Expression of mRNAs for Pro-and Anti-Apoptotic Factors in Granulosa Cells and Follicular Fluid of Women Undergoing in Vitro Fertilization. A Pilot Study

Abstract BackgroundThis observational clinical study evaluated the expression levels and predictive values of some apoptosis-related genes in granulosa cells (GCs) and follicular fluid (FF) of women undergoing in vitro fertilization (IVF).Methods GCs and FF were obtained at oocyte retrieval from 31 consecutive patients with heterogeneous infertility diagnosis (age: 34.3±5.8 years, body mass index: 24.02±3.12 kg/m2, duration of infertility: 4.2±2.1 years). mRNA expression of pro-apoptotic (BAX, CASP3, CASP8) and anti-apoptotic (BCL2, AMH, AMHR, FSHR, LHR, CYP19A1) factors was determined by quantitative RT-PCR using ROCHE LightCycler 480. Results No significant difference in GC or FF mRNA expression of pro- and anti-apoptotic factors could be demonstrated between IVF patients with (9 patients) or without (22 patients) clinical pregnancy. Each transcript investigated was detected in FF, but their levels were markedly reduced and independent of those in GCs. The number of retrieved oocytes was positively associated with GC AMHR (r=0.393, p=0.029), but the day of embryo transfer was negatively associated with GC LHR (r=-0.414, p=0.020) and GC FSHR transcripts (r=-0.535, p=0.002). When pregnancy positive group was analysed separately the impact of apoptosis- related gene expressions on some selected measures of IVF success could be observed. Strong positive relationship was found between gene expression levels of pro- and anti-apoptotic factors in GCs.ConclusionOur study provides only marginal evidences for the apoptosis dependence of IVF outcome and suggests that the apoptosis process induces adaptive increases of the anti-apoptotic gene expression to attenuate apoptosis and to protect cell survival.

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Expression of mRNAs for pro-and anti-apoptotic factors in granulosa cells and follicular fluid of women undergoing in vitro fertilization. A pilot study

BMC Pregnancy and Childbirth ◽

10.1186/s12884-021-03834-1 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

József Bódis ◽

Endre Sulyok ◽

Ákos Várnagy ◽

Viktória Prémusz ◽

Krisztina Gödöny ◽

...

Keyword(s):

Gene Expression ◽

In Vitro Fertilization ◽

Mrna Expression ◽

Granulosa Cells ◽

Follicular Fluid ◽

Vitro Fertilization ◽

Expression Levels ◽

The Impact ◽

Apoptotic Factors

Abstract Background This observational clinical study evaluated the expression levels and predictive values of some apoptosis-related genes in granulosa cells (GCs) and follicular fluid (FF) of women undergoing in vitro fertilization (IVF). Methods GCs and FF were obtained at oocyte retrieval from 31 consecutive patients with heterogeneous infertility diagnosis (age: 34.3 ± 5.8 years, body mass index: 24.02 ± 3.12 kg/m2, duration of infertility: 4.2 ± 2.1 years). mRNA expression of pro-apoptotic (BAX, CASP3, CASP8) and anti-apoptotic (BCL2, AMH, AMHR, FSHR, LHR, CYP19A1) factors was determined by quantitative RT-PCR using ROCHE LightCycler 480. Results No significant difference in GC or FF mRNA expression of pro- and anti-apoptotic factors could be demonstrated between IVF patients with (9 patients) or without (22 patients) clinical pregnancy. Each transcript investigated was detected in FF, but their levels were markedly reduced and independent of those in GCs. The number of retrieved oocytes was positively associated with GC AMHR (r = 0.393, p = 0.029), but the day of embryo transfer was negatively associated with GC LHR (r = − 0.414, p = 0.020) and GC FSHR transcripts (r = − 0.535, p = 0.002). When pregnancy positive group was analysed separately the impact of apoptosis- related gene expressions on some selected measures of IVF success could be observed. Strong positive relationship was found between gene expression levels of pro- and anti-apoptotic factors in GCs. Conclusion Our study provides only marginal evidences for the apoptosis dependence of IVF outcome and suggests that the apoptosis process induces adaptive increases of the anti-apoptotic gene expression to attenuate apoptosis and to protect cell survival.

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CALM2 gene expression levels in the cumulus cells as a marker for chromosomal abnormalities in the embryo in in vitro fertilization programs

Akusherstvo i ginekologiia ◽

10.18565/aig.2016.10.64-72 ◽

2016 ◽

Vol 10_2016 ◽

pp. 64-72

Author(s):

Safronova N.A. Safronova ◽

Kalinina E.A. Kalinina ◽

Donnikov A.E. Donnikov ◽

Burmenskaya O.V. Burmenskaya ◽

Makarova N.P. Makarova ◽

...

Keyword(s):

Gene Expression ◽

In Vitro Fertilization ◽

Chromosomal Abnormalities ◽

Cumulus Cells ◽

Vitro Fertilization ◽

Expression Levels ◽

Gene Expression Levels

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21-Hydroxylase-Derived Steroids in Follicles of Nonobese Women Undergoing Ovarian Stimulation for In Vitro Fertilization (IVF) Positively Correlate With Lipid Content of Luteinized Granulosa Cells (LGCs) as a Source of Cholesterol for Steroid Synthesis

The Journal of Clinical Endocrinology & Metabolism ◽

10.1210/jc.2013-3204 ◽

2014 ◽

Vol 99 (4) ◽

pp. 1299-1306 ◽

Cited By ~ 12

Author(s):

Marli Amin ◽

Ariel Simerman ◽

Michele Cho ◽

Prapti Singh ◽

Christine Briton-Jones ◽

...

Keyword(s):

In Vitro Fertilization ◽

Mrna Expression ◽

Granulosa Cells ◽

Lipid Content ◽

Mineralocorticoid Receptor ◽

Steroid Synthesis ◽

Vitro Fertilization ◽

Receptor Mrna

Context: Mineralocorticoid synthesis by the nonhuman primate periovulatory follicle enhances luteinization. Whether a similar event occurs in women undergoing in vitro fertilization (IVF) is unknown. Objective: The objective of the study was to determine whether human luteinized granulosa cells (LGCs) produce mineralocorticoids derived from 21-hydroxylase activity and also express mRNA for 21-hydroxylase and the mineralocorticoid receptor. Design: This was a prospective cohort study. Setting: The study was conducted at an academic center. Patients: LGC lipid content and follicle fluid (FF) hormone analysis was performed on 27 nonobese IVF women. LGCs from six additional nonobese IVF women were used for gene expression studies. Intervention: At oocyte retrieval, FF was aspirated from the first follicle (≥16 mm in size) of each ovary and pooled LGCs were collected. Main Outcome Measures: FF steroid analysis was performed by liquid chromatography-tandem mass spectrometry. LGCs were stained with lipid fluorescent dye BODIPY FL C16 to estimate lipid content by confocal microscopy as a cholesterol source for steroidogenesis in vivo. Quantitative real-time PCR was performed using LGCs to detect 21-hydroxylase and mineralocorticoid receptor mRNA expression. Pearson correlation coefficients determined associations between FF steroid levels and LGC lipid content. Results: FF levels of the 21-hydroxylase-derived steroids, 11-deoxycorticosterone [DOC, 39.97, median (13.94–63.02) ng/mL] and 11-deoxycortisol [11DOC, 2.07 (0.69–5.01) ng/mL], along with the 21-hydroxylase precursor 17-hydroxyprogesterone [1268.21 (493.26–3558.39) ng/mL], positively correlated with LGC lipid content (84 ± 43 fluorescent units/sample) (P ≤ .05, all steroids). 21-Hydroxylase and mineralocorticoid receptor mRNA expression was detected in LGCs. Conclusions: Human LGCs likely synthesize 21-hydroxylase-derived mineralocorticoids from cholesterol-containing lipid in vivo to promote postovulatory luteinization via mineralocorticoid receptor-mediated events.

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Survivin gene expression in granulosa cells from infertile patients undergoing in vitro fertilization–embryo transfer

Fertility and Sterility ◽

10.1016/j.fertnstert.2007.02.018 ◽

2008 ◽

Vol 89 (1) ◽

pp. 60-65 ◽

Cited By ~ 15

Author(s):

Kuniko Fujino ◽

Yoshiki Yamashita ◽

Atushi Hayashi ◽

Masako Asano ◽

Shoko Morishima ◽

...

Keyword(s):

Gene Expression ◽

In Vitro Fertilization ◽

Embryo Transfer ◽

Granulosa Cells ◽

Survivin Gene ◽

Vitro Fertilization ◽

Infertile Patients

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Estradiol and progesterone production by cultured granulosa cells cryopreserved from in vitro fertilization patients

Acta Endocrinologica ◽

10.1530/eje.0.1300259 ◽

1994 ◽

Vol 130 (3) ◽

pp. 259-264 ◽

Cited By ~ 5

Author(s):

Patrick M Sluss ◽

Kan-ei Lee ◽

John H Mattox ◽

Patricia C Smith ◽

Margaret C Graham ◽

...

Keyword(s):

In Vitro Fertilization ◽

Granulosa Cells ◽

Follicle Stimulating Hormone ◽

Vitro Fertilization ◽

Cells In Culture ◽

Progesterone Production ◽

Estradiol Secretion ◽

The Impact ◽

Stimulating Hormone

Sluss PM, Lee K, Mattox JH, Smith PC, Graham MC, Partridge AB. Estradiol and progesterone production by cultured granulosa cells cryopreserved from in vitro fertilization patients. Eur J Endocrinol 1994;130:259–64. ISSN 0804–4643 Gonadotropin-stimulated steroidogenesis was studied in cultured human granulosa-lutein cells obtained from patients undergoing procedures for in vitro fertilization. The impact of cryopreservation on cell function in vitro was studied. Granulosa cells obtained from in vitro fertilization patients were cultured in serum-supplemented medium or cryopreserved at −135°C for 2–22 months. Fresh (unfrozen) cells (105) produced estradiol at a rate of 1320 pmol/l (over 72 h) and progesterone at about 2500 nmol/l. Estradiol production by either fresh or cryopreserved granulosa cells in culture was unaffected by physiological concentrations of follicle-stimulating hormone (7IU/l). Adding testosterone (10−7 mol/l) to the medium increased estradiol secretion approximately sixfold. In contrast, progesterone production was not affected by follicle-stimulating hormone or testosterone. No significant differences were observed in cultures of cryopreserved granulosa cells compared to cultures of unfrozen cells with respect to estradiol secretion, the effects of follicle-stimulating hormone or testosterone on estradiol secretion, or progesterone production. Progesterone production by fresh and cryopreserved cells was stimulated by human chorionic gonadotropin. These data indicate that cryopreservation offers the potential to facilitate prospective studies utilizing large numbers of human granulosa-lutein cells in culture. Patrick M Sluss, Reproductive Endocrine Unit, Massachusetts General Hospital East, 149 13th Street, Charlestown, MA 02129, USA

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