Hormonal regulation of apoptosis in rabbit granulosa cells in vitro: evaluation by flow cytometric detection of plasma membrane phosphatidylserine externalization

Maillet, Géraldine; Breard, E; Benhaïm, Annie; Leymarie, P.; Féral, Colette

doi:10.1530/rep.0.1230243

Cited by 17 publications

(12 citation statements)

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“…Moreover, the proportion of apoptotic granulosa cells increased with the time of culture, which is in agreement with the findings of Maillet et al [45]. Lin and Rui [37] showed that the rate of cell apoptosis in porcine small follicles cultured for 24 hours was higher than that of cells isolated from large and medium ones, suggesting that follicular apoptosis may be regulated in a stage-specific manner.…”

Section: Discussionsupporting

confidence: 91%

“…In the porcine ovary, there are many follicles of different stages and sizes because of the lack of follicular waves, unlike domestic ruminants [45]. Apoptosis is induced in granulosa cells located in the inner surface of the granulosa layer but not in the cumulus cells and oocytes in the early stages of atresia [44], which was confirmed by data collected in our study.…”

Section: Discussionsupporting

confidence: 88%

See 1 more Smart Citation

Does 2-hydroxyflutamide Inhibit Apoptosis in Porcine Granulosa Cells? — An <i>In Vitro</i> Study

Duda

Durlej

Knet

et al. 2012

Journal of Reproduction and Development

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Abstract. In mammalian ovaries, the majority of follicles are lost before ovulation by atresia. This degenerative process is initiated or caused by granulosa cell apoptosis. To reveal the androgen-dependent mechanism of selective follicular atresia, the culture model system for agonism and antagonism of the androgen receptor has been established. We examined the influence of an androgen receptor antagonist, 2-hydroxyflutamide (2-Hf), on the incidence of apoptosis in cultured porcine granulosa cells. They were incubated (6 and 12-h) in the presence of testosterone (T, 10 -7 M), 2-Hf (1.7×10 -4 M) or both T and 2-Hf (T+2-Hf), and then analyzed by flow cytometry with fluorescein labelled annexin V. To better imitate in vivo conditions, the intact porcine follicles (6-8 mm in diameter) have been incubated in an organ culture system with the addition of the same factors. Sections obtained from follicles fixed after culture were stained with hematoxylin and eosin, and the presence of apoptosis-related DNA strand breaks was evaluated by the TUNEL method. Estradiol and progesterone concentrations in the culture media were measured by radioimmunoassays. The addition of T or 2-Hf to the culture media caused an increase in the number of apoptotic granulosa cells, while treatment with T+2-Hf decreased it in both in vitro and organotypic models. Follicles cultured with the addition of T or 2-Hf exhibited morphological changes indicating follicular atresia. Granulosal estradiol secretion was considerably stimulated by T+2-Hf. The highest increase in follicular estradiol secretion was observed after the anti-androgen addition. In both granulosal and follicular cultures, the production of progesterone declined in the presence of T or 2-Hf but increased after their simultaneous addition. In conclusion, androgen receptor antagonist 2-Hf attenuates induction of granulosa cell apoptosis in the presence of a high T level. The nature of this protective mechanism as yet is unknown and requires further research. Key words: Apoptosis, Granulosa cells, 2-hydroxyflutamide, Ovarian follicles, Pig (J. Reprod. Dev. 58: [438][439][440][441][442][443][444] 2012) T he ovary plays two distinct functions in female reproduction: it produces germ cells (oocytes) and provides a proper hormonal environment for ovarian follicles undergoing cycles of growth and development induced by gonadotropins [1][2][3]. Although gonadotropins are predominant regulators of follicular development, it is now clear that there is a wide range of additional regulators of follicular maturation produced within the ovarian tissue such as steroids and peptide hormones, as well as ovarian growth factors. They act by paracrine mechanisms to modulate, either to amplify or attenuate, the functions of gonadotropins [4]. Amongst the several hundred thousand primordial follicles present in the mammalian ovary, only 1% of them develop to the preovulatory stage and finally ovulate [5]. The remaining ones will be eliminated via a degenerative process called atresia [6,7]. Atresia ...

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Section: Discussionsupporting

confidence: 91%

Section: Discussionsupporting

confidence: 88%

Does 2-hydroxyflutamide Inhibit Apoptosis in Porcine Granulosa Cells? — An <i>In Vitro</i> Study

Duda

Durlej

Knet

et al. 2012

Journal of Reproduction and Development

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“…In rabbits, FSH has no effect on apoptosis of isolated granulosa cells cultured in vitro (Maillet et al, 2002), whereas in rats, pigs, and cows, FSH attenuates apoptosis in cultured isolated granulosa cells in vitro (Zhang et al, 2011).…”

Section: Effect Of Fsh In Rescuing Granulosa Cells From Apoptosis In mentioning

confidence: 91%

Rescue from dominant follicle atresia by follicle-stimulating hormone in mice

Zhou¹,

Teng²,

Cao³

et al. 2013

Genet. Mol. Res.

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ABSTRACT.We investigated the effects of follicle-stimulating hormone (FSH) on atresia of the dominant follicle and changes in relevant apoptosis genes in granulosa cells of dominant follicles regulated by FSH in vivo. Four-week-old mice were administered FSH by intraperitoneal injection to induce follicular maturation. Granulosa cells of dominant follicles were collected at 48, 72, and 96 h after the first FSH injection. Phosphate-buffered saline was injected as a control. The mRNA levels of relevant granulosa cell apoptosis genes were examined by real-time quantitative polymerase chain reaction, and apoptosis of granulosa cells in dominant ovarian follicles was determined by the terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assay. Apoptosis in granulosa cells of dominant follicles was almost TUNEL-negative at 48, 72-66, 72, and 96-90 h after the first FSH injection, but granulosa cell apoptosis in dominant follicles was clearly detected at 96, 102, and 102-96 h by TUNEL. The BIM, caspase-3, and caspase-9 mRNA expression levels were significantly lower after FSH treatment at 72-66 and 96-90 h, compared with that at 72 and 96 h (P < 0.05). Caspase-8 and FasL mRNA expressions did not respond to FSH. FSH rescued granulosa cells from apoptosis when the relevant apoptosis genes were upregulated in early atretic follicles. FSH did not rescue granulosa cells from apoptosis if the DNA was cut into fragments by endonucleases. Thus, the rescue by FSH of granulosa cells from apoptosis and dominant follicle atresia may be accomplished by inhibition of apoptosis in mitochondria.

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“…The reproductive performance of any mammalian species can be enhanced by enriching nutrition, environmental factors like photoperiod, temperature, humidity, etc., through selective breeding, endocrine manipulations, and better management practices [1][2][3][4][5][6]. The development and growth of the ovarian follicles is finely regulated by a complex interaction of extracellular and intracellular factors, any deviation thereof leads to follicular atresia [6][7][8].…”

Section: Introductionmentioning

confidence: 99%

Changes in Trace Elements during Follicular Atresia in Goat (Capra hircus) Ovary

Bhardwaj

Sharma

2010

Biol Trace Elem Res

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Quantitative analysis of follicular fluid and granulosa cells from small, medium and large antral atretic follicles of goat (Capra hircus) ovaries was conducted to study the alterations in trace elements viz zinc (Zn), copper (Cu), manganese (Mn), and iron (Fe). The zinc content was lower in the follicular fluid (0.993 ± 0.001, 0.935 ± 0.002, 1.321 ± 0.001 μg/ml) and granulosa cells (0.867 ± 0.002, 0.801 ± 0.001, 1.073 ± 0.002 μg/mg) of small, medium, and large antral atretic follicles respectively than their respective controls. Copper quantity was higher in the follicular fluid (0.113 ± 0.001, [Formula: see text], 0.224 ± 0.001 μg/ml) and granulosa cells (0.094 ± 0.001, 0.114 ± 0.001, 0.182 ± 0.001 μg/mg) from small, medium, and large antral atretic follicles respectively than their respective controls. Similarly, iron and manganese was also found higher in the follicular fluid and granulosa cells of small, medium, and large antral atretic follicles than their respective controls. The present study provides the basic data on trace elements that can be safely used as atretic marker and will find use in in vitro studies for fertility improvement plan. Thus, help in elevating the number of ovulations and screening of follicles to enhance the success rate in vivo and in vitro fertilization and embryo transfer technology.

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Hormonal regulation of apoptosis in rabbit granulosa cells in vitro: evaluation by flow cytometric detection of plasma membrane phosphatidylserine externalization

Cited by 17 publications

References 18 publications

Does 2-hydroxyflutamide Inhibit Apoptosis in Porcine Granulosa Cells? — An <i>In Vitro</i> Study

Does 2-hydroxyflutamide Inhibit Apoptosis in Porcine Granulosa Cells? — An <i>In Vitro</i> Study

Rescue from dominant follicle atresia by follicle-stimulating hormone in mice

Changes in Trace Elements during Follicular Atresia in Goat (Capra hircus) Ovary

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