Different cell death types determination in juvenile mice ovarian follicles

Document Type : Full paper (Original article)

Authors

1 Graduated from School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran

2 Department of Anatomical Sciences, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran

3 Department of Anatomical Sciences, School of Medicine, Shiraz University of Medical Sciences, Histomorphometry & Stereology Research Center, Shiraz, Iran

Abstract

Follicular atresia is a phenomenon that leads to evacuation of the ovary from the oocytes and the occurrence of menopause. The contribution of various types of cell death in atresia at different follicular developmental stages requires extensive investigation. In this study, we evaluated 3 types of programmed cell death (PCD), apoptosis, autophagy, and necrosis, in juvenile mouse ovary when we can observe all follicular stages as well as atresia. Ovaries from juvenile mice on the 21st post-natal (PN) day were prepared histologically for terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) to evaluate apoptosis and immunohistochemistry for beclin-1 to evaluate the autophagy marker. Necrotic cell death was also assessed by penetration of propidium iodide (PI). The count and percentage of the labeled follicles at different stages in the ovaries were evaluated and compared using the Kruskal-Wallis and Mann-Whitney tests. We detected TUNEL-positive granulosa cells in pre-antral and antral follicles but not in the primordial and primary follicles. Somatic cells and oocytes of primordial, primary, pre-antral and antral follicles reacted to beclin-1. The percentage of the PI-labeled primordial and primary follicles were significantly higher than the beclin-1 positive (P=0.01 and P=0.01). In conclusion, we showed that apoptosis, autophagy, and necrosis play a role in follicular atresia and the contributions of each one depends on the follicular stages. It was also demonstrated that necrosis happens particularly in the small follicles while in the large one, all three cell death types occurred with an equal ratio.

Keywords

References

Amsterdam, A; Sasson, R; Keren-Tal, I; Aharoni, D; Dantes, A; Rimon, E; Land, A; Cohen, T; Dor, Y and Hirsh, L (2003). Alternative pathways of ovarian apoptosis: death for life. Biochem. Pharmacol., 66: 1355-1362.
de Kat, AC and Broekmans, FJM (2018). Female age and reproductive chances. In: Stoop, Dominic (Ed.), Preventing age related fertility loss. (1st Edn.), Switzerland, Springer International Publishing. PP: 1-10.
D’Herde, K; De Prest, B and Roels, F (1996). Subtypes of active cell death in the granulosa of ovarian atretic follicles in the quail (Coturnix coturnix japonica). Rep. Nut. Dev., 36: 175-189.
Escobar, ML; Echeverría, OM; Ortíz, R and Vázquez-Nin, GH (2008). Combined apoptosis and autophagy, the process that eliminates the oocytes of atretic follicles in immature rats. Apoptosis. 13: 1253-1266.
Gawriluk, TR; Hale, AN; Flaws, JA; Dillon, CP; Green, DR and Rucker, EB (2011). Autophagy is a cell survival program for female germ cells in the murine ovary. Reproduction. 141: 759-765.
Gaytán, M; Morales, C; Sánchez-Criado, JE and Gaytán, F (2008). Immunolocalization of beclin 1, a bcl-2-binding, autophagy-related protein, in the human ovary: possible relation to life span of corpus luteum. Cell Tissue Res., 331: 509-517.
Hart, RJ (2016). Physiological aspects of female fertility: role of the environment, modern lifestyle, and genetics. Physiol. Rev., 96: 873-909.
Hatzirodos, N; Hummitzsch, K; Irving-Rodgers, HF; Harland, ML; Morris, SE and Rodgers, RJ (2014a). Transcriptome profiling of granulosa cells from bovine ovarian follicles during atresia. BMC Genomics. 15: 40.
Hatzirodos, N; Irving-Rodgers, HF; Hummitzsch, K and Rodgers, RJ (2014b). Transcriptome profiling of the theca interna from bovine ovarian follicles during atresia. PLoS One. 9: e99706.
Jenkins, VK; Timmons, AK and McCall, K (2013). Diversity of cell death pathways: insight from the fly ovary. Trends Cell Biol., 23: 567-574.
Kutscher, LM and Shaham, S (2017). Non-apoptotic cell death in animal development. Cell Death Differ., 24: 1326-1336.
Levine, B and Yuan, J (2005). Autophagy in cell death: an innocent convict? J. Clin. Invest., 115: 2679-2688.
Liew, SH; Nguyen, QN; Strasser, A; Findlay, JK and Hutt, KJ (2017). The ovarian reserve is depleted during puberty in a hormonally driven process dependent on the pro-apoptotic protein BMF. Cell Death Dis., 8: e2971.
Manabe, N; Goto, Y; Matsuda-Minehata, F; Inoue, N; Maeda, A; Sakamaki, K and Miyano, T (2004). Regulation mechanism of selective atresia in porcine follicles: regulation of granulosa cell apoptosis during atresia. J. Repro. Dev., 50: 493-514.
Moreno-Gonzalez, G; Vandenabeele, P and Krysko, DV (2016). Necroptosis: a novel cell death modality and its potential relevance for critical care medicine. Am. J. Respir. Crit. Care Med., 194: 415-428.
Murdoch, WJ (2000). Proteolytic and cellular death mechanisms in ovulatory ovarian rupture. Biol. Signals Recept., 9: 102-114.
Omari, S; Waters, M; Naranian, T; Kim, K; Perumalsamy, AL; Chi, M; Greenblatt, E; Moley, KH; Opferman, JT and Jurisicova, A (2015). Mcl-1 is a key regulator of the ovarian reserve. Cell Death Dis., 6: e1755.
Ouyang, L; Shi, Z; Zhao, S; Wang, FT; Zhou, TT; Liu, B and Bao, JK (2012). Programmed cell death pathways in cancer: a review of apoptosis, autophagy and programmed necrosis. Cell Proliferation. 45: 487-498.
Pru, JK and Tilly, JL (2001). Programmed cell death in the ovary: insights and future prospects using genetic technologies. Mol. Endocrinol., 15: 845-853.
Saraste, A and Pulkki, K (2000). Morphologic and biochemical hallmarks of apoptosis. Cardiovasc. Res., 45: 528-537.
Simon, CR and Ramos, RGP (2006). Death by fine tunning. Braz. J. Morphol. Sci., 23: 3-13.
Sun, YC; Sun, XF; Dyce, PW; Shen, W and Chen, H (2017). The role of germ cell loss during primordial follicle assembly: a review of current advances. Int. J. Biol. Sci., 13: 449-457.
Tingen, CM; Bristol-Gould, SK; Kiesewetter, SE; Wellington, JT; Shea, L and Woodruff, TK (2009). Prepubertal primordial follicle loss in mice is not due to classical apoptotic pathways. Biol. Rep., 81: 16-25.
Tiwari, M; Prasad, S; Tripathi, A; Pandey, AN; Ali, I; Singh, AK; Shrivastav, TG and Chaube, SK (2015). Apoptosis in mammalian oocytes: a review. Apoptosis. 20: 1019-1025.
Yu, YS; Sui, HS; Han, ZB; Li, W; Luo, MJ andTan, JH (2004). Apoptosis in granulosa cells during follicular atresia: relationship with steroids and insulin-like growth factors. Cell Res., 14: 341-346.
Zucker, RM and Jeffay, SC (2006). Confocal laser scanning microscopy of whole mouse ovaries: excellent morphology, apoptosis detection, and spectroscopy. Cytometry. 69: 930-939.

Files

Share

How to cite

Statistics