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GnRH pulse frequency and irregularity play a role in male aging

Nature Aging volume 1, pages 904–918 (2021)Cite this article

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Abstract

Gonadotropin-releasing hormone (GnRH) has a role in hypothalamic control of aging, but the underlying patterns and relationship with downstream reproductive hormones are still unclear. Here we report that hypothalamic GnRH pulse frequency and irregularity increase before GnRH pulse amplitude slowly decreases during aging. GnRH is inhibited by nuclear factor (NF)-κB, and GnRH pulses were controlled by oscillations in the transcriptional activity of NF-κB. Exposure to testosterone under pro-inflammatory conditions stimulated both NF-κB oscillations and GnRH pulses. While castration of middle-aged mice induced short-term anti-aging effects, preventing elevation of luteinizing hormone (LH) levels after castration led to long-term anti-aging effects and lifespan extension, indicating that high-frequency GnRH pulses and high-magnitude LH levels coordinately mediate aging. Reprogramming the endogenous GnRH pulses of middle-aged male mice via an optogenetic approach revealed that increasing GnRH pulses frequency causes LH excess and aging acceleration, while lowering the frequency of and stabilizing GnRH pulses can slow down aging. In conclusion, GnRH pulses are important for aging in male mice.

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Fig. 1: GnRH and LH pulses change during mouse aging.
Fig. 2: Inverse relationship between NF-κB signaling oscillation and GnRH pulses.
Fig. 3: Effects of testosterone on NF-κB cytoplasmic–nuclear oscillation.
Fig. 4: Effects of castration on GnRH pulses.
Fig. 5: Effects of castration on aging physiology.
Fig. 6: Healthspan and lifespan extension by castration in middle age in combination with LH inhibition.
Fig. 7: Optogenetic resetting of GnRH activation frequency in vivo.
Fig. 8: Effects on healthspan and lifespan of GnRH optogenetic stimulation in middle-aged mice.

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References

  1. 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 

  2. Cai, D. & Khor, S. “Hypothalamic microinflammation” paradigm in aging and metabolic diseases. Cell Metab. 30, 19–35 (2019).

    Article  CAS  PubMed  Google Scholar 

  3. Zhang, Y. L. et al. Hypothalamic stem cells control ageing speed partly through exosomal miRNAs. Nature 548, 52–57 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Hsin, H. & Kenyon, C. Signals from the reproductive system regulate the lifespan of C. elegans. Nature 399, 362–366 (1999).

    Article  CAS  PubMed  Google Scholar 

  5. Hamilton, J. B. & Mestler, G. E. Mortality and survival: comparison of eunuchs with intact men and women in a mentally retarded population. J. Gerontol. 24, 395–411 (1969).

    Article  CAS  PubMed  Google Scholar 

  6. Wilson, J. D. & Roehrborn, C. Long-term consequences of castration in men: lessons from the Skoptzy and the eunuchs of the Chinese and Ottoman courts. J. Clin. Endocrinol. Metab. 84, 4324–4331 (1999).

    Article  CAS  PubMed  Google Scholar 

  7. Min, K. J., Lee, C. K. & Park, H. N. The lifespan of Korean eunuchs. Curr. Biol. 22, R792–R793 (2012).

    Article  CAS  PubMed  Google Scholar 

  8. Veldhuis, J. D. et al. The aging male hypothalamic–pituitary–gonadal axis: pulsatility and feedback. Mol. Cell. Endocrinol. 299, 14–22 (2009).

    Article  CAS  PubMed  Google Scholar 

  9. Veldhuis, J. D. Aging and hormones of the hypothalamo–pituitary axis: gonadotropic axis in men and somatotropic axes in men and women. Ageing Res. Rev. 7, 189–208 (2008).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Tenover, J. S., Matsumoto, A. M., Plymate, S. R. & Bremner, W. J. The effects of aging in normal men on bioavailable testosterone and luteinizing hormone secretion: response to clomiphene citrate. J. Clin. Endocrinol. Metab. 65, 1118–1126 (1987).

    Article  CAS  PubMed  Google Scholar 

  11. Rosario, E. R., Carroll, J. C. & Pike, C. J. Evaluation of the effects of testosterone and luteinizing hormone on regulation of β-amyloid in male 3×Tg-AD mice. Brain Res. 1466, 137–145 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Hyde, Z. et al. Higher luteinizing hormone is associated with poor memory recall: the Health in Men Study. J. Alzheimers Dis. 19, 943–951 (2010).

    Article  CAS  PubMed  Google Scholar 

  13. Verdile, G. et al. Luteinizing hormone levels are positively correlated with plasma amyloid-β protein levels in elderly men. J. Alzheimers Dis. 14, 201–208 (2008).

    Article  CAS  PubMed  Google Scholar 

  14. Greenhill, C. Pituitary function: pulsatile GnRH therapy in CCPHD. Nat. Rev. Endocrinol. 13, 315 (2017).

    Article  PubMed  Google Scholar 

  15. Pincus, S. M., Veldhuis, J. D., Mulligan, T., Iranmanesh, A. & Evans, W. S. Effects of age on the irregularity of LH and FSH serum concentrations in women and men. Am. J. Physiol. 273, E989–E995 (1997).

    CAS  PubMed  Google Scholar 

  16. Keenan, D. M. & Veldhuis, J. D. Disruption of the hypothalamic luteinizing hormone pulsing mechanism in aging men. Am. J. Physiol. Regul. Integr. Comp. Physiol. 281, R1917–R1924 (2001).

    Article  CAS  PubMed  Google Scholar 

  17. Mulligan, T., Iranmanesh, A., Kerzner, R., Demers, L. W. & Veldhuis, J. D. Two-week pulsatile gonadotropin releasing hormone infusion unmasks dual (hypothalamic and Leydig cell) defects in the healthy aging male gonadotropic axis. Eur. J. Endocrinol. 141, 257–266 (1999).

    Article  CAS  PubMed  Google Scholar 

  18. Veldhuis, J. D., Urban, R. J., Lizarralde, G., Johnson, M. L. & Iranmanesh, A. Attenuation of luteinizing hormone secretory burst amplitude as a proximate basis for the hypoandrogenism of healthy aging in men. J. Clin. Endocrinol. Metab. 75, 707–713 (1992).

    CAS  PubMed  Google Scholar 

  19. Choe, H. K. et al. Synchronous activation of gonadotropin-releasing hormone gene transcription and secretion by pulsatile kisspeptin stimulation. Proc. Natl Acad. Sci. USA 110, 5677–5682 (2013).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Adamson, A. et al. Signal transduction controls heterogeneous NF-κB dynamics and target gene expression through cytokine-specific refractory states. Nat. Commun. https://doi.org/10.1038/ncomms12057 (2016).

  21. Ashall, L. et al. Pulsatile stimulation determines timing and specificity of NF-κB-dependent transcription. Science 324, 242–246 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Nelson, D. E. et al. Oscillations in NF-κB signaling control the dynamics of gene expression. Science 306, 704–708 (2004).

    Article  CAS  PubMed  Google Scholar 

  23. Tilbrook, A. J. & Clarke, I. J. Negative feedback regulation of the secretion and actions of gonadotropin-releasing hormone in males. Biol. Reprod. 64, 735–742 (2001).

    Article  CAS  PubMed  Google Scholar 

  24. Kelly, D. M. & Jones, T. H. Testosterone: a metabolic hormone in health and disease. J. Endocrinol. 217, R25–R45 (2013).

    Article  CAS  PubMed  Google Scholar 

  25. Holmes, S., Singh, M., Su, C. & Cunningham, R. L. Effects of oxidative stress and testosterone on pro-inflammatory signaling in a female rat dopaminergic neuronal cell line. Endocrinology 157, 2824–2835 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Bolshakova, A., Magnusson, K. E., Pinaev, G. & Petukhova, O. EGF-induced dynamics of NF-κB and F-actin in A431 cells spread on fibronectin. Histochem. Cell Biol. 1—44, 223–235 (2015).

    Article  Google Scholar 

  27. Campos, P. & Herbison, A. E. Optogenetic activation of GnRH neurons reveals minimal requirements for pulsatile luteinizing hormone secretion. Proc. Natl Acad. Sci. USA 111, 18387–18392 (2014).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Yu, B., Tang, Y. & Cai, D. Brain is an endocrine organ through secretion and nuclear transfer of parathymosin. Life Sci. Alliance https://doi.org/10.26508/lsa.202000917 (2020).

  29. Tang, Y., Zuniga-Hertz, J. P., Han, C., Yu, B. & Cai, D. Multifaceted secretion of htNSC-derived hypothalamic islets induces survival and antidiabetic effect via peripheral implantation in mice. eLife https://doi.org/10.7554/eLife.52580 (2020).

  30. Li, J. X., Tang, Y. Z. & Cai, D. S. IKKβ/NF-κB disrupts adult hypothalamic neural stem cells to mediate a neurodegenerative mechanism of dietary obesity and pre-diabetes. Nat. Cell Biol. 14, 999–1012 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Meredith, J. M., Turek, F. W. & Levine, J. E. Effects of gonadotropin-releasing hormone pulse frequency modulation on the reproductive axis of photoinhibited male Siberian hamsters. Biol. Reprod. 59, 813–819 (1998).

    Article  CAS  PubMed  Google Scholar 

  32. Cai, D. et al. IKKβ/NF-κB activation causes severe muscle wasting in mice. Cell 119, 285–298 (2004).

    Article  CAS  PubMed  Google Scholar 

  33. Oakley, A. E., Clifton, D. K. & Steiner, R. A. Kisspeptin signaling in the brain. Endocr. Rev. 30, 713–743 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Okamura, H. et al. Kisspeptin and GnRH pulse generation. Adv. Exp. Med. Biol. 784, 297–323 (2013).

    Article  CAS  PubMed  Google Scholar 

  35. Herbison, A. E. The gonadotropin-releasing hormone pulse generator. Endocrinology 159, 3723–3736 (2018).

    Article  CAS  PubMed  Google Scholar 

  36. Simonneaux, V. A Kiss to drive rhythms in reproduction. Eur. J. Neurosci. 51, 509–530 (2020).

    Article  PubMed  Google Scholar 

  37. Titolo, D., Cai, F. & Belsham, D. D. Coordinate regulation of neuropeptide Y and agouti-related peptide gene expression by estrogen depends on the ratio of estrogen receptor (ER)α to ERβ in clonal hypothalamic neurons. Mol. Endocrinol. 20, 2080–2092 (2006).

    Article  CAS  PubMed  Google Scholar 

  38. Klenke, U., Constantin, S. & Wray, S. Neuropeptide Y directly inhibits neuronal activity in a subpopulation of gonadotropin-releasing hormone-1 neurons via Y1 receptors. Endocrinology 151, 2736–2746 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Verma, S., Kirigiti, M. A., Millar, R. P., Grove, K. L. & Smith, M. S. Endogenous kisspeptin tone is a critical excitatory component of spontaneous GnRH activity and the GnRH response to NPY and CART. Neuroendocrinology 99, 190–203 (2014).

    Article  CAS  PubMed  Google Scholar 

  40. Roa, J. & Herbison, A. E. Direct regulation of GnRH neuron excitability by arcuate nucleus POMC and NPY neuron neuropeptides in female mice. Endocrinology 153, 5587–5599 (2012).

    Article  CAS  PubMed  Google Scholar 

  41. Kim, K. & Choe, H. K. Role of hypothalamus in aging and its underlying cellular mechanisms. Mech. Ageing Dev. 177, 74–79 (2019).

    Article  CAS  PubMed  Google Scholar 

  42. Kermath, B. A., Riha, P. D., Woller, M. J., Wolfe, A. & Gore, A. C. Hypothalamic molecular changes underlying natural reproductive senescence in the female rat. Endocrinology 155, 3597–3609 (2014).

    Article  PubMed  PubMed Central  Google Scholar 

  43. Zhang, X. et al. Hypothalamic IKKβ/NF-κB and ER stress link overnutrition to energy imbalance and obesity. Cell 135, 61–73 (2008).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Au, A. et al. Estrogens, inflammation and cognition. Front. Neuroendocrinol. 40, 87–100 (2016).

    Article  CAS  PubMed  Google Scholar 

  45. Zhang, Y., Reichel, J. M., Han, C., Zuniga-Hertz, J. P. & Cai, D. Astrocytic process plasticity and IKKβ/NF-κB in central control of blood glucose, blood pressure, and body weight. Cell Metab. 25, 1091–1102 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. Sieben, C. J. et al. BubR1 allelic effects drive phenotypic heterogeneity in mosaic-variegated aneuploidy progeria syndrome. J. Clin. Invest. 130, 171–188 (2020).

    Article  CAS  PubMed  Google Scholar 

  47. Johnson, M. L. et al. AutoDecon: a robust numerical method for the quantification of pulsatile events. Methods Enzymol. 454, 367–404 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  48. Pincus, S. M. & Goldberger, A. L. Physiological time-series analysis: what does regularity quantify? Am. J. Physiol. 266, H1643–H1656 (1994).

    CAS  PubMed  Google Scholar 

  49. Fix, C., Jordan, C., Cano, P. & Walker, W. H. Testosterone activates mitogen-activated protein kinase and the cAMP response element binding protein transcription factor in Sertoli cells. Proc. Natl Acad. Sci. USA 101, 10919–10924 (2004).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  50. Hatef, A. & Unniappan, S. Gonadotropin-releasing hormone, kisspeptin, and gonadal steroids directly modulate nucleobindin-2/nesfatin-1 in murine hypothalamic gonadotropin-releasing hormone neurons and gonadotropes. Biol. Reprod. 96, 635–651 (2017).

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

We thank the Cai laboratory members for technical support, A. Wolfe (Johns Hopkins University) for the Gnrh-Cre mouse model, P. Mellon (University of California, San Diego) for GT1-7 and LβT4 cells, and the IVIS facility at Einstein for in vivo imaging assistance. This study was supported directly or indirectly through Einstein resources and NIH grants R01 AG031774, DK121435 and HL147477 (all to D.C.).

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Authors and Affiliations

  1. Department of Molecular Pharmacology, Albert Einstein College of Medicine, Bronx, NY, USA

    Zhouguang Wang, Wenhe Wu, Min Soo Kim & Dongsheng Cai

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  1. Zhouguang Wang
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Contributions

Z.W. co-designed and performed experiments, including GnRH pulse imaging, optogenetic models and physiological assays, castration model follow-up, cell culture models, immunostaining, histology and biochemistry, performed data analysis, contributed to interpretation of the data and prepared the figures for the paper. W.W. co-designed and performed experiments, including castration, LH knockdown, physiology and biochemistry experiments, and performed data analysis and interpretation. M.K. contributed to the castration model, GnRH knockdown and physiology studies. D.C. conceived the idea and hypothesis, conceptualized the project, designed the whole study and specific aims, constructed the experimental framework, supervised the study, led data analysis and interpretation, and wrote the paper.

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Correspondence to Dongsheng Cai.

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The authors declare no competing interests.

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Peer review information Nature Aging thanks Cláudia Cavadas and Han Kyoung Choe for their contribution to the peer review of this work.

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

Extended data

Extended Data Fig. 1 Pulses of blood FSH concentrations in male mice of different ages.

Blood FSH concentrations were measured during 180 mins in young (3 months), early middle-aged (11–12 months) and late middle-aged (16–17 months) mice. a: Representative blood FSH concentrations. b–d: Quantitative analysis of pulse frequency (b), peak concentration (c), and AUC value (d) of blood FSH concentrations. Comparisons of all groups were performed with one-way ANOVA but without statistical significance, n = 5 mice per group (b–d). Bar graphs reflect mean ± s.e.m.

Source data

Extended Data Fig. 2 A schematic model of oscillated GnRH mRNA production and NF-κB signaling.

This schematic diagram through drawing illustrative curves (which were not experimental data) is to generalize the concept for the relationship among oscillations of NF-κB activation and the production of IκBα protein, IκBα mRNA and GnRH mRNA in GnRH cells. Based on this conceptual model, it can be deduced that when NF-κB is chronically up-activated during aging, it leads to an equilibrium of fast-frequency NF-κB nuclear oscillation and fast-frequency pulsatile GnRH mRNA production.

Extended Data Fig. 3 Late age physiology and lifespan of mice with castration at the age of 8 months.

(a) Schematic diagram of the experimental procedure. (b–i) Mice were assigned into subgroups for different behavioral assays at the age of 18–19 months for locomotion (b), treadmill (c), coordination (d), novel recognition (e), sociality (f), learning and memory (g), adhesive removal (h), and odor smelling (i). (j–k) Subgroups of mice were profiled for pulse frequencies (j) and peak concentrations (k) of blood LH. (l) Lifespan follow-up of these mice. P values are shown in figures for comparisons between groups as indicated (k, l) or labeled as non-significant (NS) (b–j), two-tailed Student’s t-test (b–k), log-rank (Mantel-Cox) test (l); n = 8 per group (b–i), n = 5 per group (j, k); n = 22 mice per group (l). Data are mean ± s.e.m.

Source data

Extended Data Fig. 4 Effects of middle-age castration plus LH inhibition on histology.

Middle-aged mice with castration and LH knockdown (KD) were described in Fig. 4, and subgroups were used for examining skeletal muscle (a) and skin (b) histology. The results present the representative H&E staining. The statistical analyses are shown in Fig. 4n–p. Scale bars: 100 μm.

Extended Data Fig. 5 GnRH-Cre in subpopulation of Sox2-positive hypothalamic cells.

GnRH-Cre mice were crossed with Rosa 26-lox-STOP-lox-GFP mice leading to the offspring of GnRHCre:Rosa GFP mice, and the hypothalamic sections of these offspring were made for co-immunostaining of Sox2 and GnRH-Cre-dependent GFP expression (labelled as GnRH-GFP). White arrows point to representative cells which are co-positive for Sox2 and GFP. Images represent 4 independent repeats per condition. Scare bar: 100 μm.

Extended Data Fig. 6 Fast optogenetic stimulation prevents NF-κB inhibition from slowing LH pluses.

GnRH-Cre mice (12-month-old male) received hypothalamic injection of AAV together with a lentivirus containing GnRH promoter-driven DN-IκBα (labelled as DNIκBα) vs. control (labeled as Control), and subsequently these animals were treated with fast optogenetic stimulation vs. sham procedure as described in Fig. 5 and profiled for blood concentrations of LH (a–d) and FSH (e–h). (a, e) Representative blood LH (a) and FSH (e) concentrations. (b–d, f–h) Quantitative analysis of pulse frequency (b, f), peak concentration (c, g) and AUC value (d, h) of blood LH (b-d) and FSH (f–h). P values are shown in figures for comparisons between groups as indicated, one-way ANOVA with Tukey’s post hoc test (b–d), n = 5 mice per group (b–d). Comparisons of all groups with one-way ANOVA for lacking statistical significance (f–h), n = 5 mice per group (f–h). Data are mean ± s.e.m.

Source data

Extended Data Fig. 7 Fast optogenetic stimulation reduces the anti-aging effect of NF-κB inhibition.

GnRH-Cre male mice (12 months old) received hypothalamic injection of AAV together with a lentivirus containing GnRH promoter-driven DNIκBα vs. Control, and subsequently these animals were treated with fast optogenetic stimulation vs. sham as described in Fig. 5 and profiled for physiology. (a–c) Longitudinal body weight (a) and food intake (b) follow-up and body composition measured at the age of about 18 months (c). (d–m) Aging-related physiology including behavioral functions assessed when 18–19 months old (d–j) and tissue histology assessed when 24 months old (k–m). P values are shown in figures for comparisons between groups as indicated, one-way ANOVA with Tukey’s post hoc test (d–m); n = 10 mice per group (a–b), n = 7 mice per group (c), n = 8 mice per group (d–m). Data are mean ± s.e.m.

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Wang, Z., Wu, W., Kim, M.S. et al. GnRH pulse frequency and irregularity play a role in male aging. Nat Aging 1, 904–918 (2021). https://doi.org/10.1038/s43587-021-00116-5

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