The Effects of Ovarian Activators, Follicle Stimulating Hormone, and Activin A on Estradiol and Prostaglandin Secretion

Abstract

ABSTRACT Objective: The ovarian follicle culture technique is the in vitro development process of an ovulatory follicle with a developmentally competent oocyte, starting from the primordial follicle. The culture of the ovarian follicles aims to preserve fertility through proper isolation and storage of the ovarian cortex, especially before the cancer treatments. This study aimed to observe the effect of follicle-stimulating hormone (FSH) and/or activin A on estradiol (E2) and prostaglandin (PGE2) secretion in bovine ovarian cortical strip culture system. Methods: Ovarian cortical strips from heifers were cultured in the presence of rhFSH (50 ng/mL), rhAct A (100 ng/mL), or a combination of these factors for 6 days. The culture media were collected and stored. The levels of estradiol and prostaglandin were determined by Enzyme-Linked Immunosorbent Assay (ELISA). Results: In the group cultured in the presence of Act A (100 ng/mL), a noticeable increase was observed in both E2 and PGE2 values compared to the other groups. This increase was significant. There was also an insignificant increase in PGE2 values in the FSH group compared to the control and FSH + Act A groups. Conclusions: The results of this study reveal changes in E2 and PGE2 secretions in ovarian follicle culture medium supplemented with activin A or FSH. In particular, activin A alone was observed to be significantly more effective in these secretions.

Keywords

• Activin A
• FSH
• Estradiol
• Prostaglandin
• Follicle Culture
• Oocyte Maturation

Ovaryum Aktivatörlerinden Folikül Stimülan Hormon ve Aktivin A'nın Östradiyol ve Prostaglandin Sekresyonu Üzerindeki Etkileri

Öz

ÖZ Amaç: Ovaryan folikül kültürü tekniği, primordiyal folikülden başlayarak gelişimsel olarak yetkin bir oosit içeren ovulatuvar folikülün in vitro gelişim sürecidir. Ovaryan folikül kültürü, özellikle kanser tedavileri öncesinde ovaryan korteksin uygun şekilde izole edilmesi ve saklanması yoluyla fertiliteyi korumayı amaçlamaktadır. Bu çalışma, sığır ovaryum kortikal şerit kültür ortamına folikül stimülan hormon (FSH) ve/veya aktivin A ilavesinin östradiyol (E2) ve prostaglandin (PGE2) sekresyonuna etkisini gözlemlemeyi amaçlamıştır. Yöntem: Dişi sığırlardan alınan ovaryum kortikal şeritleri, rhFSH (50 ng/mL), rhAct A (100 ng/mL) veya bu faktörlerin bir kombinasyonu ile 6 gün boyunca kültüre edildi. Kültür ortamları toplanarak saklandı. Östradiyol ve prostaglandin seviyeleri, Enzim Bağlı İmmünosorbent Testi (ELISA) ile belirlendi. Bulgular: Act A (100 ng/mL) varlığında kültüre edilen grupta, diğer gruplara kıyasla hem E2 hem de PGE2 değerlerinde belirgin bir artış gözlenmiştir. Bu artış istatistiksel olarak anlamlıydı. FSH grubundaki PGE2 değerlerinde de kontrol ve FSH + Act A gruplarına kıyasla anlamsız bir artış gözlenmiştir. Sonuçlar: Bu çalışmanın sonuçları, aktivin A veya FSH ilave edilmiş ovaryum folikül kültür ortamında E2 ve PGE2 sekresyonunda değişiklikler olduğunu ortaya koymaktadır. Özellikle tek başına activin A’nın bu sekresyonlarda anlamlı şekilde daha etkili olduğu gözlenmiştir.

Anahtar Kelimeler

• Aktivin A
• FSH
• Folikül kültürü
• Östradiyol
• Prostaglandin
• Oosit maturasyonu

Kaynakça

1. 1 Woodruff TK. The emergence of a new interdiscipline: Oncofertility. Cancer Treat Res 2007;138:3-11.
2. 2 Oktem O, Oktay K. Quantitative assessment of the impact of chemotherapy on ovarian follicle reserve and stromal function. Cancer 2007;110:2222-9.
3. 3 Duncan FE, Jozefik JK, Kim AM, Hirshfeld-Cytron J, Woodruff TK. The gynecologist has a unique role in providing oncofertility care to young cancer patients. US Obstet Gynecol 2011;6:24-34.
4. 4 Eppig JJ, O'Brien MJ. Development in vitro of mouse oocytes from primordial follicles. Biol Reprod 1996;54:197-207.
5. 5 Wandji SA, Eppig JJ, Fortune JE. Fsh and growth factors affect the growth and endocrine function in vitro of granulosa cells of bovine preantral follicles. Theriogenology 1996;45:817-32.
6. 6 Braw-Tal R, Yossefi S. Studies in vivo and in vitro on the initiation of follicle growth in the bovine ovary. J Reprod Fertil 1997;109:165-71.
7. 7 Wandji SA, Srsen V, Nathanielsz PW, Eppig JJ, Fortune JE. Initiation of growth of baboon primordial follicles in vitro. Hum Reprod 1997;12:1993-2001.
8. 8 Albertini DF, Akkoyunlu G. Ovarian follicle culture systems for mammals. Methods Enzymol 2010;476:107-21.

9. 9 Yang MY, Cushman RA, Fortune JE. Anti-mullerian hormone inhibits activation and growth of bovine ovarian follicles in vitro and is localized to growing follicles. Mol Hum Reprod 2017;23:282-91.
10. 10 Telfer EE, McLaughlin M, Ding C, Thong KJ. A two-step serum-free culture system supports development of human oocytes from primordial follicles in the presence of activin. Hum Reprod 2008;23:1151-8.
11. 11 Katayama T, Shiota K, Takahashi M. Activin a increases the number of follicle-stimulating hormone cells in anterior pituitary cultures. Mol Cell Endocrinol 1990;69:179-185.
12. 12 Mizunuma H, Liu X, Andoh K, Abe Y, Kobayashi J, Yamada K, Yokota H, Ibuki Y, Hasegawa Y. Activin from secondary follicles causes small preantral follicles to remain dormant at the resting stage. Endocrinology 1999;140:37-42.
13. 13 Zhao J, Taverne MAM, van der Weijden GC, Bevers MM, van den Hurk R. Effect of activin a on in vitro development of rat preantral follicles and localization of activin a and activin receptor ii. Biology of Reproduction 2001;65:967-77.
14. 14 Thomas FH, Armstrong DG, Telfer EE. Activin promotes oocyte development in ovine preantral follicles in vitro. Reprod Biol Endocrinol 2003;1:76.
15. 15 Silva JR, Tharasanit T, Taverne MA, van der Weijden GC, Santos RR, Figueiredo JR, van den Hurk R. The activin-follistatin system and in vitro early follicle development in goats. J Endocrinol 2006;189:113-25.
16. 16 McGee EA, Smith R, Spears N, Nachtigal MW, Ingraham H, Hsueh AJ. Mullerian inhibitory substance induces growth of rat preantral ovarian follicles. Biol Reprod 2001;64:293-8.
17. 17 Oktay K, Newton H, Mullan J, Gosden RG. Development of human primordial follicles to antral stages in scid/hpg mice stimulated with follicle stimulating hormone. Hum Reprod 1998;13:1133-8.
18. 18 O'Shaughnessy PJ, Dudley K, Rajapaksha WR. Expression of follicle stimulating hormone-receptor mrna during gonadal development. Mol Cell Endocrinol 1996;125:169-175.
19. 19 Yogo K, Ogawa T, Akiyama M, Ishida N, Takeya T. Identification and functional analysis of novel phosphorylation sites in cx43 in rat primary granulosa cells. FEBS Lett 2002;531:132-6.
20. 20 Findlay JK. An update on the roles of inhibin, activin, and follistatin as local regulators of folliculogenesis. Biology of Reproduction 1993;48:15-23.
21. 21 Smitz J, Dolmans MM, Donnez J, Fortune JE, Hovatta O, Jewgenow K, Picton HM, Plancha C, Shea LD, Stouffer RL, Telfer EE, Woodruff TK, Zelinski MB. Current achievements and future research directions in ovarian tissue culture, in vitro follicle development and transplantation: Implications for fertility preservation. Hum Reprod Update 2010;16:395-14.
22. 22 Tepekoy F, Akkoyunlu G. The effect of fsh and activin a on akt and mapk1/3 phosphorylation in cultured bovine ovarian cortical strips. J Ovarian Res 2016;9:13.
23. 23 McLaughlin M, Telfer EE. Oocyte development in bovine primordial follicles is promoted by activin and fsh within a two-step serum-free culture system. Reproduction 2010;139:971-8.
24. 24 Hirao Y, Itoh T, Shimizu M, Iga K, Aoyagi K, Kobayashi M, Kacchi M, Hoshi H, Takenouchi N. In vitro growth and development of bovine oocyte-granulosa cell complexes on the flat substratum: Effects of high polyvinylpyrrolidone concentration in culture medium. Biol Reprod 2004;70:83-91.
25. 25 Gosden RG, Baird DT, Wade JC, Webb R. Restoration of fertility to oophorectomized sheep by ovarian autografts stored at -196 degrees c. Hum Reprod 1994;9:597-603.
26. 26 Wang BB, Guo HX, Yu S, Qiu LL, Zhang YX, Jia HT, Chen CZ, Zhang JB, Gao F, Yuan B. Activin a promotes bovine pituitary fsh synthesis and secretion through the cooperative action of the smad signaling pathway and foxo3dagger. Biol Reprod 2025;113:1549-61.
27. 27 Jensen LB, Hojer LV, Schonhage L, Adrados CS, Botkjaer JA, Borgbo T, Cadenas J, Macklon KT, Ringholm S, Pedersen AT, Kristensen SG. Inhibitory role of activin a during luteinization of follicular and lutein granulosa cells from small antral and preovulatory human follicles. Reproductive biology 2025;26:101169.
28. 28 Xu M, Fazleabas AT, Shikanov A, Jackson E, Barrett SL, Hirshfeld-Cytron J, Kiesewetter SE, Shea LD, Woodruff TK. In vitro oocyte maturation and preantral follicle culture from the luteal-phase baboon ovary produce mature oocytes. Biol Reprod 2011;84:689-97.
29. 29 Xu M, West-Farrell ER, Stouffer RL, Shea LD, Woodruff TK, Zelinski MB. Encapsulated three-dimensional culture supports development of nonhuman primate secondary follicles. Biol Reprod 2009;81:587-94.
30. 30 Xu M, Barrett SL, West-Farrell E, Kondapalli LA, Kiesewetter SE, Shea LD, Woodruff TK. In vitro grown human ovarian follicles from cancer patients support oocyte growth. Hum Reprod 2009;24:2531-40.
31. 31 Kreeger PK, Fernandes NN, Woodruff TK, Shea LD. Regulation of mouse follicle development by follicle-stimulating hormone in a three-dimensional in vitro culture system is dependent on follicle stage and dose. Biology of Reproduction 2005;73:942-50.