.216
(15)−5.667 ± 0.6647
(15)−3.297 ± 0.3599(15) 156.7 ± 10.11
(15)10.71 ± 1.536
(15)24.08 ± 2.023
(15)−33.9 ± 1.132
(15)54.89 ± 1.13
(15)0.5633 ± 0.01703
(15) *91.8 ± 31.15
(15)RS LIV–VI
aIC-BLA
Extinction−60.93 ± 3.263
(3)7.620 ± 1.907
(3)−6.027 ± 1.062
(3)−5.797 ± 1.997
(3)224.2 ± 21.29
(3)7.515 ± 2.666
(3)28.69 ± 2.138
(3)−31.1 ± 1.372
(3)46.98 ± 4.432
(3)0.8233 ± 0.02603
(3)36.67 ± 13.33
(3)RS LIV–VI
aIC-BLA
Reinstatement−67.95 ± 1.725
(5)4.932 ± 1.893
(5)−6.532 ± 0.3344
(5)−3.228 ± 0.3214
(5)221.2 ± 18.9
(5)9.486 ± 1.846
(5)22.58 ± 2.632
(5)−33.31 ± 2.035
(5)44.58 ± 4.569
(5)0.918 ± 0.203
(5)55 ± 6.885
(5)Values are expressed in mean ± SEM. The number of cells is in parentheses. Statistical analysis was performed by One-way ANOVA post hoc Tukey’s and Dunn’s multiple comparisons. Student’s t test was performed for the comparison of two groups. RMP, resting membrane potential; fAHP, mAHP, and sAHP, fast, medium, and slow after hyperpolarization potentials, respectively; AP Thresh, action potential threshold; AP Amp, action potential amplitude; AP half-width, action potential half-width.
*RS LV/VI aIC-BLA Saccharin 1x vs. CTA Retrieval, *p < 0.05.
Figure Statistical test Results Figure 1 Figure 1B One -Way Anova
Water consumption the before the test
Water
Saccharin 1x
Quinine 1x
Saccharin 5x
Saccharin 1x (4hrs)ANOVA results:
F = 0.9766
P = 0.4424
R squared, 0.1634Figure 1D Two-way repeated measures ANOVA
Post-hoc Tukey’s multiple comparisons
LIV-VI aIC-BLA neurons
F-I curve
Cage control
Water
Saccharin 1x
Quinine 1x
Saccharin 5x
Saccharin 1x (4hr)ANOVA Results:
Treatment; p = 0.0057, F (5, 110) = 3.491
Current; p < 0.0001, F (8, 880) = 1276
Interaction; p < 0.0001, F (40, 880) = 4.141
Multiple Comparisons:
0pA
Cage control vs. Water
Mean difference = 0.000
Cage control vs. Saccharin 1x
Mean difference = 0.000
Cage control vs. Quinine 1x
Mean difference = 0.000
Cage Control vs. Saccharin 5x
Mean difference = 0.000
Cage Control vs. Saccharin 1x (4hr)
Mean difference = 0.000
Water vs. Saccharin 1x
Mean difference = 0.000
Water vs. Quinine 1x
Mean difference = 0.000
Water vs. Saccharin 5x
Mean difference = 0.000
Water vs. Saccharin 1x (4hr)
Mean difference = 0.000
Saccharin 1x vs. Quinine 1x
Mean difference = 0.000
Saccharin 1x vs. Saccharin 5x
Mean difference = 0.000
Saccharin 1x vs. Saccharin 1x (4hr)
Mean difference = 0.000
Quinine 1x vs. Saccharin 5x
Mean difference = 0.000
Quinine 1x vs. Saccharin 1x (4hr)
Mean difference = 0.000
Saccharin 5x vs. Saccharin 1x (4hr)
Mean difference = 0.000
50pA
Cage control vs. Water
p = 0.9902, q = 0.8645, df = 990.0;
Cage control vs. Saccharin 1x
p = 0.9993, q = 0.5005, df = 990.0;
Cage Control vs. Quinine 1x
p >0.9999, q = 0.2597, df = 990.00;
Cage control vs. Saccharin 5x
p = 0.9451, q = 1.281, df = 990.00;
Cage control vs. Saccharin 1x(4hr)
p = 0.9922, q = 0.8240, df = 990;
Water vs. Saccharin 1x
p = 0.9999, q = 0.3522, df = 990
Water vs. Quinine 1x
p = 0.9946, q = 0.5927, df = 990;
Water vs. Saccharin 5x
p = 0.9984, q = 0.4868, df = 990;
Water vs. Saccharin 1x (4hr)
p>0.9999, q = 0.02212, df = 990;
Saccharin 1x vs. Quinine 1x
p>0.9999, q = 0.2374, df = 990;
Saccharin 1x vs. Saccharin 5x
p = 0.9931, q = 0.8029, df = 990;
Saccharin 1x vs. Saccharin 1x (4hr)
p = 0.9999, q = 0.3478, df = 990;
Quinine 1x vs. Saccharin 5x
p = 0.9790, q = 1.024, df = 990;
Quinine 1x vs. Saccharin 1x (4hr)
p = 0.9986, q = 0.5716, df = 990;
Saccharin 5x vs. Saccharin 1x (4hr)
p = 0.9996, q = 0.4320, df = 990;
100pA
Cage control vs. Water
p = 0.8652, q = 1.610, df = 990.0;
Cage control vs. Saccharin 1x
p = 0.2233, q = 3.159, df = 990.0;
Cage Control vs. Quinine 1x
p = 0.4580, q = 2.563, df = 990.00;
Cage control vs. Saccharin 5x
p = 0.6454, q = 2.163, df = 990.00;
Cage control vs. Saccharin 1x(4hr)
p = 0.5930, q = 2.275, df = 990;
Water vs. Saccharin 1x
p = 0.8437, q = 1.677, df = 990
Water vs. Quinine 1x
p = 0.9742, q = 1.073, df = 990;
Water vs. Saccharin 5x
p = 0.9969, q = 0.6743, df = 990;
Water vs. Saccharin 1x (4hr)
p = 0.9926, q = 0.8138, df = 990;
Saccharin 1x vs. Quinine 1x
p = 0.9987, q = 0.5625, df = 990;
Saccharin 1x vs. Saccharin 5x
p = 0.9867, q = 0.9250, df = 990;
Saccharin 1x vs. Saccharin 1x (4hr)
p = 0.9945, q = 0.7652, df = 990;
Quinine 1x vs. Saccharin 5x
p = 0.9998, q = 0.3658, df = 990;
Quinine 1x vs. Saccharin 1x (4hr)
p>0.9999, q = 0.2164, df = 990;
Saccharin 5x vs. Saccharin 1x (4hr)
p>0.9999, q = 0.1422, df = 990;
150pA
Cage control vs. Water
p = 0.8024, q = 1.793, df = 990.0;
Cage control vs. Saccharin 1x
p = 0.0085, q = 4.836, df = 990.0;
Cage Control vs. Quinine 1x
p = 0.1482, q = 3.431, df = 990.00;
Cage control vs. Saccharin 5x
p = 0.2881, q = 2.970, df = 990.00;
Cage control vs. Saccharin 1x(4hr)
p = 0.5463, q = 2.374, df = 990;
Water vs. Saccharin 1x
p = 0.1962, q = 3.248, df = 990
Water vs. Quinine 1x
p = 0.8009, q = 1.797, df = 990;
Water vs. Saccharin 5x
p = 0.9345, q = 1.337, df = 990;
Water vs. Saccharin 1x (4hr)
p = 0.9953, q = 0.7397, df = 990;
Saccharin 1x vs. Quinine 1x
p = 0.9297, q = 1.361, df = 990;
Saccharin 1x vs. Saccharin 5x
p = 0.8142, q = 1.762, df = 990;
Saccharin 1x vs. Saccharin 1x (4hr)
p = 0.5843, q = 2.293, df = 990;
Quinine 1x vs. Saccharin 5x
p = 0.9997, q = 0.4145, df = 990;
Quinine 1x vs. Saccharin 1x (4hr)
p = 0.9843, q = 0. 0.9603, df = 990;
Saccharin 5x vs. Saccharin 1x (4hr)
P = 0.9989, q = 0.5448, df = 990;
200pA
Cage Control vs. Water
p = 0.9968, q = 0.6822, df = 990.0;
Cage Control vs. Saccharin 1x
p = 0.0038, q = 0.6822, df = 990.0;
Cage Control vs. Quinine 1x
p = 0.5360, q = 2.396, df = 990.0;
Cage Control vs. Saccharin 5x
p = 0.5556, q = 2.354, df = 990.0;
Cage Control vs. Saccharin 1x (4hr)
p = 0.9876, q = 0.9108, df = 990.0;
Water vs. Saccharin 1x
p = 0.0116, q = 4.708, df = 990.0;
Water vs. Quinine 1x
p = 0.7904, q = 4.708, df = 990.0;
Water vs. Saccharin 5x
p = 0.8042, q = 1.789, df = 990.0;
Water vs. Saccharin 1x (4hr)
p>0.9999, q = 0.2894, df = 990.0;
Saccharin 1x vs. Quinine 1x
p = 0.3853, q = 2.727, df = 990.0;
Saccharin 1x vs. Saccharin 5x
p = 0.3979, q = 2.698, df = 990.0;
Saccharin 1x vs. Saccharin 1x (4hr)
p = 0.0457, q = 4.083, df = 990.0;
Quinine 1x vs. Saccharin 5x
p>0.9999, q = 0.008880, df = 990.0;
Quinine 1x vs. Saccharin 1x (4hr)
p = 0.9172, q = 1.417, df = 990.0;
Saccharin 5x vs. Saccharin 1x (4hr)
p = 0.9233, q = 1.391, df = 990.0;
250pA
Cage control vs. Water
p = 0.9988, q = ,0.5590 df = 990.0;
Cage control vs. Saccharin 1x
p = 0.0011, q = 5.603, df = 990.0;
Cage Control vs. Quinine 1x
p = 0.7555, q = 1.912, df = 990.0;
Cage Control vs. Saccharin 5x
p = 0.7478, q = 1.931, df = 990.0;
Cage Control vs. Saccharin 1x (4hr)
p>0.9999, q = 0.1164, df = 990.0;
Water vs. Saccharin 1x
p = 0.0026, q = 5.304, df = 990.0;
Water vs. Quinine 1x
p = 0.9113, q = 1.442, df = 990.0;
Water vs. Saccharin 5x
p = 0.9051, q = 1.468, df = 990.0;
Water vs. Saccharin 1x (4hr)
p = 0.9972, q = 0.6633, df = 990.0;
Saccharin 1x vs. Quinine 1x
p = 0.1000, q = 3.666, df = 990.0;
Saccharin 1x vs. Saccharin 5x
p = 0.1180, q = 3.570, df = 990.0;
Saccharin 1x vs. Saccharin 1x (4hr)
p = 0.0013, q = 5.559, df = 990.0;
Quinine 1x vs. Saccharin 5x
p>0.9999, q = 0.04456, df = 990.0;
Quinine 1x vs. Saccharin 1x (4hr)
p = 0.7292, q = 1.975, df = 990.0;
Saccharin 5x vs. Saccharin 1x (4hr)
p = 0.7215, q = 1.993, df = 990.0;
300pA
Cage Control vs. Water
p = 0.9993, q = 0.4987, df = 990.0;
Cage Control vs. Saccharin 1x
p = 0.0005, q = 5.903, df = 990.0;
Cage Control vs. Quinine 1x
p = 0.9022, q = 1.479, df = 990.0;
Cage Control vs. Saccharin 5x
p = 0.7419, q = 1.945, df = 990.0;
Cage Control vs. Saccharin 1x (4hr)
p = 0.9641, q = 1.158, df = 990.0;
Water vs. Saccharin 1x
p = 0.0009, q = 5.679, df = 990.0;
Water vs. Quinine 1x
p = 0.9766, q = 1.049, df = 990.0;
Water vs. Saccharin 5x
p = 0.8854, q = 1.542, df = 990.0;
Water vs. Saccharin 1x (4hr)
p = 0.8386, q = 1.692, df = 990.0;
Saccharin 1x vs. Quinine 1x
p = 0.0232, q = 4.405, df = 990.0;
Saccharin 1x vs. Saccharin 5x
p = 0.0716, q = 3.851, df = 990.0;
Saccharin 1x vs. Saccharin 1x (4hr)
p < 0.0001, q = 6.905, df = 990.0;
Quinine 1x vs. Saccharin 5x
p = .9994, q = 0.4860, df = 990.0;
Quinine 1x vs. Saccharin 1x (4hr)
p = 0.4433, q = 2.596, df = 990.0;
Saccharin 5x vs. Saccharin 1x (4hr)
p = 0.2645, q = 3.035, df = 990.0;
350pA
Cage Control vs. Water
p = 0.9973, q = 0.6556, df = 990.0;
Cage Control vs. Saccharin 1x
p = 0.0003, q = 6.069, df = 990.0;
Cage Control vs. Quinine 1x
p = 0.9972, q = 0.6596, df = 990.0;
Cage Control vs. Saccharin 5x
p = 0.8294, q = 1.719, df = 990.0;
Cage Control vs. Saccharin 1x (4hr)
p = 0.7347, q = 1.962, df = 990.0;
Water vs. Saccharin 1x
p = 0.0009, q = 5.694, df = 990.0;
Water vs. Quinine 1x
p>0.9999, q = 0.03478, df = 990.0;
Water vs. Saccharin 5x
p = 0.9650, q = 1.151, df = 990.0;
Water vs. Saccharin 1x (4hr)
p = 0.4043, q = 2.683, df = 990.0;
Saccharin 1x vs. Quinine 1x
p = 0.0020, q = 5.401, df = 990.0;
Saccharin 1x vs. Saccharin 5x
p = 0.0328, q = 4.244, df = 990.0;
Saccharin 1x vs. Saccharin 1x (4hr)
p < 0.0001, q = 7.879, df = 990.0;
Quinine 1x vs. Saccharin 5x
p = 0.9747, q = 1.068, df = 990.0;
Quinine 1x vs. Saccharin 1x (4hr)
p = 0.4400, q = 2.603, df = 990.0;
Saccharin 5x vs. Saccharin 1x (4hr)
p = 0.1105, q = 3.609, df = 990.0;
400pA
Cage Control vs. Water
p = 0.9988, q = 0.5513, df = 990.0;
Cage Control vs. Saccharin 1x
p = 0.0004, q = 5.939, df = 990.0;
Cage Control vs. Quinine 1x
p = 0.9987, q = 0.5623, df = 990.0;
Cage Control vs. Saccharin 5x
p = 0.9132, q = 1.435, df = 990.0;
Cage Control vs. Saccharin 1x (4hr)
p = 0.3765, q = 2.748, df = 990.0;
Water vs. Saccharin 1x
p = 0.0009, q = 5.663, df = 990.0;
Water vs. Quinine 1x
p>0.9999, q = 0.03710, df = 990.0;
Water vs. Saccharin 5x
p = 0.9845, q = 0.9564, df = 990.0;
Water vs. Saccharin 1x (4hr)
p = 0.1551, q = 3.402, df = 990.0;
Saccharin 1x vs. Quinine 1x
p = 0.0021, q = 5.369, df = 990.0;
Saccharin 1x vs. Saccharin 5x
p = 0.0233, q = 4.403, df = 990.0;
Saccharin 1x vs. Saccharin 1x (4hr)
p < 0.0001, q = 8.548, df = 990.0;
Quinine 1x vs. Saccharin 5x
p = 0.9894, q = 0.8800, df = 990.0;
Quinine 1x vs. Saccharin 1x (4hr)
p = 0.1832, q = 3.294, df = 990.0;
Saccharin 5x vs. Saccharin 1x (4hr)
p = 0.0435, q = 4.108, df = 990.0.Figure 1H One-way ANOVA
Kruskal-Wallis test
Post-hoc Dunn's multiple comparisons test
LIV-VI aIC-BLA neurons
fAHP
Cage control
Water
Saccharin 1x
Quinine 1x
Saccharin 5x
Saccharin 1x (4hr)ANOVA results:
Kruskal-Wallis test, p < 0.0001;
Kruskal-Wallis statistic, 29.91.
Multiple Comparisons:
Cage Control vs. Water
p >0.9999, z = 0.2306;
Cage Control vs. Saccharin 1x
p = 0.0136, z = 3.318;
Cage Control vs. Quinine 1x
p >0.9999, z = 1.809;
Cage Control vs. Saccharin 5x
p >0.9999, z = 0.4824;
Cage Control vs. Saccharin 1x (4hr)
p>0.9999, z = 1.648;
Water vs. Saccharin 1x
p = 0.0177, z = 3.243;
Water vs. Quinine 1x
p = 0.5054, z = 2.124;
Water vs. Saccharin 5x
p>0.9999, z = 0.2771;
Water vs. Saccharin 1x (4hr)
p>0.9999, z = 1.497;
Saccharin 1x vs. Quinine 1x
p < 0.0001, z = 5.150;
Saccharin 1x vs. Saccharin 5x
p = 0.0807, z = 2.783;
Saccharin 1x vs. Saccharin 1x (4hr)
p>0.9999, z = 1.554;
Quinine 1x vs. Saccharin 5x
p = 0.3511 z = 2.267;
Quinine 1x vs. Saccharin 1x (4hr)
p = 0.0099, z = 3.406;
Saccharin 5x vs. Saccharin 1x (4hr)
p>0.9999, z = 1.1583Figure 1I One-way ANOVA
Kruskal-Wallis test
Post-hoc Dunn's multiple comparisons test
LIV-VI aIC-BLA neurons
Action Potential Half-width
Cage control
Water
Saccharin 1x
Quinine 1x
Saccharin 5x
Saccharin 1x (4hr)ANOVA results:
Kruskal-Wallis test, p = 0.0125; Kruskal-Wallis statistic,14.54.
Multiple Comparisons:
Cage Control vs. Water
p >0.9999, z = 0.7692;
Cage Control vs. Saccharin 1x
p>0.9999, z = 1.627;
Cage Control vs. Quinine 1x
p >0.9999, z = 0.9868 ;
Cage Control vs. Saccharin 5x
p >0.9999, z = 0.6540;
Cage Control vs. Saccharin 1x (4hr)
P = 0.8292, z = 1.917;
Water vs. Saccharin 1x
p >0.9999, z = 0.9244;
Water vs. Quinine 1x
p>0.9999, z = 0.2636;
Water vs. Saccharin 5x
p>0.9999, z = 0.07420;
Water vs. Saccharin 1x (4hr)
p = 0.0905, z = 2.746;
Saccharin 1x vs. Quinine 1x
p>0.9999, z = 0.6271
Saccharin 1x vs. Saccharin 5x
p = >0.9999, z = 0.9418;
Saccharin 1x vs. Saccharin 1x (4hr)
p = 0.0065, z = 3.519;
Quinine 1x vs. Saccharin 5x
p>0.9999z = 0.3194;
Quinine 1x vs. Saccharin 1x (4hr)
p = 0.0605, z = 2.876;
Saccharin 5x vs. Saccharin 1x (4hr)
p = 0.1721, z = 2.528Figure 1J One-way ANOVA
Post-hoc Tukey’s multiple comparisons
LIV-VI aIC-BLA neurons
Membrane Time Constant
Cage control
Water
Saccharin 1x
Quinine 1x
Saccharin 5x
Saccharin 1x (4hr)ANOVA results:
Treatment; p < 0.0001, F (5, 110) = 6.094;
R squared, 0.2169;
Multiple Comparisons:
Cage Control vs. Water
p = 0.4608, q = 2.566, df = 110;
Cage Control vs. Saccharin 1x
p >0.9999, q = 0.1233, df = 110;
Cage Control vs. Quinine 1x
p = 0.0864, q = 3.798, df = 110;
Cage Control vs. Saccharin 5x
p = 0.9398, q = 1.306, df = 110;
Cage Control vs. Saccharin 1x (4hr)
p = 0.0003, q = 6.326, df = 110;
Water vs. Saccharin 1x
p = 0.3890, q = 2.731, df = 110;
Water vs. Quinine 1x
p = 0.9184, q = 1.408, df = 110;
Water vs. Saccharin 5x
p = 0.9628, q = 1.163, df = 110;
Water vs. Saccharin 1x (4hr)
p = 0.0488, q = 4.115, df = 110;
Saccharin 1x vs. Quinine 1x
p = 0.0639, q = 3.969, df = 110;
Saccharin 1x vs. Saccharin 5x
p = 0.9101, q = 1.443, df = 110;
Saccharin 1x vs. Saccharin 1x (4hr)
p = 0.0002, q = 6.521, df = 110;
Quinine 1x vs. Saccharin 5x
p = 0.5180, q = 2.440, df = 110;
Quinine 1x vs. Saccharin 1x (4hr)
p = 0.4302, q = 2.635, df = 110;
Saccharin 5x vs. Saccharin 1x (4hr)
p = 0.0081, q = 4.975, df = 110;Figure 2 Figure 2B Two-tailed Unpaired t-test
Saccharin consumption on the test day
Saccharin 2x
CTA retrievalt-test Results:
Mann-Whitney test, p = 0.0085,
Mann-Whitney U, 2.500Figure 2D Two-way repeated measures ANOVA
Post-hoc Šídák's multiple comparisons test
LIV-VI aIC-BLA neurons
F-I curve
Saccharin 2x
CTA retrievalANOVA Results:
Treatment; p < 0.0014, F (1, 45) = 11.60
Current; p < 0.0001, F (8, 360) = 483.3
Interaction; p < 0.0001, F (8, 360) = 9.398
Multiple Comparisons:
0pA
Saccharin 2x vs. CTA retrieval
Mean difference = 0.000
50pA
Saccharin 2x vs. CTA retrieval
p>0.9999, t = 0.1045, df = 405.0;
100pA
Saccharin 2x vs. CTA retrieval
p = 0.5860, t = 1.682, df = 405.0;
150pA
Saccharin 2x vs. CTA retrieval
p = 0.0286, t = 2.964, df = 405.0;
200pA
Saccharin 2x vs. CTA retrieval
p = 0.0019, t = 3.738, df = 405.0;
250pA
Saccharin 2x vs. CTA retrieval
p = 0.0005, t = 4.090, df = 405.0;
300pA
Saccharin 2x vs. CTA retrieval
p = 0.0002, t = 4.280, df = 405.0;
350pA
Saccharin 2x vs. CTA retrieval
p < 0.0001, t = 4.517, df = 405.0;
400pA
Saccharin 2x vs. CTA retrieval
p = 0.0003, t = 4.161, df = 405.0Figure 2G Two-tailed Unpaired t-test
LIV-VI aIC-BLA neurons
Action Potential Amplitude
Saccharin 2x
CTA retrievalt-test Results:
p = 0.0002
t = 3.983
df = 45
Difference between means = 7.080±1.777
R squared, 0.2607Figure 2H Two-tailed Unpaired t-test
LIV-VI aIC-BLA neurons
Input Resistance
Saccharin 2x
CTA retrievalt-test Results:
p = 0.0036
t = 3.072
df = 45
Difference between means = 44.75±14.57
R squared = 0.1734Figure 2I Two-tailed Unpaired t-test
LIV-VI aIC-BLA neurons
SAG Ratio
Saccharin 2x
CTA retrievalt-test Results:
p = 0.0037
t = 3.060
df = 45
Difference between means = 5.597±1.829
R squared = 0.1723Figure 3 Figure 3B Two-way ANOVA
Post-hoc Šídák's multiple comparisons testANOVA Results:
Treatment, P = 0.068, F (1, 54) = 3.466;
Interaction, P = 0.9697, F (8, 54) = 0.2803.Figure 3C Two-tailed Unpaired t-test
Saccharin consumption on the test day
Extinction
Reinstatementt-test Results:
Mann-Whitney test, p = 0.0179;
Mann-Whitney U, 0Figure 3E Two-way repeated measures ANOVA
Post-hoc Šídák’s multiple comparisons test
LIV-VI aIC-BLA neurons
F-I curveANOVA Results:
Treatment; p = 0.0013, F (3, 72) = 5.837
Current; p < 0.0001, F (1.959, 141.0) = 802.5
Interaction; p < 0.0001, F (24, 567) = 6.468
Multiple Comparisons:
0pA
Extinction vs. Reinstatement
Mean difference = 0.000;
50pA
Extinction vs. Reinstatement
p >0.9999, t = 0.1718, df = 243.0;
100pA
Extinction vs. Reinstatement
p = 0.8899, t = 1.237, df = 243.0;
150pA
Extinction vs. Reinstatement
p >0.9999, t = 0.2492, df = 243.0;
200pA
Extinction vs. Reinstatement
p = 0.5553, t = 1.723, df = 243.0;
250pA
Extinction vs. Reinstatement
p = 0.0341, t = 2.919, df = 243.0;
300pA
Extinction vs. Reinstatement
p = 0.0030, t = 3.636, df = 243.0;
350pA
Extinction vs. Reinstatement
p = 0.0003, q = 4.203, df = 243.0;
400pA
Extinction vs. Reinstatement
p < 0.0001, t = 4.578, df = 243.0.Figure 3F Two-tailed Unpaired t-test
LIV-VI aIC-BLA neurons
Action Potential Threshold
Extinction
Reinstatementt-test Results:
p = 0.0076
t = 2.887
df = 27
Difference between means; 6.621 ± 2.293
R squared, 0.2359Figure 3G Two-tailed Unpaired t-test
LIV-VI aIC-BLA neurons
Membrane Time Constant
Extinction
Reinstatementt-test Results:
p = 0.0153
t = 2.589
df = 27
Difference between means ;7.931 ± 3.064
R squared; 0.1988Figure 4 Figure 4A Two-way repeated measures ANOVA
Post-hoc Tukey’s multiple comparisons
LIV-VI aIC-BLA neurons
F-I Curve
Saccharin 1x
Quinine 1x
Saccharin 2x
CTA retrieval
Extinction
ReinstatementANOVA results:
Treatment; p = 0.0014, F (5, 109) = 4.281
Current; p < 0.0001, F (1.990, 216.9) = 1218
Interaction; p < 0.0001, F (40, 872) = 4.978
Multiple Comparisons:
0pA
Saccharin 1x vs. Quinine 1x
Mean difference = 0.000
Saccharin 1x vs. Saccharin 2x
Mean difference = 0.000
Saccharin 1x vs. CTA Retrieval
Mean difference = 0.000
Saccharin 1x vs. Extinction
Mean difference = 0.000
Saccharin 1x vs. Reinstatement
Mean difference = 0.000
Quinine 1x vs. Saccharin 2x
Mean difference = 0.000
Quinine 1x vs. CTA Retrieval
Mean difference = 0.000
Quinine 1x vs. Extinction
Mean difference = 0.000
Quinine 1x vs. Reinstatement
Mean difference = 0.000
Saccharin 2x vs. CTA Retrieval
Mean difference = 0.000
Saccharin 2x vs. Extinction
Mean difference = 0.000
Saccharin 2x vs. Reinstatement
Mean difference = 0.000
CTA Retrieval vs. Extinction
Mean difference = 0.000
CTA Retrieval vs. Reinstatement
Mean difference = 0.000
Extinction vs. Reinstatement
Mean difference = 0.000
50pA
Saccharin 1x vs. Quinine 1x
p>0.9999q = 0.2195, df = 981.0;
Saccharin 1x vs. Saccharin 2x
p = 0.9993, q = 0.5023, df = 981.0;
Saccharin 1x vs. CTA Retrieval
p = 0.9998, q = 0.3831, df = 981.0;
Saccharin 1x vs. Extinction
p = 0.9986, q = 0.5691, df = 981.0;
Saccharin 1x vs. Reinstatement
p = 0.9999q = 0.3539, df = 981.0;
Quinine 1x vs. Saccharin 2x
p = 0.9960, q = 0.7153, df = 981.0;
Quinine 1x vs. CTA Retrieval
p = 0.9981, q = 0.6123, df = 981.0;
Quinine 1x vs. Extinction
p = 0.9945, q = 0.7626, df = 981.0;
Quinine 1x vs. Reinstatement
p = 0.9988, q = 0.5535, df = 981.0;
Saccharin 2x vs. CTA Retrieval
p>0.9999, q = 0.1553, df = 981.0;
Saccharin 2x vs. Extinction
p>0.9999, q = 0.1132, df = 981.0;
Saccharin 2x vs. Reinstatement
p>0.9999, q = 0.1112, df = 981.0;
CTA Retrieval vs. Extinction
p>0.9999, q = 0.2589, df = 981.0;
CTA Retrieval vs. Reinstatement
p>0.9999, q = 0.02435, df = 981.0;
Extinction vs. Reinstatement
p>0.9999, q = 0.2084, df = 981.0;
100pA
Saccharin 1x vs. Quinine 1x
p = 0.9991, q = 0.5199, df = 981.0;
Saccharin 1x vs. Saccharin 2x
p = 0.8659, q = 1.608, df = 981.0;
Saccharin 1x vs. CTA Retrieval
p = 0.9941, q = 0.7759, df = 981.0;
Saccharin 1x vs. Extinction
p = 0.9933, q = 0.7975, df = 981.0;
Saccharin 1x vs. Reinstatement
p = 0.9924, q = 0.8188, df = 981.0;
Quinine 1x vs. Saccharin 2x
p = 0.6709, q = 2.107, df = 981.0;
Quinine 1x vs. CTA Retrieval
p >0.9999, q = 0.2082, df = 981.0;
Quinine 1x vs. Extinction
p >0.9999, q = 0.3161, df = 981.0;
Quinine 1x vs. Reinstatement
p = 0.9431, q = 1.292, df = 981.0;
Saccharin 2x vs. CTA Retrieval
p = 0.4875, q = 2.499, df = 981.0;
Saccharin 2x vs. Extinction
p = 0.6016, q = 2.257, df = 981.0;
Saccharin 2x vs. Reinstatement
p = 0.9970, q = 0.6698, df = 981.0;
CTA Retrieval vs. Extinction
p >0.9999, q = 0.1487, df = 981.0;
CTA Retrieval vs. Reinstatement
p = 0.8745, q = 1.579, df = 981.0;
Extinction vs. Reinstatement
p = 0.8967, q = 1.500, df = 981.0
150pA
Saccharin 1x vs. Quinine 1x
p = 0.9491, q = 1.258, df = 981.0;
Saccharin 1x vs. Saccharin 2x
p = 0.8741, q = 1.258, df = 981.0;
Saccharin 1x vs. CTA Retrieval
p = 0.3926, q = 2.71, df = 981.0;
Saccharin 1x vs. Extinction
p>0.9999, q = 0.2459, df = 981.0;
Saccharin 1x vs. Reinstatement
p = 0.9985, q = 0.5798, df = 981.0;
Quinine 1x vs. Saccharin 2x
p = 0.5798, q = 2.818, df = 981.0;
Quinine 1x vs. CTA Retrieval
p = 0.937, q = 1.324, df = 981.0;
Quinine 1x vs. Extinction
p = 0.9882, q = 0.9008, df = 981.0;
Quinine 1x vs. Reinstatement
p = 0.9983, q = 0.5934, df = 981.0;
Saccharin 2x vs. CTA Retrieval
p = 0.0233, q = 4.404, df = 981.0;
Saccharin 2x vs. Extinction
p = 0.8426, q = 0.8426, df = 981.0;
Saccharin 2x vs. Reinstatement
p = 0.6995, q = 0.6995, df = 981.0;
CTA Retrieval vs. Extinction
p = 0.6431, q = 2.168, df = 981.0;
CTA Retrieval vs. Reinstatement
p = 0.7735, q = 1.868, df = 981.0;
Extinction vs. Reinstatement
p>0.9999, q = 0.3023, df = 981.0
200pA
Saccharin 1x vs. Quinine 1x
p = 0.4777, q = 2.521, df = 981.0;
Saccharin 1x vs. Saccharin 2x
p = 0.9511, q = 1.246, df = 981.0;
Saccharin 1x vs. CTA Retrieval
p = 0.0346, q = 4.219, df = 981.0;
Saccharin 1x vs. Extinction
p>0.9999, q = 0.07085, df = 981.0;
Saccharin 1x vs. Reinstatement
p = 0.627, q = 2.202, df = 981.0;
Quinine 1x vs. Saccharin 2x
p = 0.0862, q = 3.75, df = 981.0;
Quinine 1x vs. CTA Retrieval
p = 0.907, q = 1.46, df = 981.0;
Quinine 1x vs. Extinction
p = 0.5516, q = 2.363, df = 981.0;
Quinine 1x vs. Reinstatement
p>0.9999, q = 0.1599, df = 981.0;
Saccharin 2x vs. CTA Retrieval
p = 0.0013, q = 5.554, df = 981.0;
Saccharin 2x vs. Extinction
p = 0.9756, q = 1.06, df = 981.0;
Saccharin 2x vs. Reinstatement
p = 0.1668, q = 3.356, df = 981.0;
CTA Retrieval vs. Extinction
p = 0.0712, q = 3.854, df = 981.0;
CTA Retrieval vs. Reinstatement
p = 0.8889, q = 1.529, df = 981.0;
Extinction vs. Reinstatement
p = 0.6782, q = 2.091, df = 981.0
250pA
Saccharin 1x vs. Quinine 1x
p = 0.1585, q = 3.389, df = 981.0;
Saccharin 1x vs. Saccharin 2x
p = 0.9907, q = 0.8555, df = 981.0;
Saccharin 1x vs. CTA Retrieval
p = 0.0038, q = 5.16, df = 981.0;
Saccharin 1x vs. Extinction
p >0.9999, q = 0.3001, df = 981.0;
Saccharin 1x vs. Reinstatement
p = 0.1227, q = 3.547, df = 981.0;
Quinine 1x vs. Saccharin 2x
p = 0.0336, q = 4.233, df = 981.0;
Quinine 1x vs. CTA Retrieval
p = 0.9074, q = 1.459, df = 981.0;
Quinine 1x vs. Extinction
p = 0.1609, q = 3.379, df = 981.0;
Quinine 1x vs. Reinstatement
p = 0.9998, q = 0.3641, df = 981.0;
Saccharin 2x vs. CTA Retrieval
p = 0.0003, q = 6.078, df = 981.0;
Saccharin 2x vs. Extinction
p = 0.9994, q = 0.4763, df = 981.0;
Saccharin 2x vs. Reinstatement
p = 0.0268, q = 4.339, df = 981.0;
CTA Retrieval vs. Extinction
p = 0.0066, q = 4.94, df = 981.0;
CTA Retrieval vs. Reinstatement
p = 0.9838, q = 0.9659, df = 981.0;
Extinction vs. Reinstatement
p = 0.1238, q = 3.541, df = 981.0
300pA
Saccharin 1x vs. Quinine 1x
p = 0.0468, q = 4.071, df = 981.0;
Saccharin 1x vs. Saccharin 2x
p = 0.9991, q = 0.5264, df = 981.0;
Saccharin 1x vs. CTA Retrieval
p = 0.0006, q = 5.795, df = 981.0;
Saccharin 1x vs. Extinction
p = 0.9998, q = 0.3808, df = 981.0;
Saccharin 1x vs. Reinstatement
p = 0.023, q = 4.41, df = 981.0;
Quinine 1x vs. Saccharin 2x
p = 0.0153, q = 4.591, df = 981.0;
Quinine 1x vs. CTA Retrieval
p = 0.9311, q = 1.354, df = 981.0;
Quinine 1x vs. Extinction
p = 0.046, q = 4.08, df = 981.0;
Quinine 1x vs. Reinstatement
p = 0.9985, q = 0.585, df = 981.0;
Saccharin 2x vs. CTA Retrieval
p = 0.0001, q = 6.36, df = 981.0;
Saccharin 2x vs. Extinction
p >0.9999, q = 0.09689, df = 981.0;
Saccharin 2x vs. Reinstatement
p = 0.0073, q = 4.898, df = 981.0;
CTA Retrieval vs. Extinction
p = 0.0011, q = 5.594, df = 981.0;
CTA Retrieval vs. Reinstatement
p = 0.9978, q = 0.6315, df = 981.0;
Extinction vs. Reinstatement
p = 0.0229, q = 4.411, df = 981.0
350pA
Saccharin 1x vs. Quinine 1x
p = 0.0058, q = 4.992, df = 981.0;
Saccharin 1x vs. Saccharin 2x
p = 0.9998, q = 0.4001, df = 981.0;
Saccharin 1x vs. CTA Retrieval
p = 0.0001, q = 6.284, df = 981.0;
Saccharin 1x vs. Extinction
p >0.9999, q = 0.2699, df = 981.0;
Saccharin 1x vs. Reinstatement
p = 0.0028, q = 5.272, df = 981.0;
Quinine 1x vs. Saccharin 2x
p = 0.0021, q = 5.387, df = 981.0;
Quinine 1x vs. CTA Retrieval
p = 0.9909, q = 0.8504, df = 981.0;
Quinine 1x vs. Extinction
p = 0.0092, q = 4.807, df = 981.0;
Quinine 1x vs. Reinstatement
p = 0.9985, q = 0.5836, df = 981.0;
Saccharin 2x vs. CTA Retrieval
p < 0.0001, q = 6.713, df = 981.0;
Saccharin 2x vs. Extinction
p >0.9999, q = 0.09312, df = 981.0;
Saccharin 2x vs. Reinstatement
p = 0.001, q = 5.642, df = 981.0;
CTA Retrieval vs. Extinction
p = 0.0005, q = 5.914, df = 981.0;
CTA Retrieval vs. Reinstatement
p >0.9999, q = 0.1648, df = 981.0;
Extinction vs. Reinstatement
p = 0.0044, q = 5.099, df = 981.0
400pA
Saccharin 1x vs. Quinine 1x
p = 0.0062, q = 4.963, df = 981.0;
Saccharin 1x vs. Saccharin 2x
p >0.9999, q = 0.24, df = 981.0;
Saccharin 1x vs. CTA Retrieval
p = 0.0004, q = 5.925, df = 981.0;
Saccharin 1x vs. Extinction
p = 0.9991, q = 0.5192, df = 981.0;
Saccharin 1x vs. Reinstatement
p = 0.0014, q = 5.513, df = 981.0;
Quinine 1x vs. Saccharin 2x
p = 0.0034, q = 5.2, df = 981.0;
Quinine 1x vs. CTA Retrieval
p = 0.9991, q = 0.5284, df = 981.0;
Quinine 1x vs. Extinction
p = 0.0053, q = 5.027, df = 981.0;
Quinine 1x vs. Reinstatement
p = 0.991, q = 0.8492, df = 981.0;
Saccharin 2x vs. CTA Retrieval
p = 0.0002, q = 6.183, df = 981.0;
Saccharin 2x vs. Extinction
p >0.9999, q = 0.3014, df = 981.0;
Saccharin 2x vs. Reinstatement
p = 0.0008, q = 5.736, df = 981.0;
CTA Retrieval vs. Extinction
p = 0.0005, q = 5.857, df = 981.0;
CTA Retrieval vs. Reinstatement
p = 0.9997, q = 0.4195, df = 981.0;
Extinction vs. Reinstatement
p = 0.0013, q = 5.554, df = 981.0Figure 4B One-way ANOVA
Post-hoc Tukey’s multiple comparisons
LIV-VI aIC-BLA neurons
fAHP
Saccharin 1x
Quinine 1x
Saccharin 2x
CTA retrieval
Extinction
ReinstatementANOVA results:
Treatment; p < 0.0001, F (5, 109) = 10.64;
R squared, 0.3283;
Multiple Comparisons:
Saccharin 1x vs. Quinine 1x
p < 0.0001, q = 9.380, df = 109;
Saccharin 1x vs. Saccharin 2x
p = 0.7249, q = 1.985, df = 109;
Saccharin 1x vs. CTA Retrieval
p = 0.0127, q = 4.774, df = 109;
Saccharin 1x vs. Extinction
p = 0.9204, q = 1.399, df = 109;
Saccharin 1x vs. Reinstatement
p = 0.5239, q = 2.428, df = 109;
Quinine 1x vs. Saccharin 2x
p < 0.0001, q = 7.421, df = 109;
Quinine 1x vs. CTA Retrieval
p = 0.0035, q = 5.331, df = 109;
Quinine 1x vs. Extinction
p < 0.0001, q = 7.147, df = 109;
Quinine 1x vs. Reinstatement
p = 0.0003, q = 6.299, df = 109;
Saccharin 2x vs. CTA Retrieval
p = 0.4251, q = 2.647, df = 109;
Saccharin 2x vs. Extinction
p = 0.9997, q = 0.4017, df = 109;
Saccharin 2x vs. Reinstatement
p = 0.9983, q = 0.5902, df = 109;
CTA Retrieval vs. Extinction
p = 0.3621, q = 2.796, df = 109;
CTA Retrieval vs. Reinstatement
p = 0.7995, q = 1.799, df = 109;
Extinction vs. Reinstatement
p = 0.9868, q = 0.9191, df = 109.Figure 4C One-way ANOVA
Post-hoc Tukey’s multiple comparisons
LIV-VI aIC-BLA neurons
Input Resistance
Saccharin 1x
Quinine 1x
Saccharin 2x
CTA retrieval
Extinction
ReinstatementANOVA results:
Treatment; p = 0.0213, F (5, 109) = 2.775;
R squared, 0.1129;
Multiple Comparisons:
Saccharin 1x vs. Quinine 1x
p >0.9999, q = 0.03668, df = 109;
Saccharin 1x vs. Saccharin 2x
p = 0.2331, q = 3.152, df = 109;
Saccharin 1x vs. CTA Retrieval
p = 0.9876, q = 0.9065, df = 109;
Saccharin 1x vs. Extinction
p = 0.9997, q = 0.4256, df = 109;
Saccharin 1x vs. Reinstatement
p = 0.3953, q = 2.716, df = 109;
Quinine 1x vs. Saccharin 2x
p = 0.2582, q = 3.075, df = 109;
Quinine 1x vs. CTA Retrieval
p = 0.9859, q = 0.9323, df = 109;
Quinine 1x vs. Extinction
p = 0.9998, q = 0.3877, df = 109;
Quinine 1x vs. Reinstatement
p = 0.4229, q = 2.652, df = 109;
Saccharin 2x vs. CTA Retrieval
p = 0.0352, q = 4.286, df = 109;
Saccharin 2x vs. Extinction
p = 0.5204, q = 2.435, df = 109;
Saccharin 2x vs. Reinstatement
p >0.9999, q = 0.2024, df = 109;
CTA Retrieval vs. Extinction
p = 0.9475, q = 1.262, df = 109;
CTA Retrieval vs. Reinstatement
p = 0.1001, q = 3.711, df = 109;
Extinction vs. Reinstatement
p = 0.6757, q = 2.098, df = 109.Figure 4D One-way ANOVA
Post-hoc Tukey’s multiple comparisons
LIV-VI aIC-BLA neurons
Sag ratio
Saccharin 1x
Quinine 1x
Saccharin 2x
CTA retrieval
Extinction
ReinstatementANOVA results:
Treatment; p = 0.0286, F (5, 109) = 2.610;
R squared, 0.1069;
Multiple Comparisons:
Saccharin 1x vs. Quinine 1x
p = 0.9862, q = 0.9280, df = 109;
Saccharin 1x vs. Saccharin 2x
p = 0.5707, q = 2.327, df = 109;
Saccharin 1x vs. CTA Retrieval
p = 0.6972, q = 2.049, df = 109;
Saccharin 1x vs. Extinction
p = 0.9794, q = 1.015, df = 109;
Saccharin 1x vs. Reinstatement
p = 0.9643, q = 1.152, df = 109;
Quinine 1x vs. Saccharin 2x
p = 0.2112, q = 3.225, df = 109;
Quinine 1x vs. CTA Retrieval
p = 0.9784, q = 1.026, df = 109;
Quinine 1x vs. Extinction
p>0.9999, q = 0.1598, df = 109;
Quinine 1x vs. Reinstatement
p = 0.7184, q = 2.000, df = 109;
Saccharin 2x vs. CTA Retrieval
p = 0.0209, q = 4.543, df = 109;
Saccharin 2x vs. Extinction
p = 0.2415, q = 3.126, df = 109;
Saccharin 2x vs. Reinstatement
p = 0.9805q = 1.002, df = 109;
CTA Retrieval vs. Extinction
p = 0.9944, q = 0.7617, df = 109;
CTA Retrieval vs. Reinstatement
p = 0.2504, q = 3.099, df = 109;
Extinction vs. Reinstatement
p = 0.7140, q = 2.010, df = 109.Figure 4E One-way ANOVA
Post-hoc Tukey’s multiple comparisons
LIV-VI aIC-BLA neurons
Action Potential Amplitude
Saccharin 1x
Quinine 1x
Saccharin 2x
CTA retrieval
Extinction
ReinstatementANOVA results:
Treatment; p = 0.0054, F (5, 109) = 3.526;
R squared, 0.1392;
Multiple Comparisons:
Saccharin 1x vs. Quinine 1x
p = 0.2342, q = 3.149, df = 109;
Saccharin 1x vs. Saccharin 2x
p = 0.7922, q = 1.818, df = 109;
Saccharin 1x vs. CTA Retrieval
p = 0.3531, q = 2.818, df = 109;
Saccharin 1x vs. Extinction
p = 0.8190, q = 1.746, df = 109;
Saccharin 1x vs. Reinstatement
p = 0.9979, q = 0.6222df = 109;
Quinine 1x vs. Saccharin 2x
p = 0.0087, q = 4.944, df = 109;
Quinine 1x vs. CTA Retrieval
p = 0.9983, q = 0.5921, df = 109;
Quinine 1x vs. Extinction
p = 0.9662, q = 1.137, df = 109;
Quinine 1x vs. Reinstatement
p = 0.5806, q = 2.305, df = 109;
Saccharin 2x vs. CTA Retrieval
p = 0.0129, q = 4.768, df = 109;
Saccharin 2x vs. Extinction
p = 0.1650, q = 3.396, df = 109;
Saccharin 2x vs. Reinstatement
p = 0.5804, q = 2.306, df = 109;
CTA Retrieval vs. Extinction
p = 0.9968, q = 0.6777, df = 109;
CTA Retrieval vs. Reinstatement
p = 0.7511, q = 1.922, df = 109;
Extinction vs. Reinstatement
p = 0.9746, q = 1.065, df = 109.Figure 4F One-way ANOVA
Kruskal-Wallis test
Post-hoc Dunn's multiple comparisons test
LIV-VI aIC-BLA neurons
Action Potential Half-width
Saccharin 1x
Quinine 1x
Saccharin 2x
CTA retrieval
Extinction
ReinstatementANOVA results:
Kruskal-Wallis test; p = 0.0002; Kruskal-Wallis statistic,24.03
Multiple Comparisons:
Saccharin 1x vs. Quinine 1x
p >0.9999, z = 0.6106
Saccharin 1x vs. Saccharin 2x
p >0.9999, z = 0.2586;
Saccharin 1x vs. CTA Retrieval
p >0.9999, z = 0.04096;
Saccharin 1x vs. Extinction
p = 0.0485, z = 2.944;
Saccharin 1x vs. Reinstatement
p = 0.0200, z = 3.208;
Quinine 1x vs. Saccharin 2x
p >0.9999, q = 0.8658;
Quinine 1x vs. CTA Retrieval
p >0.9999, z = 0.6129;
Quinine 1x vs. Extinction
p = 0.2759, z = 2.358;
Quinine 1x vs. Reinstatement
p = 0.1372, z = 2.607;
Saccharin 2x vs. CTA Retrieval
p >0.9999, z = 0.3181;
Saccharin 2x vs. Extinction
p = 0.0222, z = 3.179;
Saccharin 2x vs. Reinstatement
p = 0.0085, z = 3.448;
CTA Retrieval vs. Extinction
p = 0.0312, z = 3.079;
CTA Retrieval vs. Reinstatement
p = 0.0115, z = 3.366;
Extinction vs. Reinstatement
p >0.9999, z = 0.1880.Figure 4G One-way ANOVA
Post-hoc Tukey’s multiple comparisons
LIV-VI aIC-BLA neurons
Membrane Time Constant
Saccharin 1x
Quinine 1x
Saccharin 2x
CTA retrieval
Extinction
ReinstatementANOVA results:
Treatment; p = 0.0047, F (5, 109) = 0.1419;
R squared, 0.1419;
Multiple Comparisons:
Saccharin 1x vs. Quinine 1x
p = 0.0987, q = 3.720, df = 109;
Saccharin 1x vs. Saccharin 2x
p = 0.4932, q = 2.495, df = 109;
Saccharin 1x vs. CTA Retrieval
p = 0.1046, q = 3.685, df = 109;
Saccharin 1x vs. Extinction
p = 0.9230, q = 1.388, df = 109;
Saccharin 1x vs. Reinstatement
p = 0.0022, q = 5.525, df = 109;
Quinine 1x vs. Saccharin 2x
p = , 0.9484 q = 1.257, df = 109;
Quinine 1x vs. CTA Retrieval
p = 0.9999, q = 0.3489, df = 109;
Quinine 1x vs. Extinction
p = 0.7139, q = 2.010, df = 109;
Quinine 1x vs. Reinstatement
p = 0.7124, q = 2.014, df = 109;
Saccharin 2x vs. CTA Retrieval
p = 0.9798, q = 1.011, df = 109;
Saccharin 2x vs. Extinction
p = 0.9894, q = 0.8761, df = 109;
Saccharin 2x vs. Reinstatement
p = 0.2138, q = 3.216, df = 109;
CTA Retrieval vs. Extinction
p = 0.7866, q = 1.833, df = 109;
CTA Retrieval vs. Reinstatement
p = 0.4978, q = 2.484, df = 109;
Extinction vs. Reinstatement
p = 0.0896, q = 3.777, df = 109.
Extended Data Figure 1-1
Histological verification of rAAV-mCherry virus expression and locations of whole-cell patch-clamp recordings. A, A representative image showing the distribution of retrograde injections into the BLA and aIC-BLA projection neuron at aIC. B, Locations showing the retroviral injections sites in the BLA. C, Mean localization of BLA projecting neurons of the agranular aIC used for electrophysiological whole-cell recordings. Download Figure 1-1, TIF file.
Extended Data Figure 1-2
The ratio of burst spiking and regular spiking LIV–VI aIC-BLA projecting neurons changes in relation to the uncertainty associated with taste experiences. A, Representative traces from Burst (BS) and Regular (RS) spiking LIV–VI aIC-BLA projecting neurons in response to rheobase current injections. The neurons showing doublets or triplets in response to rheobase current injection were considered BS. The neurons showing single spike in response to rheobase current injection considered RS. Scale bars: 20 mV and 100 ms. B, Pie charts showing the change in the ratio of BS versus RS LIV–VI aIC-BLA projection neurons, expressed as a percentage of the sampled population across the Saccharin 1x, Saccharin 2x, Saccharin 5x, CTA Retrieval, Extinction, and Reinstatement groups. C, Heat map summary of the change in the ratio of BS versus RS LIV–VI aIC-BLA projection neurons, expressed as a percentage of the sampled population across the six treatment groups. Download Figure 1-2, TIF file.
Extended Data Figure 1-3
Appetitive novel taste alters the intrinsic properties of burst spiking LIV–VI aIC-BLA neurons. We compared the intrinsic properties of BS and RS LIV–VI aIC-BLA neurons among the Cage Control (n = 13 cells), Water (n = 11cells), Saccharin 1x (n = 17 cells), Quinine 1x (n = 9 cells), Saccharin 5x (n = 10 cells), and Saccharin 1x (4 h, n = 6 cells). A, Excitability in BS LIV–VI aIC-BLA was not significantly different among the treatment groups. Two-way repeated measures ANOVA, Current × Treatment: p < 0.0001, Group interaction p = 0.0666. B, fAHP was significantly enhanced in Quinine 1x (13.67 ± 2.681 mV) and Saccharin 5x (11.30 ± 1.727 mV) BS neurons compared to Saccharin 1x BS neurons (2.870 ± 1.044 mV). One-way ANOVA, p = 0.0004. C, Action potential amplitude was significantly different between the groups. Cage Controls (56.27 ± 1.147 mV), Water (54.21 ± 1.572 mV), Saccharin 1x (51.64 ± 1.473 mV), Quinine 1x (58.86 ± 2.003 mV), Saccharin 5x (58.40 ± 1.812 mV), and Saccharin 1x (4 h) (46.79 ± 4.359 mV). One-way ANOVA, p = 0.0097. D, Action potential half-width in BS LIV–VI aIC-BLA neurons of the Saccharin 1x (4 h) group (0.8850 ± 0.05943ms) was increased compared to the Saccharin 1x (1 h) group, 0.5976 ± 0.03555 ms. One-way ANOVA, p = 0.0139. E, Action potential threshold was not significantly different between the groups. Cage Control (−31.83 ± 2.971 mV), Water (−29.27 ± 2.060 mV), Saccharin 1x (−30.73 ± 2.385 mV), Quinine 1x (−29.35 ± 3.071 mV), Saccharin 5x (−30.38 ± 2.493 mV), and Saccharin 1x (4 h) (−34.61 ± 2.174 mV). One-way ANOVA, p = 0.7652. F, Input resistance was similar among the different treatment groups. Cage Control (118.4 ± 9.771 MΩ), Water (136.5 ± 14.40 MΩ), Saccharin 1x (146.6 ± 14.22 MΩ), Quinine 1x (139.2 ± 16.86 MΩ), Saccharin 5x (156.1 ± 22.85 MΩ), and Saccharin 1x (4 h) (154.9 ± 22.41 MΩ). One-way ANOVA, p = 0.6304. G, SAG ratio was not significantly different between the groups. Cage Control (14.91 ± 2.195), Water (8.751 ± 2.021), Saccharin 1x (11.67 ± 1.790), Quinine 1x (14.15 ± 2.159), Saccharin 5x (11.92 ± 3.395), and Saccharin 1x (4 h) (14.99 ± 2.770). One-way ANOVA, p = 0.2232. H, Membrane time constant was significantly different among the treatment groups. Cage Control (14.71 ± 1.944 ms), Water (18.03 ± 2.309 ms), Saccharin 1x (14.27 ± 1.666 ms), Quinine 1x (23.21 ± 2.717 ms), Saccharin 5x (17.11 ± 2.296 ms), and Saccharin 1x (4 h) (26.09 ± 5.331 ms). One-way ANOVA, p = 0.0321. Data are shown as mean ± SEM. *p < 0.05, **p < 0.01. Download Figure 1-3, TIF file.
Extended Data Figure 2-1
Learned aversive taste memory retrieval decreases the excitability of burst spiking LIV–VI aIC-BLA neurons. We compared the intrinsic properties of BS and RS LIV–VI aIC-BLA neurons following Saccharin 2xs (BS = 13, RS = 7, cells) and CTA memory retrieval (BS = 12, RS = 15, cells). A, Excitability in BS LIV–VI aIC-BLA neurons was significantly reduced in the CTA Retrieval group compared to Saccharin 2x. Two-way repeated measures ANOVA, Current × Treatment: p < 0.0001. B, Input resistance in BS LIV–VI aIC-BLA neurons was significantly enhanced in the Saccharin 2x (180.3 ± 15.15 MΩ) compared to CTA Retrieval (110.9 ± 12.98 MΩ). Unpaired t test, p = 0.0022. C, Action potential amplitude in BS LIV–VI aIC-BLA neurons was significantly increased in the CTA Retrieval group compared to Saccharin 2x (46.18 ± 1.666 mV) and CTA Retrieval (57.87 ± 1.678 mV). Mann–Whitney test, p < 0.0001. D, SAG ratio in BS LIV–VI aIC-BLA neurons was significantly decreased in the Saccharin 2x (7.017 ± 1.317) compared to CTA Retrieval (16.8 ± 1.869). Mann–Whitney test, p = 0.0005. E, Representative traces of RS LIV–VI aIC-BLA neurons firing from the two treatments. Scale bars: 20 mV vertical and 50ms horizontal in response to 150-pA step current. F, Excitability in RS LIV–VI aIC-BLA neurons was similar in the CTA Retrieval and Saccharin 2x. Two-way repeated measures ANOVA, Current × Treatment: p = 0.0953. G, Input resistance in RS LIV–VI aIC-BLA neurons was not significantly different in between the groups. Saccharin 2x (182.6 ± 19.62 MΩ), and CTA Retrieval (156.7 ± 10.11 MΩ). Mann–Whitney test, p > 0.9999. H, SAG ratio in RS LIV–VI aIC-BLA neurons was not significantly different between the groups. Saccharin 2x (9.297 ± 2.347), and CTA Retrieval (10.71 ± 1.536). Mann–Whitney test, p = 0.5815. I, Action potential amplitude in RS LIV–VI aIC-BLA neurons was not significantly different between the groups. Saccharin 2x (54.62 ± 2.058 mV), and CTA Retrieval (54.89 ± 1.13 mV). Mann–Whitney test, p > 0.9999. J, AP half-width in RS LIV–VI aIC-BLA neurons was significantly reduced following CTA memory retrieval (0.5633 ± 0.01703 ms) compared to the Saccharin 2x (0.6614 ± 0.04149 ms). Mann–Whitney test, p = 0.0200. K, Membrane time constant was similar in both treatment groups. Saccharin 2x RS (18.36 ± 2.842ms), and CTA memory retrieval RS (24.08 ± 2.023 ms). Mann–Whitney test, p = 0.0777. Data are shown as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Download Figure 2-1, TIF file.
Extended Data Figure 3-1
Extinction of CTA enhances, excitability of burst spiking LIV–VI aIC-BLA projecting neurons We compared the intrinsic properties of BS and RS LIV–VI aIC-BLA neurons following the Extinction (BS = 11, RS = 3, cells) and Reinstatement (BS = 10, RS = 5, cells). A, Excitability in BS LIV–VI aIC-BLA was significantly enhanced in Extinction group comparing to Reinstatement. Two-way repeated measures ANOVA, Current × Treatment: p < 0.0001. B, sAHP in BS LIV–VI aIC-BLA neurons was significantly enhanced in the Extinction group (−2.104 ± 0.4466 mV) compared to Reinstatement (−3.804 ± 1.339 mV) neurons. Mann–Whitney test, p = 0.0230. C, Action potential threshold in BS LIV–VI aIC-BLA neurons was significantly reduced in the Extinction group (−37.41 ± 1.636 mV) compared to Reinstatement (−27.5 ± 2.195 mV). Unpaired t test, p = 0.0016. D, Input resistance in BS LIV–VI aIC-BLA neurons was similar in the two treatment groups. Extinction (131.1 ± 13.93 MΩ) and Reinstatement BS (157.4 ± 10.56 MΩ). Mann–Whitney test, p = 0.1321. E, SAG ratio in BS LIV–VI aIC-BLA neurons was enhanced following Extinction (13.69 ± 1.541) neurons compared to Reinstatement BS (9.124 ± 1.03). Unpaired t test, p = 0.0262. F, Membrane time constant in BS LIV–VI aIC-BLA neurons was significantly reduced in the Extinction group (14.52 ± 2.714 ms) compared to Reinstatement (26.93 ± 1.893) neurons. Mann–Whitney test, p = 0.0062. G, Representative traces of RS LIV–VI aIC-BLA firing from two treatment groups. Scale bars: 20 mV vertical and 50 ms horizontal in response to 150-pA current step. H, Excitability of RS LIV–VI aIC-BLA neurons in both treatment groups. I, Input resistance in RS LIV–VI aIC-BLA neurons was similar in the Extinction (224.2 ± 21.29 MΩ) and Reinstatement (221.2 ± 18.9 MΩ) groups. J, SAG ratio in RS LIV–VI aIC-BLA neurons was not different between the Extinction (7.515 ± 2.666) and Reinstatement (9.486 ± 1.846) groups. K, Membrane time constant in RS LIV–VI aIC-BLA neurons was not different between the Extinction (28.69 ± 2.138 ms) and Reinstatement groups (22.58 ± 2.632 ms). Download Figure 3-1, TIF file.
Extended Data Figure 5-2
PCA variable contributions and component loadings of burst-spiking and regular-spiking LIV–VI aIC-BLA projecting neurons. A, Column chart demonstrating the individual and cumulative proportion of the variance accounted by principal components following PCA of BS LIV–VI aIC-BLA projecting neurons in the two groups of treatments (Saccharin 1x, Saccharin 2x, Extinction vs CTA Retrieval, 5x Saccharin, Reinstatement). B, Table summarizing the contribution of individual variables (loadings) to the coordinate value of the principal components segregating the two groups (score). C, Communalities table, demonstrating the amount of variance in each variable that is accounted for by the extraction of principal components. Initial communalities are estimates of the variance in each variable accounted for by all components or factors (=1.00). Download Figure 5-2, TIF file.
Extended Data Figure 5-1
PCA showing Burst versus Regular spiking LIV–VI aIC-BLA neurons all range of excitability versus 350 pA only. A, PCA of BS and RS LIV–VI aIC-BLA neurons all range of excitability (50–350 pA and all other intrinsic properties measured). Sampled population across six treatment groups (Saccharin 1x, Saccharin 2x, Saccharin 5x, CTA Retrieval, Extinction, Reinstatement). B, PCA of BS and RS LIV–VI aIC-BLA neurons excitability of 350 pA only and all other intrinsic properties measured. Sampled population across six treatment groups (Saccharin 1x, Saccharin 2x, Saccharin 5x, CTA Retrieval, Extinction, Reinstatement). Download Figure 5-1, TIF file.
Extended Data 1
Source data files for figures. Download Extended Data 1, ZIP file.
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