CoChat
Sign Up
Cited by 48Neuroendocrine regulation and behaviorStress Responses and CortisolEstrogen and related hormone effects

The Androgen Metabolite, 5α-Androstane-3β,17β-Diol (3β-Diol), Activates the Oxytocin Promoter Through an Estrogen Receptor-β Pathway

Open access
Hiroi, Ryoko, Lacagnina, Anthony F., Hinds, Laura R., Carbone, David G., Uht, Rosalie M., Handa, Robert J.·2013·Endocrinology
DOI 10.1210/en.2012-2253PMC PMC3628024
Open this paper in CoChat — free accountOpen PDF

Abstract

Testosterone has been shown to suppress the acute stress-induced activation of the hypothalamic-pituitary-adrenal axis; however, the mechanisms underlying this response remain unclear. The hypothalamic-pituitary-adrenal axis is regulated by a neuroendocrine subpopulation of medial parvocellular neurons in the paraventricular nucleus of the hypothalamus (PVN). These neurons are devoid of androgen receptors (ARs). Therefore, a possibility is that the PVN target neurons respond to a metabolite in the testosterone catabolic pathway via an AR-independent mechanism. The dihydrotestosterone metabolite, 5α-androstane-3β,17β-diol (3β-diol), binds and activates estrogen receptor-β (ER-β), the predominant ER in the PVN. In the PVN, ER-β is coexpressed with oxytocin (OT). Therefore, we tested the hypothesis that 3β-diol regulates OT expression through ER-β activation. Treatment of ovariectomized rats with estradiol benzoate or 3β-diol for 4 days increased OT mRNA selectively in the midcaudal, but not rostral PVN compared with vehicle-treated controls. 3β-Diol treatment also increased OT mRNA in the hypothalamic N38 cell line in vitro. The functional interactions between 3β-diol and ER-β with the human OT promoter were examined using an OT promoter-luciferase reporter construct (OT-luc). In a dose-dependent manner, 3β-diol treatment increased OT-luc activity when cells were cotransfected with ER-β, but not ER-α. The 3β-diol–induced OT-luc activity was reduced by deletion of the promoter region containing the composite hormone response element (cHRE). Point mutations of the cHRE also prevented OT-luc activation by 3β-diol. These results indicate that 3β-diol induces OT promoter activity via ER-β–cHRE interactions.

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1210/en.2012-2253

Cites

Statistics

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • The effects of ERβ agonists on Ot gene expression are tracked to an ERE half-site in the Ot proximal promoter.

    “These effects of ERβ agonists were tracked to an ERE half-site in the Ot proximal promoter and thereby verify some molecular mechanisms for Ot gene expression regulation by ERβ.”

    From Hypothalamic–pituitary–adrenal and hypothalamic–pituitary–gonadal axes: sex differences in regulation of stress responsivity (2017) · cites this paper as Hiroi et al., 2013

    In the section on OT promoter activation (page unavailable), mutating the proximal promoter’s cHRE eliminated reporter responses to E2, DPN and 3β-diol with ERβ. The paper identifies that element as a complex hormone response element, not specifically an ERE half-site, and does not establish that ERβ acts directly at such a half-site. (LLM, conf=0.86)

    Experimental — accuracy not yet measured

  • 3β-diol increases Ot mRNA expression in the midcaudal, but not the rostral, portion of the rat PVN.

    “Consistent with these findings, 3β-diol also increases Ot mRNA expression in the midcaudal, but not the rostral portion, of the rat PVN (Hiroi et al., 2013).”

    From Hypothalamic–pituitary–adrenal and hypothalamic–pituitary–gonadal axes: sex differences in regulation of stress responsivity (2017) · cites this paper as Hiroi et al., 2013

  • 3β-diol increases Ot mRNA in hypothalamic N38 cells.

    “This study also reported an increase in Ot mRNA in hypothalamic N38 cells following 3β-diol treatment, indicating it effectiveness in regulating OT in vivo and in vitro.”

    From Hypothalamic–pituitary–adrenal and hypothalamic–pituitary–gonadal axes: sex differences in regulation of stress responsivity (2017) · cites this paper as Hiroi et al., 2013

  • ERβ has comparatively higher promiscuity (relative to ERα) for ligands including multiple phytoestrogens including GEN, and even the androgen metabolite 5α-androstane-3β,17β-diol (3β-diol).

    “Given the comparatively higher promiscuity of ERβ (relative to ERα) for ligands including multiple phytoestrogens including GEN (82), and even the androgen metabolite 5α-androstane-3β,17β-diol (3β-diol) (141), this is a strikingly understudied but likely avenue for endocrine disruption.”

    From Endocrine Disruption of Vasopressin Systems and Related Behaviors (2017) · cites this paper as [141]

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

The citation lists haven’t been fetched yet

References 62 · Cited by 48

Research works printed but not linked: 53

  1. Handa RJ, Burgess LH, Kerr JE, O'Keefe JA. Gonadal steroid hor- mone receptors and sex differences in the hypothalamo-pituitary- adrenal axis. Horm Behav. 1994;28:464 -476.
  2. Bingaman EW, Baeckman LM, Yracheta JM, Handa RJ, Gray TS. Localization of androgen receptor within peptidergic neurons of the rat forebrain. Brain Res Bull. 1994;35:379 -382.
  3. Bingham B, Myung C, Innala L, Gray M, Anonuevo A, Viau V. Androgen receptors in the posterior bed nucleus of the stria termi- nalis increase neuropeptide expression and the stress-induced acti- vation of the paraventricular nucleus of the hypothalamus. Neuro- psychopharmacology. 2011;36:1433-1443.
  4. Williamson M, Bingham B, Gray M, Innala L, Viau V. The medial preoptic nucleus integrates the central influences of testosterone on the paraventricular nucleus of the hypothalamus and its extended circuitries. J Neurosci. 2010;30:11762-11770.
  5. Lund TD, Hinds LR, Handa RJ. The androgen 5␣-dihydrotestos- terone and its metabolite 5␣-androstan-3␤,17␤-diol inhibit the hy- pothalamo-pituitary-adrenal response to stress by acting through estrogen receptor ␤-expressing neurons in the hypothalamus. J Neu- rosci. 2006;26:1448 -1456.
  6. Gangloff A, Shi R, Nahoum V, Lin SX. Pseudo-symmetry of C19 steroids, alternative binding orientations, and multispecificity in hu- man estrogenic 17␤-hydroxysteroid dehydrogenase. FASEB J. 2003;17:274 -276.
  7. Jin Y, Penning TM. Steroid 5␣-reductases and 3␣-hydroxysteroid dehydrogenases: key enzymes in androgen metabolism. Best Pract Res Clin Endocrinol Metab. 2001;15:79 -94.
  8. Steckelbroeck S, Jin Y, Gopishetty S, Oyesanmi B, Penning TM. Human cytosolic 3␣-hydroxysteroid dehydrogenases of the aldo- keto reductase superfamily display significant 3␤-hydroxysteroid dehydrogenase activity: implications for steroid hormone metabo- lism and action. J Biol Chem. 2004;279:10784 -10795.
  9. Torn S, Nokelainen P, Kurkela R, et al. Production, purification, and functional analysis of recombinant human and mouse 17␤- hydroxysteroid dehydrogenase type 7. Biochem Biophys Res Com- mun. 2003;305:37-45.
  10. Guerini V, Sau D, Scaccianoce E, et al. The androgen derivative 5␣-androstane-3␤,17␤-diol inhibits prostate cancer cell migration through activation of the estrogen receptor ␤ subtype. Cancer Res. 2005;65:5445-5453.
  11. Kuiper GG, Shughrue PJ, Merchenthaler I, Gustafsson JA. The es- trogen receptor ␤ subtype: a novel mediator of estrogen action in neuroendocrine systems. Front Neuroendocrinol. 1998;19:253- 286.
  12. Oliveira AG, Coelho PH, Guedes FD, Mahecha GA, Hess RA, Ol- iveira CA. 5␣-Androstane-3␤,17␤-diol (3␤-diol), an estrogenic me- tabolite of 5␣-dihydrotestosterone, is a potent modulator of estro- gen receptor ER␤ expression in the ventral prostrate of adult rats. Steroids. 2007;72:914 -922.
  13. Weihua Z, Lathe R, Warner M, Gustafsson JA. An endocrine path- way in the prostate, ER␤, AR, 5␣-androstane-3␤,17␤-diol, and CYP7B1, regulates prostate growth. Proc Natl Acad Sci U S A. 2002;99:13589 -13594.
  14. Alves SE, Lopez V, McEwen BS, Weiland NG. Differential colocal- ization of estrogen receptor ␤ (ER␤) with oxytocin and vasopressin in the paraventricular and supraoptic nuclei of the female rat brain: an immunocytochemical study. Proc Natl Acad Sci U S A. 1998; 95:3281-3286.
  15. Hrabovszky E, Kallo I, Hajszan T, Shughrue PJ, Merchenthaler I, Liposits Z. Expression of estrogen receptor-␤ messenger ribonucleic acid in oxytocin and vasopressin neurons of the rat supraoptic and paraventricular nuclei. Endocrinology. 1998;139:2600 -2604.
  16. Nomura M, McKenna E, Korach KS, Pfaff DW, Ogawa S. Estrogen receptor-␤ regulates transcript levels for oxytocin and arginine va- sopressin in the hypothalamic paraventricular nucleus of male mice. Brain Res Mol Brain Res. 2002;109:84 -94.
  17. Patisaul HB, Scordalakes EM, Young LJ, Rissman EF. Oxytocin, but not oxytocin receptor, is regulated by oestrogen receptor ␤ in the female mouse hypothalamus. J Neuroendocrinol. 2003;15:787- 793.
  18. Amico JA, Crowley RS, Insel TR, Thomas A, O'Keefe JA. Effect of gonadal steroids upon hypothalamic oxytocin expression. Adv Exp Med Biol. 1995;395:23-35.
  19. Blyth BJ, Hollingshead DJ, Amico JA. Time course of induction of oxytocin messenger ribonucleic acid levels in the hypothalamic paraventricular nucleus of ovariectomized rats following gonadal steroid administration. Life Sci. 1997;60:2427-2433.
  20. Shughrue PJ, Dellovade TL, Merchenthaler I. Estrogen modulates oxytocin gene expression in regions of the rat supraoptic and para- ventricular nuclei that contain estrogen receptor-␤. Prog Brain Res. 2002;139:15-29.
  21. Thomas A, Amico JA. Sequential estrogen and progesterone (P) followed by P withdrawal increases the level of oxytocin messenger ribonucleic acid in the hypothalamic paraventricular nucleus of the male rat. Life Sci. 1996;58:1615-1620.
  22. Thomas A, Crowley RS, Amico JA. Effect of progesterone on hy- pothalamic oxytocin messenger ribonucleic acid levels in the lactat- ing rat. Endocrinology. 1995;136:4188 -4194.
  23. Sharma D, Handa RJ, Uht RM. The ER␤ ligand 5␣-androstane, 3␤,17␤-diol (3␤-diol) regulates hypothalamic oxytocin (Oxt) gene expression. Endocrinology. 2012;153:2353-2361.
  24. Burbach JP, Adan RA, Tol HH, et al. Regulation of the rat oxytocin gene by estradiol. J Neuroendocrinol. 1990;2:633-639.
  25. Stedronsky K, Telgmann R, Tillmann G, Walther N, Ivell R. The affinity and activity of the multiple hormone response element in the proximal promoter of the human oxytocin gene. J Neuroendocrinol. 2002;14:472-485.
  26. Adan RA, Cox JJ, Beischlag TV, Burbach JP. A composite hormone response element mediates the transactivation of the rat oxytocin gene by different classes of nuclear hormone receptors. Mol Endo- crinol. 1993;7:47-57.
  27. Osterlund MK, Gustafsson JA, Keller E, Hurd YL. Estrogen receptor ␤ (ER␤) messenger ribonucleic acid (mRNA) expression within the human forebrain: distinct distribution pattern to ER␣ mRNA. J Clin Endocrinol Metab. 2000;85:3840 -3846.
  28. Hrabovszky E, Kallo I, Steinhauser A, et al. Estrogen receptor-␤ in oxytocin and vasopressin neurons of the rat and human hypothal- amus: Immunocytochemical and in situ hybridization studies. J Comp Neurol. 2004;473:315-333.
  29. Windle RJ, Shanks N, Lightman SL, Ingram CD. Central oxytocin administration reduces stress-induced corticosterone release and anxiety behavior in rats. Endocrinology. 1997;138:2829 -2834.
  30. Carbone DL, Zuloaga DG, Hiroi R, Foradori CD, Legare ME, Handa RJ. Prenatal dexamethasone exposure potentiates diet-in- duced hepatosteatosis and decreases plasma IGF-I in a sex-specific fashion. Endocrinology. 2012;153:295-306.
  31. Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987;162:156 -159.
  32. Paxinos G, Watson C. The rat brain in stereotaxic coordinates. New York, NY: Academic Press; 1998.
  33. Pak TR, Chung WC, Lund TD, Hinds LR, Clay CM, Handa RJ. The androgen metabolite, 5␣-androstane-3␤,17␤-diol, is a potent mod- ulator of estrogen receptor-␤1-mediated gene transcription in neu- ronal cells. Endocrinology. 2005;146:147-155.
  34. Behl C, Widmann M, Trapp T, Holsboer F. 17-␤-Estradiol protects neurons from oxidative stress-induced cell death in vitro. Biochem Biophys Res Commun. 1995;216:473-482.
  35. Green PS, Gridley KE, Simpkins JW. Nuclear estrogen receptor- independent neuroprotection by estratrienes: a novel interaction with glutathione. Neuroscience. 1998;84:7-10.
  36. Kim H, Bang OY, Jung MW, et al. Neuroprotective effects of es- trogen against ␤-amyloid toxicity are mediated by estrogen recep- tors in cultured neuronal cells. Neurosci Lett. 2001;302:58 -62.
  37. Fleming JG, Spencer TE, Safe SH, Bazer FW. Estrogen regulates transcription of the ovine oxytocin receptor gene through GC-rich SP1 promoter elements. Endocrinology. 2006;147:899 -911.
  38. Safe S. Transcriptional activation of genes by 17 ␤-estradiol through estrogen receptor-Sp1 interactions. Vitam Horm. 2001;62:231- 252.
  39. Handa RJ, Weiser MJ, Zuloaga DG. A role for the androgen me- tabolite, 5␣-androstane-3␤,17␤-diol, in modulating oestrogen re- ceptor ␤-mediated regulation of hormonal stress reactivity. J Neu- roendocrinol. 2009;21:351-358.
  40. Saijo K, Collier JG, Li AC, Katzenellenbogen JA, Glass CK. An ADIOL-ER␤-CtBP transrepression pathway negatively regulates microglia-mediated inflammation. Cell. 2011;145:584 -595.
  41. Ochedalski T, Subburaju S, Wynn PC, Aguilera G. Interaction be- tween oestrogen and oxytocin on hypothalamic-pituitary-adrenal axis activity. J Neuroendocrinol. 2007;19:189 -197.
  42. Windle RJ, Gamble LE, Kershaw YM, Wood SA, Lightman SL, Ingram CD. Gonadal steroid modulation of stress-induced hypo- thalamo-pituitary-adrenal activity and anxiety behavior: role of cen- tral oxytocin. Endocrinology. 2006;147:2423-2431.
  43. Windle RJ, Kershaw YM, Shanks N, Wood SA, Lightman SL, In- gram CD. Oxytocin attenuates stress-induced c-fos mRNA expres- sion in specific forebrain regions associated with modulation of hy- pothalamo-pituitary-adrenal activity. J Neurosci. 2004;24:2974 - 2982.
  44. Crowley RS, Insel TR, O'Keefe JA, Kim NB, Amico JA. Increased accumulation of oxytocin messenger ribonucleic acid in the hypo- thalamus of the female rat: induction by long term estradiol and progesterone administration and subsequent progesterone with- drawal. Endocrinology. 1995;136:224 -231.
  45. Chung SK, McCabe JT, Pfaff DW. Estrogen influences on oxytocin mRNA expression in preoptic and anterior hypothalamic regions studied by in situ hybridization. J Comp Neurol. 1991;307:281- 295.
  46. Koohi MK, Ivell R, Walther N. Transcriptional activation of the oxytocin promoter by oestrogens uses a novel non-classical mech- anism of oestrogen receptor action. J Neuroendocrinol. 2005;17: 197-207.
  47. Koohi MK, Walther N, Ivell R. A novel molecular assay to discrim- inate transcriptional effects caused by xenoestrogens. Mol Cell En- docrinol. 2007;276:45-54.
  48. Richard S, Zingg HH. The human oxytocin gene promoter is reg- ulated by estrogens. J Biol Chem. 1990;265:6098 -6103.
  49. Adan RA, Burbach JP. Regulation of vasopressin and oxytocin gene expression by estrogen and thyroid hormone. Prog Brain Res. 1992; 92:127-136.
  50. Richard S, Zingg HH. Identification of cis-acting regulatory ele- ments in the human oxytocin gene promoter. Mol Cell Neurosci. 1991;2:501-510.
  51. Belsham DD, Cai F, Cui H, Smukler SR, Salapatek AM, Shkreta L. Generation of a phenotypic array of hypothalamic neuronal cell models to study complex neuroendocrine disorders. Endocrinology. 2004;145:393-400.
  52. Miller L, Foradori CD, Lalmansingh AS, Sharma D, Handa RJ, Uht RM. Histone deacetylase 1 (HDAC1) participates in the down-reg- ulation of corticotropin releasing hormone gene (crh) expression. Physiol Behav. 2011;104:312-320.
  53. Osterlund MK, Grandien K, Keller E, Hurd YL. The human brain has distinct regional expression patterns of estrogen receptor ␣ mRNA isoforms derived from alternative promoters. J Neurochem. 2000;75:1390 -1397.

Authors
Hiroi, R., Lacagnina, A. F., Hinds, L. R., Carbone, D. G., Uht, R. M., & Handa, R. J.
Affiliations
  • University of Arizona College of Medicine- Phoenix (US)
  • University of North Texas Health Science Center (US)
Corresponding author
Handa, Robert J.
ORCID
Venue
Endocrinology
Publisher
The Endocrine Society
Published
2013-03-20
Volume
154
Issue
5
Pages
1802-1812
ISSN
  • 0013-7227 (ISSN-L)
  • 1945-7170
DOI
10.1210/en.2012-2253
PMC
PMC3628024
OpenAlex
W2040981352
Cited by
48
References
62
Citations by year
2026: 3 · 2025: 1 · 2024: 3 · 2023: 4 · 2022: 1 · 2021: 6 · 2020: 2 · 2019: 4 · 2018: 4 · 2017: 8 · 2016: 1 · 2015: 6 · 2014: 4 · 2013: 1
FWCI
6.00
Type
journal article
Language
en
Open access
Locations
  • Endocrinology (publishedVersion)
  • Europe PMC (PubMed Central) (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
Funders
  • Arizona State University
  • Frances McClelland Institute for Children, Youth, and Families
  • National Institutes of Health (NS039951)
  • National Institute of Mental Health (F32MH093145)
  • National Institute of Neurological Disorders and Stroke (R01NS039951)
Topics
  • Social Sciences › Psychology › Social Psychology › Neuroendocrine regulation and behavior
  • Life Sciences › Neuroscience › Behavioral Neuroscience › Stress Responses and Cortisol
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Estrogen and related hormone effects
Keywords
5α-androstane-3β,17β-diol, estrogen receptor beta, paraventricular nucleus, androgen metabolism, hypothalamic-pituitary-adrenal axis, acute stress, estradiol benzoate
MeSH terms
Androgens, Animals, Cells, Cultured, Female, Humans, Ovariectomy, Oxytocin, Promoter Regions, Genetic, Androstane-3,17-diol, Signal Transduction, Transcriptional Activation, Rats, Sprague-Dawley, Response Elements, Estrogen Receptor beta, Mice, Rats

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review