Fear conditioning biases olfactory sensory neuron frequencies across generations
- Mortimer B. Zuckerman Mind Brain and Behavior Institute, Columbia University, United States
- Department of Neuroscience, Columbia University, United States
- Howard Hughes Medical Institute, Columbia University, United States
- Department of Psychology, Columbia University, United States
Figures
Figure 1 with 2 supplements
Olfactory fear conditioning leads to an increase in conditioned-odor-responsive cells in parents (F0) that is heritable (F1).
(A) Schematic representation of the mouse main olfactory epithelium and olfactory bulb. MOE: main olfactory epithelium. OB: olfactory bulb. (B) Timeline of olfactory fear conditioning, breeding for the F1 generation, and MOE collection. (C) Experimental paradigms for olfactory fear conditioning groups. Mice in the paired condition received a foot shock that co-terminated with odor presentation, while mice in the unpaired condition received a foot shock 60 s after odor presentation. (D) Schematic demonstrating the process by which cells of interest in the MOE were quantified. Epithelia from both Olfr151IRES-tauGFP/IRES-tauGFP (M71-GFP) and Olfr16IRES-tauGFP/IRES-tauGFP (MOR23-GFP) adult mice were cleared using the iDISCO+ tissue-clearing protocol. Samples were imaged on a light sheet microscope and analyzed using Imaris spot detection software. (E) Images of the MOE before (left) and after (right) optical tissue clearing. (F) Example images of M71 OSNs in zone 1 of cleared MOE from both the unpaired (left) and paired (middle) conditions. Example image of an MOE with the counted cells represented by colored dots (right). Each set of colors represents a distinct counting cube. Scale bar: 200 µm. (G) The average number of M71 OSNs in a 3503 µm3 cube of epithelium of naive (gray), acetophenone unpaired (lighter green), and acetophenone paired (darker green) conditions in F0 and F1 (Error bars are standard error. One-way ANOVA. p<0.0001. Tukey’s multiple comparisons. Naive vs. F0 paired p<0.0001. F0 unpaired vs. paired p<0.0001. Naive vs. F1 paired p<0.0001. F1 unpaired vs. paired p<0.0001. n=12,11,12,12,14.). Squares indicate males, triangles indicate females. (H) Example images of MOR23 OSNs in zone 1 of cleared MOE from both the unpaired (left) and paired (middle) conditions. Example image of an MOE with the counted cells represented by colored dots (right). Scale bar: 200 µm. (I) The average number (+/- standard error) of MOR23 OSNs in a 3503 µm3 cube of epithelium in naive (gray), lyral unpaired (lighter purple), and lyral paired (darker purple) conditions in F0 and F1 (Error bars are standard error. One-way ANOVA. p<0.0001. Tukey’s multiple comparisons. Naive vs. F0 paired p=<0.0001. F0 unpaired vs. paired p<0.0001. F1 unpaired vs. paired p=0.0368. n=7,9,9,6,6.).
Figure 1—figure supplement 1
Conditioned odor-responsive OSN counts by sex.
(A) The average number (+/- standard error) of M71 OSNs in a 3503 µm3 cube of epithelium of naive (gray), acetophenone unpaired (lighter green), and acetophenone paired (darker green) conditions in F0 and F1, separated by sex (Two-way ANOVA. Treatment factor p<0.0001. Sex factor p=0.0425. n=5,7,9,2,12,0,5,7,9,5.). (B) The average number (+/- standard error) of MOR23 OSNs in a 3503 µm3 cube of epithelium of naive (gray), lyral unpaired (lighter purple), and lyral paired (darker purple) conditions in F0 and F1, separated by sex (Two-way ANOVA. Treatment factor p<0.0001. Sex factor p=0.7206. n=6,1,7,2,7,2,2,4,1,5.).
Figure 1—figure supplement 2
F1 conditioned odor-responsive OSN counts by litter.
(A) The average number (+/- standard error) of M71 OSNs in a 3503 µm3 cube of epithelium of the F1 offspring of acetophenone-unpaired (lighter green), and acetophenone-paired (darker green) fathers, separated by litter (One-way ANOVA. p<0.0001. Tukey’s multiple comparisons. UP_A vs. P_B p=0.0329. UP_A vs. P_C p=0.0041. UP_A vs. P_E p=0.0001. UP_B vs. P_B p=0.038. UP_B vs. P_C p=0.0043. UP_B vs. P_E p=0.0002. UP_C vs. P_A p=0.046. UP_C vs. P_B p=0.006. UP_C vs. P_C p=0.0015. UP_C vs. P_D p=0.0241. UP_C vs. P_E p<0.0001. P_A vs. P_E p=0.0257. n=4,3,5,4,4,1,1,4.). (B) The average number (+/- standard error) of MOR23 OSNs in a 3503 µm3 cube of epithelium of the F1 offspring of lyral-unpaired (lighter green), and lyral-paired (darker green) fathers, separated by litter (One-way ANOVA. p=0.0004. UP_A vs. P_A p=0.0008. UP_A vs. P_B p=0.0133. UP_A vs. P_C p=0.002. n=6,3,1,2.).
Figure 2
Increased OSN abundance is specific to OSN subtypes responsive to the conditioned odor.
(A) Timeline of olfactory fear conditioning and MOE collection for both experiments. (B) Olfr151IRES-tauRFP2/IRES-tauRFP2 (M71-RFP) targeted mutation. (C) Representative images showing DAPI (blue, top left), endogenous RFP in M71 OSNs (red, top right), endogenous GFP in MOR23 OSNs (green, bottom left), and the merged channels (bottom right) in a homozygous M71-RFP;MOR23-GFP animal. Scale bar: 50 µm. (D) The average number of M71 olfactory sensory neurons in a 3503 µm3 cube of the epithelium in the propanol unpaired (light blue) and propanol paired (dark blue) conditions (Error bars are standard error. Student’s unpaired t-test. Unpaired vs. paired p=0.3009. n=6,7.). Squares indicate males, triangles indicate females. (E) The ratio of M71 OSNs to 100 MOR23 OSNs in homozygous M71-RFP;MOR23-GFP mice following no conditioning (naive, gray), unpaired (light green), or paired (dark green) olfactory fear conditioning with acetophenone (Error bars are standard error. One-way ANOVA. p=0.0062. Tukey’s multiple comparisons. Naive vs. paired p=0.0097. Unpaired vs. paired p=0.0163. n=2,4,4.).
Figure 3
Olfactory fear conditioning biases olfactory receptor choice toward conditioned-odor-responsive cell-specific identities.
(A) Timeline of olfactory fear conditioning, EdU injections, and MOE collection. (B) Schematic representation of the layers of the MOE, showing the stem cell, immature OSN, and mature OSN populations (left). Representative image of the MOE from a homozygous MOR23-GFP mouse showing EdU-positive cells (magenta) and a newborn (EdU+) MOR23 OSN (green and magenta). Scale bar: 20 µm. (C) Representative images showing MOE staining of EdU (red, first column), endogenous GFP (green, second column), DAPI (blue, third column), and the merged channels (fourth column) in homozygous M71-GFP and MOR23-GFP mice. Scale bar: 40 µm. (D) Percentage of EdU-positive M71 OSNs in naive, unpaired, and paired groups (Error bars are standard error. Kruskal-Wallis test. p<0.0001. Dunn’s multiple comparisons. Naive vs. paired p=0.0009. Unpaired vs. paired p=0.0799. n=6,6,6.). Squares indicate males, triangles indicate females. (E) Percentage of EdU-positive MOR23 OSNs in naive, unpaired, and paired groups (Error bars are standard error. Kruskal-Wallis test. p=0.0003. Dunn’s multiple comparisons. Naive vs. paired p=0.0040. Unpaired vs. paired p=0.0725. n=4,6,8.).
Figure 4 with 2 supplements
Conditioned-odor-responsive cell increase is sustained through at least 9 weeks of cell turnover.
(A) Timeline of olfactory fear conditioning and extended MOE collection time points. (B) Example image of M71 OSNs in zone 1 of cleared MOE collected 42 days post-conditioning. Scale bar: 200 µm. (C) The average number of M71 OSNs in a 3503 µm3 cube of epithelium of unpaired (light green) and paired (dark green) mice, 42- or 63 days post-conditioning (Error bars are standard error. One-way ANOVA. p=0.0033. Tukey’s multiple comparisons. 42d unpaired vs. paired p=0.0476. 63d unpaired vs. paired p=0.0203. n=8,8,4,6.). Squares indicate males, triangles indicate females. (D) Schematic of the trichamber behavioral approach-avoidance assay and equation for the approach-avoid index. Positive values indicate approach to the conditioned odor (acetophenone), while negative values indicate avoidance. (E) The approach-avoid indices of unpaired and paired F0 mice at day 42 (Error bars are standard error. Student’s unpaired t-test. 42d unpaired vs. paired p=0.2248. n=10,11.). Squares indicate males, triangles indicate females. (F) The approach-avoid indices of unpaired and paired F0 mice at day 63 (Error bars are standard error. Student’s unpaired t-test. 63d unpaired vs paired p=0.8987. n=6,8.).
Figure 4—figure supplement 1
Extended timepoint OSN counts by sex.
(A) The average number (+/- standard error) of M71 OSNs in a 3503 µm3 cube of epithelium of unpaired (light green) and paired (dark green) mice, 42- or 63 days post-conditioning, separated by sex (Two-way ANOVA. Treatment factor p=0.0123. Sex factor p=0.9889. n=5,3,6,2,2,2,3,3.).
Figure 4—figure supplement 2
Extended timepoint trichamber assay by sex.
(A) The approach-avoid indices of female unpaired and paired F0 mice at day 42 (Error bars are standard error. Student’s unpaired t-test. p=0.0055. n=4,4.). (B) The approach-avoid indices of male unpaired and paired F0 mice at day 42 (Error bars are standard error. Student’s unpaired t-test. p=0.6534. n=6,7.). (C) The approach-avoid indices of female unpaired and paired F0 mice at day 63 (Error bars are standard error. Student’s unpaired t-test. p=0.8762. n=3,4.). (D) The approach-avoid indices of male unpaired and paired F0 mice at day 63 (Error bars are standard error. Student’s unpaired t-test. p=0.9859. n=3,4.).
Figure 5 with 3 supplements
Olfactory fear conditioning leads to nuanced behavioral differences in F1 offspring.
(A) Schematic of the trichamber assay showing the three conditioning odors and control odors. Propanol was the control odor for acetophenone and lyral, and acetophenone was the control odor for propanol. (B) Timeline of olfactory fear conditioning, behavior testing in F0, F0 breeding for the F1 generation, and F1 behavior testing. Conditioned F0 males used to breed for F1s did not undergo behavioral testing to prevent any extinction effects. (C) The approach-avoid indices of acetophenone-conditioned naive, unpaired, and paired F0 mice, and F1 mice bred from unpaired and paired F0 fathers (Error bars are standard error. One-way ANOVA. p<0.0001. Tukey’s multiple comparisons. Naive vs. F0 Paired p<0.0001. F0 Unpaired vs. F0 Paired p<0.0001. F0 Paired vs. F1 Unpaired p<0.0001. F0 Paired vs. F1 Paired p<0.0001. n=25,10,15,22,18.). Squares indicate males, triangles indicate females. (D) Group-averaged heat maps for acetophenone-conditioned F0 mice and F1 offspring, with the acetophenone chamber on the left and the control (propanol) chamber on the right. (E) Distance traveled in the trichamber assay for acetophenone-conditioned F0 mice and F1 offspring (Error bars are standard error. One-way ANOVA. p=0.0041. Tukey’s multiple comparisons. Naive vs. F0 Paired p=0.0032. F0 Paired vs. F1 Unpaired p=0.0141. F0 Paired vs. F1 Paired p=0.0103. n=25,10,15,22,18). (F) Time freezing for acetophenone-conditioned F0 mice and F1 offspring (Error bars are standard error. Kruskal-Wallis test. p=0.0022. Dunn’s multiple comparisons. F0 Unpaired vs. F1 Unpaired p=0.0406. F0 Paired vs. F1 Unpaired p=0.0262. n=25,10,15,22,18.). (G) The approach-avoid indices of lyral-conditioned naive, unpaired, and paired F0 mice, and F1 mice bred from unpaired and paired F0 fathers (Error bars are standard error. One-way ANOVA. p<0.0001. Tukey’s multiple comparisons. Naive vs. F0 Paired p=0.0004. F0 Unpaired vs. F0 Paired p<0.0001. F0 Paired vs. F1 Unpaired p=0.0013. F0 Paired vs. F1 Paired p<0.0001. n=10,17,20,6,13.). (H) Group-averaged heat maps for lyral-conditioned F0 mice and F1 offspring, with the lyral chamber on the left and the control (propanol) chamber on the right. (I) Distance traveled in the trichamber assay for lyral-conditioned F0 mice and F1 offspring (Error bars are standard error. One-way ANOVA. p<0.0001. Tukey’s multiple comparisons. Naive vs. F1 Paired p<0.0001. F0 Unpaired vs. F1 Unpaired p=0.037. F0 Unpaired vs. F1 Paired p<0.0001. F0 Paired vs. F1 Unpaired p=0.0121. F0 Paired vs. F1 Paired p<0.0001. F1 Unpaired vs. F1 Paired p=0.0325. n=10,17,20,6,13.). (J) Time freezing for lyral-conditioned F0 mice and F1 offspring (Error bars are standard error. Kruskal-Wallis test. p<0.0001. Dunn’s multiple comparisons. Naive vs. F0 Unpaired p=0.0001. Naive vs. F0 Paired p=0.0005. F0 Unpaired vs. F1 Paired p=0.0012. F0 Paired vs. F1 Paired 0.0041. n=10,17,20,6,13.). (K) The approach-avoid indices of propanol-conditioned naive, unpaired, and paired F0 mice, and F1 mice bred from unpaired and paired F0 fathers (Error bars are standard error. One-way ANOVA. p<0.0001. Tukey’s multiple comparisons. Naive vs. F0 Paired p<0.0001. F0 Unpaired vs. F0 Paired p<0.0001. F0 Paired vs. F1 Unpaired p<0.0001. F0 Paired vs. F1 Paired p<0.0001. n=25,13,18,13,13.). (L) Group-averaged heat maps for propanol-conditioned F0 mice and F1 offspring, with the propanol chamber on the left and the control (acetophenone) chamber on the right. (M) Distance traveled in the trichamber assay for propanol-conditioned F0 mice and F1 offspring (Error bars are standard error. One-way ANOVA. p<0.0001. Tukey’s multiple comparisons. Naive vs. F0 Unpaired p=0.0304. Naive vs. F0 Paired p<0.0001. F0 Unpaired vs. F1 Unpaired p=0.0014. F0 Paired vs. F1 Unpaired p<0.0001. F1 Unpaired vs. F1 Paired p=0.0045. n=25,13,18,13,13.). (N) Time freezing for propanol-conditioned F0 mice and F1 offspring (Error bars are standard error. Kruskal-Wallis test. p<0.0001. Dunn’s multiple comparisons. Naive vs. F0 Unpaired p=0.0074. Naive vs. F0 Paired p<0.0001. F0 Paired vs. F1 Unpaired p=0.0011. F0 Paired vs. F1 Paired p=0.028. n=25,13,18,13,13.).
Figure 5—figure supplement 1
Trichamber assay metrics by sex.
(A) The approach-avoid indices of acetophenone-conditioned naive, unpaired, and paired F0 mice, and F1 mice bred from unpaired and paired F0 fathers, separated by sex (Error bars are standard error. Two-way ANOVA. Treatment factor p<0.0001. Sex factor p=0.8137. n=25,0,10,0,15,0,10,12,15,3.). (B) Distance traveled in the trichamber assay for acetophenone-conditioned F0 mice and F1 offspring, separated by sex (Error bars are standard error. Two-way ANOVA. Treatment factor p=0.0049. Sex factor p=0.0261. n=25,0,10,0,15,0,10,12,15,3.). (C) Time freezing for acetophenone-conditioned F0 mice and F1 offspring, separated by sex (Error bars are standard error. Two-way ANOVA. Treatment factor p=0.0008. Sex factor p=0.5149. n=25,0,10,0,15,0,10,12,15,3.). (D) The approach-avoid indices of lyral-conditioned naive, unpaired, and paired F0 mice, and F1 mice bred from unpaired and paired F0 fathers, separated by sex (Error bars are standard error. Two-way ANOVA. Treatment factor p<0.0001. Sex factor p=0.9573. n=5,5,9,8,7,13,4,2,6,7.). (E) Distance traveled in the trichamber assay for lyral-conditioned F0 mice and F1 offspring, separated by sex (Error bars are standard error. Two-way ANOVA. Treatment factor p<0.0001. Sex factor p=0.016. n=5,5,9,8,7,13,4,2,6,7.). (F) Time freezing for lyral-conditioned F0 mice and F1 offspring, separated by sex (Error bars are standard error. Two-way ANOVA. Treatment factor p=0.0013. Sex factor p=0.0938. n=5,5,9,8,7,13,4,2,6,7.). (G) The approach-avoid indices of propanol-conditioned naive, unpaired, and paired F0 mice, and F1 mice bred from unpaired and paired F0 fathers, separated by sex (Error bars are standard error. Two-way ANOVA. Treatment factor p<0.0001. Sex factor p=0.4375. n=25,0,12,1,17,1,8,8,16,4.). (H) Distance traveled in the trichamber assay for propanol-conditioned F0 mice and F1 offspring, separated by sex (Error bars are standard error. Two-way ANOVA. Treatment factor p<0.0001. Sex factor p=0.0005. Tukey’s multiple comparisons. F0 Unpaired male vs. female p=0.0165. F0 Paired male vs. female p=0.0165. F1 Unpaired male vs. female p=0.0165. F1 Paired male vs. female p=0.0165. n=25,0,12,1,17,1,8,8,16,4.). (I) Time freezing for propanol-conditioned F0 mice and F1 offspring, separated by sex (Error bars are standard error. Two-way ANOVA. Treatment factor p<0.0001. Sex factor = 0.3945. n=25,0,12,1,17,1,8,8,16,4.).
Figure 5—figure supplement 2
Trichamber assay avoidance index by litter.
(A) The approach-avoid indices (+/- standard error) of acetophenone-conditioned naive, unpaired, and paired F0 mice, and F1 mice bred from unpaired and paired F0 fathers, separated by litter (One-way ANOVA. p=0.487. n=4,4,9,5,3,2,7,6.). (B) The approach-avoid indices (+/- standard error) of lyral-conditioned naive, unpaired, and paired F0 mice, and F1 mice bred from unpaired and paired F0 fathers, separated by litter (One-way ANOVA. p=0.9939. n=1,5,9,4.). (C) The approach-avoid indices (+/- standard error) of propanol-conditioned naive, unpaired, and paired F0 mice, and F1 mice bred from unpaired and paired F0 fathers, separated by litter One-way ANOVA. p=0.0199. Tukey’s multiple comparisons. UP_A vs. UP_C p=0.0306. UP_A vs. P_B p=0.0088. UP_A vs. P_C p=0.041.
Figure 5—figure supplement 3
Additional trichamber assay metrics.
(A) The mean speed (+/- standard error) of acetophenone-conditioned naive, unpaired, and paired F0 mice, and F1 mice bred from unpaired and paired F0 fathers (One-way ANOVA. p=0.003. Tukey’s multiple comparisons. Naive vs. F0 Paired p=0.0024. F0 Paired vs. F1 Unpaired p=0.0121. F0 Paired vs. F1 Paired p=0.0081. n=25,10,15,22,18.). (B) The number of entries (+/- standard error) into the propanol (control) and acetophenone (conditioned odor) chambers in naive, unpaired, and paired F0 mice, and F1 mice bred from unpaired and paired F0 fathers (Two-way ANOVA. Treatment factor p=0.0174. Odor factor p=0.1339. p-Values from Tukey’s multiple comparisons tests in Supplementary file 2. n=25,10,15,22,18.). (C) Time spent (+/- standard error) in the propanol (control) and acetophenone (conditioned odor) chambers in naive, unpaired, and paired F0 mice, and F1 mice bred from unpaired and paired F0 fathers (Two-way ANOVA. Treatment factor p=0.0022. Odor factor p=0.0053. p-Values from Tukey’s multiple comparisons tests in Supplementary file 2. n=25,10,15,22,18.). (D) The mean speed (+/- standard error) of lyral-conditioned naive, unpaired, and paired F0 mice, and F1 mice bred from unpaired and paired F0 fathers (One-way ANOVA. p<0.0001. Tukey’s multiple comparisons. Naive vs. F1 Paired p<0.0001. F0 Unpaired vs. F1 Unpaired p=0.0438. F0 Unpaired vs. F1 Paired p<0.0001. F0 Paired vs. F1 Unpaired p=0.0128. F0 Paired vs. F1 Paired p<0.0001. F1 Unpaired vs. F1 Paired p=0.0315. n=10,17,20,6,13.). (E) The number of entries (+/- standard error) into the propanol (control) and lyral (conditioned odor) chambers in naive, unpaired, and paired F0 mice, and F1 mice bred from unpaired and paired F0 fathers (Two-way ANOVA. Treatment factor p<0.0001. Odor factor p=0.6465. p-Values from Tukey’s multiple comparisons tests in Supplementary file 2. n=10,17,20,6,13.). (F) Time spent (+/- standard error) in the propanol (control) and lyral (conditioned odor) chambers in naive, unpaired, and paired F0 mice, and F1 mice bred from unpaired and paired F0 fathers (Two-way ANOVA. Treatment factor p=0.0007. Odor factor p=0.0914. p-Values from Tukey’s multiple comparisons tests in Supplementary file 2. n=10,17,20,6,13.). (G) The mean speed (+/- standard error) of propanol-conditioned naive, unpaired, and paired F0 mice, and F1 mice bred from unpaired and paired F0 fathers (One-way ANOVA. p<0.0001. Tukey’s multiple comparisons. Naive vs. F0 Unpaired p=0.0295. Naive vs. F0 Paired p<0.0001. F0 Unpaired vs F1 Unpaired p=0.0017. F0 Paired vs. F1 Unpaired p<0.0001. F1 Unpaired vs. F1 Paired p=0.0051. n=25,13,18,13,13.). (H) The number of entries (+/- standard error) into the acetophenone (control) and propanol (conditioned odor) chambers in naive, unpaired, and paired F0 mice, and F1 mice bred from unpaired and paired F0 fathers (Two-way ANOVA. Treatment factor p<0.0001. Odor factor p=0.0435. p-Values from Tukey’s multiple comparisons tests in Supplementary file 2. n=25,13,18,13,13.). (I) Time spent (+/- standard error) in the acetophenone (control) and propanol (conditioned odor) chambers in naive, unpaired, and paired F0 mice, and F1 mice bred from unpaired and paired F0 fathers (Two-way ANOVA. Treatment factor p=0.4521. Odor factor p=0.0016. p-Values from Tukey’s multiple comparisons tests in Supplementary file 2. n=25,13,18,13,13.).
Figure 6 with 1 supplement
Unsupervised machine learning analysis identifies behavioral differences in F1.
(A) Schematic of the trichamber assay showing the three conditioning odors and control odors. Propanol was the control odor for acetophenone and lyral, and acetophenone was the control odor for propanol. (B) Analysis pipeline for Keypoint-MoSeq. Eight key points were tracked across the entire 10-min trichamber assay for 99 videos, and the tracking data was used to train a Keypoint-MoSeq model. (C) Trajectory plots of the 21 most frequently used syllables across the dataset. (D) The relative usage frequencies of syllables in the whole trichamber arena in the F1 offspring of unpaired (light pink) and paired (dark pink) fathers. Syllables with significantly different usage between groups are underlined in the x-axis and denoted with asterisks above the data points (Error bars are standard error. Kruskal-Wallis test with Dunn’s multiple comparisons. Syllable 3 F1 unpaired vs. F1 paired p=0.00696. Syllable 5 p=0.0054. Syllable 11 p=0.02505. Syllable 22 p=0.0184. n=35,34.). (E) Histogram of the frequency (number of observations divided by the bin width) of each animal’s center point along the x-axis (x distance from the conditioned odor port). The two vertical bars indicate the divisions between the three chambers (conditioned odor chamber left, control odor chamber right). (F) The relative usage frequencies of syllables in the conditioned odor chamber in F1 unpaired (light pink) and F1 paired (dark pink) (Error bars are standard error. Kruskal-Wallis test with Dunn’s multiple comparisons. Syllable 5 F1 unpaired vs. F1 paired p=0.0406. Syllable 17 P=0.046695. n=35,34.). (G) The relative usage frequencies of syllables in the whole arena in the F1 offspring of acetophenone-unpaired (light green) and acetophenone-paired (dark green) fathers (Error bars are standard error. n=18.13.). (H) The relative usage frequencies of syllables in the whole arena in the F1 offspring of lyral-unpaired (light purple) and lyral-paired (dark purple) fathers (Error bars are standard error. Kruskal-Wallis test with Dunn’s multiple comparisons. Syllable 2 lyral F1 unpaired vs. F1 paired p=0.0016. Syllable 3 p=0.02724. Syllable 5 p=0.003. Syllable 7 p=0.00252. Syllable 15 p=0.0208. Syllable 20 p=0.00015. n=6,13.). (I) The relative usage frequencies of syllables in the whole arena in the F1 offspring of propanol-unpaired (light blue) and propanol-paired (dark blue) fathers (Error bars are standard error. Kruskal-Wallis test with Dunn’s multiple comparisons. Syllable 6 propanol F1 unpaired vs. F1 paired p=0.0206. Syllable 12 p=0.04068. Syllable 14 p=0.0072. Syllable 15 p=0.03204. Syllable 20 p=0.0148. n=11,8.).
Figure 6—figure supplement 1
Density of syllable usage across space in F1 offspring.
(A) Kernel density estimate (KDE) plots of syllable usage as a function of the animal’s center point along the x-axis (x distance from the conditioned odor port). The two vertical bars indicate the divisions between the three chambers (conditioned odor chamber left, control odor chamber right). n=35 (F1 unpaired), 34 (F1 paired).
Videos
Video 1
Representative video of zone 1 of a cleared MOE from a homozygous M71-GFP acetophenone unpaired mouse.
M71 OSNs are visualized in white. Scale indicated in video.
Video 2
Representative video zooming into a nasal turbinate (zone 1) of a cleared MOE from a homozygous M71-GFP acetophenone unpaired mouse.
M71 OSNs are visualized in white. Scale indicated in video.
Video 3
Representative video of a cleared MOE from a homozygous M71-GFP acetophenone unpaired mouse, showing coronal slices of a section of zone 1 from anterior to posterior.
The top of the video is dorsal, and the bottom is ventral. Scale indicated in video.
Video 4
Representative video of zone 1 of a cleared MOE from a homozygous M71-GFP acetophenone paired mouse.
M71 OSNs are visualized in white. Scale indicated in video.
Video 5
Representative video zooming into a nasal turbinate (zone 1) of a cleared MOE from a homozygous M71-GFP acetophenone paired mouse.
M71 OSNs are visualized in white. Scale indicated in video.
Video 6
Representative video of a cleared MOE from a homozygous M71-GFP acetophenone paired mouse, showing coronal slices of a section of zone 1 from anterior to posterior.
The top of the video is dorsal, and the bottom is ventral. Scale indicated in video.
Video 7
Representative videos of odor preference behavior assay.
Left chamber: propanol. Right chamber: lyral. In the left video, the mouse has undergone unpaired conditioning with lyral. In the right video, the mouse has undergone paired conditioning with lyral. Both videos are at 20X playback speed.
Additional files
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Supplementary file 1
- https://cdn.elifesciences.org/articles/92882/elife-92882-supp1-v1.xlsx
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Supplementary file 2
Spreadsheet with all statistics for all figure supplements.
- https://cdn.elifesciences.org/articles/92882/elife-92882-supp2-v1.xlsx
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MDAR checklist
- https://cdn.elifesciences.org/articles/92882/elife-92882-mdarchecklist1-v1.pdf
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Fear conditioning biases olfactory sensory neuron frequencies across generations
eLife 12:RP92882.
https://doi.org/10.7554/eLife.92882.3
