Letrozole treatment of pubertal female mice results in activational effects on reproduction, metabolism and the gut microbiome

Autoři: Pablo Arroyo aff001;  Bryan S. Ho aff002;  Lillian Sau aff002;  Scott T. Kelley aff001;  Varykina G. Thackray aff002
Působiště autorů: Department of Biology, San Diego State University, San Diego, CA, United States of America aff001;  Department of Obstetrics, Gynecology and Reproductive Sciences, University of California, San Diego, La Jolla, CA, United States of America aff002
Vyšlo v časopise: PLoS ONE 14(9)
Kategorie: Research Article
doi: 10.1371/journal.pone.0223274


Polycystic ovary syndrome (PCOS) is a common endocrine disorder in reproductive-aged women that is comprised of two out of the following three features: hyperandrogenism, oligo- or amenorrhea, or polycystic ovaries. In addition to infertility, many women with PCOS have metabolic dysregulation that increases the risk of developing type 2 diabetes, hypertension, and non-alcoholic fatty liver disease. Changes in the gut microbiome are associated with PCOS and gut microbes may be involved in the pathology of this disorder. Since PCOS often manifests in the early reproductive years, puberty is considered to be a critical time period for the development of PCOS. Exposure to sex steroid hormones during development results in permanent, organizational effects, while activational effects are transient and require the continued presence of the hormone. Androgens exert organizational effects during prenatal or early post-natal development, but it is unclear whether androgen excess results in organizational or activational effects during puberty. We recently developed a letrozole-induced PCOS mouse model that recapitulates both reproductive and metabolic phenotypes of PCOS. In this study, we investigated whether letrozole treatment of pubertal female mice exerts organizational or activational effects on host physiology and the gut microbiome. Two months after letrozole removal, we observed recovery of reproductive and metabolic parameters, as well as diversity and composition of the gut microbiome, indicating that letrozole treatment of female mice during puberty resulted in predominantly activational effects. These results suggest that if exposure to excess androgens during puberty leads to the development of PCOS, reduction of androgen levels during this time may improve reproductive and metabolic phenotypes in women with PCOS. These results also imply that continuous letrozole exposure is required to model PCOS in pubertal female mice since letrozole exerts activational rather than organizational effects during puberty.

Klíčová slova:

Androgens – Insulin – Microbiome – Mouse models – Ovaries – Testosterone – Polycystic ovary syndrome – Puberty


1. Fauser BC, Tarlatzis BC, Rebar RW, Legro RS, Balen AH, Lobo R, et al. Consensus on women's health aspects of polycystic ovary syndrome (PCOS): the Amsterdam ESHRE/ASRM-Sponsored 3rd PCOS Consensus Workshop Group. Fertil Steril. 2012;97(1):28–38 e25. doi: 10.1016/j.fertnstert.2011.09.024 22153789

2. Rotterdam EA-SPCWG. Revised 2003 consensus on diagnostic criteria and long-term health risks related to polycystic ovary syndrome. Fertil Steril. 2004;81(1):19–25. doi: 10.1016/j.fertnstert.2003.10.004 14711538

3. Teede HJ, Misso ML, Deeks AA, Moran LJ, Stuckey BG, Wong JL, et al. Assessment and management of polycystic ovary syndrome: summary of an evidence-based guideline. Med J Aust. 2011;195(6):S65–112. doi: 10.5694/mja11.10915 21929505

4. Yao K, Bian C, Zhao X. Association of polycystic ovary syndrome with metabolic syndrome and gestational diabetes: Aggravated complication of pregnancy. Exp Ther Med. 2017;14(2):1271–6. doi: 10.3892/etm.2017.4642 28810587

5. Barthelmess EK, Naz RK. Polycystic ovary syndrome: current status and future perspective. Frontiers in bioscience. 2014;6:104–19.

6. Barber TM, Franks S. Genetics of polycystic ovary syndrome. Front Horm Res. 2013;40:28–39. doi: 10.1159/000341682 24002403

7. Legro RS, Driscoll D, Strauss JF, Fox J 3rd, Dunaif A. Evidence for a genetic basis for hyperandrogenemia in polycystic ovary syndrome. Proc Natl Acad Sci U S A. 1998;95(25):14956–60. doi: 10.1073/pnas.95.25.14956 9843997

8. Vink JM, Sadrzadeh S, Lambalk CB, Boomsma DI. Heritability of polycystic ovary syndrome in a Dutch twin-family study. J Clin Endocrinol Metab. 2006;91(6):2100–4. doi: 10.1210/jc.2005-1494 16219714

9. Abbott DH, Bacha F. Ontogeny of polycystic ovary syndrome and insulin resistance in utero and early childhood. Fertil Steril. 2013;100(1):2–11. doi: 10.1016/j.fertnstert.2013.05.023 23809624

10. Anderson AD, Solorzano CM, McCartney CR. Childhood obesity and its impact on the development of adolescent PCOS. Semin Reprod Med. 2014;32(3):202–13. doi: 10.1055/s-0034-1371092 24715515

11. Abbott DH, Nicol LE, Levine JE, Xu N, Goodarzi MO, Dumesic DA. Nonhuman primate models of polycystic ovary syndrome. Mol Cell Endocrinol. 2013;373(1–2):21–8. doi: 10.1016/j.mce.2013.01.013 23370180

12. Padmanabhan V, Veiga-Lopez A. Animal models of the polycystic ovary syndrome phenotype. Steroids. 2013;78(8):734–40. doi: 10.1016/j.steroids.2013.05.004 23701728

13. Walters KA, Bertoldo MJ, Handelsman DJ. Evidence from animal models on the pathogenesis of PCOS. Best Pract Res Clin Endocrinol Metab. 2018;32(3):271–81. doi: 10.1016/j.beem.2018.03.008 29779581

14. Walters KA, Allan CM, Handelsman DJ. Rodent models for human polycystic ovary syndrome. Biol Reprod. 2012;86(5):149, 1–12. doi: 10.1095/biolreprod.111.097808 22337333

15. van Houten EL, Visser JA. Mouse models to study polycystic ovary syndrome: a possible link between metabolism and ovarian function? Reprod Biol. 2014;14(1):32–43. doi: 10.1016/j.repbio.2013.09.007 24607253

16. Caldwell AS, Middleton LJ, Jimenez M, Desai R, McMahon AC, Allan CM, et al. Characterization of reproductive, metabolic, and endocrine features of polycystic ovary syndrome in female hyperandrogenic mouse models. Endocrinology. 2014;155(8):3146–59. doi: 10.1210/en.2014-1196 24877633

17. Franks S. Animal models and the developmental origins of polycystic ovary syndrome: increasing evidence for the role of androgens in programming reproductive and metabolic dysfunction. Endocrinology. 2012;153(6):2536–8. doi: 10.1210/en.2012-1366 22610962

18. Arnold AP, Breedlove SM. Organizational and activational effects of sex steroids on brain and behavior: A reanalysis. Hormones and Behavior. 1985;19(4):469–98. 3910535

19. Sullivan SD, Moenter SM. Prenatal androgens alter GABAergic drive to gonadotropin-releasing hormone neurons: implications for a common fertility disorder. Proc Natl Acad Sci USA. 2004;101(18):7129–34. doi: 10.1073/pnas.0308058101 15096602

20. Moore AM, Prescott M, Marshall CJ, Yip SH, Campbell RE. Enhancement of a robust arcuate GABAergic input to gonadotropin-releasing hormone neurons in a model of polycystic ovarian syndrome. Proc Natl Acad Sci USA. 2015;112(2):596–601. doi: 10.1073/pnas.1415038112 25550522

21. Caldwell ASL, Middleton LJ, Jimenez M, Desai R, McMahon AC, Allan CM, et al. Characterization of Reproductive, Metabolic, and Endocrine Features of Polycystic Ovary Syndrome in Female Hyperandrogenic Mouse Models. Endocrinology. 2014;155(8):3146–59. doi: 10.1210/en.2014-1196 24877633

22. Moore AM, Prescott M, Campbell RE. Estradiol negative and positive feedback in a prenatal androgen-induced mouse model of polycystic ovarian syndrome. Endocrinology. 2013;154(2):796–806. doi: 10.1210/en.2012-1954 23254197

23. Roland AV, Nunemaker CS, Keller SR, Moenter SM. Prenatal androgen exposure programs metabolic dysfunction in female mice. J Endocrinol. 2010;207(2):213–23. doi: 10.1677/JOE-10-0217 20713501

24. Witham EA, Meadows JD, Shojaei S, Kauffman AS, Mellon PL. Prenatal exposure to low levels of androgen accelerates female puberty onset and reproductive senescence in mice. Endocrinology. 2012;153(9):4522–32. doi: 10.1210/en.2012-1283 22778229

25. van Houten EL, Kramer P, McLuskey A, Karels B, Themmen AP, Visser JA. Reproductive and metabolic phenotype of a mouse model of PCOS. Endocrinology. 2012;153(6):2861–9. doi: 10.1210/en.2011-1754 22334715

26. Kauffman AS, Thackray VG, Ryan GE, Tolson KP, Glidewell-Kenney CA, Semaan SJ, et al. A novel letrozole model recapitulates both the reproductive and metabolic phenotypes of Polycystic Ovary Syndrome in female mice. Biol Reprod. 2015;93(3):69. doi: 10.1095/biolreprod.115.131631 26203175

27. Skarra DV, Hernandez-Carretero A, Rivera AJ, Anvar AR, Thackray VG. Hyperandrogenemia Induced by Letrozole Treatment of Pubertal Female Mice Results in Hyperinsulinemia Prior to Weight Gain and Insulin Resistance. Endocrinology. 2017;158(9):2988–3003. doi: 10.1210/en.2016-1898 28911175

28. Kelley ST, Skarra DV, Rivera AJ, Thackray VG. The Gut Microbiome Is Altered in a Letrozole-Induced Mouse Model of Polycystic Ovary Syndrome. PLoS One. 2016;11(1):e0146509. doi: 10.1371/journal.pone.0146509 26731268

29. Marcondes FK, Bianchi FJ, Tanno AP. Determination of the estrous cycle phases of rats: some helpful considerations. Braz J Biol. 2002;62(4A):609–14. doi: 10.1590/s1519-69842002000400008 12659010

30. Caporaso JG, Lauber CL, Walters WA, Berg-Lyons D, Huntley J, Fierer N, et al. Ultra-high-throughput microbial community analysis on the Illumina HiSeq and MiSeq platforms. ISME J. 2012;6(8):1621–4. doi: 10.1038/ismej.2012.8 22402401

31. Fierer N, Hamady M, Lauber CL, Knight R. The influence of sex, handedness, and washing on the diversity of hand surface bacteria. Proc Natl Acad Sci U S A. 2008;105(46):17994–9. doi: 10.1073/pnas.0807920105 19004758

32. Caporaso JG, Kuczynski J, Stombaugh J, Bittinger K, Bushman FD, Costello EK, et al. QIIME allows analysis of high-throughput community sequencing data. Nat Methods. 2010;7(5):335–6. doi: 10.1038/nmeth.f.303 20383131

33. Faith DP. Conservation evaluation and phylogenetic diversity. Biological Conservation. 1992;61(1):1–10.

34. Watson M, McMurdie PJ, Holmes S. phyloseq: An R Package for Reproducible Interactive Analysis and Graphics of Microbiome Census Data. PLoS ONE. 2013;8(4).

35. Lozupone C, Lladser ME, Knights D, Stombaugh J, Knight R. UniFrac: an effective distance metric for microbial community comparison. ISME J. 2011;5(2):169–72. doi: 10.1038/ismej.2010.133 20827291

36. Love MI, Huber W, Anders S. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol. 2014;15(12):550. doi: 10.1186/s13059-014-0550-8 25516281

37. Lindheim L, Bashir M, Munzker J, Trummer C, Zachhuber V, Leber B, et al. Alterations in Gut Microbiome Composition and Barrier Function Are Associated with Reproductive and Metabolic Defects in Women with Polycystic Ovary Syndrome (PCOS): A Pilot Study. PLoS One. 2017;12(1):e0168390. doi: 10.1371/journal.pone.0168390 28045919

38. Liu R, Zhang CH, Shi Y, Zhang F, Li LX, Wang XJ, et al. Dysbiosis of Gut Microbiota Associated with Clinical Parameters in Polycystic Ovary Syndrome. Frontiers in Microbiology. 2017;8. doi: 10.3389/fmicb.2017.00008

39. Torres PJ, Siakowska M, Banaszewska B, Pawelczyk L, Duleba AJ, Kelley ST, et al. Gut Microbial Diversity in Women with Polycystic Ovary Syndrome Correlates with Hyperandrogenism. J Clin Endocrinol Metab. 2018;103(4):1502–1511. doi: 10.1210/jc.2017-02153 29370410

40. Torres PJ, Ho BS, Arroyo P, Sau L, Chen A, Kelley ST, et al. Exposure to a Healthy Gut Microbiome Protects Against Reproductive and Metabolic Dysregulation in a PCOS Mouse Model. Endocrinology. 2019; 160(5):1193–1204. doi: 10.1210/en.2019-00050 30924862

41. Torres PJ, Skarra DV, Ho BS, Sau L, Anvar AR, Kelley ST, et al. Letrozole treatment of adult female mice results in a similar reproductive phenotype but distinct changes in metabolism and the gut microbiome compared to pubertal mice. BMC Microbiol. 2019;19(1):57. doi: 10.1186/s12866-019-1425-7 30871463

42. Hu M, Richard JE, Maliqueo M, Kokosar M, Fornes R, Benrick A, et al. Maternal testosterone exposure increases anxiety-like behavior and impacts the limbic system in the offspring. Proceedings of the National Academy of Sciences of the United States of America. 2015;112(46):14348–53. doi: 10.1073/pnas.1507514112 26578781

43. Torres Fernandez ED, Adams KV, Syed M, Maranon RO, Romero DG, Yanes Cardozo LL. Long-Lasting Androgen-Induced Cardiometabolic Effects in Polycystic Ovary Syndrome. Journal of the Endocrine Society. 2018;2(8):949–64. doi: 10.1210/js.2018-00131 30087950

44. Thaiss CA, Itav S, Rothschild D, Meijer M, Levy M, Moresi C, et al. Persistent microbiome alterations modulate the rate of post-dieting weight regain. Nature. 2016;540(7634):544–551. doi: 10.1038/nature20796 27906159

45. Thackray VG. Sex, Microbes, and Polycystic Ovary Syndrome. Trends in Endocrinology & Metabolism. 2019;30(1):54–65.

46. Schulz KM, Molenda-Figueira HA, Sisk CL. Back to the future: The organizational-activational hypothesis adapted to puberty and adolescence. Horm Behav. 2009;55(5):597–604. doi: 10.1016/j.yhbeh.2009.03.010 19446076

47. Guo YJ, Qi Y, Yang XF, Zhao LH, Wen S, Liu YH, et al. Association between Polycystic Ovary Syndrome and Gut Microbiota. Plos One. 2016;11(4).

48. Rajan RK, M SS, Balaji B. Soy isoflavones exert beneficial effects on letrozole-induced rat polycystic ovary syndrome (PCOS) model through anti-androgenic mechanism. Pharm Biol. 2017;55(1):242–51. doi: 10.1080/13880209.2016.1258425 27927075

49. Karateke A, Dokuyucu R, Dogan H, Ozgur T, Tas ZA, Tutuk O, et al. Investigation of Therapeutic Effects of Erdosteine on Polycystic Ovary Syndrome in a Rat Model. Med Princ Pract. 2018;27(6):515–22. doi: 10.1159/000494300 30293079

50. Patel R, Shah G. Evaluation of ovarian and metabolic effects of GnRH modulators in two rat models of polycystic ovary syndrome. Molecular reproduction and development. 2018;85(10):778–89. doi: 10.1002/mrd.23059 30203897

Článek vyšel v časopise


2019 Číslo 9

Nejčtenější v tomto čísle

Tomuto tématu se dále věnují…


Zvyšte si kvalifikaci online z pohodlí domova

Antiseptika a prevence ve stomatologii
nový kurz
Autoři: MUDr. Ladislav Korábek, CSc., MBA

Citikolin v neuroprotekci a neuroregeneraci: od výzkumu do klinické praxe nejen očních lékařů
Autoři: MUDr. Petr Výborný, CSc., FEBO

Zánětlivá bolest zad a axiální spondylartritida – Diagnostika a referenční strategie
Autoři: MUDr. Monika Gregová, Ph.D., MUDr. Kristýna Bubová

Diagnostika a léčba deprese pro ambulantní praxi
Autoři: MUDr. Jan Hubeňák, Ph.D

Význam nemocničního alert systému v době SARS-CoV-2
Autoři: doc. MUDr. Helena Lahoda Brodská, Ph.D., prim. MUDr. Václava Adámková

Všechny kurzy
Kurzy Doporučená témata