Article Figures & Data
Figures
- Figure 1.
AIA interneurons are required to regulate forgetting of diacetyl olfactory adaptation. A, A simplified neural network for olfactory sensing in C. elegans (White et al., 1986; Chalasani et al., 2010; Larsch et al., 2015; Dobosiewicz et al., 2019). B–D, Behavioral assays of animals with ablation of AIA, AIB, and AIY. Chemotaxis of naive, adapted, and 4 h recovered animals was analyzed. Boxes, First to third quartiles (Q1 25th to Q3 75th percentile) of each dataset; black line in the boxes, medians; black dots, mean; whiskers, minimum and maximum, excluding outliers (beyond 1.5-fold interquartile range from Q3 and Q1); x, outliers (AIB–, AIY–, and AIB–; AIY–: n = 6, two-way ANOVA, Fstrain(3,60) = 12.11, p = 2.66e−6, η2 = 0.3772; AIA–: n = 6, two-way ANOVA, Fstrain(5,90) = 21.07, p = 6.86e−14, η2 = 0.5393)a,b. E, Dose dependency of chemotaxis to diacetyl in AIA– animals (1:10−2 diacetyl n = 6, t(5) = 3.3608, p = 0.0201; 1:10−3 diacetyl t(5) = 0.6986, p = 0.5159; 1:10−4 diacetyl t(5) = 1.2714, p = 0.2595; 1:10−5 diacetyl t(5) = 0.7888, p = 0.466; mean ± SEM)c. B–D, Post hoc t test with Bonferroni’s correction; E, Student’s t test. Error bars represent the SEM.
- Figure 2.
Time course of olfactory adaptation recovery. Time course of chemotaxis recovery after adaptation in tir-1(tm3036) and AIA– (unc-103 gf) animals. Chemotaxis in naive animals, and after adaptation (0 h), and after 4, 8, and 24 h of recovery was analyzed (n ≥ 6, two-way ANOVA, Fstrain(2,9) = 61.26, p = 4.84e−18, η2 = 0.5531; mean ± SEM)d. Post hoc t test with Bonferroni’s correction. Error bars represent the SEM.
- Figure 3.
The Ca2+ responses to diacetyl of AWA neurons. A, Ca2+ responses of AWAs in wild-type, tir-1(tm3036), and AIA– animals in naive, adaptation, and recovery phases (n ≥ 21). The black line represents the application of odor stimulation (1:10−7 dilution of diacetyl). B, Relative Ca2+ responses of AWAs in wild-type, tir-1(tm3036), and AIA– animals. Values are normalized to the average naive value in respective animals. Boxes, First to third quartiles (Q1 25th to Q3 75th percentile) of each dataset; black line in the boxes, medians; black dots, mean; whiskers, minimum and maximum, excluding outliers (beyond 1.5-fold interquartile range from Q3 and Q1); x, outliers (n ≥ 21, two-way ANOVA, Fstrain(2,247) = 3.6626, p = 0.0271, η2 = 0.0288; mean ± SEM)e. Post hoc t test with Bonferroni’s correction. A, Error bars represent the SEM.
- Figure 4.
The Ca2+ responses to diacetyl of AIA interneurons. A–C, Left, Ca2+ responses of AIAs in wild-type animals in naive, adaptation, and recovery phases (n = 36; mean ± SEM). Black line represents the application of odor stimulation (1:10−7 dilution of diacetyl). Right, Ca2+ responses of AIAs before (color; 2.5–5 s) and after (pattern; 7.5–10 s) stimulation in wild-type animals in naive, adaptation, and recovery phases. Boxes, First to third quartiles (Q1 25th to Q3 75th percentile) of each dataset; black line in the boxes, medians; black dots, mean; whiskers, minimum and maximum, excluding outliers (beyond 1.5-fold interquartile range from Q3 and Q1); x, outliers (naive: n = 36, t(35) = 2.2336, p = 0.032; adaptation: t(35) = 1.5505, p = 0.13; recovery: t(35) = 2.5616, p = 0.0149)f. Student’s t test. A–C, Left, Error bars represent the SEM.
- Figure 5.
Genetic epistasis between TIR-1 and AIA interneurons. Chemotaxis to diacetyl was analyzed in tir-1(gf), AIA–, and tir-1(gf) animals with no functional AIA in naive, adaptation, and recovery phases. Boxes, first to third quartiles (Q1 25th to Q3 75th percentile) of each dataset; black line in the boxes, medians; black dots, mean; whiskers, minimum and maximum, excluding outliers (beyond 1.5-fold interquartile range from Q3 and Q1); x, outliers (n = 8, two-way ANOVA, Fstrain(5,126) = 38.4268, p = 8.41e−24, η2 = 0.6039)g. Post hoc t test with Bonferroni’s correction.
- Figure 6.
Chemotaxis to AWC-sensed isoamyl alcohol in AIA– animals. Chemotaxis to isoamyl alcohol was analyzed in two AIA– transgenic animals, AIAp::mec-4(d) and AIAp::unc-103 (gf), in naive, adapted, and 4 h recovery phases. Boxes, First to third quartiles (Q1 25th to Q3 75th percentile) of each dataset; black line in the boxes, medians; black dots, mean; whiskers, minimum and maximum, excluding outliers (beyond 1.5-fold interquartile range from Q3 and Q1); x, outliers (n = 8, two-way ANOVA, Fstrain(3,82) = 11.4853, p = 2.31e−6, η2 = 0.2959)h. Post hoc t test with Bonferroni’s correction.
- Figure 7.
AIAs may regulate forgetting of diacetyl olfactory adaptation via circuit plasticity. A, B, Two hypothetical neural circuits of AIA-dependent behavioral plasticity in the forgetting of olfactory adaptation to diacetyl. In the two models, naive chemotactic behavior might be regulated along with (A) or independent from (B) an AIA-dependent functional neural circuit. In both models, after conditioning, the AIA-dependent functional neural circuit is required to regulate the corresponding behavioral output after the animal recovers from adaptation.
Tables
Strain name Genotype Source N2 CGC tir-1(tm3036) National Bioresource Project RB1085 tir-1(ok1052) CGC JN578 peIs578[npr-9p::casp1, npr-9p::venus, unc-122p::mCherry] (AIB-) Satoh et al., 2014 JN579 peIs579[ttx-3p::casp1, ttx-3p::venus, lin-44p::gfp] (AIY-) Satoh et al., 2014 JN580 peIs580[ins-1(short)p::casp1, ins-1(short)p::venus, unc-122p::gfp] (AIA–) Satoh et al., 2014 QD155 qjEx3[gcy-28.dp::mec-4(d), gcy-28::gfp, lin-44p::gfp] Shinkai et al., 2011 QD156 qjEx4[gcy-28.dp::unc-103(gf), myo-3p::gfp] Shinkai et al., 2011 QD139 lin-15(n765ts); qjEx39[odr-10p::YC3.60, pBLH98] Inoue et al., 2013 QD140 tir-1(tm3036); lin-15(n765ts); qjEx39 [odr-10p::YC3.60, pBLH98] Inoue et al., 2013 QD157 peIs578[npr-9p::casp1, npr-9p::venus, unc-122p::mCherry];
peIs579[ttx-3p::casp1, ttx-3p::venus, lin-44p::gfp] (AIB-; AIY-)This article QD165 tir-1(ok1052); qjEx4 [gcy-28.dp::unc-103(gf), myo-3p::gfp] This article QD164 tir-1(tm3036); qjEx3 [gcy-28.dp::mec-4(d), gcy-28::gfp, lin-44p::gfp] This article QD166 tir-1(tm3036); qjEx4[gcy-28.dp::unc-103(gf), myo-3p::gfp] This article QD153 lin-15(n765ts); qjEx39[odr-10p::YC3.60, pBLH98]; peIs580[ins-1(short)p::casp1,
ins-1(short)p::venus, unc-122p::gfp]]This article QD272 qjEx52[gcy-28.dp::GCaMP6f, gcy-28.dp::paQuasAr3-citrine, lin-44p::gfp] This article CGC, Caenorhabditis Genetics Center.
Data structure Type of test Power (α = 0.05) a Figure 1B,C Normal distribution Two-way ANOVA Strains: <0.0001
Conditions: <0.0001b Fig. 1D Normal distribution Two-way ANOVA Strains: <0.0001
Conditions: <0.0001c Fig. 1E Normal distribution Student’s t test 1:10−2: 0.0201
1:10−3: 0.5159
1:10−4: 0.2595
1:10−5: 0.466d Fig. 2 Normal distribution Two-way ANOVA Strains: <0.0001
Conditions: <0.0001e Fig. 3B Normal distribution Two-way ANOVA Strains: 0.0271
Conditions: <0.0001f Fig. 4B Normal distribution Student’s t test Naive: 0.032
Adaptation: 0.13
Recovery: 0.0149g Fig. 5 Normal distribution Two-way ANOVA Strains: <0.0001
Conditions: <0.0001h Fig. 6 Normal distribution Two-way ANOVA Strains: <0.0001
Conditions: <0.0001
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