1. Neuroscience

Adult-born granule cells modulate CA2 network activity during retrieval of developmental memories of the mother

  1. Blake J Laham
  2. Isha R Gore
  3. Casey J Brown
  4. Elizabeth Gould  Is a corresponding author
  1. Princeton Neuroscience Institute, Princeton University, United States
https://doi.org/10.7554/eLife.90600.3
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  1. Blake J Laham
  2. Isha R Gore
  3. Casey J Brown
  4. Elizabeth Gould
(2024)
Adult-born granule cells modulate CA2 network activity during retrieval of developmental memories of the mother
eLife 12:RP90600.
https://doi.org/10.7554/eLife.90600.3
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4 figures, 1 table and 1 additional file

Figures

Figure 1 with 2 supplements
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Adult-born granule cells support retrieval of developmental remote memories of the mother.

(a-aiii) Confocal images of dentate gyrus granule cells, both mature (ZnT3+) and adult-born (3R-Tau+), and their robust projections to CA2. Confocal images of adult-born granule cell (abGC) 3R-Tau+axons (arrows) (b) near proximal dendrite of a CA2 PCP4 + pyramidal cell and (c) near proximal dendrite of a CA2 mCherry + PV+ interneuron. (d) abGC fibers (3R-Tau+) in the CA2 are more abundant near proximal dendrites (n=7; paired t-test: t6=3.163, p=0.0195) of PV + interneurons (mCherry+) than those of pyramidal cells (PCP4+). (e) Timeline for behavioral experiment. (f) Schematic demonstrating direct social interaction assay used at three experimental time points. (g) Ablation of abGCs abolishes difference in investigation time between mother and novel mother (CD1: n=17; TK: n=14; two-way RM ANOVA: Genotype x Drug: F2,58 = 5.352, p=0.0074; Šídák’s test p=0.0021). After removing valganciclovir (VGCV) from rodent chow and allowing adult neurogenesis to recover over the course of 6 weeks, mice were able to discriminate between mother and novel mother again (p>0.999). (h-hii) Confocal images of abGCs in the DG at three different drug time points. VGCV administration produces a dramatic loss of (h-hii) abGCs (3R-Tau + cells) in DG (VGCV-: n=8; VGCV+: n=8; unpaired t-test: t14=6.588, p<0.0001) and (i-iii) 3R-Tau+ fibers in CA2 (VGCV: n=8; VGCV+: n=8; unpaired t-test: t14=3.488, p=0.0036). 6 weeks after removing VGCV from the rodent chow, DG abGC number (j, k) undergoes a 70% recovery (CD1: n=8; TK: n=8; unpaired t-test: t14=3.218, p=0.0062), and (l, m) 3R-Tau+ fibers present in CA2 undergo a complete recovery (CD1: n=8; TK: n=8; unpaired t-test: t14=0.7271, p=0.4791). *p<0.05, bars represent mean + SEM. Scale bars = 200 μm for a, h, i; 2 μm for b, c. ns = not significant.

© 2024, BioRender Inc. Figure 1f was created using BioRender, and is published under a CC BY-NC-ND 4.0. Further reproductions must adhere to the terms of this license

Figure 1—source data 1

Adult-born granule cells support retrieval of developmental remote memories of the mother.

https://cdn.elifesciences.org/articles/90600/elife-90600-fig1-data1-v1.xlsx
Figure 1—figure supplement 1
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Total investigation times during social interaction.

(a) Total investigation times of the mother and a novel mother were assessed for CD1 and GFAP-TK mice before (VGCV-), during (VGCV+), and after valganciclovir (VGCV) administration (VGCV- Recovery) (CD1: n=17; TK: n=14; RM ANOVA: Genotype x Drug: F6,116 = 1.672, p=0.1339; Drug: F1.173 = 5.515, p=0.0072). (b) Total investigation times of the mother and a novel mother were quantified for Nestin-cre mice and Nestin-cre:Gi mice with a 4–6-week-old population of inhibitory DREADD + adult-born granule cells (abGCs) (Nestin-cre: n=31; Nestin-cre:Gi: n=33; Mixed-effects ANOVA: Genotype x Drug x Stimulus: F1,56 = 4.290, p=0.0430; Tukey’s test: Nestin-cre Veh Mother vs Novel p=0.0004, Nestin-cre CNO Mother vs Novel p=0.0274, Nestin-cre:Gi Veh Mother vs Novel p=0.0326, Nestin-cre:Gi CNO Mother vs Novel p=0.9904). (c) Total investigation times of the mother and a novel mother were quantified for Nestin-cre mice and Nestin-cre:Gi mice with a 10–12-week-old population of inhibitory DREADD + abGCs (Nestin-cre: n=21; Nestin-cre:Gi: n=17; Mixed-effects ANOVA: Genotype x Drug x Stimulus: F1,12 = 1.204, p=0.2941; Drug: F1,40 = 60.46, p<0.0001). Bars represent mean + SEM. This figure contains data that relates to both main Figures 1 and 2.

Figure 1—figure supplement 1—source data 1

Total investigation times during social interaction.

https://cdn.elifesciences.org/articles/90600/elife-90600-fig1-figsupp1-data1-v1.xlsx
Figure 1—figure supplement 2
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Valganciclovir (VGCV) or clozapine-N-oxide (CNO) administration does not alter locomotion.

(a) GFAP-TK mice display normal locomotion during social interaction after 6 weeks of VGCV administration (CD1: n=17; TK: n=14; three-way ANOVA: Genotype x Drug x Stimulus: F1,29 = 0.01998, p=0.8886). CNO administration has no effect on locomotion during (b) baseline (Nestin-cre: n=7; Nestin-cre:Gi: n=9; two-way RM ANOVA: Genotype x Drug: F1,14 = 0.02291, p=0.8818) or (c) social stimulus trials (Nestin-cre: n=7; Nestin-cre:Gi: n=9; three-way ANOVA: Genotype x Drug x Stimulus: F1,14 = 0.06126, p=0.8081) in Nestin-cre and Nestin-cre:Gi mice. A significant two-way interaction was observed during social trials, but all post hoc tests failed to reach significance (two-way ANOVA: Genotype x Drug: F1,14 = 7.180, p=0.0180). Bars represent mean + SEM. ns = not significant. This figure contains data that relates to both main Figures 1 and 2.

Figure 1—figure supplement 2—source data 1

Valganciclovir (VGCV) or clozapine-N-oxide (CNO) administration does not alter locomotion.

https://cdn.elifesciences.org/articles/90600/elife-90600-fig1-figsupp2-data1-v1.xlsx
Figure 2
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Inhibiting 4–6-week-old abGC projections to CA2 prevents discrimination between mothers and novel mothers.

(a) Schematic of breeding to produce Nestin-cre:Gi double transgenic offspring. (b) Timeline outlining tamoxifen administration, cannula implantation, and behavioral testing. (c) Schematic demonstrating direct social interaction testing. (d-dii) Confocal images depicting 4–6-week-old Gi-DREADD + abGCs in the DG. (e) Inhibiting 4–6-week-old adult-born granule cell (abGC) projections to CA2 with clozapine-N-oxide (CNO) prevents discrimination between mother and novel mother (Nestin-cre: n=31; Nestin-cre:Gi: n = 33; two-way RM ANOVA: Genotype x Drug: F1,62 = 4.042, p=0.0487; Šídák’s test p=0.0057). (f) Inhibiting 10–12-week old abGC projections to CA2 does not influence discrimination between mother and novel mother Nestin-cre: n=21 (Veh), n=17 (CNO); Nestin-cre:Gi: n=18 (Veh), n=15 (CNO); Mixed-effects ANOVA: Genotype x Drug:1,67 = 1.669, Pp=0.2008. (g) Schematic for experiment assessing abGC contributions to social memory consolidation. (h) Systemic inhibition of abGCs during memory consolidation has no significant effect on social memory (Nestin-cre: n=21; Nestin-cre:Gi: n=18; two-way RM ANOVA: Genotype x Drug: F1,37 = 1.238, p=0.2730). *p<0.05, bars represent mean + SEM. ns = not significant. Scale bars in d=200 μm.

© 2024, BioRender Inc. Figure 2a, c and g were created using BioRender, and is published under a CC BY-NC-ND 4.0. Further reproductions must adhere to the terms of this license

Figure 2—source data 1

Inhibiting 4–6-week old abGC projections to CA2 prevents discrimination between mothers and novel mothers.

https://cdn.elifesciences.org/articles/90600/elife-90600-fig2-data1-v1.xlsx
Figure 3 with 3 supplements
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Adult-born neurons influence CA2 sharp wave ripple (SWR) changes associated with discrimination between novel mother and mother.

(a) Timeline of experiment demonstrating tamoxifen injection schedule and electrode implantation in CA2. (b) Confocal image demonstrating accuracy of electrode placement. (c) Example of SWR trace and (d) filtered trace recorded from CA2. (e) CA2 SWR frequency is increased during social interactions, regardless of whether the stimulus is the novel mother or mother (Veh: n=14; CNO: n=14; 2-way RM ANOVA: Stimulus: F1,13 = 46.40, p<0.0001). (f) Inhibiting 4–6-weekold adult-born granule cells (abGCs) with clozapine-N-oxide (CNO) prevents characteristic SWR production patterns present during exposure to novel mothers vs mothers (Nestin-cre: n=7; Nestin-cre:Gi: n=9; Mixed-effects ANOVA: Genotype x Drug: F1,110 = 18.6529, p<0.0001; Tukey’s test p<0.0001). (g) Inhibiting 4–6-week old abGCs significantly reduces social trial SWR production (Nestin-cre: n=7; Nestin-cre:Gi; n=9; Mixed-effects ANOVA: Genotype x Drug: F1,228 = 16.2465, P<0.0001; Tukey’s test p=0.0018). (h) Inhibiting 4–6 week old abGCs increases SWR peak amplitude during social interaction trials (Nestin-cre: n=7; Nestin-cre:Gi: n=9; Mixed-effects ANOVA: Genotype x Drug: F1,238 = 3.69, p=0.0559) and (j) baseline (Nestin-cre: n=7; Nestin-cre:Gi: n=9; Mixed-effects ANOVA: Genotype x Drug: F1,237 = 6.0322, p=0.01477; Tukey’s test p=0.0133). (i) Changes in baseline SWR production did not reach significance (Nestin-cre: n=7; Nestin-cre:Gi: n=9; Mixed-effects ANOVA: Genotype x Drug: F1,238 = 3.5101, p=0.0622). (k) At the 10–12 week abGC time point, CA2 SWR frequency is increased during social interactions, regardless of stimulus (Veh: n=9; CNO: n=11; Mixed-effects ANOVA: Stimulus: F1,10 = 133.1, p<0.0001). (l) Inhibiting 10–12-week old abGCs has no influence on characteristic SWR production patterns present during exposure to the novel mother vs mother (Nestin-cre: n=4 (Veh), n=5 (CNO); Nestin-cre:Gi: n=7 (Veh and CNO); Mixed-effects ANOVA: Genotype x Drug: F1,82 = 1.7431, p=0.1904), nor on (m) SWR production during social trials (Nestin-cre: n=4 (Veh), n=5 (CNO) Nestin-cre:Gi: n=8 (Veh and CNO); Mixed-effects ANOVA: Genotype x Drug: F1,170 = 0.4496, p=0.503), or (o) during baseline recording (Nestin-cre: n=4 (Veh), n=5 (CNO); Nestin-cre:Gi: n=8 (Veh and CNO); Mixed-effects ANOVA: Genotype x Drug: F1,170 = 2.0552, p=0.15352). (n) Inhibiting 10–12-week old adult-born granule cells (abGCs) has no influence on SWR peak amplitude social interaction trials (Nestin-cre: n=4 (Veh), n=5 (CNO); Nestin-cre:Gi: n=8 (Veh and CNO); Mixed-effects ANOVA: Genotype x Drug: F1,18 = 0.9744, p=0.3250) or (p) during baseline (Nestin-cre: n=4 (Veh), n=5 (CNO); Nestin-cre:Gi: n=8 (Veh and CNO); Mixed-effects ANOVA: Genotype x Drug: F1,172 = 0.540, p=0.8165), *p<0.05, bars represent mean + SEM. Scale bar in b=1500 μm. ns = not significant.

© 2024, BioRender Inc. Figure 3a was created using BioRender, and is published under a CC BY-NC-ND 4.0. Further reproductions must adhere to the terms of this license

Figure 3—source data 1

Adult-born neurons influence CA2 SWR changes associated with discrimination between novel mother and mother.

https://cdn.elifesciences.org/articles/90600/elife-90600-fig3-data1-v1.xlsx
Figure 3—figure supplement 1
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4–6-week-old a adult-born granule cells (bGCs) influence multiple features of CA2 sharp wave ripples (SWRs).

(a) Schematic of electrophysiological recordings during baseline and social trials during the 4–6-week-old abGC timepoint. (b) Silencing 4–6-week old abGCs reduces average CA2 SWR integral (Nestin-cre: n=7; Nestin-cre:Gi: n=9; Mixed-effects ANOVA: Genotype x Drug: F1,236 = 12.83, p=0.0004017; Tukey’s test p=0.0099). (c) Silencing 4–6-week old abGCs reduces average CA2 SWR duration (Nestin-cre: n=7; Nestin-cre:Gi: n=9; Mixed-effects ANOVA: Genotype x Drug: F1,223 = 24.077, p<0.0001; Tukey’s test p<0.0001). (d) Silencing 10–12-week old abGCs has no effect on average CA2 SWR integral but a significant interaction is observed (Nestin-cre: n=4 (Veh), n=5 (CNO); Nestin-cre:Gi: n=8 (Veh and CNO); Mixed-effects ANOVA: Genotype x Drug: F1,172 = 4.0061, p=0.0469; Tukey’s test p=0.7457). (e) Silencing 10–12-week old abGCs does not reduce average CA2 SWR duration but a significant interaction is observed (Nestin-cre: n=4 (Veh), n=5 (CNO); Nestin-cre:Gi: n=8 (Veh and CNO); Mixed-effects ANOVA: Genotype x Drug: F1172 = 8.5609, p=0.003898; Tukey’s test p=0.1439). *p<0.05, bars represent mean + SEM. ns = not significant.

© 2024, BioRender Inc. Figure 3—figure supplement 1a was created using BioRender, and is published under a CC BY-NC-ND 4.0. Further reproductions must adhere to the terms of this license

Figure 3—figure supplement 1—source data 1

4–6-weekold abGCs influence multiple features of CA2 SWRs.

https://cdn.elifesciences.org/articles/90600/elife-90600-fig3-figsupp1-data1-v1.xlsx
Figure 3—figure supplement 2
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Normalized sharp wave ripple (SWR) generation.

(a) Silencing 4–6-week-old adult-born granule cells (bGCs) alters CA2 SWR generation (Nestin-cre: n=7; Nestin-cre:Gi: n=9; Mixed-effects ANOVA: Genotype x Drug x Stimulus: F1,234 = 13.3621, p=0.000317; Tukey’s test: Nestin-cre Veh Mother vs Novel p=0.0042, Nestin-cre CNO Mother vs Novel p< 0.0001, Nestin-cre:Gi Veh Mother vs Novel p0.0003, Nestin-cre:Gi CNO Mother vs Novel p=0.7190). (b) Silencing 10–12 week-old abGCs has no influence on CA2 SWR generation (Nestin-cre: n=5; Nestin-cre:Gi: n=7; Mixed-effects ANOVA: Genotype x Drug x Stimulus: F1,161 = 2.1093, p=0.14834; Stimulus: F1,161 = 33.0013, p<0.0001). *p<0.05, bars represent mean + SEM.

Figure 3—figure supplement 2—source data 1

Normalized sharp wave ripple (SWR) generation.

https://cdn.elifesciences.org/articles/90600/elife-90600-fig3-figsupp2-data1-v1.xlsx
Figure 3—figure supplement 3
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Nonsocial stimuli suppress CA2 sharp wave ripple (SWR) frequency.

(a) Schematic of electrophysiological recordings during baseline and exposure to nonsocial object (plastic toy animal) trials during the 4–6-week old abGC timepoint. (b) Nonsocial stimuli slightly inhibit CA2 SWR frequency (Veh: n=14; CNO: n=16; 2-way RM ANOVA: Stimulus: F1,28 = 7.760, p=0.0095). (c) Silencing 4–6-week old adult-born granule cells (abGCs) has no influence on CA2 SWR frequency during object exposure (Nestin-cre: n=5 (Veh), n=7 (CNO); Nestin-cre:Gi: n=8 (Veh), n=9 (CNO); Mixed-effects ANOVA: Genotype x Drug: F1,95 = 0.7761, p=0.3806), (d) average integral (Nestin-cre: n=5, n=7 (CNO); Nestin-cre:Gi: n=9 (Veh and CNO); Mixed-effects ANOVA: Genotype x Drug: F1,108 = 0.1653, p=0.2027). (e) Silencing 4–6-week old abGCs reduces average SWR duration during object trials (Nestin-cre: n=5 (Veh), n=7 (CNO); Nestin-cre:Gi: n=9 (Veh and CNO); Mixed-effects ANOVA: Genotype x Drug: F1,106 = 7.9118, p=0.005849; Tukey’s test p=0.0094). (f) Silencing 4–6-week old abGCs has no effect on SWR peak z-score (Nestin-cre: n=5 (Veh), n=7 (CNO); Nestin-cre:Gi: n=9 (Veh and CNO); Mixed-effects ANOVA: Genotype x Drug: F1,111 = 0.0003, p=0.9874). *p<0.05, bars represent mean + SEM. ns = not significant.

© 2024, BioRender Inc. Figure 3—figure supplement 3a was created using BioRender, and is published under a CC BY-NC-ND 4.0. Further reproductions must adhere to the terms of this license

Figure 3—figure supplement 3—source data 1

Nonsocial stimuli suppress CA2 sharp wave ripple (SWR) frequency.

https://cdn.elifesciences.org/articles/90600/elife-90600-fig3-figsupp3-data1-v1.xlsx
Figure 4 with 1 supplement
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Adult-born neurons influence CA2 phase-amplitude coupling (PAC) changes associated with retrieval of developmental memories of the mother.

(a) Example trace of raw local field potential (LFP) (black), 4–12 Hz theta oscillation (magenta), and 50–100 Hz gamma oscillation (green). (b,c) Graphs demonstrating mid-gamma amplitude modulation across theta phases in Nestin-cre:Gi mice. (d) Inhibiting 4–6-week old adult-born granule cells (abGCs) abolishes the increase in theta-mid-gamma PAC present during exposure to the mother (Nestin-cre: n=7; Nestin-cre:Gi: n=8; Mixed-effects ANOVA: Genotype x Drug: F1,103 = 4.6729, p=0.03296; Šídák’s test p=0.0219). (e,f) Graphs demonstrating mid-gamma amplitude modulation across theta phases in Nestin-cre:Gi mice. (g) Inhibiting 10–12-week old abGCs has no influence on theta-mid-gamma PAC during exposure to the mother (Nestin-cre: n=4 (Veh), n=5 (CNO); Nestin-cre:Gi: n=8 (Veh), n=7 (CNO); Mixed-effects ANOVA: Genotype x Drug: F1,18 = 0.2616, p=0.6153). *p<0.05, bars represent mean + SEM.

Figure 4—source data 1

Adult-born neurons influence CA2 phase-amplitude coupling (PAC) changes associated with retrieval of developmental memories of the mother.

https://cdn.elifesciences.org/articles/90600/elife-90600-fig4-data1-v1.xlsx
Figure 4—figure supplement 1
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4–6-week old adult-born granule cell (abGC) inhibition does not influence theta and gamma power during social interaction.

(a,ai,c,ci,e,ei) Normalized power spectra graphs (Social stimulus/baseline). (b) Silencing 4–6-week old abGCs has no influence on theta power (Nestin-cre: n=7; Nestin-cre:Gi: n=9; Mixed-effects ANOVA: Genotype x Drug: F1,14 = 0.2303, p=0.6387). (d) Silencing 4–6-week old abGCs has no influence on low-gamma power (Nestin-cre: n=7; Nestin-cre:Gi: n=9; Mixed effects ANOVA: Genotype x Drug: F1,14 = 0.7979, p=0.3868). (f) Silencing 4–6-week old abGCs has no influence on mid-gamma power (Nestin-cre: n=7; Nestin-cre:Gi: n=9; Mixed-effects ANOVA: Genotype x Drug: F1,28 = 0.3702, p=0.5478). Bars represent mean + SEM for a-ai,cci,e-ei; + SEM for b,d,e. ns = not significant.

Figure 4—figure supplement 1—source data 1

4–6-week old adult-born granule cell (abGC) inhibition does not influence theta and gamma power during social interaction.

https://cdn.elifesciences.org/articles/90600/elife-90600-fig4-figsupp1-data1-v1.xlsx

Tables

Key resources table
Reagent type (species) or resourceDesignationSource or referenceIdentifiersAdditional information
Strain, strain background
(Mus musculus - males and females)		C57Bl/6-Tg(Nes-cre/ERT2)KEisc/J,Jackson labs016261
Strain, strain background
(Mus musculus - males and females)		B6.129-Gt(ROSA)26Sortm1(CAG-CHRM4*,-mCitrine)Ute/J,Jackson labs026219
Strain, strain background
(Mus musculus - males and females)		C57Bl/6,Jackson labs000644
Strain, strain background
(Mus musculus - males and females)		B6.129P2-Pvalbtm1(cre)Arbr/J,Jackson labs017320
Antibody3R-Tau, mouse, monoclonalMillipore Sigma05–8031:500
AntibodyZnt3, rabbit, polyclonalAlomone LabsAZT-0131:500
AntibodyRGS14, mouse, monoclonalUC Davis/NIH Neuromab75–1701:500
AntibodyPCP4, rabbit, polyclonalSigma-AldrichHPA0057921:500
Software, algorithmFIJIImageJNIH
Software, algorithmPrism 9.2.0GraphpadGraphPad Software
Software, algorithmNeuroExplorerNEX5NEX Technologies
OtherCre-dependent mCherry AAV2/5, pAAV-hSyn-DIO-mCherryAddgene50459Virus – see surgical procedures in Methods

Additional files

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  1. Blake J Laham
  2. Isha R Gore
  3. Casey J Brown
  4. Elizabeth Gould
(2024)
Adult-born granule cells modulate CA2 network activity during retrieval of developmental memories of the mother
eLife 12:RP90600.
https://doi.org/10.7554/eLife.90600.3