International Journal of Reproductive BioMedicine

ISSN: 2476-3772

The latest discoveries in all areas of reproduction and reproductive technology.

 

Prenatal exposure to a single dose of testosterone adversely affects the oocyte and embryo quality in rats during adulthood: An experimental study

Published date: Sep 21 2025

Journal Title: International Journal of Reproductive BioMedicine

Issue title: International Journal of Reproductive BioMedicine (IJRM): Volume 23, Issue No. 8

Pages: 647 – 658

DOI: 10.18502/ijrm.v23i8.19814

Authors:

Elahe Sadeghian BakhiDepartment of Biology, School of Basic Science, Science and Research Branch, Islamic Azad University, Tehran

Nasim Hayati RoodbariDepartment of Biology, School of Basic Science, Science and Research Branch, Islamic Azad University, Tehran

Morteza AnvariResearch and Clinical Center for Infertility, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd

Fahimeh Ramezani Tehraniramezani@endocrine.ac.irReproductive Endocrinology Research Center, Research Institute for Endocrine Molecular Biology, Research Institute for Endocrine Sciences, Shahid Beheshti University of Medical Sciences, Tehran

Abstract:

Background: Prenatal exposure to excess androgen can adversely affect the hypothalamic-pituitary-ovarian axis in the developing fetus, potentially leading to long-term reproductive system dysfunction in later life.

Objective: We aimed to investigate whether prenatal exposure to a single dose of testosterone can affect the reproductive system, especially oocyte and embryo quality, and the ovarian expression of growth differentiation factor-9 (GDF-9) gene in rats during adulthood.

Materials and Methods: In this experimental study, pregnant Wistar rats (12–13 wk, 185 ± 10 gr) in the experimental group (n = 5) subcutaneously received free testosterone (5 mg) on the 20th day of pregnancy, whereas controls received only solvent (n = 5). The offspring were evaluated for oocyte and embryo quality (zona pellucida thickness, number of normal fertilization, number of 2-cell embryos, and embryo arrest rate) and ovarian expression of GDF-9 gene during adulthood.

Results: A significant increase was observed in the zona pellucida thickness (p = 0.02) in prenatally-androgenized (PNA) rats compared to controls. However, reductions in the number of normal fertilization and 2-cell embryos (p = 0.007, p = 0.01, respectively) in PNA rats compared to controls were observed. Furthermore, embryo arrest rate in PNA rats was significantly higher than in controls (p = 0.004). No significant difference was observed in the ovarian expression of GDF-9 gene in PNA rats compared to controls.

Conclusion: Prenatal exposure to a single dose of testosterone during a critical window of fetal development significantly impairs oocyte and embryo quality in adult rats. Further studies are needed to validate these findings and to elucidate the underlying molecular and physiological mechanisms.

Keywords: Androgen, Oocyte, Embryo, Growth differentiation factor-9, Rat

References:

[1] Daan NMP, Koster MPH, Steegers-Theunissen RP, Eijkemans MJC, Fauser BCJM. Endocrine and cardiometabolic cord blood characteristics of offspring born to mothers with and without polycystic ovary syndrome. Fertil Steril 2017; 107: 261–268.

[2] Andræ F, Abbott D, Stridsklev S, Schmedes AV, Odsæter IH, Vanky E, et al. Sustained maternal hyperandrogenism during PCOS pregnancy reduced by metformin in non-obese women carrying a male fetus. J Clin Endocrinol Metab 2020; 105: 3762–3770.

[3] Al-Omary HL, Alawad ZM. Cord blood sex hormones concentration: Relation to birth weight and pregnancy complications. J Contem Med Sci 2019; 5: 106–111.

[4] Barrett ES, Hoeger KM, Sathyanarayana S, Abbott DH, Redmon JB, Nguyen RHN, et al. Anogenital distance in newborn daughters of women with polycystic ovary syndrome indicates fetal testosterone exposure. J Dev Orig Health Dis 2018; 9: 307–314.

[5] Ramezani Tehrani F, Noroozzadeh M, Zahediasl S, Piryaei A, Azizi F. Introducing a rat model of prenatal androgen-induced polycystic ovary syndrome in adulthood. Exp Physiol 2014; 99: 792–801.

[6] Gurule S, Sustaita-Monroe J, Padmanabhan V, Cardoso R. Developmental programming of the neuroendocrine axis by steroid hormones: Insights from the sheep model of PCOS. Front Endocrinol (Lausanne) 2023; 14: 1096187.

[7] Chappell NR, Zhou B, Hosseinzadeh P, Schutt A, Gibbons WE, Blesson CS. Hyperandrogenemia alters mitochondrial structure and function in the oocytes of obese mouse with polycystic ovary syndrome. F S Sci 2021; 2: 101–112.

[8] Chappell NR, Zhou B, Schutt AK, Gibbons WE, Blesson CS. Prenatal androgen induced lean PCOS impairs mitochondria and mRNA profiles in oocytes. Endocr Connect 2020; 9: 261–270.

[9] Zhou Y, Zhang Q, Ding R, Fan X, Li Q, Li Z. Effects of high levels of androgens on oocyte maturation and potential regulatory role of retinoic acid. Life Sci 2025; 365: 123463.

[10] Han S, Lv J, Sun X, Xie Y, Shi Y. Transcriptome analysis of early embryonic development in a mouse model of polycystic ovary syndrome. Front Cell Dev Biol 2025; 13: 1554437.

[11] Chappell NR, Barsky M, Shah J, Peavey M, Yang L, Sangi-Haghpeykar H, et al. Embryos from polycystic ovary syndrome patients with hyperandrogenemia reach morula stage faster than controls. F S Rep 2020; 1: 125–132.

[12] Stener-Victorin E, Padmanabhan V, Walters KA, Campbell RE, Benrick A, Giacobini P, et al. Animal models to understand the etiology and pathophysiology of polycystic ovary syndrome. Endocr Rev 2020; 4: bnaa010.

[13] Barsky M, Merkison J, Hosseinzadeh P, Yang L, Bruno-Gaston J, Dunn J, et al. Fetal programming of polycystic ovary syndrome: Effects of androgen exposure on prenatal ovarian development. J Steroid Biochem Mol Biol 2021; 207: 105830.

[14] Zhang Y, Wang H, Li T, Zhang N, Chen J, Yang H, et al. Association of BMP15 and GDF9 gene polymorphisms with litter size in Hu sheep. Genes (Basel) 2025; 16: 168.

[15] Patnaik SS, Kotipalli R, Jerald MK, Muralidharan K. Combination treatment of recombinant growth differentiation factor-9 and cetrorelix improves gestational origin of the polycystic ovarian syndrome in female rats. Life Sci 2023; 321: 121638.

[16] Karagul MI, Aktaş S, Coşkun Yılmaz B, Yılmaz M, Orekici Temel G. GDF9 and BMP15 expressions and fine structure changes during folliculogenesis in polycystic ovary syndrome. Balkan Med J 2018; 35: 43–54.

[17] Ye W, Xie T, Song Y, Zhou L. The role of androgen and its related signals in PCOS. J Cell Moll Med 2021; 25: 1825–1837.

[18] Cai J, Luo X, Wang Z, Chen Z, Huang D, Cao H, et al. Comparing GDF9 in mature follicles and clinical outcomes across different PCOS phenotype. Heliyon 2024; 10: e29879.

[19] Al-Mutar H, Younis L. Effect of point mutation in the growth differentiation factor 9 gene of oocytes on the sterility and fertility of Awassi sheep. Arch Razi Inst 2020; 75: 101–108.

[20] Navid Sh, Saadatian Z, Talebi A. Assessment of developmental rate of mouse embryos yielded from in vitro fertilization of the oocyte with treatment of melatonin and vitamin C simultaneously. BMC Womens Health 2023; 23: 525.

[21] Mohammadzadeh M, Fesahat F, Khoradmehr A, Khalili MA. Influential effect of age on oocyte morphometry, fertilization rate and embryo development following IVF in mice. Middle East Fertil Soc J 2018; 23: 117–120.

[22] Petersen CG, Mauri AL, Vagnini LD, Renzi A, Petersen B, Matilla MC, et al. Randomized comparison of two commercial culture media (cook and vitrolife) for embryo culture after IMSI. JBRA Assist Reprod 2019; 23: 33–36.

[23] Lemseffer Y, Terret ME, Campillo C, Labrune E. Methods for assessing oocyte quality: A review of literature. Biomedicines 2022; 10: 2184.

[24] Sfakianoudis K, Maziotis E, Karantzali E, Kokkini G, Grigoriadis S, Pantou A, et al. Molecular drivers of developmental arrest in the human preimplantation embryo: A systematic review and critical analysis leading to mapping future research. Int J Mol Sci 2021; 22: 8353.

[25] Liu D, Chen W, Zhong Q. Multisystem health consequences of prenatal hyperandrogenism in offspring. Clin Exp Obstet Gynecol 2024; 51: 223.

[26] Wang Z, Shen M, Xue P, DiVall SA, Segars J, Wu S. Female offspring from chronic hyperandrogenemic dams exhibit delayed puberty and impaired ovarian reserve. Endocrinology 2018; 159: 1242–1252.

[27] Danfeng D, Ke D, Dengxuan F, Xuelian L, Congjian X. Oocyte quality is impaired in a hyperandrogenic PCOS mouse model by increased Foxo1 expression. Reprod Biol 2023; 23: 100812.

[28] Abruzzese GA, Silva AF, Velazquez ME, Ferrer M-J, Motta AB. Hyperandrogenism and polycystic ovary syndrome: Effects in pregnancy and offspring development. WIREs Mech Dis 2022; 14: e1558.

[29] Butikofer A, Figlio DN, Karbownik K, Kuzawa CW, Salvanes KG. Evidence that prenatal testosterone transfer from male twins reduces the fertility and socioeconomic success of their female co-twins. Proc Natl Acad Sci U S A 2019; 116: 6749–6753.

[30] Eini F, Kutenaei MA, Foroutan T, Salehi E. High levels of follicular fluid testosterone could impair oocyte developmental competency via affecting aryl hydrocarbon receptor pathway in PCOS patients. BMC Mol Cell Biol 2022; 23: 47.

[31] Liao B, Qi X, Yun C, Qiao J, Pang Y. Effects of androgen excess-related metabolic disturbances on granulosa cell function and follicular development. Front Endocrinol (Lausanne) 2022; 13: 815968.

[32] Derakhshan Z, Bahmanpour S, Nasr-Esfahani MH, Masjedi F, Mirani M, Dara M, et al. Alpha-lipoic acid ameliorates impaired steroidogenesis in human granulosa cells induced by advanced glycation end-products. Iran J Med Sci 2024; 49: 515–527.

[33] Gao Y, Zou Y, Wu G, Zheng L. Oxidative stress and mitochondrial dysfunction of granulosa cells in polycystic ovarian syndrome. Front Med (Lausanne) 2023; 10: 1193749.

[34] Voros C, Athanasiou D, Papapanagiotou I, Mavrogianni D, Varthaliti A, Bananis K, et al. Cracking the code of oocyte quality: The oxidative stress link to IVF success. Int J Mol Sci 2025; 26: 6377.

[35] Li X, Luan T, Wei Y, Zhang J, Zhao C, Ling X. The association between triglyceride glucose-body mass index and in vitro fertilization outcomes in women with polycystic ovary syndrome: A cohort study. J Ovarian Res 2024; 17: 90.

[36] Zhang Y, Liu L, Yin T-L, Yang J, Xiong C-L. Follicular metabolic changes and effects on oocyte quality in polycystic ovary syndrome patients. Oncotarget 2017; 8: 80472–80480.

[37] Lei R, Chen S, Li W. Advances in the study of the correlation between insulin resistance and infertility. Front Endocrinol (Lausanne) 2024; 15: 1288326.

[38] Sanfins A, Rodrigues P, Albertini DF. GDF-9 and BMP-15 direct the follicle symphony. J Assist Reprod Genet 2018; 35: 1741–1750.