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Medial preoptic area antagonistically mediates stress-induced anxiety and parental behavior

Nature Neuroscience volume 24, pages 516–528 (2021)Cite this article

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Abstract

Anxiety is a negative emotional state that is overly displayed in anxiety disorders and depression. Although anxiety is known to be controlled by distributed brain networks, key components for its initiation, maintenance and coordination with behavioral state remain poorly understood. Here, we report that anxiogenic stressors elicit acute and prolonged responses in glutamatergic neurons of the mouse medial preoptic area (mPOA). These neurons encode extremely negative valence and mediate the induction and expression of anxiety-like behaviors. Conversely, mPOA GABA-containing neurons encode positive valence and produce anxiolytic effects. Such opposing roles are mediated by competing local interactions and long-range projections of neurons to the periaqueductal gray. The two neuronal populations antagonistically regulate anxiety-like and parental behaviors: anxiety is reduced, while parenting is enhanced and vice versa. Thus, by evaluating negative and positive valences through distinct but interacting circuits, the mPOA coordinates emotional state and social behavior.

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Fig. 1: mPOA glutamatergic neurons are activated by physical stress.
Fig. 2: Activating mPOA glutamatergic neurons enhances anxiety-like behaviors.
Fig. 3: Silencing mPOA glutamatergic neurons reduces stress-induced anxiety-like behaviors.
Fig. 4: mPOA glutamatergic neurons antagonistically regulate social stress-induced anxiety and parental behavior.
Fig. 5: The mPOA-to-PAG pathway primarily accounts for the role of the mPOA in regulating anxiety-like behaviors.
Fig. 6: GABAergic neurons play opposing roles in regulating anxiety state and parental behavior.
Fig. 7: Monosynaptic inputs to mPOA glutamatergic neurons.

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Data availability

All data that support the findings of this study are available from the corresponding authors upon request.

Code availability

The Python code for animal detection is available at https://github.com/GuangWei-Zhang/TraCon-Toolbox. The custom pupil size detection Python code and custom MATLAB code for fiber photometry or electrophysiological data analysis will be provided upon request to the corresponding authors.

References

  1. Calhoon, G. G. & Tye, K. M. Resolving the neural circuits of anxiety. Nat. Neurosci. 18, 1394–1404 (2015).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Craske, M. G. & Stein, M. B. Anxiety. Lancet 388, 3048–3059 (2016).

    Article  PubMed  Google Scholar 

  3. Tovote, P., Fadok, J. P. & Lüthi, A. Neuronal circuits for fear and anxiety. Nat. Rev. Neurosci. 16, 317–331 (2015).

    Article  CAS  PubMed  Google Scholar 

  4. Fenster, R. J., Lebois, L. A. M., Ressler, K. J. & Suh, J. Brain circuit dysfunction in post-traumatic stress disorder: from mouse to man. Nat. Rev. Neurosci. 19, 535–551 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Merikangas, K. R. et al. Longitudinal trajectories of depression and anxiety in a prospective community study: the Zurich Cohort Study. Arch. Gen. Psychiatry 60, 993–1000 (2003).

    Article  PubMed  Google Scholar 

  6. Adhikari, A. Distributed circuits underlying anxiety. Front. Behav. Neurosci. 8, 112 (2014).

    Article  PubMed  PubMed Central  Google Scholar 

  7. Tye, K. M. et al. Amygdala circuitry mediating reversible and bidirectional control of anxiety. Nature 471, 358–362 (2011).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Jennings, J. H. et al. Distinct extended amygdala circuits for divergent motivational states. Nature 496, 224–228 (2013).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Kim, S. Y. et al. Diverging neural pathways assemble a behavioural state from separable features in anxiety. Nature 496, 219–223 (2013).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Anthony, T. E. et al. Control of stress-induced persistent anxiety by an extra-amygdala septohypothalamic circuit. Cell 156, 522–536 (2014).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Padilla-Coreano, N. et al. Hippocampal–prefrontal theta transmission regulates avoidance behavior. Neuron 104, 601–610 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Felix-Ortiz, A. C., Burgos-Robles, A., Bhagat, N. D., Leppla, C. A. & Tye, K. M. Bidirectional modulation of anxiety-related and social behaviors by amygdala projections to the medial prefrontal cortex. Neuroscience 321, 197–209 (2016).

    Article  CAS  PubMed  Google Scholar 

  13. Adhikari, A., Topiwala, M. A. & Gordon, J. A. Synchronized activity between the ventral hippocampus and the medial prefrontal cortex during anxiety. Neuron 65, 257–269 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Wichmann, R. et al. Acute stress induces long-lasting alterations in the dopaminergic system of female mice. Preprint at bioRxiv https://doi.org/10.1101/168492 (2017).

  15. Wei, Y. C. et al. Medial preoptic area in mice is capable of mediating sexually dimorphic behaviors regardless of gender. Nat. Commun. 9, 279 (2018).

    Article  PubMed  PubMed Central  Google Scholar 

  16. Xu, X. et al. Modular genetic control of sexually dimorphic behaviors. Cell 148, 596–607 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Dulac, C., O’Connell, L. A. & Wu, Z. Neural control of maternal and paternal behaviors. Science 345, 765–770 (2014).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Fang, Y. Y., Yamaguchi, T., Song, S. C., Tritsch, N. X. & Lin, D. A hypothalamic midbrain pathway essential for driving maternal behaviors. Neuron 98, 192–207 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Wu, Z., Autry, A. E., Bergan, J. F., Watabe-Uchida, M. & Dulac, C. G. Galanin neurons in the medial preoptic area govern parental behaviour. Nature 509, 325–330 (2014).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. McHenry, J. A. et al. Hormonal gain control of a medial preoptic area social reward circuit. Nat. Neurosci. 20, 449–458 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Carola, V., D’Olimpio, F., Brunamonti, E., Mangia, F. & Renzi, P. Evaluation of the elevated plus-maze and open-field tests for the assessment of anxiety-related behaviour in inbred mice. Behav. Brain Res. 134, 49–57 (2002).

    Article  PubMed  Google Scholar 

  22. Ahrens, S. et al. A central extended amygdala circuit that modulates anxiety. J. Neurosci. 38, 5567–5583 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Xiu, J. et al. Visualizing an emotional valence map in the limbic forebrain by TAI–FISH. Nat. Neurosci. 17, 1552–1559 (2014).

    Article  CAS  PubMed  Google Scholar 

  24. Luo, L., Callaway, E. M. & Svoboda, K. Genetic dissection of neural circuits: a decade of progress. Neuron 98, 256–281 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Liu, Z. et al. Dorsal raphe neurons signal reward through 5-HT and glutamate. Neuron 81, 1360–1374 (2014).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Hu, H. Reward and aversion. Annu. Rev. Neurosci. 39, 297–324 (2016).

    Article  CAS  PubMed  Google Scholar 

  27. Zhang, G. W. et al. Transforming sensory cues into aversive emotion via septal–habenular pathway. Neuron 99, 1016–1028 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Breton-Provencher, V. & Sur, M. Active control of arousal by a locus coeruleus GABAergic circuit. Nat. Neurosci. 22, 218–228 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Neumann, I. D., Veenema, A. H. & Beiderbeck, D. I. Aggression and anxiety: social context and neurobiological links. Front. Behav. Neurosci. 4, 12 (2010).

    PubMed  PubMed Central  Google Scholar 

  30. Bluett, R. J. et al. Central anandamide deficiency predicts stress-induced anxiety: behavioral reversal through endocannabinoid augmentation. Transl. Psychiatry 4, e408 (2014).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Wang, L. et al. The coding of valence and identity in the mammalian taste system. Nature 558, 127–131 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Zingg, B. et al. AAV-mediated anterograde transsynaptic tagging: mapping corticocollicular input-defined neural pathways for defense behaviors. Neuron 93, 33–47 (2017).

    Article  CAS  PubMed  Google Scholar 

  33. Zhu, Y., Wienecke, C. F. R., Nachtrab, G. & Chen, X. A thalamic input to the nucleus accumbens mediates opiate dependence. Nature 530, 219–222 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Moffitt, J. R. et al. Molecular, spatial, and functional single-cell profiling of the hypothalamic preoptic region. Science 362, eaau5324 (2018).

    Article  PubMed  PubMed Central  Google Scholar 

  35. Callaway, E. M. & Luo, L. Monosynaptic circuit tracing with glycoprotein-deleted rabies viruses. J. Neurosci. 35, 8979–8985 (2015).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Kim, J. et al. Rapid, biphasic CRF neuronal responses encode positive and negative valence. Nat. Neurosci. 22, 576–585 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Davis, M., Walker, D. L., Miles, L. & Grillon, C. Phasic vs sustained fear in rats and humans: role of the extended amygdala in fear vs anxiety. Neuropsychopharmacology 35, 105–135 (2010).

    Article  PubMed  Google Scholar 

  38. Parfitt, G. M. et al. Bidirectional control of anxiety-related behaviors in mice: role of inputs arising from the ventral hippocampus to the lateral septum and medial prefrontal cortex. Neuropsychopharmacology 42, 1715–1728 (2017).

    Article  PubMed  PubMed Central  Google Scholar 

  39. McHenry, J. A., Rubinow, D. R. & Stuber, G. D. Maternally responsive neurons in the bed nucleus of the stria terminalis and medial preoptic area: putative circuits for regulating anxiety and reward. Front. Neuroendocrinol. 38, 65–72 (2015).

    Article  PubMed  PubMed Central  Google Scholar 

  40. Oh, S. W. et al. A mesoscale connectome of the mouse brain. Nature 508, 207–214 (2014).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Park, S. G. et al. Medial preoptic circuit induces hunting-like actions to target objects and prey. Nat. Neurosci. 21, 364–372 (2018).

    Article  CAS  PubMed  Google Scholar 

  42. Chung, S. et al. Identification of preoptic sleep neurons using retrograde labelling and gene profiling. Nature 545, 477–481 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Zhao, Z. D. et al. A hypothalamic circuit that controls body temperature. Proc. Natl Acad. Sci. USA 114, 2042–2047 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Papadimitriou, G. N. & Linkowski, P. Sleep disturbance in anxiety disorders. Int. Rev. Psychiatry 17, 229–236 (2005).

    Article  PubMed  Google Scholar 

  45. Tye, K. M. Neural circuit motifs in valence processing. Neuron 100, 436–452 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. Glasheen, C., Richardson, G. A. & Fabio, A. A systematic review of the effects of postnatal maternal anxiety on children. Arch. Women’s Ment. Health 13, 61–74 (2010).

    Article  Google Scholar 

  47. Hård, E. & Hansen, S. Reduced fearfulness in the lactating rat. Physiol. Behav. 35, 641–643 (1985).

    Article  PubMed  Google Scholar 

  48. Lonstein, J. S. Regulation of anxiety during the postpartum period. Front. Neuroendocrinol. 28, 115–141 (2007).

    Article  PubMed  Google Scholar 

  49. Kohl, J. et al. Functional circuit architecture underlying parental behaviour. Nature 556, 326–331 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  50. Dong, H. W., Petrovich, G. D. & Swanson, L. W. Topography of projections from amygdala to bed nuclei of the stria terminalis. Brain Res. Rev. 38, 192–246 (2001).

    Article  CAS  PubMed  Google Scholar 

  51. Tervo, D. G. R. et al. A designer AAV variant permits efficient retrograde access to projection neurons. Neuron 92, 372–382 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  52. Zhang, G., Shen, L., Li, Z., Tao, H. W. & Zhang, L. I. Track-Control, an automatic video-based real-time closed-loop behavioral control toolbox. Preprint at bioRxiv https://doi.org/10.1101/2019.12.11.873372 (2019).

Download references

Acknowledgements

This work was supported by grants from the US National Institutes of Health to H.W.T. (EY019049, MH116990) and L.I.Z. (R01DC008983, RF1MH114112, MH116990). We thank Y. Xiong and J. Wei at Southern Medical University for generous help with behavioral experiments.

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Authors and Affiliations

  1. Zilkha Neurogenetic Institute, University of Southern California, Los Angeles, CA, USA

    Guang-Wei Zhang, Li Shen, Can Tao, A-Hyun Jung, Bo Peng, Zhong Li, Li I. Zhang & Huizhong Whit Tao

  2. Graduate Program in Neuroscience, University of Southern California, Los Angeles, CA, USA

    A-Hyun Jung & Bo Peng

  3. Department of Physiology and Neuroscience, Keck School of Medicine, University of Southern California, Los Angeles, CA, USA

    Li I. Zhang & Huizhong Whit Tao

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Contributions

G.-W.Z. performed behavioral and photometry experiments and data analysis. L.S. performed optrode recording experiments. A.-H.J. contributed to immunohistochemistry experiments and image analysis. C.T. and B.P. performed slice recording experiments. Z.L. helped with photometry experiments. L.I.Z. and H.W.T supervised the project. H.W.T., G.-W.Z. and L.I.Z. wrote the manuscript.

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Correspondence to Li I. Zhang or Huizhong Whit Tao.

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Peer review information Nature Neuroscience thanks the anonymous reviewers for their contribution to the peer review of this work.

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Extended data

Extended Data Fig. 1 Quantification pipeline for c-fos staining and Vglut2 expression in mPOA. (Associated with Fig. 1).

a, Illustration of force swimming application. The bottom of the test chamber was a metal mesh. For the control experiment, the animal was placed in the same context without being submerged into water (condition on the left). b, Illustration of heat plate application. For the control experiment, the animal was placed in the same context without being touched by the heat plate (condition on the left). c, Protocol for c-fos staining and imaging. Animals were exposed to one of the stressors for 5 min and were sacrificed 3 hours after the treatment. Scale, 100 μm. d, Pipeline for image processing and cell counting. e, Spatial distribution of c-fos+ cells under treatments of three different stressors. LPO, lateral preoptic area; VLPO, ventral lateral preoptic area. Scale, 200 μm. f, Left, Nissl staining; right, tdTomato expression in the same coronal brain section. Images were obtained from transgenic mice by crossing Ai75 (Cre-dependent nucleus-targeted tdTomato reporter) and Vglut2-Cre. g, A more posterior section. BAC, bed nucleus of the anterior commissure; aco, anterior commissure. Scale, 500 µm. Images in c,d are representative of n = 9animals, Images in e,f,g are representative of n=3 animals.

Extended Data Fig. 2 Quality control for fiber photometry. (Associated with Fig. 1).

a, Illustration of the fiber photometry setup. A protective cover helps to prevent the optic fiber from bumping against the wall of the test box/chamber. Neurons express Cre-dependent GCaMP6s. b, Example full trace of calcium signals in the control condition for forced swimming. Dashed line indicates the presumptive operation time (no operation was actually applied). c, Example full trace of calcium signals for forced swimming application. Bar represents the exposure duration. d, Example full trace of calcium signals for heat plate application. e, Plot of calcium transients (blue) and concurrent locomotion speed (red, freely moving) in an open field. Z-score = 3 was used as the detection threshold. f, Plot of locomotion speed vs. amplitude of calcium transients. g, Spearman r calculated for each mouse. N= 8 animals. Bars represent mean ± s.d. h, Fluorescence signals in a control animal expressing GFP only. Black bar marks duration of forced swimming exposure. i, Peak calcium transients during the baseline period and stressor application for GFP control animals (n = 4). Statistics can be found in Fig.1. FST, forced swimming test. j, Heat plate exposure in GFP control animals (n = 4). k, Electric shock exposure in GFP control animals (n = 4). (see Supplementary Table 1 for detailed statistics).

Extended Data Fig. 3 Optrode recording. (Associated with Fig. 1).

a, Top, example full trace of fiber photometry recording (left, each black dot represents one application of foot shocks) and average Ca2+ response (averaged over trials) to a foot shock application (right). b, Top, raster plot of LED-induced spike responses for an example mPOA glutamatergic neuron. Blue dots indicate the duration of LED pulse (5ms). Bottom, corresponding peri-stimulus spike time histogram (PSTH). Raster plot and PSTH for spikes induced by a single LED pulse (blue line). Thick black line indicates the duration of LED stimulation. Only cells show 1st spike latency shorter than 4ms were considered as valid optogenetically-identified Vglut2+ neurons and included for analysis. c, Example full trace of single-unit responses to repeated foot shock stimulation. Spontaneous spikes before the first and last electric shock application are shown on top for visualization. d, Spontaneous firing rates of mPOA opto-IDed Vglut2+ neurons at different time points after exposure to foot shocks. N = 19 cells from 2 animals. e, Center time in OFT performed at different time points after exposure to foot shocks. N = 7 animals. f, Heatmaps of single-cell spike responses to heat (top) and electric shocks (bottom) of opto-IDed Vglut2+ neurons. The same cells are shown to demonstrate multimodal responses. g, Population average of PSTHs from cells shown in (f). h, Heatmap of single-cell spike responses to electric shocks for non-optotagged (presumably Vglut2-) neurons. Pie chart shows the percentage of presumably Vglut2- neurons that shows activated, no, or suppressive responses to electric shocks, respectively. Spontaneous firing rates of non-optotagged neurons at different time points after exposure to foot shocks. N = 35 cells from 2 animals.

Extended Data Fig. 4 Locations of somas with viral expression. (Associated with Figs. 2–4).

Schematic coronal sections ranging from 0.98 mm anterior to 1.06 mm posterior to Bregma. a, Left two, Cre-dependent GFP expression at the injection site in a Vglut2-Cre mouse and spatial distribution of expressing cell bodies. Right three, axons in more posterior sections. Same image as in Fig. 5a. Blue, Nissl staining. Scale, 300 µm. (b) a, Representative images of GFP-labeled mPOA glutamatergic neurons (left) and their axons in PAG (right). Scale bar: 500 µm. c, Superimposed ChR2-EYFP expressing cell locations for all mice from anterior to posterior sections. Each small red dot represents one cell. d, Superimposed hM3Dq-mCherry expressing cell locations for all mice. e, Superimposed hM4Di-mCherry expressing cell locations for all mice. f, Superimposed ArchT-GFP expressing cell locations for all mice. Images in a are representative of n=6 animals.

Extended Data Fig. 5 Activation of mPOA Vglut2 neurons. (Associated with Fig. 2).

a, Experimental setup for conditioned place preference test. During conditioning, the animal was subjected to LED stimulation whenever it was in the conditioned chamber. b, Conditioned place aversion tested 24 hours after paring photo-stimulation with one chamber. Time spent in the conditioned chamber (Cond) or unconditioned chamber (Un-cond) was quantified. **P < 0.01, Mann–Whitney test, n = 5 animals. c, Quantification of center time in OFT under different light stimulation frequencies (Kolmogorov–Smirnov test with Bonferroni correction, P < 0.001, n = 5 and 5 animals for GFP control and ChR2 groups respectively). Each animal was tested for one session per day with stimulation frequencies randomly selected. d, Quantification of open-arm time in EPM (Kolmogorov–Smirnov test with Bonferroni correction, P < 0.001, n = 5 and 5 animals for GFP control and ChR2 groups respectively). e, The OFT arena was divided into 16 subregions and locomotion speed were calculated for each specific subregion. f, Locomotion speed in center vs peripheral subregions. Each dot is one animal. N.S., non-significant; paired t test. N=12. g, The EPM arena was divided into 9 subregions. h, Locomotion speed in closed-arm, open-arm and center subregions. N=12. i, Expressing hM3Dq receptors in mPOA glutamatergic neurons. j, Raw recorded trace of the membrane potential of a hM3Dq-expessing mPOA glutamatergic neuron in response to CNO application in slice recording. k, Subthreshold membrane potential voltages before and after perfusion of CNO as well as after washing out CNO. **P < 0.01, one-way repeated-measures ANOVA, n = 5 cells from 2 animals. l, Quantification of center time in OFT for mCherry control and hM3Dq expressing animals. **P < 0.01, Mann–Whitney test, n = 6 animals for each group. m, Quantification of open-arm time in EPM for mCherry control and hM3Dq expressing animals. **P < 0.01, Mann–Whitney test, n = 6 animals for each group. n, Experimental timeline: expressing ChR2 in mPOA glutamatergic neurons, optogenetic stimulation for 5-min (20 Hz) and anxiety-related behavioral test one hour later. o, Quantification of center time in OFT for GFP control and ChR2-expressing groups. **P < 0.01, Mann–Whitney test, n = 7 animals for each group. Quantification of open arm time in EPM for GFP control and ChR2-expressing groups. **P < 0.01, Mann–Whitney test, n = 7 animals for each group. (see Supplementary Table 1 for detailed statistics).

Extended Data Fig. 6 Strong activation of mPOA glutamatergic neurons. (Associated with Fig. 2).

a, Illustration of conflict tests. b, Movement tracing of a GFP control animal (left) and a ChR2-expressing animal (right) in a conflict test. Photostimulation at 15 Hz was applied whenever the mouse was in the light gray marked zone. c, Percentage time spent in the physically harmful side for control and ChR2 animals. *P < 0.05; **P < 0.01, Mann–Whitney test, n = 6 animals for each group. d, Photo of mouse rearing during LED activation of mPOA glutamatergic neurons at 15 Hz. e, Frequency of rearing in GFP control (n = 7) and ChR2 expressing (n = 5) animals. **P < 0.01, Mann–Whitney test with Bonferroni correction. f, Photo of mouse jumping during LED activation of nPOA glutamatergic neurons at 15 Hz. g, Frequency of jumping in GFP control (n = 7) and ChR2-expressing (n = 5) animals. **P < 0.01, Mann–Whitney test with Bonferroni correction. h, Food intake within 2 hours after being food-deprived for 24 hours. During the 2h test, mPOA glutamatergic neurons were photo-stimulated continuously. **P < 0.01, Mann–Whitney test, n = 5 animals for each group. i, Frequency of rearing in mCherry control (n = 7) and hM3Dq-expressing (n = 6) animals. **P < 0.01, Mann–Whitney test with Bonferroni correction. j, Comparison of rearing frequency between mCherry control (n = 7) and hM3Dq- expressing (n = 6) animals. **P < 0.01, Mann–Whitney test with Bonferroni correction. k, Food intake within 2 hours after being food-deprived for 24 hours. CNO injection (i.p.) was performed 20 min before the test. **P < 0.01, Mann–Whitney test with Bonferroni correction, n = 5 for each group. l, Strategy of viral injection. m, Movement tracing of a GFP control animal (upper) and an ArchT-expressing animal (lower) in a two-chamber place preference test. Continuous LED stimulation was applied whenever the animal stayed in the light gray marked chamber. n, Percentage time spent in the LED-on chamber. **P<0.01, Mann-Whitney test, n = 9 (5 males) for each group. o, Upper, experimental time line. LED light was applied only during the electric shocks. Center time in OFT for GFP control (n = 7) and ArchT-expressing (n = 7; 4 males) animals. **P < 0.01, Mann–Whitney test. p, Open-arm time in EPM for GFP control (n = 7) and ArchT-expressing (n = 7; 4 males) animals. **P < 0.05, Mann–Whitney test, n = 7 animals for each group. (see Supplementary Table 1 for detailed statistics).

Extended Data Fig. 7 Anxiety tests after exposure to social stress. (Associated with Fig. 4).

a, Anxiety-like behaviors of virgin males with or without exposure to male intruders. **P < 0.01, Mann–Whitney test, n = 8 animals for each group. b, Anxiety-like behaviors for virgin males with or without exposure to foreign pups. **P < 0.01, Mann–Whitney test, n = 10 animals for each group. c, Duration of pup grooming for virgin males in LED-off and LED-on conditions. **P < 0.01, Mann–Whitney test with Bonferroni correction, n = 10 animals for each group. d, Anxiety-like behaviors of virgin females with or without exposure to foreign pups. ‘n.s.’, not significant, Mann–Whitney test, n = 10 animals for each group. e, Anxiety-related tests in virgin males/females not exposed to stress with (green) and without (grey) optogenetic silencing of mPOA Vglut2 neurons. *P < 0.05, **P < 0.01, Mann–Whitney test with Bonferroni correction, n = 7 animals for each group. f, Anxiety-related tests in virgin females not exposed to stress with (green) and without (grey) chemogenetic silencing of mPOA Vglut2 neurons. **P < 0.01, Mann–Whitney test with Bonferroni correction, n =8 animals for each group. g, Left, schematic open field test with a shelter. Right, total time spent in shelter within a 5-min test session. **P < 0.01, Mann–Whitney test, n = 6 (3 males) animals. h, Total time spent in the light side of a light-dark box. **P < 0.01, Mann–Whitney test, n = 7 (4 males) animals for each group. (see Supplementary Table 1 for detailed statistics).

Extended Data Fig. 8 Potential targets of mPOA glutamatergic neurons. (Associated with Fig. 5).

a, Imaging area (top) and a confocal image (bottom) showing EYPF labeling in mPOA (left) and different downstream structures. Scale, 500 µm. VMH, ventral medial hypothalamic nucleus; MM, medial mammillary nucleus. b, Schematic terminal stimulation in a potential mPOA target. c, Frequency of jumping induced by activating axonal terminals of mPOA glutamatergic neurons in different target areas. **P < 0.01, One-way ANOVA test, n = 5 animals for each group. GFP control group is for animals expressing GFP in mPOA with fiber implantation in mPOA. d, Two-chamber place preference test when activating axonal terminals of mPOA glutamatergic neurons in different target areas. **P < 0.01, One-way ANOVA test, n = 5 animals for each group. e, Left, viral injection strategy to express ChR2 in PAG-projecting mPOA neurons in Ai14 mice. Right, images showing tdTomato- and ChR2-EYFP labeled neurons in mPOA. Scale bar, 500 µm. f, Left, viral injection strategy to transsynaptically label mPOA-recipient PAG neurons. Right, image showing labeled neurons in PAG (with a blow-up image on the right). Scale bar, 500 µm. Images in a,e are representative of n=5 animals. Images in f are representative of n =3 animals. (see Supplementary Table 1 for detailed statistics).

Extended Data Fig. 9 Manipulation of the mPOA to VTA pathway. (Associated with Fig. 5).

a, Top, viral injection strategy. Optic cannula was implanted above VTA. Bottom, labeling of dopamine neurons (red) by crossing DAT-Cre and Ai14 mice (left) and mPOA glutamatergic axons in VTA and surrounding regions (right). Scale, 400 µm. b, Percentage time spent in the LED-on chamber in PPT for GFP control and ChR2-expressing animals. **P<0.01, Mann-Whitney test, n = 7 (4 males) for each group. c, Center time in OFT. N.S., no statistical difference, Mann–Whitney test, n = 7 animals for each group. (d) Open-arm time in EPM. Mann–Whitney test, n = 7 animals for each group. e, Double retrograde dye injection in PAG (green) and VTA (red). f, Representative image showing retrogradely labelled neurons in mPOA. g, Quantification of singly and doubly labeled neurons in mPOA. N=4 animals. h, Viral strategy to label axon collaterals of PAG-projecting mPOA neurons in VTA. i, Images showing many labeled axons in PAG (left two), but extremely sparse axons in VTA (right two). Injection site is shown in Fig. 5b (shared blue channel as reference). Scale, 500 µm. Images in a,i are representative of n =3 animals, images in f are representative of n = 4 animals. (see Supplementary Table 1 for detailed statistics).

Extended Data Fig. 10 Control experiments for manipulating neurons in mPOA and proposed circuit model. (Associated with Fig. 6).

a, Open-arm time for GFP control animals in LED-off and LED-on conditions. ‘n.s.’, non-significant, Mann–Whitney test, n = 10 animals for each group. b, Schematic pup exposure test for virgin males or females. c, Percentage of trials with pup retrieval for GFP control virgin females in LED-on and LED-off conditions respectively. ‘n.s.’, non-significant, Fisher’s exact test, n = 7 animals for each group. Each animal was subjected to 2–4 trials and all trials were pooled together. d, Duration of pup grooming for GFP control virgin females. ‘n.s.’, non-significant, Mann–Whitney test, n = 9 animals for each group. e, Percentage of trials with pup attack for GFP control virgin males in LED-on and LED-off conditions respectively. ‘n.s.’, non-significant, Fisher’s exact test, n = 7 animals for each group. Each animal was subjected to 2–4 trials and all trials were pooled together. f, Left, viral injection strategy and implantation of the optic fiber above PAG. Right, image showing axons of mPOA GABAergic neurons in PAG. Scale, 500 µm. g, Representative images showing injection sites in VLPO (top) and MnPO (bottom). Scale, 500 µm. h, Core body temperature measured after 10-Hz LED stimulation for 30 min in animals expressing ChR2 in mPOA, ventral lateral preoptic area (vLPO) and median preoptic area (MnPO) respectively in Vglut2-Cre animals. **P < 0.01, two-tailed t-test, n = 5 animals for each group. bar, s.d. i, Left, photo of a freely moving mouse in an open arena, with a 2x2x2cm 3D object placed in the center. The behavioral test consisted of 3 blocks: LED-off, LED-on, and then LED-off, with each lasting 3 min. 10-Hz LED stimulation was applied during the LED-on block. Right, dislocation of the 3D object by the mouse. No statistical significance was observed between blocks; two-way repeated-measures ANOVA, n = 5 animals. j, CTb488 injection in PAG of either Vglut2-Cre::Ai14 or Vgat-Cre::Ai14 mice. k, Images showing overlap between CTb-labeled and Vglut2+ (top) / Vgat+ (bottom) neurons in mPOA. Scale, 200 µm. l, Quantification of the percentage of PAG-projecting mPOA neurons that are Vglut2+ or Vgat+. **P<0.01, Mann-Whitney test with Bonferroni correction, n = 3 animals for each group. Bar, s.d. m, Representative image showing the CTb injection site in medial PAG. n, Quantification of the percentage of Vglut2+ or Vgat+ mPOA neurons that were labeled by CTb injected in PAG. **P<0.01, Mann-Whitney test with Bonferroni correction, n = 3 animals for each group. Bar, s.d. o, Illustration of the proposed circuit model. Note that due to potentially different inputs, glutamatergic neurons in mPOA respond to physical and social stressors but not to social rewards, while GABAergic neurons are activated by social rewards (for example during parenting) but not stressors. Images in f are representative of n = 8 animals. Images in g are representative of n = 5 animals. Images in k, m are representative of n = 3 animals. (see Supplementary Table 1 for detailed statistics).

Supplementary information

Supplementary Information

Supplementary Table 1.

Reporting Summary

Supplementary Video 1

An example video for control group animals undergoing the PPT. Light stimulation was delivered to the mPOA, with GFP expressed in glutamatergic neurons in the mPOA.

Supplementary Video 2

An example video for test group animals undergoing the PPT. Light stimulation was delivered to the mPOA, with ChR2 expressed in glutamatergic neurons in the mPOA.

Supplementary Video 3

An example video for control group animals undergoing the conflict test. Light stimulation was delivered to the mPOA, with GFP expressed in glutamatergic neurons in the mPOA.

Supplementary Video 4

An example video for test group animals undergoing the conflict test. Light stimulation was delivered to the mPOA, with ChR2 expressed in glutamatergic neurons in the mPOA.

Supplementary Data

Statistical resources for all related figures.

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Zhang, GW., Shen, L., Tao, C. et al. Medial preoptic area antagonistically mediates stress-induced anxiety and parental behavior. Nat Neurosci 24, 516–528 (2021). https://doi.org/10.1038/s41593-020-00784-3

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