Abstract
Humans and other animals can sense the negative states of other individuals and respond with prosocial helping behaviour to improve their conditions1,2. Although prosocial helping behaviour is proposed to have an evolutionary root in caring for vulnerable newborn offspring1,3, whether the neural substrates underlying parenting may contribute to adult-directed helping behaviours remains largely unclear. Here we show that mice with higher levels of parenting show more prosocial allogrooming towards stressed adults. The medial preoptic area (MPOA), a brain area involved in parenting behaviour, bidirectionally regulates allogrooming towards stressed conspecifics. Allogrooming and parenting behaviours recruit a partially overlapping neuronal ensemble in the MPOA, are both controlled by an MPOA-to-ventral tegmental area pathway and are associated with dopamine release in the nucleus accumbens. Using activity-dependent labelling, we demonstrate that MPOA neuronal ensembles engaged during parenting behaviours are functionally required for allogrooming. Conversely, MPOA neurons activated during prosocial behaviour are functionally required for pup grooming. Collectively, these findings uncover a neural circuit mechanism of prosocial helping behaviour and reveal partially shared neural substrates between parenting and helping behaviours, suggesting that the neural systems evolved for offspring care may have provided a scaffold for the emergence of broader prosocial support between adults.
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Data availability
All data and analyses necessary to understand the conclusions of the paper are presented in the main text and in Extended Data. Source data are provided with this paper.
Code availability
Code for behavioural analysis is available via GitHub at https://github.com/hongw-lab/Behavior_Annotator, animal pose tracking is available via GitHub at https://github.com/murthylab/sleap/releases/tag/v1.2.9 and microendoscopic imaging data analysis is available via GitHub at https://github.com/etterguillaume/MiniscopeAnalysis, https://github.com/zhoupc/CNMF_E, https://github.com/flatironinstitute/NoRMCorre and https://github.com/hongw-lab/CScreener.
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Acknowledgements
We thank L. McLeod, D. Fillet and S. Chaudhry for technical assistance, and members of the Hong laboratory for valuable comments. Schematics in Figs. 1a,i,j, 2b,n,u, 3a,e, 4a,g and 5b, Extended Data Figs. 3i and 9a,p,u were created in BioRender. Brain region outlines in Figs. 1j, 2a,m,u, 4f,p,s and 5b and Extended Data Figs. 5j, 6a,d,g, 7a,i,t and 9a were adapted from the Allen Mouse Brain Reference Atlas (https://atlas.brain-map.org). This work was supported in part by the National Institutes of Health (grant nos. R01 MH130941, R01 NS113124, R01 MH132736 and RF1 NS132912 to W.H. and F31 MH134495 to K.Y.L.), a Vallee Scholar Award, a Mallinckrodt Scholar Award (to W.H.), a Marion Bowen Postdoctoral grant (to F.S.) and Brain and Behaviour Research Foundation Young Investigator grants (to Y.E.W. and to F.S.).
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Contributions
F.S. performed microendoscopic imaging, fibre photometry and chemogenetic experiments including activity-dependent manipulation (TRAP) and photo-inhibition of VTA-projecting MPOA neurons, and analysed the data. K.Y.L. and J.D. performed prosocial and pup interaction assays, optogenetic manipulations of MPOA neurons, photo-activation of MPOA projections to the VTA and analysed the data. Y.E.W. performed computational analysis of microendoscopic imaging data. W.H., Y.E.W., F.S. and K.Y.L. wrote the paper with the input from L.I.Z. W.H. supervised the entire study.
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Extended data figures and tables
Extended Data Fig. 1 Correlation between prosocial and parenting behaviors.
a–c, f–h, Correlation between the duration of pup grooming behavior and the increase in allogrooming toward stressed adult conspecifics (a, f), the duration of baseline adult investigation (b, g), and the duration of baseline self-grooming (c, h) in females (a-c) and males (f-h). We note that Fig. 1f-h and Extended Data Fig. 1f-h appear to be similar but present different data: in Fig. 1f-h, we present the total duration of parenting behaviors on the Y-axis, and in Extended Data Fig. 1f-h, we present the total duration of pup grooming on the Y-axis. Given that parenting behaviors in virgin males predominantly consist of pup grooming (99.4%), the differences between the figures appear minimal. d, e, i, j, Correlation between the duration of parenting behaviors (d, i) or pup grooming behavior (e, j) and the duration of baseline allogrooming toward unstressed adult conspecifics in females (d, e) and males (i, j). k, l, Example ethograms of different behaviors displayed by mother (k) or father (l) mice during interactions with stressed adults (left) or pups (right). m–r, Correlation between the duration of all parenting behaviors (m-o) or pup grooming behavior (p-r) and the increase in allogrooming toward stressed adult conspecifics (m, p), the duration of baseline adult investigation (n, q), and the duration of baseline self-grooming (o, r) in mothers and fathers. Opposite-sex mates of the parental animals were used as demonstrators. In (m), both sexes showed a positive correlation between these behaviors: R² = 0.3605, p = 0.0139 for mothers, and R² = 0.2728, p = 0.0458 for fathers. s–v, No correlation between the duration of allogrooming toward stressed adult conspecifics and the duration of self-grooming in the prosocial interactions in females (s), males (t), mothers (u) and fathers (v). w, x, Duration of allogrooming (w) and investigation (x) displayed by subject animals during interaction with stressed adults in darkness or under light illumination. y, Number of USV syllables produced by the subjects during interactions with unstressed partners, stressed partners, and pups. (w-y), The center line in the boxplots indicates the median, the box limits indicate the upper and lower quartiles, and the whiskers indicate the range from the minimum to the maximum value. (a-j, m-v), Linear regression, the linear regression lines (solid lines) and 95% confidence intervals (shaded areas) are shown. (w, x), Two-sided Wilcoxon signed-rank test. (y), Kruskal-Wallis test with post hoc Dunn’s multiple comparisons test. n = 12 mice in (a-e, s), 14 mice in (f-j, t), 31 mice in (m-r), 16 mice in (u), 15 mice in (v), 8 mice in (w, x), 6 mice in each group in (y). See Supplementary Table 1 for details of statistical analyses. *P < 0.05. **P < 0.01. ns, not significant.
Extended Data Fig. 2 MPOA representations of the stress state of others and allogrooming behavior.
a, Performance of decoders in differentiating between investigation events toward stressed versus unstressed conspecifics in male and female subject animals. b–d, Performance of decoders in discriminating between allogrooming and baseline (b), between allogrooming and investigation toward stressed conspecifics (c), and between allogrooming and self-grooming (d) in male and female subject animals. e, Performance of 4-way decoders in classifying investigation toward stressed conspecifics, allogrooming, self-grooming, and baseline. f, The magnitude of the coefficients of investigation and allogrooming toward stressed conspecifics in generalized linear models between the activities of individual neurons and various types of behaviors (investigation toward unstressed animals, investigation toward stressed conspecifics, allogrooming, self-grooming, and baseline). g, The fraction of cells with significant, positive coefficients for investigation or allogrooming toward stressed conspecifics in the generalized linear models. h, The magnitude of the coefficients of investigation and allogrooming toward stressed conspecifics in linear mixed-effects models of the activities of individual neurons that included various types of behaviors as fixed effects and the identity of the interacting partners as a random effect. i, The fraction of cells with significant, positive coefficients for investigation or allogrooming toward stressed conspecifics in the linear mixed-effects models. j–m, Performance of decoders in discriminating between investigation events toward stressed versus unstressed conspecifics (j), between allogrooming and baseline (k), between allogrooming and investigation toward stressed conspecifics (l), and between allogrooming and self-grooming (m) when using different numbers of randomly sampled cells. n, o, The fraction of cells significantly activated (determined by ROC analysis) during different types of behaviors among all cells (n) or subsets of cells with an auROC value > 0.7 (o). (a-e, g, i, j-o), The center line in the boxplots indicates the median, the box limits indicate the upper and lower quartiles, and the whiskers indicate the range from the minimum to the maximum value. (f, h), The center line in the boxplots indicates the median, the box limits indicate the upper and lower quartiles, and the whiskers indicate data within 1.5× interquartile range. (a-e, f-i), Two-sided Wilcoxon signed-rank test. n = 10 male mice and 11 female mice in (a, g, i, n, o), 9 male mice and 9 female mice in (b-d, e, j-m), 1772 cells from 21 mice (10 male mice and 11 female mice) in (f, h). See Supplementary Table 1 for details of statistical analyses. **P < 0.01. ***P < 0.001. ****P < 0.0001.
Extended Data Fig. 3 Functional manipulations of the MPOA in pup grooming.
a, d, Example ethograms showing pup-directed behaviors exhibited by hM4Di-expressing (a) or mCherry-expressing (d) animals injected with either saline or CNO. b, c, e–h, Duration of pup grooming (b, e), duration of pup sniffing (c, f), percentages of time that animals engaged in self-grooming behavior during periods when they did not display social behavior (social investigation or allogrooming) in the prosocial interaction assay (g, h) following saline or CNO injection in hM4Di-expressing (b, c, g) or mCherry-expressing (e, f, h) animals. i, Schematic of optogenetic inhibition of MPOAVgat+ neurons during pup interactions (top) and example ethograms showing pup grooming during 5-s real (light on) and sham (light off) photo-inhibition in GtACR-expressing animals or during light delivery in mCherry-expressing control animals. j–l, Probability of pup grooming at various time points (j), average pup grooming duration across all trials within individual mice (k), and pup grooming duration in individual trials (l) during the 5-s real (light on) and sham (light off) photo-inhibition in GtACR-expressing mice and during light delivery in mCherry-expressing control mice. m, n, Average pup grooming duration during the 15-s real (light on) and sham (light off) photo-activation in individual mice expressing ChR2 (m) or EYFP (n). o-q, Pup grooming probability (o), pup grooming duration in individual trials (p), and pup grooming onset latency in individual trials (q) during the 15-s real (light on) and sham (light off) photo-activation in ChR2-expressing mice that showed any pup grooming during stimulations. (b, c, e, f, g, h, l, p, q), The center line in the boxplots indicates the median, the box limits indicate the upper and lower quartiles, and the whiskers indicate the range from the minimum to the maximum value. (b, c, e, f, g, h, k, m, n), Two-sided Wilcoxon signed-rank test. (l), Kruskal-Wallis test with post hoc Dunn’s multiple comparisons test. (p, q), Two-sided Wilcoxon rank-sum test. n = 7 mice in (b, c), 4 mice in (e, f), 10 mice in (g), 6 mice in (h), 13 mice in (k), 89-115 trials in 10-13 mice in (l), 31 mice in (m), 10 mice in (n), and 107-171 trials from 17 mice in (p, q). See Supplementary Table 1 for details of statistical analyses. *P < 0.05. ****P < 0.0001. ns, not significant. Schematic in i created in BioRender; Sun, F. https://biorender.com/pn3jiby, https://biorender.com/9fr13q3 (2026).
Extended Data Fig. 4 MPOA representations of parenting and prosocial behaviors.
a, Calcium traces of example MPOA neurons that are significantly activated in response to unstressed adult conspecifics, stressed adult conspecifics, or pups. b, Performance of decoders in differentiating between investigation events toward stressed adults versus pups in all animals, males alone, or females alone. c, Venn diagram of MPOA neurons that are significantly activated during investigation toward stressed adults or pups when these two types of social targets are presented simultaneously. d, Performance of decoders trained using MPOA population activities in differentiating between investigation events toward stressed adults versus pups during simultaneous presentation. e, Principal component (PC) projections of MPOA population activity associated with episodes (individual dots) of investigation toward unstressed adults, stressed adults, and pups from example animals. Each plot represents an individual subject animal. a.u., arbitrary units. Cross lines, mean ± SD. f, Average Mahalanobis distances between investigation events toward pups and unstressed adults (“Pup–Unstressed adult”) and between investigation events toward pups and stressed adults (“Pup–Stressed adult”) in males and females. The distances are calculated using the first 3 PCs of MPOA population activities. g, Venn diagram of neurons significantly activated during allogrooming, pup grooming, or self-grooming. Total neurons imaged: 1,772; non-responsive neurons: 820. h, Example field of view showing the spatial distribution of neurons activated during allogrooming, pup grooming, or self-grooming. i, j, Fraction of cells with significant, positive coefficients for both allogrooming and pup grooming in generalized linear models (i) and linear mixed-effects models (j) between single-neuron activities and behaviors. k, l, Performance of models trained to decode allogrooming versus baseline in predicting pup grooming (k) and models trained to decode pup grooming versus baseline in predicting allogrooming (l) in male and female subjects. m, n, Performance of models trained to decode allogrooming versus baseline in predicting self-grooming (m) and models trained to decode self-grooming versus baseline in predicting allogrooming (n) in male and female subjects. o, p, Performance of mutual decoding between allogrooming and self-grooming when models are trained using events from the same (o) or different (p) sessions. To predict each self-grooming event, the models were trained using allogrooming events from the same (o) or different (p) sessions. To predict each allogrooming event, the models were trained using self-grooming events from the same (o) or different (p) sessions. Each presentation of an adult conspecific or pup is considered a “session”. q, Performance of mutual decoding between allogrooming and pup grooming after removing cells that are activated during both behaviors. r, Venn diagram of neurons that are significantly activated during investigation toward stressed adults or pups in mothers and fathers. s, Performance of decoders in differentiating between investigation events toward stressed adults versus pups in mothers and fathers. Same-sex, familiar non-cage mates of the parental animals were used as demonstrators. t, Average Mahalanobis distances between investigation events toward pups and unstressed adults (“Pup–Unstressed adult”) and between investigation events toward pups and stressed adults (“Pup–Stressed adult”) in mothers and fathers. The distances are calculated using the first 3 PCs. u, v, Performance of models trained to decode allogrooming versus baseline in predicting pup grooming (u) and self-grooming (v) in parents. (b, d, i-q, s), The center line in the boxplots indicates the median, the box limits indicate the upper and lower quartiles, and the whiskers indicate the range from the minimum to the maximum value. (b, d, f, i-q, s-v), Two-sided Wilcoxon signed-rank test. n = 10 male mice and 11 female mice in (b, f, i, j), 6 male mice in (d), 9 male mice and 9 female mice in (k-n, q), 6 male mice and 7 female mice in (o), 5 male mice and 7 female mice in (p), 13 mice (9 mothers and 4 fathers) in (s, t) and 6 male mice in (u, v). See Supplementary Table 1 for details of statistical analyses. *P < 0.05. **P < 0.01. ***P < 0.001. ****P < 0.0001. ns, not significant.
Extended Data Fig. 5 Dopamine release during allogrooming and pup grooming.
a, Average ∆F/F0 signals within 2 seconds around the onset of investigation toward stressed adults, pups and during sucrose consumption in the NAc of GRABDA-expressing mice b, Venn diagram of neurons that are significantly activated during investigation toward pups, stressed adults, or sucrose consumption. Total neurons imaged: 401; non-responsive neurons: 258. c, Example traces of neurons that significantly activated only during investigation toward stressed adults, investigation toward pups, or sucrose consumption, but not during the other two events. d,e, Average MPOAVgat+ neural activity (mean ± SEM) during investigation toward stressed adults (d) or pup (e). Time 0 indicates behavior onset. f,g, Average NAc dopamine signal (mean ± SEM) during investigation toward stressed adults (f) or pup (g). Time 0 indicates behavior onset. h, PCC between MPOAVgat+ neural activity and NAc dopamine signal during 15-s periods around the onset of self-grooming events (from 5 s before to 10 s after). PCC in the “shuffle” group is calculated using temporally permutated calcium traces. i, PCC between MPOAVgat+ neural activity and NAc dopamine signal during 5-min non-social sessions and 8-min prosocial interaction sessions. The PCC of prosocial interaction sessions is re-plotted from Fig. 4k. j, Schematic of fiber photometry recording in the NAc combined with optogenetic activation of MPOAVgat+ or MPOAVglut2+ neurons. k, Example micrographs showing GRABDA expression and optic fiber placement in the NAc as well as ChrimsonR expression and optic fiber placement in the MPOA in Vgat-Cre and Vglut2-Cre animals. Scale bar, 500 μm. l–o, Average responses of NAc dopamine signals (l, n) and mean ∆F/F0 (m, o) during 5-s photo-activation of MPOAVgat+ neurons (l, m) or MPOAVglut2+ neurons (n, o) in ChrimsonR-expressing mice and mCherry-expressing control mice. (n, o), The center line in the boxplots indicates the median, the box limits indicate the upper and lower quartiles, and the whiskers indicate the range from the minimum to the maximum value. (a, d-i, l, n), Data are presented as the mean ± SEM. (a), Kruskal-Wallis test with post hoc Dunn’s multiple comparisons test. (h, i), Two-sided Wilcoxon signed-rank test. (m, n), Two-sided Wilcoxon rank-sum test. n = 6 mice in (a), 7 mice in (h, i), 7 mice for the ChrimsonR group and 4 mice for the mCherry group in (m), 6 mice for the ChrimsonR group and 4 mice for the mCherry group in (o). See Supplementary Table 1 for details of statistical analyses. *P < 0.05. **P < 0.01. ns, not significant. Schematic in j adapted from the Allen Mouse Brain Reference Atlas (https://atlas.brain-map.org).
Extended Data Fig. 6 Activation of the MPOAVgat+–VTA pathway is associated with positive valence.
a–c, Schematic of stimulation strategy (a), population median heatmaps (b), and the fraction of time spent on the stimulation side (c) in ChR2-expressing mice and EYFP-expressing control mice in the real-time place preference test (RTPP) during photo-activation of MPOAVgat+ neurons. d–f, Schematic of stimulation strategy (d), population median heatmaps (e), and the fraction of time spent on the stimulation side (f) in ChR2-expressing mice and EYFP-expressing control mice in the RTPP test during photo-activation of VTA-projecting MPOAVgat+ neurons. g–i, Schematic of stimulation strategy (g), population median heatmaps (h), and the fraction of time spent on the stimulation side (i) in ChR2-expressing mice and EYFP-expressing control mice in the RTPP test during photo-activation of the VTA projections of MPOAVgat+ neurons. (c, f, i), The center line in the boxplots indicates the median, the box limits indicate the upper and lower quartiles, and the whiskers indicate the range from the minimum to the maximum value. (c, f, i), Two-sided Wilcoxon rank-sum test. n = 13 mice in the ChR2 group and 6 mice in the control group in (c), 18 mice in the ChR2 group and 9 mice in the control group in (f), and 10 mice in the ChR2 group and 5 mice in the control group in (i). See Supplementary Table 1 for details of statistical analyses. **P < 0.01, ***P < 0.001, ****P < 0.0001. Schematics in a, d and g adapted from the Allen Mouse Brain Reference Atlas (https://atlas.brain-map.org).
Extended Data Fig. 7 Functional manipulation of the MPOA-VTA pathway in allogrooming and pup grooming behaviors.
a, Schematic of labeling of MPOAVgat+ neurons and retrograde labeling of VTA-projecting MPOA neurons. b, Percentage of VTA-projecting MPOAVgat+ neurons within the MPOAVgat+ neurons. c, d, Average duration of pup grooming across all trials within individual mice (c) and the duration of pup grooming in individual trials (d) during the 5-s real (light on) and sham (light off) photo-inhibition in GtACR-expressing mice and during light delivery in EYFP-expressing control mice. e–h, Probability of pup grooming at various time points (e), average pup grooming duration across all trials within individual mice (f), duration of pup grooming in individual trials (g), and onset latency of pup grooming in individual trials (h) during the 15-s real (light on) and sham (light off) photo-activation in ChR2-expressing mice and during light delivery in EYFP-expressing control mice. i, Schematic of optogenetic activation of VTA-projecting MPOAVgat+ neurons. j–q, Average allogrooming (j, n) or pup grooming (l, p) duration across all trials within individual mice and duration of allogrooming (k, o) or pup grooming (m, q) in individual trials during the 15-s real (light on) and sham (light off) photo-activation in ChR2-expressing mice and during light delivery in EYFP-expressing control mice. In (j-m), subject mice are virgin males or females; in (n-q), subject mice are fathers or mothers. r, s, Fraction of animals showing optogenetically induced allogrooming (r) or pup grooming (s) during stimulation of the general MPOAVgat+ population, selective activation of the VTA-projecting subpopulation of MPOAVgat+ neurons, or MPOAVgat+-VTA projection in several experiments. Subject mice are virgin males or females. t, Schematic of optogenetic activation of the MPOAVgat+-VTA projection with concurrent lidocaine infusion in the MPOA. u, v, Average allogrooming (u) or pup grooming (v) duration across all trials within individual mice during the 15-s real (light on) and sham (light off) photo-activation of the MPOAVgat+-VTA projection with the infusion of lidocaine or saline in the MPOA. w, Average allogrooming duration across all trials within individual mice during the 15-s real (light on) and sham (light off) photo-activation of VTA-projecting MPOAVgat+ neurons in ChR2-expressing mice when they are engaged in the interaction with unstressed partners and stressed partners. (b, d, g, h, k, m, o, q, w), The center line in the boxplots indicates the median, the box limits indicate the upper and lower quartiles, and the whiskers indicate the range from the minimum to the maximum value. (c, f, j, l, n, p), Two-sided Wilcoxon signed-rank test. (d, g, h, k, m, o, q), Kruskal-Wallis test with post hoc Dunn’s multiple comparisons test. (u, v, w), Two-way ANOVA with Tukey’s multiple comparisons test. n = 6 independently injected hemispheres (3 brain sections per hemisphere) in (b). n = 11 mice in (c), 64-91 trials from 7-11 mice in each group in (d), 10 mice in (f), 44-82 trials from 6-10 mice in each group in (g, h), 18 mice in (j), 93-202 trials from 10-18 mice in each group in (k), 17 mice in (l), 96-182 trials from 10-17 mice in each group in (m), 6 mice in (n, p), 34-71 trials from 5-6 mice in each group in (o), 38-69 trials from 5-6 mice in each group in (q), 8 mice in each group in (u, v), and 5 mice in each group in (w). See Supplementary Table 1 for details of statistical analyses. *P < 0.05. **P < 0.01. ***P < 0.001. ****P < 0.0001. ns, not significant. Schematics in a, i and t adapted from the Allen Mouse Brain Reference Atlas (https://atlas.brain-map.org).
Extended Data Fig. 8 Functional manipulation of the VTA-projecting MPOA neurons in male and female mice.
a, d, Average duration of allogrooming (a) or pup grooming (d) across all trials within individual mice during the 5-s real (light on) and sham (light off) photo-inhibition in GtACR-expressing male and female mice. b, c, e, f, The duration of allogrooming (b, c) or pup grooming (e, f) in individual trials during the 5-s real (light on) and sham (light off) photo-inhibition in GtACR-expressing mice and during light delivery in EYFP-expressing control mice. Data from male mice (b, e) and female mice (c, f) are separately presented. g, j, Average allogrooming (g) or pup grooming (j) duration across all trials within individual mice during the 15-s real (light on) and sham (light off) photo-activation in ChR2-expressing male and female mice. h, i, k, l, Allogrooming (h, i) or pup grooming (k, l) duration in individual trials during the 15-s real (light on) and sham (light off) photo-activation in ChR2-expressing mice and during light delivery in EYFP-expressing control mice. Data from male mice (h, k) and female mice (i, l) are separately presented. (a-l), The center line in the boxplots indicates the median, the box limits indicate the upper and lower quartiles, and the whiskers indicate the range from the minimum to the maximum value. (a, d, g, j), Two-way ANOVA with Fisher’s LSD test. (b, c, e, f, h, i, k, l), Kruskal-Wallis test with post hoc Dunn’s multiple comparisons test. n = 4 males and 4 females in (a), 6 males and 5 females in (d), 8 males and 10 females in (g), 8 males and 9 females in (j). n = 41-47 trials from 4 GtACR-expressing male mice, 35 trials from 3 control male mice in (b). n = 21-24 trials from 4 GtACR-expressing female mice, 30 trials from 3 control female mice in (c). n = 26-27 trials from 6 GtACR-expressing male mice, 34 trials from 3 control male mice in (e). n = 37-47 trials from 5 GtACR-expressing female mice, 57 trials from 4 control female mice in (f). n = 52-83 trials from 8 ChR2-expressing male mice, 49 trials from 5 control male mice in (h). n = 80-119 trials from 10 ChR2-expressing female mice, 44 trials from 5 control female mice in (i). n = 61-91 trials from 8 ChR2-expressing male mice, 56 trials from 5 control male mice in (k). n = 69-91 trials from 9 ChR2-expressing female mice, 40 trials from 5 control female mice in (l). See Supplementary Table 1 for details of statistical analyses. *P < 0.05. **P < 0.01. ****P < 0.0001. ns, not significant.
Extended Data Fig. 9 Functional manipulation of MPOA neurons activated during pup-TRAP, non-social-TRAP, sucrose-TRAP, and pup-sniffing-TRAP.
a, Schematic of the experimental timeline, including virus injection, labeling of non-social-TRAP neurons, and behavioral testing. b, e, Example ethograms showing behaviors exhibited by non-social-TRAP animals injected with either saline (control) or CNO during interactions with stressed adults (b) or pups (e). c, d, Duration of social investigation (c) and allogrooming (d) exhibited by non-social-TRAP animals injected with either saline (control) or CNO during prosocial interactions. f, g, Duration of pup investigation (f) and pup grooming (g) exhibited by non-social-TRAP animals injected with either saline (control) or CNO during pup interactions. h, i, l, m, Bout of investigation (h, l) and allogrooming (i, m) exhibited by pup-TRAP animals (h, i), or non-social-TRAP animals (l, m) injected with either saline (control) or CNO during interactions with stressed animals. j, k, n, o, Bout of pup investigation (j, n) and pup grooming (k, o) exhibited by pup-TRAP animals (j, k) or non-social-TRAP animals (n, o) injected with either saline (control) or CNO during interactions with pups. p, u, Schematic of the behavior paradigm used to label pup-sniffing-TRAP neurons (p) and sucrose-TRAP neurons (u). q, v, Example images showing hM4Di-mCherry expression in pup-sniffing-TRAP+ neurons (q) and sucrose-TRAP+ neurons (v) in the MPOA. Scale bars, 500 μm. r, w, Example ethograms showing behaviors exhibited by pup-sniffing-TRAP animals (r), or sucrose-TRAP animals (w), injected with either saline (control) or CNO during interactions with stressed adults. s, t, x, y, Duration of social investigation (s, x) and allogrooming (t, y) exhibited by pup-sniffing-TRAP animals (s, t), or sucrose-TRAP animals (x, y), injected with either saline (control) or CNO during prosocial interactions. (c, d, f, g, h-o, s, t, x, y), The center line in the boxplots indicates the median, the box limits indicate the upper and lower quartiles, and the whiskers indicate the range from the minimum to the maximum value. (c, d, f, g, h-o, s, t, x, y), Two-sided Wilc–oxon signed-rank test. n = 12 mice in (c, d, f, g, l-o), 13 mice in (h-k), 16 mice in (s, t, x, y). See Supplementary Table 1 for details of statistical analyses. *P < 0.05. **P < 0.01. ns, not significant. Schematic in a adapted from the Allen Mouse Brain Reference Atlas (https://atlas.brain-map.org). Schematics in a, p and u created in BioRender; Sun, F. https://biorender.com/u651o8b, https://biorender.com/pn3jiby, https://biorender.com/06frs8k, https://biorender.com/9fr13q3 (2026).
Extended Data Fig. 10 Characterizations of TRAP+ neurons in the MPOA.
a-g, Characterizations of TRAP+ neurons. a1, b1, c1, d1, Example images showing hM4Di-mCherry expression in pup-TRAP+ neurons (a1), non-social-TRAP+ neurons (b1), stressed-TRAP+ neurons (c1), and unstressed-TRAP+ neurons (d1). Scale bars, 500 μm. a2, b2, Example images showing pup-TRAP+ neurons (a2) or non-social-TRAP+ neurons (b2) expressing hM4Di-mCherry (red), cells expressing Fos (green) following pup interactions, and the overlap between the two populations in the MPOA. Scale bar, 50 μm. c2, d2, Example images showing stressed-TRAP+ neurons (c2) or unstressed-TRAP+ neurons (d2) expressing hM4Di-mCherry (red), cells expressing Fos (green) following interactions with stressed adults, and the overlap between the two populations in the MPOA. Scale bar, 50 μm. e, Percentages of pup-TRAP+ and non-social-TRAP+ neurons that overlapped with Fos+ cells. f, Percentages of stressed-TRAP+ and unstressed-TRAP+ neurons that overlapped with Fos+ cells. g, Percentage of hM4Di-expressing neurons within the NeuN+ neurons in the MPOA of TRAP animals. h, i, j, k, Percentages of time that pup-TRAP animals (h), non-social-TRAP animals (i), stressed-TRAP animals (j), and unstressed-TRAP animals (k) engaged in self-grooming behavior during periods when they did not display social behavior (social investigation or allogrooming) in the prosocial interaction assay following saline or CNO injection. l, o, r, u, Example locomotion trajectories of pup-TRAP animals (l), non-social-TRAP animals (o), stressed-TRAP animals (r), and unstressed-TRAP animals (u) in the open field test following saline (control) or CNO injection. m, n, p, q, s, t, v, w, Time spent in the centers and travel distance of pup-TRAP animals (m, n), non-social-TRAP animals (p, q), stressed-TRAP animals (s, t), and unstressed-TRAP animals (v, w) in the open field test following saline (control) or CNO injection. (e-k, m, n, s, t, p, q, v, w), The center line in the boxplots indicates the median, the box limits indicate the upper and lower quartiles, and the whiskers indicate the range from the minimum to the maximum value. (e, f), Two-sided Wilcoxon rank-sum test. (g), Kruskal-Wallis test with post hoc Dunn’s multiple comparison test. (h-k, m, n, s, t, p, q, v, w), Two-sided Wilcoxon signed-rank test. n = 8 independently injected hemispheres (3 brain sections per hemisphere) in pup-TRAP group and 6 independently injected hemispheres (3 brain sections per hemisphere) in non-social-TRAP group in (e). n = 8 independently injected hemispheres (3 brain sections per hemisphere) in each group in (f, g). n = 13 mice in (h, m, n), 12 mice in (i, p, q), 20 mice in (j), 19 mice in (s, t), 14 mice in (k, v, w). ns, not significant. See Supplementary Table 1 for details of statistical analyses. *P < 0.05. **P < 0.01. ***P < 0.001. ns, not significant.
Supplementary information
Supplementary Information (download PDF )
This file includes Supplementary Table 1, which contains detailed statistical information, and Supplementary Notes 1–7, containing further notes and discussions.
Supplementary Video 1 (download MP4 )
Example annotated video showing prosocial interaction between a subject animal (a virgin female) and a same-sex stressed adult conspecific. Annotated behaviours of the subject animal are shown both in real-time within each frame and in the embedded ethogram, including social investigation and allogrooming episodes.
Supplementary Video 2 (download MP4 )
Example annotated video showing the interaction between a subject animal (a virgin female) and pups. Annotated behaviours of the subject animal are shown both in real-time within each frame and in the embedded ethogram, including pup investigation and pup grooming episodes.
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Sun, F., Lim, K.Y., Dang, J. et al. Shared neural substrates of prosocial and parenting behaviours. Nature (2026). https://doi.org/10.1038/s41586-026-10327-8
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DOI: https://doi.org/10.1038/s41586-026-10327-8
