Growth and differentiation factor – 9 (GDF-9) increases the in vitro growth rates of isolated goat early antral follicles

André Luiz da Conceição Santos; Anna Clara Ferreira; Naiza Sá; Renato Silva; Gaby Quispe-Palomino; Everton Lopes; Jesús Cadenas; Benner Alves; Juliana Celestino; Ana Paula Rodrigues; José Ricardo Figueiredo

doi:10.1590/1809-6891v24e-74980E

Authors

André Luiz da Conceição Santos Universidade Estadual do Ceará (UECE), Fortaleza, Ceará, Brasil https://orcid.org/0000-0002-5251-9598
Anna Clara Ferreira Universidade Estadual do Ceará (UECE), Fortaleza, Ceará, Brasil https://orcid.org/0000-0002-2125-9471
Naiza Sá Universidade Estadual do Ceará (UECE), Fortaleza, Ceará, Brasil https://orcid.org/0000-0003-4745-0108
Renato Silva Universidade Estadual do Ceará (UECE), Fortaleza, Ceará, Brasil https://orcid.org/0000-0003-0001-2960
Gaby Quispe-Palomino Universidade Estadual do Ceará (UECE), Fortaleza, Ceará, Brasil https://orcid.org/0000-0003-2957-0536
Everton Lopes Universidade Estadual do Ceará (UECE), Fortaleza, Ceará, Brasil https://orcid.org/0000-0002-1250-4818
Jesús Cadenas University Hospital of Copenhagen, Rigshospitalet, Copenhaguen, Denmark https://orcid.org/0000-0001-8164-3697
Benner Alves Universidade Federal de GoiásConception Biosciences, Berkeley, California, United States https://orcid.org/0000-0001-9465-8054
Juliana Celestino Universidade da Integração Internacional da Lusofonia Afro Brasileira (UNILAB), Redenção, Ceará, Brasil https://orcid.org/0000-0002-9930-7541
Ana Paula Ribeiro Rodrigues Universidade Estadual do Ceará (UECE), Fortaleza, Ceará, Brasil https://orcid.org/0000-0001-9786-1250
José Ricardo de Figueiredo Universidade Estadual do Ceará (UECE), Fortaleza, Ceará, Brasil https://orcid.org/0000-0002-5139-0964

DOI:

https://doi.org/10.1590/1809-6891v24e-74980E

Abstract

This study aimed to investigate the effect of growth and differentiation factor 9 (GDF-9) during the in vitro culture of isolated caprine early antral follicles. The isolated and selected early antral follicles were individually cultured for 18 days, and the following treatments were tested: α-MEM+ (control treatment) or α-MEM+ supplemented with 200 ng/mL GDF-9. The following endpoints were evaluated: follicular growth and morphology, estradiol production, oocyte nuclear maturation, and relative expression of key genes related to steroidogenesis (CYP19A1, CYP17, and insulin receptor) and basement membrane remodeling (MMP-9 and TIMP-2). In both treatments, a decrease was observed in the percentage of morphologically intact follicles with a concomitant increase in the rates of extruded and degenerated follicles (P < 0.05). The GDF-9 treatment showed higher rates of extruded follicles only on day 6 of culture (P < 0.05). Follicle diameter increased progressively throughout the culture period (P < 0.05) with similar diameters between treatments at all culture times (P > 0.05). Growth and differentiation factor 9 increased the daily growth rate from the first to the second third of culture, with higher values (P < 0.05) than control in the second third. Oocyte maturation rate as well as estradiol levels and relative mRNA expression for CYP19A1, CYP17, MMP-9, TIMP-2, and insulin receptor genes were similar between treatments (P > 0.05). This study shows for the first time that GDF-9 added to a culture medium increased the follicle growth rate of goat early antral follicles cultured in vitro.
Keywords: in vitro culture; antral follicle; goat; GDF-9

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References

Sá NAR, Araújo VR, Correia HHV, Ferreira ACA, Guerreiro DD, Sampaio AM, et al. Anethole improves the in vitro development of isolated caprine secondary follicles. Theriogenology. 2017;1(89):226–34. https://doi.org/10.1016/j.theriogenology.2015.12.014

Sá NAR, Ferreira ACA, Sousa FGC, Duarte ABG, Paes VM, Cadenas J, et al. First pregnancy after in vitro culture of early antral follicles in goats: Positive effects of anethole on follicle development and steroidogenesis. Mol Reprod Dev. 2020;87(9):966–77. https://doi.org/10.1002/mrd.23410

Elvin JA, Clark AT, Wang P, Wolfman NM, Matzuk MM. Paracrine actions of growth differentiation factor-9 in the mammalian ovary. Mol Endocrinol. 1999;13(6):1035–48. https://doi.org/10.1210/mend.13.6.0310

Silva JRV, Van Den Hurk R, Van Tol HTA, Roelen BAJ, Figueiredo JR. Expression of growth differentiation factor 9 (GDF9), bone morphogenetic protein 15 (BMP15), and BMP receptors in the ovaries of goats. Mol Reprod Dev. 2005;70(1):11–9. https://doi.org/10.1002/mrd.20127

Hreinsson JG, Scott JE, Rasmussen C, Swahn ML, Hsueh AJW, Hovatta O. Growth differentiation factor-9 promotes the growth, development, and survival of human ovarian follicles in organ culture. J Clin Endocrinol Metab. 2002;87(1):316–21. https://doi.org/10.1210/jcem.87.1.8185

Martins FS, Celestino JJH, Saraiva MVA, Matos MHT, Bruno JB, Rocha CMC, et al. Growth and differentiation factor-9 stimulates activation of goat primordial follicles in vitro and their progression to secondary follicles. Reprod Fertil Dev. 2008;20(8):916–24. https://doi.org/10.1071/RD08108

Cook-Andersen H, Curnow KJ, Su HI, Chang RJ, Shimasaki S. Growth and differentiation factor 9 promotes oocyte growth at the primary but not the early secondary stage in three-dimensional follicle culture. J Assist Reprod Genet. 2016;33(8):1067–77. https://doi.org/10.1007/s10815-016-0719-z

Almeida AP, Saraiva MVA, Araújo VR, Magalhães DM, Duarte ABG, Frota IMA, et al. Expression of growth and differentiation factor 9 (GDF-9) and its effect on the in vitro culture of caprine preantral ovarian follicles. Small Rumin Res. 2011;100(2–3):169–76. https://doi.org/10.1016/j.smallrumres.2011.06.001

Chaves RN, Martins FS, Saraiva MVA, Celestino JJH, Lopes CAP, Correia JC, et al. Chilling ovarian fragments during transportation improves viability and growth of goat preantral follicles cultured in vitro. Reprod Fertil Dev. 2008;20(5):640–7. https://doi.org/10.1071/RD07195

Cadenas J, Leiva-Revilla J, Vieira LA, Apolloni LB, Aguiar FLN, Alves BG, et al. Caprine ovarian follicle requirements differ between preantral and early antral stages after IVC in medium supplemented with GH and VEGF alone or in combination. Theriogenology. 2017;87:321–32. https://doi.org/10.1016/j.theriogenology.2016.09.008

Cadenas J, Maside C, Ferreira ACA, Vieira LA, Leiva-Revilla J, Paes VM, et al. Relationship between follicular dynamics and oocyte maturation during in vitro culture as a non-invasive sign of caprine oocyte meiotic competence. Theriogenology. 2018;107:95–103. https://doi.org/10.1016/j.theriogenology.2017.10.038

Jiatsa Donfack N, Alves KA, Alves BG, Pedrosa Rocha RM, Bruno JB, Lobo CH, et al. Xenotransplantation of goat ovary as an alternative to analyse follicles after vitrification. Reprod Domest Anim. 2019;54(2):216–24. https://doi.org/10.1111/rda.13340

Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods. 2001;25(4):402–8. https://doi.org/10.1006/meth.2001.1262

Ferreira ACA, Cadenas J, Sá NAR, Correia HHV, Guerreiro DD, Lobo CH, et al. In vitro culture of isolated preantral and antral follicles of goats using human recombinant FSH: Concentration-dependent and stage-specific effect. Anim Reprod Sci. 2018;196:120–9. https://doi.org/10.1016/j.anireprosci.2018.07.004

Campos LB, Silva AM, Praxedes ÉCG, Bezerra LGP, Freitas JLS, Melo LM, et al. Effect of growth differentiation factor 9 (GDF-9) on in vitro development of collared peccary preantral follicles in ovarian tissues. Anim Reprod Sci. 2021;226:106717. https://doi.org/10.1016/j.anireprosci.2021.106717

Dipaz-Berrocal DJ, Sá NAR, Guerreiro DD, Celestino JJH, Leiva-Revilla J, Alves BG, et al. Refining insulin concentrations in culture medium containing growth factors BMP15 and GDF9: An in vitro study of the effects on follicle development of goats. Anim Reprod Sci. 2017;185:118–27. https://doi.org/10.1016/j.anireprosci.2017.08.011

Chaves RN, Duarte ABG, Rodrigues GQ, Celestino JJH, Silva GM, Lopes CAP, et al. The Effects of Insulin and Follicle-Simulating Hormone (FSH) During In vitro Development of Ovarian Goat Preantral Follicles and the Relative mRNA Expression for Insulin and FSH Receptors and Cytochrome P450 Aromatase in Cultured Follicles1. Biol Reprod. 2012;87(3):1–11. https://doi.org/10.1095/biolreprod.112.099010

Ferreira ACA, Maside C, Sá NAR, Guerreiro DD, Correia HHV, Leiva-Revilla J, et al. Balance of insulin and FSH concentrations improves the in vitro development of isolated goat preantral follicles in medium containing GH. Anim Reprod Sci. 2016;165:1–10. https://doi.org/10.1016/j.anireprosci.2015.10.010

Peralta MB, Baravalle ME, Belotti EM, Stassi AF, Salvetti NR, Ortega HH, et al. Involvement of Matrix Metalloproteinases and their Inhibitors in Bovine Cystic Ovarian Disease. J Comp Pathol. 2017;156(2–3):191–201. https://doi.org/10.1016/j.jcpa.2016.10.012

Huang Q, Cheung AP, Zhang Y, Huang HF, Auersperg N, Leung PCK. Effects of growth differentiation factor 9 on cell cycle regulators and ERK42/44 in human granulosa cell proliferation. Am J Physiol Endocrinol Metab. 2009;296(6):1344-53. https://doi.org/10.1152/ajpendo.90929.2008

Farkas S, Szabó A, Hegyi AE, Török B, Fazekas CL, Ernszt D, Kovács T, Zelena D. Estradiol and Estrogen-like Alternative Therapies in Use: The Importance of the Selective and Non-Classical Actions. Biomedicines. 2022;10(4):861. https://doi.org/10.3390/biomedicines10040861