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Morphological study of apoptosis in granulosa cells and ovulation in a model of atresia in rat preovulatory follicles

Published online by Cambridge University Press:  02 October 2018

Frederico da Silva Bitecourt ,
Carina Oliveira Dumont Horta ,
Karen Santos Lima ,
Bruno Bastos Godoi ,
Fernanda Luiza Menezes Bello ,
Cíntia Maria Rodrigues ,
Luana Pereira Leite Schetino  and
Kinulpe Honorato-Sampaio Open the ORCID record for Kinulpe Honorato-Sampaio [Opens in a new window]
Frederico da Silva Bitecourt
Affiliation:
Faculdade de Medicina, Campus JK, Universidade Federal dos Vales do Jequitinhonha e Mucuri, Diamantina, Brasil
Carina Oliveira Dumont Horta
Affiliation:
Departamento de Fisiologia e Biofísica, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil
Karen Santos Lima
Affiliation:
Faculdade de Medicina, Campus JK, Universidade Federal dos Vales do Jequitinhonha e Mucuri, Diamantina, Brasil
Bruno Bastos Godoi
Affiliation:
Faculdade de Medicina, Campus JK, Universidade Federal dos Vales do Jequitinhonha e Mucuri, Diamantina, Brasil
Fernanda Luiza Menezes Bello
Affiliation:
Programa Multicêntrico de Pós-Graduação em Ciências Fisiológicas, Sociedade Brasileira de Fisiologia, São Paulo, Brasil
Cíntia Maria Rodrigues
Affiliation:
Programa Multicêntrico de Pós-Graduação em Ciências Fisiológicas, Sociedade Brasileira de Fisiologia, São Paulo, Brasil
Luana Pereira Leite Schetino
Affiliation:
Faculdade de Medicina, Campus JK, Universidade Federal dos Vales do Jequitinhonha e Mucuri, Diamantina, Brasil
Kinulpe Honorato-Sampaio*
Affiliation:
Faculdade de Medicina, Campus JK, Universidade Federal dos Vales do Jequitinhonha e Mucuri, Diamantina, Brasil Programa Multicêntrico de Pós-Graduação em Ciências Fisiológicas, Sociedade Brasileira de Fisiologia, São Paulo, Brasil
*
Author for correspondence: Kinulpe Honorato-Sampaio, Faculdade de Medicina, Universidade Federal dos Vales do Jequitinhonha e Mucuri, Campus JK, Rodovia MGT 367, Km 583, no. 5000, Alto da Jacuba, Diamantina/MG, Brazil. 39100–000. Tel: +55 31 3532 1201. E-mail: kinulpe@yahoo.com.br
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Summary

Previous studies have established a model of atresia in preovulatory follicles after stimulation of immature rats with equine chorionic gonadotropin (eCG). This gonadotropin recruits a follicular pool and the deprivation of preovulatory luteinizing hormone (LH) surge induces the atresia in preovulatory follicles. The present study investigated the occurrence of ovulation and provided some morphological features of granulosa cell (GC) apoptosis of atretic follicles at 0, 48, 72 and 120 h after eCG stimulation. Histological sections of ovaries from untreated animals (0 h) showed primordial, primary, secondary and early antral follicles. After 48 h ovaries showed large antral follicles. Preovulatory follicles were observed at 72 h, and two out of five rats displayed cumulus–oocyte complexes (COCs) in the oviducts. All animals exhibited corpora lutea after 120 h. We observed increased estradiol (E2) levels 48 h after eCG treatment that might trigger an endogenous preovulatory gonadotropin surge. Higher progesterone (P4) level, which is the hallmark of a functional corpus luteum, was observed at 120 h. Atresia in secondary and antral follicles was observed by pyknotic granulosa cell nuclei in histology and positive immunolabelling for cleaved caspase 3. We also observed macrophages in secondary and antral follicles in atresia. Transmission electron microscopy revealed GCs with compacted chromatin against the nuclear envelope, nuclear fragmentation, cell shrinkage and fragmentation. No preovulatory follicles showed apoptosis of GCs. In conclusion, our results suggested the occurrence of an endogenous gonadotropin surge, promoting ovulation and preventing atresia of preovulatory follicles.

Keywords

AtresiaGonadotropinGranulosa cell apoptosisOvulationPreovulatory follicle
Type
Short Communication
Information
Zygote , Volume 26 , Issue 4 , August 2018 , pp. 336 - 341
Copyright
© Cambridge University Press 2018 

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References

Asselin, E, Wang, Y and Tsang, BK (2001) X-linked inhibitor of apoptosis protein activates the phosphatidylinositol 3-kinase/Akt pathway in rat granulosa cells during follicular development. Endocrinology 142, 2451–2457.Google Scholar
Boone, DL, Carnegie, JA, Rippstein, PU and Tsang, BK (1997) Induction of apoptosis in equine chorionic gonadotropin (eCG)-primed rat ovaries by anti-eCG antibody. Biol Reprod 57, 420–427.Google Scholar
Braw, RH and Tsafriri, A (1980) Effect of PMSG on follicular atresia in the immature rat ovary. J Reprod Fertil 59, 267–272.Google Scholar
Caligaris, L, Astrada, JJ and Taleisnik, S (1971) Release of luteinizing hormone induced by estrogen injection into ovariectomized rats. Endocrinology 88, 810–815.Google Scholar
Choi, JY, Jo, MW, Lee, EY, Yoon, B-K and Choi, DS (2010) The role of autophagy in follicular development and atresia in rat granulosa cells. Fertil Steril 93, 2532–2537.Google Scholar
Dhanasekaran, N and Moudgal, NR (1989) Biochemical and histological validation of a model to study follicular atresia in rats. Endocrinol Exp 23, 155–166.Google Scholar
Erickson, GF and Danforth, DR (1995) Ovarian control of follicle development. Am J Obstet Gynecol 172(Pt 2), 736–747.Google Scholar
Franke, TF, Kaplan, DR and Cantley, LC (1997) PI3K: downstream AKT ion blocks apoptosis. Cell 88, 435–437.Google Scholar
Friese, K, Jarrin-Franco, MC, Lenhard, M, Mayerhofer, A and Jeschke, U (2014) The G-protein-coupled estrogen receptor (GPER/GPR30) in ovarian granulosa cell tumors. Int J Mol Sci 15, 15161–15172.Google Scholar
Gavrieli, Y, Sherman, Y and Ben-Sasson, SA (1992) Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J Cell Biol 119, 493–501.Google Scholar
Gonzaga, ACR, Campolina-Silva, GH, Werneck-Gomes, H and Oliveira, CA (2017) Profile of cell proliferation and apoptosis activated by the intrinsic and extrinsic pathways in the prostate of aging rats. Prostate 77, 937–948.Google Scholar
Grasl-Kraupp, B, Ruttkay-Nedecky, B, Koudelka, H, Bukowska, K, Bursch, W and Schulte-Hermann, R (1995) In situ detection of fragmented DNA (TUNEL assay) to discriminate among apoptosis necrosis and auto cell death: a cautionary note. Hepatology 21, 1465–1468.Google Scholar
Hillier, SG (2009) Paracrine support of ovarian stimulation. Mol Hum Reprod 15, 843–850.Google Scholar
Hu, C-L, Cowan, RG, Harman, RM and Quirk, SM (2004) Cell cycle progression and activation of Akt kinase are required for insulin-like growth factor I-mediated suppression of apoptosis in granulosa cells. Mol Endocrinol 18, 326–338.Google Scholar
Hughes, FM and Gorospe, WC (1991) Biochemical identification of apoptosis (programmed cell death) in granulosa cells: evidence for a potential mechanism underlying follicular atresia. Endocrinology 129, 2415–2422.Google Scholar
Hunzicker-Dunn, M and Mayo, K (2015) Gonadotropin signaling in the ovary In Knobil and Neill’s Physiology of Reproduction. 4th edn Elsevier pp. 895–945.Google Scholar
Hussein, MR (2005) Apoptosis in the ovary: molecular mechanisms Hum Reprod Update 11, 162–177.Google Scholar
Johnson, AL and Bridgham, JT (2002) Caspase-mediated apoptosis in the vertebrate ovary Reproduction 124, 19–27.Google Scholar
Katabuchi, H, Fukumatsu, Y, Araki, M, Suenaga, Y, Ohtake, H and Okamura, H (1996) Role of macrophages in ovarian follicular develpment. Horm Res 46, 45–51.Google Scholar
Kim, JM, Yoon, YD and Tsang, BK (1999) Involvement of the Fas/Fas ligand system in p53-mediated granulosa cell apoptosis during follicular development and atresia. Endocrinology 140, 2307–2317.Google Scholar
Krysko, DV, Diez-Fraile, A, Criel, G, Svistunov, A, Vandenabeele, P and D’Herde, K (2008) Life and death of female gametes during oogenesis and folliculogenesis. Apoptosis 13, 1065–1087.Google Scholar
Levine, JE (2015) Neuroendocrine Control of the Ovarian Cycle of the Rat Knobil and Neill’s Physiology of Reproduction. 4th edn Elsevier pp. 1199–257.Google Scholar
Luo, W and Zhu, C (2000) Expression and regulation of mRNAs for insulin-like growth factor-I receptor and LH receptor in corpora lutea. Sci China C Life Sci 43, 183–190.Google Scholar
Matsuda-Minehata, F, Inoue, N, Goto, Y and Manabe, N (2006) The regulation of ovarian granulosa cell death by pro- and anti-apoptotic molecules. J Reprod Dev 52, 695–705.Google Scholar
Matsuda, F, Inoue, N, Manabe, N and Ohkura, S (2012) Follicular growth and atresia in mammalian ovaries: regulation by survival and death of granulosa cells. J Reprod Dev 58, 44–50.Google Scholar
Mirzaei, M, Razi, M and Sadrkhanlou, R (2017) Nanosilver particles increase follicular atresia: correlation with oxidative stress and aromatization. Environ Toxicol 32, 2244–2255.Google Scholar
Pavlik, R, Wypior, G, Hecht, S, Papadopoulos, P, Kupka, M, Thaler, C, Wiest, I, Pestka, A, Friese, K and Jeschke, U (2011) Induction of G protein-coupled estrogen receptor (GPER) and nuclear steroid hormone receptors by gonadotropins in human granulosa cells. Histochem Cell Biol 136, 289–299.Google Scholar
Pangas, SA, Rajkovic, A, Vinci, D and Carpi, B (2015) Follicular development: mouse sheep and human models. In Knobil and Neill’s Physiology of Reproduction. 4th edn Elsevier pp. 947–95.Google Scholar
Pereira, VM, Honorato-Sampaio, K, Martins, AS, Reis, FM and Reis, AM (2014) Downregulation of natriuretic peptide system and increased steroidogenesis in rat polycystic ovary. Peptides 60, 80–85.Google Scholar
Quirk, SM, Cowan, RG, Harman, RM, Hu, CL, and Porter, DA (2004) Ovarian follicular growth and atresia: the relationship between cell proliferation and survival. J Anim Sci 82, E40–E52.Google Scholar
Revelli, A, Piane, LD, Casano, S, Molinari, E, Massobrio, M and Rinaudo, P (2009) Follicular fluid content and oocyte quality: from single biochemical markers to metabolomics. Reprod Biol Endocrinol 7, 40.Google Scholar
Richards, JS, Liu, Z and Shimada, M (2015) Ovulation. In Knobil and Neill’s Physiology of Reproduction 4th edn. Elsevier pp. 997–1021.Google Scholar
Sengupta, P (2014) The laboratory rat: relating its age with human’s. Int J Prev Med 4, 624–630.Google Scholar
Wang, XL, Wu, Y, Tan, L and Bin Zeng, SM (2012) Follicle-stimulating hormone regulates pro-apoptotic protein Bcl-2-interacting mediator of cell death-extra long (BimEL)-induced porcine granulosa cell apoptosis. J Biol Chem 287, 10166–10177.Google Scholar
Wu, R, Van der Hoek, KH, Ryan, NK, Norman, RJ and Robker, RL (2004) Macrophage contributions to ovarian function. Hum Reprod Update 10, 119–133.Google Scholar
Zwain, I and Amato, P (2000) Clusterin protects granulosa cells from apoptotic cell death during follicular atresia. Exp Cell Res 257, 101–110.Google Scholar