Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

A GnRH neuronal population in the olfactory bulb translates socially relevant odors into reproductive behavior in male mice

Nature Neuroscience volume 27, pages 1758–1773 (2024)Cite this article

Subjects

Abstract

Hypothalamic gonadotropin-releasing hormone (GnRH) neurons regulate fertility and integrate hormonal status with environmental cues to ensure reproductive success. Here we show that GnRH neurons in the olfactory bulb (GnRHOB) of adult mice can mediate social recognition. Specifically, we show that GnRHOB neurons extend neurites into the vomeronasal organ and olfactory epithelium and project to the median eminence. GnRHOB neurons in males express vomeronasal and olfactory receptors, are activated by female odors and mediate gonadotropin release in response to female urine. Male preference for female odors required the presence and activation of GnRHOB neurons, was impaired after genetic inhibition or ablation of these cells and relied on GnRH signaling in the posterodorsal medial amygdala. GnRH receptor expression in amygdala kisspeptin neurons appear to be required for GnRHOB neurons’ actions on male mounting behavior. Taken together, these results establish GnRHOB neurons as regulating fertility, sex recognition and mating in male mice.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on SpringerLink
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Fig. 1: A significant proportion of GnRH neurons is located in the OBs of mice and is connected with the OE and VNO.
Fig. 2: Male GnRHOB neurons respond to female urine.
Fig. 3: GnRHOB neurons mediate LH secretion in response to opposite-sex odors.
Fig. 4: Connectivity between GnRHOB and GnRHPOA neurons.
Fig. 5: GnRHOB neurons drive olfactory preference in male mice.
Fig. 6: GnRHOB neurons recognize opposite-sex odors and initiate mating via the pdMeA.
Fig. 7: GnRHR expression on kisspeptin neurons in the pdMeA mediates mating behavior in male mice.
Fig. 8: GnRHMOB and GnRHAOB neuronal projections and neuroendocrine/behavioral responses to odors in male mice.

Similar content being viewed by others

Data availability

All data generated or analyzed in the current study are included in the article and its Extended Data. The raw snRNA-seq data conducted in this study have been deposited in Gene Expression Omnibus under accession code GSE268138. For snRNA-seq experiments, raw reads were aligned to Ensembl genome GRCm39 release 105_cr6.1.2. Source data are provided with this paper.

References

  1. Dulac, C. & Torello, A. T. Molecular detection of pheromone signals in mammals: from genes to behaviour. Nat. Rev. Neurosci. 4, 551–562 (2003).

    Article  CAS  PubMed  Google Scholar 

  2. Belluscio, L., Koentges, G., Axel, R. & Dulac, C. A map of pheromone receptor activation in the mammalian brain. Cell 97, 209–220 (1999).

    Article  CAS  PubMed  Google Scholar 

  3. Rodriguez, I., Feinstein, P. & Mombaerts, P. Variable patterns of axonal projections of sensory neurons in the mouse vomeronasal system. Cell 97, 199–208 (1999).

    Article  CAS  PubMed  Google Scholar 

  4. Lehman, M. N., Winans, S. S. & Powers, J. B. Medial nucleus of the amygdala mediates chemosensory control of male hamster sexual behavior. Science 210, 557–560 (1980).

    Article  CAS  PubMed  Google Scholar 

  5. Baird, A. D., Wilson, S. J., Bladin, P. F., Saling, M. M. & Reutens, D. C. The amygdala and sexual drive: insights from temporal lobe epilepsy surgery. Ann. Neurol. 55, 87–96 (2004).

    Article  PubMed  Google Scholar 

  6. Bayless, D. W. et al. A neural circuit for male sexual behavior and reward. Cell 186, 3862–3881 (2023).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Keller, M., Pillon, D. & Bakker, J. Olfactory systems in mate recognition and sexual behavior. Vitam. Horm. 83, 331–350 (2010).

    Article  CAS  PubMed  Google Scholar 

  8. Keverne, E. B. Importance of olfactory and vomeronasal systems for male sexual function. Physiol. Behav. 83, 177–187 (2004).

    Article  CAS  PubMed  Google Scholar 

  9. Aoki, M. et al. Prolactin-sensitive olfactory sensory neurons regulate male preference in female mice by modulating responses to chemosensory cues. Sci. Adv. 7, eabg4074 (2021).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Mandiyan, V. S., Coats, J. K. & Shah, N. M. Deficits in sexual and aggressive behaviors in Cnga2 mutant mice. Nat. Neurosci. 8, 1660–1662 (2005).

    Article  CAS  PubMed  Google Scholar 

  11. Wang, Z. et al. Pheromone detection in male mice depends on signaling through the type 3 adenylyl cyclase in the main olfactory epithelium. J. Neurosci. 26, 7375–7379 (2006).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Duittoz, A. H. et al. Development of the gonadotropin-releasing hormone system. J. Neuroendocrinol. 34, e13087 (2022).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Herbison, A. E. Control of puberty onset and fertility by gonadotropin-releasing hormone neurons. Nat. Rev. Endocrinol. 12, 452–466 (2016).

    Article  CAS  PubMed  Google Scholar 

  14. Bronson, F. H. The regulation of luteinizing hormone secretion by estrogen: relationships among negative feedback, surge potential, and male stimulation in juvenile, peripubertal, and adult female mice. Endocrinology 108, 506–516 (1981).

    Article  CAS  PubMed  Google Scholar 

  15. Dluzen, D. E., Ramirez, V. D., Carter, C. S. & Getz, L. L. Male vole urine changes luteinizing hormone-releasing hormone and norepinephrine in female olfactory bulb. Science 212, 573–575 (1981).

    Article  CAS  PubMed  Google Scholar 

  16. Boehm, U., Zou, Z. & Buck, L. B. Feedback loops link odor and pheromone signaling with reproduction. Cell 123, 683–695 (2005).

    Article  CAS  PubMed  Google Scholar 

  17. Yoon, H., Enquist, L. W. & Dulac, C. Olfactory inputs to hypothalamic neurons controlling reproduction and fertility. Cell 123, 669–682 (2005).

    Article  CAS  PubMed  Google Scholar 

  18. Casoni, F. et al. Development of the neurons controlling fertility in humans: new insights from 3D imaging and transparent fetal brains. Development 143, 3969–3981 (2016).

    Article  CAS  PubMed  Google Scholar 

  19. Boehm, U. et al. Expert consensus document: European Consensus Statement on congenital hypogonadotropic hypogonadism–pathogenesis, diagnosis and treatment. Nat. Rev. Endocrinol. 11, 547–564 (2015).

    Article  PubMed  Google Scholar 

  20. Messina, A. et al. A microRNA switch regulates the rise in hypothalamic GnRH production before puberty. Nat. Neurosci. 19, 835–844 (2016).

    Article  CAS  PubMed  Google Scholar 

  21. Belle, M. et al. Tridimensional visualization and analysis of early human development. Cell 169, 161–173 (2017).

    Article  CAS  PubMed  Google Scholar 

  22. Renier, N. et al. iDISCO: a simple, rapid method to immunolabel large tissue samples for volume imaging. Cell 159, 896–910 (2014).

    Article  CAS  PubMed  Google Scholar 

  23. Spergel, D. J., Kruth, U., Hanley, D. F., Sprengel, R. & Seeburg, P. H. GABA- and glutamate-activated channels in green fluorescent protein-tagged gonadotropin-releasing hormone neurons in transgenic mice. J. Neurosci. 19, 2037–2050 (1999).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Wang, J. & Hamill, O. P. Piezo2-peripheral baroreceptor channel expressed in select neurons of the mouse brain: a putative mechanism for synchronizing neural networks by transducing intracranial pressure pulses. J. Integr. Neurosci. 20, 825–837 (2021).

    Article  PubMed  Google Scholar 

  25. Zeppilli, S. et al. Molecular characterization of projection neuron subtypes in the mouse olfactory bulb. eLife 10, e65445 (2021).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Jammal Salameh, L., Bitzenhofer, S. H., Hanganu-Opatz, I. L., Dutschmann, M. & Egger, V. Blood pressure pulsations modulate central neuronal activity via mechanosensitive ion channels. Science 383, eadk8511 (2024).

    Article  CAS  PubMed  Google Scholar 

  27. Galliano, E. et al. Embryonic and postnatal neurogenesis produce functionally distinct subclasses of dopaminergic neuron. eLife 7, e32373 (2018).

    Article  PubMed  PubMed Central  Google Scholar 

  28. Castle, M. J., Gershenson, Z. T., Giles, A. R., Holzbaur, E. L. & Wolfe, J. H. Adeno-associated virus serotypes 1, 8, and 9 share conserved mechanisms for anterograde and retrograde axonal transport. Hum. Gene Ther. 25, 705–720 (2014).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Vardy, E. et al. A new DREADD facilitates the multiplexed chemogenetic interrogation of behavior. Neuron 86, 936–946 (2015).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Johnston, R. E. & Bronson, F. Endocrine control of female mouse odors that elicit luteinizing hormone surges and attraction in males. Biol. Reprod. 27, 1174–1180 (1982).

    Article  CAS  PubMed  Google Scholar 

  31. Maruniak, J. A. & Bronson, F. H. Gonadotropic responses of male mice to female urine. Endocrinology 99, 963–969 (1976).

    Article  CAS  PubMed  Google Scholar 

  32. Chu, Z. & Moenter, S. M. Endogenous activation of metabotropic glutamate receptors modulates GABAergic transmission to gonadotropin-releasing hormone neurons and alters their firing rate: a possible local feedback circuit. J. Neurosci. 25, 5740–5749 (2005).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Wen, S. et al. Genetic identification of GnRH receptor neurons: a new model for studying neural circuits underlying reproductive physiology in the mouse brain. Endocrinology 152, 1515–1526 (2011).

    Article  CAS  PubMed  Google Scholar 

  35. Wen, S. et al. Functional characterization of genetically labeled gonadotropes. Endocrinology 149, 2701–2711 (2008).

    Article  CAS  PubMed  Google Scholar 

  36. Kang, N., Baum, M. J. & Cherry, J. A. A direct main olfactory bulb projection to the ‘vomeronasal’ amygdala in female mice selectively responds to volatile pheromones from males. Eur. J. Neurosci. 29, 624–634 (2009).

    Article  PubMed  PubMed Central  Google Scholar 

  37. Kevetter, G. A. & Winans, S. S. Connections of the corticomedial amygdala in the golden hamster. II. Efferents of the “olfactory amygdala”. J. Comp. Neurol. 197, 99–111 (1981).

    Article  CAS  PubMed  Google Scholar 

  38. Pineda, R., Plaisier, F., Millar, R. P. & Ludwig, M. Amygdala kisspeptin neurons: putative mediators of olfactory control of the gonadotropic axis. Neuroendocrinology 104, 223–238 (2017).

    Article  CAS  PubMed  Google Scholar 

  39. Seminara, S. B. et al. The GPR54 gene as a regulator of puberty. N. Engl. J. Med. 349, 1614–1627 (2003).

    Article  CAS  PubMed  Google Scholar 

  40. Topaloglu, A. K. et al. Inactivating KISS1 mutation and hypogonadotropic hypogonadism. N. Engl. J. Med. 366, 629–635 (2012).

    Article  CAS  PubMed  Google Scholar 

  41. de Roux, N. et al. Hypogonadotropic hypogonadism due to loss of function of the KiSS1-derived peptide receptor GPR54. Proc. Natl Acad. Sci. USA 100, 10972–10976 (2003).

    Article  PubMed  PubMed Central  Google Scholar 

  42. Stephens, S. B. Z. & Kauffman, A. S. Regulation and possible functions of kisspeptin in the medial amygdala. Front. Endocrinol. 8, 191 (2017).

    Article  Google Scholar 

  43. Lin, D. Y., Zhang, S. Z., Block, E. & Katz, L. C. Encoding social signals in the mouse main olfactory bulb. Nature 434, 470–477 (2005).

    Article  CAS  PubMed  Google Scholar 

  44. Shani-Narkiss, H. et al. Young adult-born neurons improve odor coding by mitral cells. Nat. Commun. 11, 5867 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Hellier, V. et al. Female sexual behavior in mice is controlled by kisspeptin neurons. Nat. Commun. 9, 400 (2018).

    Article  PubMed  PubMed Central  Google Scholar 

  46. Manfredi-Lozano, M. et al. GnRH replacement rescues cognition in Down syndrome. Science 377, eabq4515 (2022).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  47. Schang, A. L. et al. GnRH receptor gene expression in the developing rat hippocampus: transcriptional regulation and potential roles in neuronal plasticity. Endocrinology 152, 568–580 (2011).

    Article  CAS  PubMed  Google Scholar 

  48. Skrapits, K. et al. The cryptic gonadotropin-releasing hormone neuronal system of human basal ganglia. eLife 10, e67714 (2021).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  49. Hurst, J. L. Female recognition and assessment of males through scent. Behav. Brain Res 200, 295–303 (2009).

    Article  CAS  PubMed  Google Scholar 

  50. Choi, J. M. et al. Development of the main olfactory system and main olfactory epithelium-dependent male mating behavior are altered in Go-deficient mice. Proc. Natl Acad. Sci. USA 113, 10974–10979 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  51. Adekunbi, D. A. et al. Kisspeptin neurones in the posterodorsal medial amygdala modulate sexual partner preference and anxiety in male mice. J. Neuroendocrinol. 30, e12572 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  52. Lehman, M. N. & Winans, S. S. Vomeronasal and olfactory pathways to the amygdala controlling male hamster sexual behavior: autoradiographic and behavioral analyses. Brain Res. 240, 27–41 (1982).

    Article  CAS  PubMed  Google Scholar 

  53. Wood, R. I. & Newman, S. W. Integration of chemosensory and hormonal cues is essential for mating in the male Syrian hamster. J. Neurosci. 15, 7261–7269 (1995).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  54. Gomez, D. M. & Newman, S. W. Differential projections of the anterior and posterior regions of the medial amygdaloid nucleus in the Syrian hamster. J. Comp. Neurol. 317, 195–218 (1992).

    Article  CAS  PubMed  Google Scholar 

  55. Kevetter, G. A. & Winans, S. S. Connections of the corticomedial amygdala in the golden hamster. I. Efferents of the “vomeronasal amygdala”. J. Comp. Neurol. 197, 81–98 (1981).

    Article  CAS  PubMed  Google Scholar 

  56. Meredith, M. Human vomeronasal organ function: a critical review of best and worst cases. Chem. Senses 26, 433–445 (2001).

    Article  CAS  PubMed  Google Scholar 

  57. Frasnelli, J., Lundstrom, J. N., Boyle, J. A., Katsarkas, A. & Jones-Gotman, M. The vomeronasal organ is not involved in the perception of endogenous odors. Hum. Brain Mapp. 32, 450–460 (2011).

    Article  PubMed  Google Scholar 

  58. Zhang, G. et al. Hypothalamic programming of systemic ageing involving IKK-β, NF-κB and GnRH. Nature 497, 211–216 (2013).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  59. Mayer, C. et al. Timing and completion of puberty in female mice depend on estrogen receptor alpha-signaling in kisspeptin neurons. Proc. Natl Acad. Sci. USA 107, 22693–22698 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  60. Zheng, G. X. et al. Massively parallel digital transcriptional profiling of single cells. Nat. Commun. 8, 14049 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  61. Imamura, F., Ito, A. & LaFever, B. J. Subpopulations of projection neurons in the olfactory bulb. Front. Neural Circuits 14, 561822 (2020).

    Article  PubMed  PubMed Central  Google Scholar 

  62. Tepe, B. et al. Single-cell RNA-seq of mouse olfactory bulb reveals cellular heterogeneity and activity-dependent molecular census of adult-born neurons. Cell Rep. 25, 2689–2703 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  63. Steyn, F. J. et al. Development of a methodology for and assessment of pulsatile luteinizing hormone secretion in juvenile and adult male mice. Endocrinology 154, 4939–4945 (2013).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  64. Brock, O., Bakker, J. & Baum, M. J. Assessment of urinary pheromone discrimination, partner preference, and mating behaviors in female mice. Methods Mol. Biol. 1068, 319–329 (2013).

    Article  CAS  PubMed  Google Scholar 

  65. Chachlaki, K. et al. NOS1 mutations cause hypogonadotropic hypogonadism with sensory and cognitive deficits that can be reversed in infantile mice. Sci. Transl. Med. 14, eabh2369 (2022).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  66. Silva, M. S. B. et al. Female sexual behavior is disrupted in a preclinical mouse model of PCOS via an attenuated hypothalamic nitric oxide pathway. Proc. Natl Acad. Sci. USA 26, e2203503119 (2022).

    Article  Google Scholar 

Download references

Acknowledgements

We thank all staff of the Lille Neuroscience & Cognition research center for their constant assistance. We thank C. Laloux and Lille In vivo Imaging and Functional Exploration platform (Univ. Lille, CNRS, Inserm, CHU Lille, Institut Pasteur of Lille, US41-UMS2014 PLBS, France) for technical support during behavioral assessment experiments. We also thank M. Tardivel and A. Bongiovanni (microscopy core facility; The BioImaging Center Lille, BICeL, US41-UMS2014 PLBS, France), N. Jouy (cytometry core facility, BICeL, US41-UMS2014 PLBS, France) and J. Devassine (Animal facility, BICeL, US41-UMS2014 PLBS, France) for expert technical assistance. This work was supported by the European Research Council under the European Union’s Horizon 2020 Research and Innovation program (ERC-2016-CoG to P.G., grant agreement no. 725149/REPRODAMH), by the Agence Nationale de la Recherche (ANR-18-CE14-0017, ANR-19-CE16-0021-02 to P.G.) and INSERM-France (grant no. U1172 to P.G. and V.P.). Schematics and drawings were created with BioRender.com.

Author information

Author notes

  1. Sara Trova

    Present address: Centro CMP3VdA, Istituto Italiano di Tecnologia (IIT), Aosta, Italy

  2. Mauro S. B. Silva

    Present address: Division of Endocrinology, Diabetes and Hypertension, Department of Medicine, Brigham and Women’s Hospital and Harvard Medical School, Boston, MA, USA

  3. These authors contributed equally: Laurine Decoster, Sara Trova, Mauro S. B. Silva, Paolo Giacobini.

Authors and Affiliations

  1. Laboratory of Development and Plasticity of the Neuroendocrine Brain, FHU 1000 Days for Health, School of Medicine, Lille, France

    Laurine Decoster, Sara Trova, Gaetan Ternier, Tori Lhomme, Amandine Legrand, Sarah Gallet, Sowmyalakshmi Rasika, Vincent Prevot, Mauro S. B. Silva & Paolo Giacobini

  2. Univ. Lille, Inserm, CHU Lille, Lille Neuroscience & Cognition, UMR-S 1172, Labex DistAlz, Lille, France

    Laurine Decoster, Sara Trova, Gaetan Ternier, Tori Lhomme, Amandine Legrand, Sarah Gallet, Sowmyalakshmi Rasika, Vincent Prevot, Mauro S. B. Silva & Paolo Giacobini

  3. Department of Life Sciences and Systems Biology, University of Torino, Torino, Italy

    Stefano Zucca, Federico Luzzati, Marco Fogli, Paolo Peretto & Serena Bovetti

  4. Neuroscience Institute Cavalieri Ottolenghi, Orbassano, Italy

    Stefano Zucca, Federico Luzzati, Giulia Nato, Marco Fogli, Paolo Peretto & Serena Bovetti

  5. Max Planck Institute for Metabolism Research, Max Planck Research Group Neurocircuit Wiring and Function, Cologne, Germany

    Janice Bulk, Ayden Gouveia & Sophie M. Steculorum

  6. Experimental Pharmacology, Center for Molecular Signaling (PZMS), Center for Gender-specific Biology and Medicine (CGBM), Saarland University School of Medicine, Homburg, Germany

    Ulrich Boehm, Amanda Wyatt, Vanessa Wahl & Philipp Wartenberg

  7. Laboratory of Reproductive Neurobiology, Hun-Ren Institute of Experimental Medicine, Budapest, Hungary

    Erik Hrabovszky

  8. Department of Pathology and Experimental Cancer Research, Semmelweis University, Budapest, Hungary

    Gergely Rácz

  9. Department of Neuroscience “Rita Levi Montalcini”, University of Turin, Turin, Italy

    Giulia Nato

  10. German Center for Diabetes Research (DZD), Neuherberg, Germany

    Sonja C. Schriever, Miriam Bernecker, Paul T. Pfluger & Sophie M. Steculorum

  11. Research Unit Neurobiology of Diabetes, Institute for Diabetes and Obesity, Helmholtz Munich, Neuherberg, Germany

    Sonja C. Schriever, Miriam Bernecker & Paul T. Pfluger

  12. Division of Neurobiology of Diabetes, TUM School of Medicine, Technical University of Munich, Munich, Germany

    Miriam Bernecker & Paul T. Pfluger

  13. Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases (CECAD), University of Cologne, Cologne, Germany

    Sophie M. Steculorum

Authors
  1. Laurine Decoster
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Sara Trova
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Stefano Zucca
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Janice Bulk
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ayden Gouveia
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Gaetan Ternier
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Tori Lhomme
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Amandine Legrand
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Sarah Gallet
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Ulrich Boehm
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Amanda Wyatt
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Vanessa Wahl
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Philipp Wartenberg
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Erik Hrabovszky
    View author publications

    You can also search for this author in PubMed Google Scholar

  15. Gergely Rácz
    View author publications

    You can also search for this author in PubMed Google Scholar

  16. Federico Luzzati
    View author publications

    You can also search for this author in PubMed Google Scholar

  17. Giulia Nato
    View author publications

    You can also search for this author in PubMed Google Scholar

  18. Marco Fogli
    View author publications

    You can also search for this author in PubMed Google Scholar

  19. Paolo Peretto
    View author publications

    You can also search for this author in PubMed Google Scholar

  20. Sonja C. Schriever
    View author publications

    You can also search for this author in PubMed Google Scholar

  21. Miriam Bernecker
    View author publications

    You can also search for this author in PubMed Google Scholar

  22. Paul T. Pfluger
    View author publications

    You can also search for this author in PubMed Google Scholar

  23. Sophie M. Steculorum
    View author publications

    You can also search for this author in PubMed Google Scholar

  24. Serena Bovetti
    View author publications

    You can also search for this author in PubMed Google Scholar

  25. Sowmyalakshmi Rasika
    View author publications

    You can also search for this author in PubMed Google Scholar

  26. Vincent Prevot
    View author publications

    You can also search for this author in PubMed Google Scholar

  27. Mauro S. B. Silva
    View author publications

    You can also search for this author in PubMed Google Scholar

  28. Paolo Giacobini
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

L.D., S.T. and M.S.B.S. designed and performed the study and were involved in all steps of the paper preparation. J.B. and A.G. performed the bioinformatic analyses. G.T. performed the whole-mount immunohistochemistry experiments. T.L. performed the electrophysiological experiments. A.L. performed the FACS isolation experiments. S.G. performed the RT–qPCR experiments. U.B. provided (GRIC)/eR26-τGFP and the Kiss1-IRES-Cre mouse lines and suggestions on the experiments, supervised the CRISPR genome editing experiments, and edited the paper. A.W. and V.W. designed and produced the AAV2/5-CAG-mCherry_U6-Gnrhr gRNA. P.W. performed the stereotaxic injections for the in vivo CRISPR genome editing experiments and was involved in the analysis. E.H and G.R. provided human biological resources and edited the paper. F.L., G.N. and M.F. performed retrobeads tracing experiments and data analyses. S.Z. and S.B. designed and performed the in vivo calcium experiments and wrote the paper. P.P. contributed to experimental designs for the olfactory behavior experiments and edited the paper. S.C.S., M.B. and P.T.P. designed and performed snRNA-seq experiments and curated the data. S.M.S. supervised the bioinformatic analysis and edited the paper. S.R. and V.P. edited the paper. P.G. conceived this study, designed and managed the experimental studies, prepared the figures, and wrote the paper with input from all authors.

Corresponding author

Correspondence to Paolo Giacobini.

Ethics declarations

Competing interests

The authors declare no competing interests.

Peer review

Peer review information

Nature Neuroscience thanks Julie Bakker, Dayu Lin and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Extended data

Extended Data Fig. 1 Distribution of GnRH neurons within the olfactory bulbs of humans.

a, Top: Lateral and inferior views of a human brain. The olfactory bulb (OB) and the olfactory tract (OT) are depicted in red. Bottom: schematic representation of solvent-based tissue clearing and light-sheet imaging protocol of the human post-mortem olfactory bulbs (OBs). b, iDisco+ and light-sheet microscopy imaging showing the presence of GnRH neurons (arrows) and fibers (arrowheads) in the OB and OT of a 37-year-old postmortem brain (male). Representative pictures of n = 3 postmortem brains of men and women. Scale bars: (b) left Fig. 1 mm, right upper figure 20 µm, right lower figure 50 µm.

Extended Data Fig. 2 GnRHOB neurons express odorant and pheromone receptors.

a, Schematic representing single-nucleus RNA sequencing protocol. Two biological replicates were analyzed and each replicate comprised three olfactory bulbs (males, 18-week-old). b, UMAP visualization of the OB, colored by major cell types. c, UMAP visualization of Gnrh1 expressing cells in the OB (black dots; other cell types in grey). d, Transcriptomic landscape of Gnrh1-positive cells compared with Gnrh1-negative cells in the OB. e, f, Heatmaps indicating the relative expression of olfactory and vomeronasal receptors within each neuronal cluster for Gnrh1-positive cells and Gnrh1-negative cells of the olfactory bulb. g, Percentage expression profile of olfactory receptors in Gnrh1-positive cells and Gnrh1-negative cells of the olfactory bulb. h, Percentage expression profile of vomeronasal receptors in Gnrh1-positive cells and Gnrh1-negative cells of the olfactory bulb. Abbreviations: Oligo, Oligodendrocytes; PG, Periglomerular cells; GC, Granule cells; M/T, Mitral/Tufted cells; OEC, Olfactory ensheathing cells; IN, Inhibitory neurons; sSA, Superficial short axon cells; ET, endothelial cells; MES, mesenchymal cells; dSA, Deep short axon cells.

Extended Data Fig. 3 GnRHOB neurons express olfactory/vomeronasal receptors and mechanosensitive ion channels.

a, Schematic strategy for sorting GnRHOB and GnRHPOA cells from Gnrh::Gfp naïve male mice. b, mRNA expression level of Olfr113, Olfr323, Vmn1r209, Vmn2r1 in GnRH-positive (GFP + ) and GnRH-negative (GFP-) cells isolated from the OB. Values are mean ± s.e.m (n = 4; 12-week-old males; *P = 0.0286; ns P > 0.05; two-sided paired Mann-Whitney U test). c, Schematic of the murine olfactory epithelium and vomeronasal organ. d, Representative coronal section of a VNO from an adult male mouse (P90) showing expression of Vmn2r1 mRNA (white staining) by smFISH. The red dotted-line indicates the boundary between the basal and apical layer of the VNO. e, Representative coronal section of an OE from an adult male mouse (P90) showing expression of Olfr323 mRNA (red) by smFISH. f, Schematic representation of the OB. GnRH cells located in the OB granular layer (GrL) are depicted in green. g-i, Representative coronal section of the OB from adult male mice (P90) showing expression of Gnrh1 (green), Vmn2r1 (white) and Olfr323 (red) in the MOB granular layer (MOB GrL). Arrows point to a Gnrh1-positive neuron co-expressing Vmn2r1 and Olfr323, while the arrowhead in j point to a GnRH neuron which does not express Vmn2r1 nor Olfr323. n = 3 mice used for the smFISH. Scale bars: d, 50 µm; e, 100 µm; g-j, 10 µm; inset in i, 5 µm. k, Schematic representing single-nucleus RNA sequencing protocol. l, UMAP visualization of the OB representing the distribution of Piezo1 and Piezo2 across the different cell clusters. m, Heatmaps indicating the relative expression of Piezo1 and Piezo2 within each neuronal cluster for Gnrh1-positive and Gnrh1-negative cells of the OB. n, Percentage expression profile of Piezo receptors in Gnrh1+ compared to Gnrh1- of the OB. Abbreviations: Oligo, Oligodendrocytes; PG, Periglomerular cells; GC, Granule cells; M/T, Mitral/Tufted cells; OEC, Olfactory ensheathing cells; IN, Inhibitory neurons; sSA, Superficial short axon cells; ET, endothelial cells; MES, mesenchymal cells; dSA, Deep short axon cells.

Source data

Extended Data Fig. 4 GnRHOB neurons are preferentially activated by female urine.

a, Representative in vivo calcium-imaging responses of a cell to female urine (FU), saline (Sal) and male urine (MU). Three consecutive trials for each stimulus are represented. Dashed lines indicate stimulus onset. The displayed cell (cell ID = 38) was classified as a responsive cell to FU but not for Sal and MU. b, Quantification of cell responsiveness to Sal (n = 29 cells recorded from 13 mice; paired two-sided t test). Left: average fluorescent signal across all recorded cells in response to Saline (dark gray) or fresh air (gray). Right: quantification of peak fluorescence in the 10 s after stimulus onset for Sal or Air. Gray dots and lines indicate single recorded cells. Black dots and line show the mean and the associated standard error (paired two-sided t test, n = 29 cells). c, Same as in b but for responses to MU (n = 19 cells recorded from 9 mice) (paired two-sided t test). d, Same as in b but for responses to methyl butyrate, n = 7 cells (paired two-sided t test). P values are shown in the figure.

Source data

Extended Data Fig. 5 Hypothalamic GnRH neurons are not infected by DREADDshM3Dq/hKORD viruses injected in the OB.

a-d, Representative pictures showing immunostainings for GnRH, mCherry and mCitrine across the septal-hypothalamic area, performed in three biological replicates. rPOA: rostral preoptic area; OVLT: organum vasculosum laminae terminalis. Scale bars: a-c, 100 µm; d, 20 µm.

Extended Data Fig. 6 Male mice exposed to the same-sex smell do not exhibit alterations in their levels of luteinizing hormone (LH).

LH pulsatility in male mice (n = 6, 16-week-old) following the exposure to either male urine or saline. Mice are exposed to male urine 1 min before T0. Statistics by two-way ANOVA with Holm–Šídák’s multiple-comparison test (P > 0.05). Values are represented as mean ± s.e.m.

Source data

Extended Data Fig. 7 Sex-steroid receptor expression in the GnRHOB neurons.

Representative coronal section of the OB from an adult male mouse (P90) showing expression of a-c, GnRH and ERα; d-f, GnRH and PR; g-i, GnRH and AR by immunohistochemistry; n = 3 biological replicates. Scale bars: 20 µm.

Extended Data Fig. 8 Distribution of GnRH receptor expressing cells in the mouse brain.

Coronal sections and GFP immunostaining (green) reveal the presence of Gnrhr-expressing cells along different areas of the male mouse’s brain (twelve-week-old): a, AOB; b, septum; c, cortex; d, pririform area; e, olfactory tubercle; f, thalamus; g, hypothalamus; h, hippocampus; i, medial amygdala; j, cortical amygdala. k, Schematics showing the distribution of Gnrhr-expressing cells in male mice brains. Lower number indicates the distance from bregma. The experiments performed in at least three biological replicates. AOB: accessory olfactory bulbs; MOB: main olfactory bulbs; SVZ: subventricular zone. Scale bar: 50 μm.

Extended Data Fig. 9 The Inhibition of the GnRHOB neurons projecting to the medial amygdala does not modify the exploratory behavior or stress in male mice.

a, Schematic representation of the bilateral AAV-hSyn-DIO-hM4D(Gi)-mCherry stereotaxic injection in the pdMeA. b, Percentage of GnRH neurons infected by the viral tracer in the OB and rPOA (n = 5 male mice, age: P150). c, Number of explorations and latency to seek for female and male smells after CNO or saline injections (two-sided paired Wilcoxon signed rank test). d, Analysis of copulatory behavior after CNO or saline injections. The graphs represent the investigation duration, the intromission latency, the number of intromissions and the latency to first mount (two-sided paired Mann-Whitney U test). e, Graphs showing the different parameters evaluated during the elevated plus maze test (EPM) and the open field test after the chemogenetic inhibition of GnRHOB neurons projecting to the pdMeA (two-sided paired Mann-Whitney U test). Values in all graphs are represented as mean ± s.e.m; n = 9, 12-24-week-old males; ns P > 0.05.

Source data

Extended Data Fig. 10 Number of intromissions and ejaculation latency in CRISPRKiss1-Gnrhr male mice.

a, Number of intromission and b, ejaculation latency during the copulatory behavior test following or not CRISPR genome editing of pdMeAKiss1 neurons in vivo (n = 5 controls male mice; n = 9 CRISPRKiss1-Gnrhr male mice; 16-20-week-old males; ns P > 0.05; two-sided Mann-Whitney U test for both a and b). Values are represented as mean ± s.e.m.

Source data

Supplementary information

Supplementary Information

Supplementary Fig. 1.

Reporting Summary

Supplementary Video 1

In vivo two-photon functional imaging showing the response of a GnRH-OB neuron to female urine based on change in fluorescence after stimulus onset.

Source data

Source Data Fig. 1

Statistical source data.

Source Data Fig. 2

Statistical source data.

Source Data Fig. 3

Statistical source data.

Source Data Fig. 4

Statistical source data.

Source Data Fig. 5

Statistical source data.

Source Data Fig. 6

Statistical source data.

Source Data Fig. 7

Statistical source data.

Source Data Extended Data Fig. 3

Statistical source data.

Source Data Extended Data Fig. 4

Statistical source data.

Source Data Extended Data Fig. 6

Statistical source data.

Source Data Extended Data Fig. 9

Statistical source data.

Source Data Extended Data Fig. 10

Statistical source data.

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Decoster, L., Trova, S., Zucca, S. et al. A GnRH neuronal population in the olfactory bulb translates socially relevant odors into reproductive behavior in male mice. Nat Neurosci 27, 1758–1773 (2024). https://doi.org/10.1038/s41593-024-01724-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41593-024-01724-1

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing