Letter | Published:

Scalable control of mounting and attack by Esr1+ neurons in the ventromedial hypothalamus

Nature volume 509, pages 627632 (29 May 2014) | Download Citation

Subjects

Abstract

Social behaviours, such as aggression or mating, proceed through a series of appetitive and consummatory phases1 that are associated with increasing levels of arousal2. How such escalation is encoded in the brain, and linked to behavioural action selection, remains an unsolved problem in neuroscience. The ventrolateral subdivision of the murine ventromedial hypothalamus (VMHvl) contains neurons whose activity increases during male–male and male–female social encounters. Non-cell-type-specific optogenetic activation of this region elicited attack behaviour, but not mounting3. We have identified a subset of VMHvl neurons marked by the oestrogen receptor 1 (Esr1), and investigated their role in male social behaviour. Optogenetic manipulations indicated that Esr1+ (but not Esr1) neurons are sufficient to initiate attack, and that their activity is continuously required during ongoing agonistic behaviour. Surprisingly, weaker optogenetic activation of these neurons promoted mounting behaviour, rather than attack, towards both males and females, as well as sniffing and close investigation. Increasing photostimulation intensity could promote a transition from close investigation and mounting to attack, within a single social encounter. Importantly, time-resolved optogenetic inhibition experiments revealed requirements for Esr1+ neurons in both the appetitive (investigative) and the consummatory phases of social interactions. Combined optogenetic activation and calcium imaging experiments in vitro, as well as c-Fos analysis in vivo, indicated that increasing photostimulation intensity increases both the number of active neurons and the average level of activity per neuron. These data suggest that Esr1+ neurons in VMHvl control the progression of a social encounter from its appetitive through its consummatory phases, in a scalable manner that reflects the number or type of active neurons in the population.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1.

    in Physiological Mechanisms in Animal Behaviour Vol. IV Symposia of the Society for Experimental Biology 305–312 (Academic Press, 1950)

  2. 2.

    , , & CNS arousal mechanisms bearing on sex and other biologically regulated behaviors. Physiol. Behav. 88, 283–293 (2006)

  3. 3.

    et al. Functional identification of an aggression locus in the mouse hypothalamus. Nature 470, 221–226 (2011)

  4. 4.

    et al. Genome-wide atlas of gene expression in the adult mouse brain. Nature 445, 168–176 (2007)

  5. 5.

    , , & Mapping patterns of c-fos expression in the central nervous system after seizure. Science 237, 192–197 (1987)

  6. 6.

    et al. Cloning of the human estrogen receptor cDNA. Proc. Natl Acad. Sci. USA 82, 7889–7893 (1985)

  7. 7.

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

  8. 8.

    , , , & Millisecond-timescale, genetically targeted optical control of neural activity. Nature Neurosci. 8, 1263–1268 (2005)

  9. 9.

    et al. An optical neural interface: in vivo control of rodent motor cortex with integrated fiberoptic and optogenetic technology. J. Neural Eng. 4, S143–S156 (2007)

  10. 10.

    & Ethoexperimental approaches to the biology of emotion. Annu. Rev. Psychol. 39, 43–68 (1988)

  11. 11.

    et al. Sexually dimorphic neurons in the ventromedial hypothalamus govern mating in both sexes and aggression in males. Cell 153, 896–909 (2013)

  12. 12.

    , , , & Differential effects of site-specific knockdown of estrogen receptor alpha in the medial amygdala, medial pre-optic area, and ventromedial nucleus of the hypothalamus on sexual and aggressive behavior of male mice. Eur. J. Neurosci. 37, 1308–1319 (2013)

  13. 13.

    et al. Molecular and cellular approaches for diversifying and extending optogenetics. Cell 141, 154–165 (2010)

  14. 14.

    , & Problems in the study of rodent aggression. Horm. Behav. 44, 161–170 (2003)

  15. 15.

    et al. Ultrasensitive fluorescent proteins for imaging neuronal activity. Nature 499, 295–300 (2013)

  16. 16.

    et al. Reversible silencing of neuronal excitability in behaving mice by a genetically targeted, ivermectin-gated Cl channel. Neuron 54, 35–49 (2007)

  17. 17.

    & Ventromedial hypothalamic neurons in the mediation of long-lasting effects of estrogen on lordosis behavior. Prog. Neurobiol. 38, 423–453 (1992)

  18. 18.

    , , , & RNAi-mediated silencing of estrogen receptor α in the ventromedial nucleus of hypothalamus abolishes female sexual behaviors. Proc. Natl Acad. Sci. USA 103, 10456–10460 (2006)

  19. 19.

    et al. Estrogen-induced sexual incentive motivation, proceptivity and receptivity depend on a functional estrogen receptor alpha in the ventromedial nucleus of the hypothalamus but not in the amygdala. Neuroendocrinology 91, 142–154 (2010)

  20. 20.

    et al. The role of the estrogen receptor alpha in the medial amygdala and ventromedial nucleus of the hypothalamus in social recognition, anxiety and aggression. Behav. Brain Res. 210, 211–220 (2010)

  21. 21.

    Wired for reproduction: organization and development of sexually dimorphic circuits in the mammalian forebrain. Annu. Rev. Neurosci. 25, 507–536 (2002)

  22. 22.

    , & Sexual differentiation of the vertebrate nervous system. Nature Neurosci. 7, 1034–1039 (2004)

  23. 23.

    & Control of masculinization of the brain and behavior. Curr. Opin. Neurobiol. 21, 116–123 (2011)

  24. 24.

    , & Acute estrogen potentiates excitatory responses of neurons in rat hypothalamic ventromedial nucleus. Brain Res. 10, 124–131 (2005)

  25. 25.

    & Neural control of behavior. Annu. Rev. Neurosci. 1, 35–59 (1978)

  26. 26.

    et al. Discriminant analysis of the localization of aggression-inducing electrode placements in the hypothalamus of male rats. Brain Res. 260, 61–79 (1983)

  27. 27.

    Optogenetics, sex, and violence in the brain: implications for psychiatry. Biol. Psychiatry 71, 1081–1089 (2011)

  28. 28.

    Neuronal decision-making circuits. Curr. Biol. 18, R928–R932 (2008)

  29. 29.

    et al. Genetic dissection of an amygdala microcircuit that gates conditioned fear. Nature 468, 270–276 (2010)

  30. 30.

    , , , & Genetic identification of C fibres that detect massage-like stroking of hairy skin in vivo. Nature 493, 669–673 (2013)

  31. 31.

    , , , & High-efficient FLPo deleter mice in C57BL/6J background. PLoS ONE 4, e8054 (2009)

  32. 32.

    et al. Functional identification of an aggression locus in the mouse hypothalamus. Nature 470, 221–226 (2011)

  33. 33.

    , & Adeno-associated viral vectors for anterograde axonal tracing with fluorescent proteins in nontransgenic and cre driver mice. Curr Protoc Neurosci Chapter 1, Unit 1 20 21-18. (2012)

  34. 34.

    , , , & Development of a high resolution three-dimensional surgical atlas of the murine head for strains 129S1/SvImJ and C57Bl/6J using magnetic resonance imaging and micro-computed tomography. Neuroscience 144, 604–615 (2007)

  35. 35.

    et al. Genome-wide atlas of gene expression in the adult mouse brain. Nature 445, 168–176 (2007)

  36. 36.

    et al. Genetic dissection of an amygdala microcircuit that gates conditioned fear. Nature 468, 270–276 (2010)

  37. 37.

    , , & Deconstruction of a neural circuit for hunger. Nature 488, 172–177 (2012)

  38. 38.

    et al. Ultrasensitive fluorescent proteins for imaging neuronal activity. Nature 499, 295–300 (2013)

  39. 39.

    , , , & Genetic identification of C fibres that detect massage-like stroking of hairy skin in vivo. Nature 493, 669–673 (2013)

  40. 40.

    , , , & Neuron-type-specific signals for reward and punishment in the ventral tegmental area. Nature 482, 85–88 (2012)

  41. 41.

    et al. Distinct behavioural and network correlates of two interneuron types in prefrontal cortex. Nature 498, 363–366 (2013)

Download references

Acknowledgements

We thank C. Park for behavioural scoring, R. Robertson for behavioural scoring and MATLAB programming, L. Lo for testing Cre-mediated recombination in Esr1cre/+ male mice, C. Chiu and X. Wang for histology, M. McCardle for genotyping, J. S. Chang for technical assistance, S. Pease for generation of knock-in mice, H. Cai for training in slice electrophysiology, A. Wong for assistance with two-photon imaging, K. Deisseroth and J. Harris for AAV constructs, E. Boyden for advice on ferrule fibre fabrication, D. Lin and M. Boyle for their contributions to early stages of this project, W. Hong and R. Axel for comments on the manuscript, C. Chiu for laboratory management and G. Mancuso for administrative assistance. D.J.A. is an Investigator of the Howard Hughes Medical Institute and a Paul G. Allen Distinguished Investigator. This work was supported in part by NIH grant no. R01MH070053, and grants from the Gordon Moore Foundation and Ellison Medical Research Foundation. H.L. was supported by the NIH Pathway to Independence Award 1K99NS074077. T.E.A. was supported by NIH NRSA postdoctoral fellowship grant 1F32HD055198-01 and a Beckman Fellowship.

Author information

Affiliations

  1. Division of Biology and Biological Engineering 156-29, California Institute of Technology, Pasadena, California 91125, USA

    • Hyosang Lee
    • , Ryan Remedios
    • , Todd E. Anthony
    • , Angela Chang
    •  & David J. Anderson
  2. Howard Hughes Medical Institute, Pasadena, California 91125, USA

    • Hyosang Lee
    •  & David J. Anderson
  3. Computation and Neural Systems, California Institute of Technology, Pasadena, California 91125, USA

    • Dong-Wook Kim
    •  & David J. Anderson
  4. Allen Institute for Brain Science, Seattle, Washington 98103, USA

    • Linda Madisen
    •  & Hongkui Zeng

Authors

  1. Search for Hyosang Lee in:

  2. Search for Dong-Wook Kim in:

  3. Search for Ryan Remedios in:

  4. Search for Todd E. Anthony in:

  5. Search for Angela Chang in:

  6. Search for Linda Madisen in:

  7. Search for Hongkui Zeng in:

  8. Search for David J. Anderson in:

Contributions

H.L. characterized Esr1cre mice, designed and performed optogenetic behavioural experiments and co-wrote the manuscript; D.-W.K. performed slice electrophysiology and imaging experiments; R.R. performed in vivo electrophysiology; T.E.A. generated the Esr1cre targeting construct and AAV vectors; A.C. carried out some behavioural experiments; L.M. and H.Z. performed in situ hybridization experiments; D.J.A. supervised experiments and co-wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to David J. Anderson.

Extended data

Supplementary information

PDF files

  1. 1.

    Supplementary Information

    This file contains Supplementary Notes 1-3.

Videos

  1. 1.

    Optogenetic activation of VMHvl Esr1+ neurons in an Esr1cre/+ male mouse evokes attack towards an intact C57BL/6 female (part 1) and a castrated BALB/c male intruders (part 2).

    The ChR2-expressing Esr1cre/+ male mice connected to a fiber-optic cable were photostimulated during the period indicated by either "Light on" or the LED light at the bottom right corner.

  2. 2.

    Acute silencing of VMHvl Esr1+ neurons in an Esr1cre/+ male mouse inhibits naturally occurring attack towards an intact BALB/c male intruder.

    The eNpHR3.0-expressing Esr1cre/+ male mouse (black) was photostimulated during the period indicated by either "Light on" or the LED light at the bottom right corner.

  3. 3.

    Acute silencing of VMHvl Esr1+ neurons in an Esr1cre/+ male mouse inhibits aggressive approach towards an intact BALB/c male intruder.

    The eNpHR3.0-expressing Esr1cre/+ male mouse (black) was photostimulated during the period indicated by "Light on".

  4. 4.

    Optogenetic activation of VMHvl Esr1+ neurons in an Esr1cre/+ female mouse elicits close investigation towards a castrated BALB/c male intruder.

    The ChR2-expressing Esr1cre/+ female mouse (black) was photostimulated during the period indicated by either "Light on" or the LED light at the bottom right corner.

  5. 5.

    Optogenetic activation of VMHvl Esr1+ neurons in an Esr1cre/+ male mouse evokes aggressive sniffing towards a castrated BALB/c male intruder.

    The ChR2-expressing Esr1cre/+ male mouse (black) was photostimulated during the period indicated by either "Light on" or the LED light at the bottom right corner.

  6. 6.

    Optogenetic activation of VMHvl Esr1+ neurons in an Esr1cre/+ male mouse evokes mounting behavior towards an unreceptive intact BALB/c female intruder.

    The ChR2-expressing Esr1cre/+ male mouse connected to a fiber-optic cable was photostimulated during the period indicated by either "Light on" or the LED light at the bottom right corner at indicated photostimulation intensities.

  7. 7.

    Optogenetic activation of VMHvl Esr1+ neurons in an Esr1cre/+ male mouse evokes mounting behavior towards a castrated BALB/c male (part 1) and an intact BALB/c male intruders (part 2).

    The ChR2-expressing Esr1cre/+ male mouse (black) was photostimulated during the period indicated by either "Light on" or the LED light at the bottom right corner.

  8. 8.

    Activation of VMHvl Esr1+ neurons in an Esr1cre/+ male mouse shifts behavioral responses from mounting, to a mixture of mounting and attack, to attack towards an unreceptive intact BALB/c female intruder as the photostimulation intensity is increased.

    The ChR2-expressing Esr1cre/+ male mouse (black) was photostimulated during the period indicated by either "Light on" or the LED light at the bottom right corner at indicated photostimulation intensities.

  9. 9.

    Activation of VMHvl Esr1+ neurons in an Esr1cre/+ male mouse shifts behavioral responses from mounting, to a mixed behavior of mounting and attack, to attack towards a castrated BALB/c male intruder as the photostimulation intensity is increased.

    The ChR2-expressing Esr1cre/+ male mouse (black) was photostimulated during the period indicated by either "Light on" or the LED light at the bottom right corner at indicated photostimulation intensities.

  10. 10.

    Optogenetic activation of VMHvl Esr1+ neurons in an Esr1cre/+ male mouse at intermediate photostimulation intensities evokes repeated, interspersed attempts at mounting and attack towards an unreceptive intact BALB/c female intruder.

    The ChR2-expressing Esr1cre/+ male mouse connected to a fiber-optic cable was photostimulated during the period indicated by either "Light on" or the LED light at the bottom right corner.

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/nature13169

Further reading

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.