scholarly journals Ovulation Involves the Luteinizing Hormone-Dependent Activation of Gq/11 in Granulosa Cells

2013 ◽  
Vol 27 (9) ◽  
pp. 1483-1491 ◽  
Author(s):  
Shawn M. Breen ◽  
Nebojsa Andric ◽  
Tai Ping ◽  
Fang Xie ◽  
Stefan Offermans ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Target Genes ◽  
Inositol Phosphate ◽  
Conclusive Evidence ◽  
Corpora Lutea ◽  
Heterotrimeric G Proteins ◽  
Lh Receptor ◽  
Physiological Importance ◽  
Preovulatory Follicles ◽  
Inositol Phosphate Accumulation

The LH receptor (LHR) activates several families of heterotrimeric G proteins, but only the activation of Gs and subsequent generation of cAMP are universally accepted as important mediators of LH actions. To examine the involvement of the Gq/11 family on the actions of LH, we crossed Cyp19Cre and Gαqf/f;Gα11−/− mice to generate mice with a granulosa cell-specific deletion of Gαq in the context of a global deletion of Gα11. Granulosa cells from Gαqf/f;Gα11−/−;Cre+ mice have barely detectable levels of Gαq/11, have a normal complement of LHR, and respond to LHR activation with a transient increase in cAMP accumulation, but they fail to respond with increased inositol phosphate accumulation, an index of the activation of Gαq/11. The LHR-provoked resumption of meiosis, cumulus expansion, and luteinization are normal. However, the Gαqf/f;Gα11−/−;Cre+ mice display severe subfertility because many of the oocytes destined for ovulation become entrapped in preovulatory follicles or corpora lutea. Because follicular rupture is known to be dependent on the expression of the progesterone receptor (Pgr), we examined the LHR-induced expression of Pgr and 4 of its target genes (Adamts-1, Ctsl1, Edn2, and Prkg2). These actions of the LHR were impaired in the ovaries of the Gαqf/f;Gα11−/−;Cre+ mice. We conclude that the defect in follicular rupture is secondary to the failure of the LHR to fully induce the expression of the Pgr. This is the first conclusive evidence for the physiological importance of the activation of Gq/11 by the LHR and for the involvement of Gαq/11 in ovulation.

scholarly journals Ovarian characteristics in sheep with multiple fecundity genes

Reproduction ◽  
2017 ◽  
Vol 153 (2) ◽  
pp. 233-240 ◽  
Author(s):  
Kenneth P McNatty ◽  
Derek A Heath ◽  
Zaramasina Clark ◽  
Karen Reader ◽  
Jennifer L Juengel ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Total Mass ◽  
Total Cell ◽  
Ovulation Rate ◽  
Corpora Lutea ◽  
Preovulatory Follicles ◽  
Luteal Tissue ◽  
In The Wild ◽  
The Mean ◽  
The One

Ewes heterozygous for combinations of the Inverdale (FecXI; I+), Booroola (FecB; B+) and Woodlands (FecX2W; W+) mutations have ovulation rates higher than each mutation separately. The aims of the experiments described herein were to examine the ovarian phenotypes in I+B+ and I+B+W+ ewes and to compare these with the appropriate ++ (controls), I+ and BB animals available for this study. The mean ± s.e.m. ovulation rates in the ++ (n = 23), I+ (10), I+B+ (7), I+B+W+ (10) and BB (3) animals were 1.8 ± 0.1, 2.5 ± 0.2, 6.6 ± 1.0, 9.6 ± 0.9 and 9.7 ± 0.9 respectively. The maximum number of granulosa cells per follicle in the ++ and I+ genotypes was accumulated after exceeding 5 mm diameter, whereas in I+B+, I+B+W+ and BB animals, this was achieved when follicles reached >2–3 mm. The number of putative preovulatory follicles, as assessed from those with LH-responsive granulosa cells, 24 h after the induction of luteolysis, was higher (P < 0.01) in the I+B+ and I+B+W+ compared to the ++ and I+ genotypes. The median follicular diameters of these follicles in the ++, I+, I+B+, I+B+W+ and BB genotypes were 6, 5, 3, 3 and 3 mm respectively. The total number of granulosa cells in the putative preovulatory follicles when added together, and total mass of luteal tissue, did not differ between the genotypes. Thus, despite large ovulation rate differences between animals with one or more fecundity genes, the total cell compositions over all preovulatory follicles and corpora lutea, when added together, are similar to that from the one or two such follicles in the wild types.

scholarly journals Relationship between FoxO1 protein levels and follicular development, atresia, and luteinization in the rat ovary

2003 ◽  
Vol 179 (2) ◽  
pp. 195-203 ◽  
Author(s):  
F Shi ◽  
PS LaPolt
Keyword(s):  
Chorionic Gonadotropin ◽  
Granulosa Cells ◽  
Cell Cycle Progression ◽  
Follicular Development ◽  
Immunohistochemical Analysis ◽  
Female Rats ◽  
Corpora Lutea ◽  
Protein Levels ◽  
Physiological Processes ◽  
Preovulatory Follicles

FoxO1 is a transcription factor implicated in a growing number of physiological processes, including apoptosis, cell cycle progression, and insulin signaling. Recent findings indicate that FSH and growth factors influence ovarian functions in part through regulation of FoxO1. The present study utilized immunohistochemical analysis to determine the ovarian localization and regulation of FoxO1 protein levels in neonatal rats, immature rats during gonadotropin-induced follicular development, ovulation, and luteinization, and in spontaneously developing ovarian cysts of aging rats. In postnatal rats, FoxO1 immunoreactivity was very faint in ovaries of 5- and 10-day-old females. In contrast, strong immunoreactivity was observed in granulosa cells of larger developing follicles at 25 days of age. To stimulate follicle development, immature female rats received equine chorionic gonadotropin (eCG) followed 52 h later by an ovulatory dose of human chorionic gonadotropin (hCG). Prior to gonadotropin treatment, moderate FoxO1 immunoreactivity was observed in granulosa cells of small follicles. Subsequently, treatment with eCG markedly decreased FoxO1 protein levels in granulosa cells of healthy antral and preovulatory follicles. Interestingly, FoxO1 staining was observed in cumulus and antral, but not mural granulosa cells of preovulatory follicles. Induction of ovulation and luteinization with hCG further decreased ovarian FoxO1 levels, with no staining evident in corpora lutea. At all time points, the most intensive FoxO1 staining was observed in granulosa cells of atretic follicles, with predominantly nuclear localization. Similarly, while FoxO1 levels were low in granulosa cells of preovulatory follicles in proestrous rats, FoxO1 staining was intense in granulosa cells of spontaneously developing cystic follicles in aged, acyclic females. Together, these findings indicate that FoxO1 is expressed in a regulated, cell-specific manner during ovarian follicular development, atresia and luteinization, suggesting roles in these physiological processes.

scholarly journals Identification of Differentially Expressed Non-coding RNA Networks With Potential Immunoregulatory Roles During Salmonella Enteritidis Infection in Ducks

2021 ◽  
Vol 8 ◽  
Author(s):  
Yu Zhang ◽  
Xiaoqian Dong ◽  
Lie Hou ◽  
Zhengfeng Cao ◽  
Guoqiang Zhu ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Regulatory Networks ◽  
Egg Production ◽  
Salmonella Enteritidis ◽  
Time Course ◽  
Target Genes ◽  
Differentially Expressed ◽  
Receptor Interaction ◽  
Circular Rnas ◽  
Preovulatory Follicles

Salmonella enterica serovar Enteritidis (S. Enteritidis) is a pathogen that can colonize the preovulatory follicles of poultry, thereby causing both reduced egg production and an elevated risk of foodborne salmonellosis in humans. Although a few studies have revealed S. Enteritidis preferentially invades the granulosa cell layer within these follicles, it can readily persist and proliferate through mechanisms that are not well-understood. In this study, we characterized competing endogenous RNA (ceRNA) regulatory networks within duck granulosa cells following time-course of S. Enteritidis challenge. The 8108 long non-coding RNAs (lncRNAs), 1545 circular RNAs (circRNAs), 542 microRNAs (miRNAs), and 4137 mRNAs (fold change ≥2; P &lt; 0.01) were differentially expressed during S. Enteritidis challenge. Also, eight mRNAs, eight lncRNAs and five circRNAs were selected and the consistent expression trend was found between qRT-PCR detection and RNA-seq. Moreover, the target genes of these differentially expressed ncRNAs (including lncRNAs, circRNAs and miRNAs) were predicted, and significantly enriched in the innate immune response and steroidogenesis pathways. Then, the colocalization and coexpression analyses were conducted to investigate relationships between ncRNAs and mRNAs. The 16 differentially expressed miRNAs targeting 60 differentially expressed mRNAs were identified in granulosa cells at 3 and 6 h post-infection (hpi) and enriched in the MAPK, GnRH, cytokine-cytokine receptor interaction, Toll-like receptor, endocytosis, and oxidative phosphorylation signaling pathways. Additionally, underlying lncRNA-miRNA-mRNA and circRNA-miRNA-mRNA ceRNA networks were then constructed to further understand their interaction during S. Enteritidis infection. Lnc_012227 and novel_circ_0004892 were identified as ceRNAs, which could compete with miR-let-7g-5p and thereby indirectly modulating map3k8 expression to control S. Enteritidis infection. Together, our data thus identified promising candidate ncRNAs responsible for regulating S. Enteritidis infection in the preovulatory follicles of ducks, offering new insights regarding the ovarian transmission of this pathogen.

435. Follicle differentiation and luteinisation in the mouse is associated with hypoxia inducible factor activity

2008 ◽  
Vol 20 (9) ◽  
pp. 115
Author(s):  
K. Tam ◽  
D. Russell ◽  
K. Kind ◽  
J. Thompson
Keyword(s):  
Granulosa Cells ◽  
Target Genes ◽  
Ovarian Function ◽  
Egfp Expression ◽  
Corpora Lutea ◽  
Limited Information ◽  
Antral Follicles ◽  
Follicle Differentiation

Hypoxia inducible factors (HIF) are transcription factors that mediate the response to hypoxic stress. Under hypoxic conditions, HIF is stabilised, translocates to the nucleus, and binds to the Hypoxia Response Elements (HRE) upstream of numerous target genes involved in angiogenesis and glycolysis, including Vegf, Glut-1 and Ldha. Little is known about the role of HIFs in regulating ovarian function. In rat granulosa cells, FSH stimulates HIF 1α via the PI3K/Akt pathway, demonstrating a role for HIFs during follicular development. In contrast, there is limited information regarding the role of HIFs during corpus luteum formation. In this study we investigated whether HIFs play a role in follicle differentiation and luteinisation. Prepubertal C57Bl6 females were stimulated with eCG (5 IU) followed 46 h later by hCG (5 IU). Mice were sacrificed at 0, 4, 8, 12, 16 and 24 h post hCG and granulosa cells were collected for Western analysis of HIF-1a protein. To investigate HIF activation in the ovary, a transgenic reporter mouse line was developed by lenti-viral incorporation of an HRE (4)-SV40-eGFP construct. Ovaries were collected from mice plugged day 1, 4 and 8 for CL analysis in vivo.A time- dependent increase of HIF 1α protein levels in granulosa cells, maximal around time of ovulation, was observed. Ovaries from cycling HRE-eGFP transgenic mice exhibited no eGFP in primordial, primary or preantral follicles. Upon antrum formation, eGFP was evident in occasional sections in antral follicles but HIF signalling was restricted within the theca. In contrast, corpora lutea on pregnancy day 1, 4 and 8 readily expressed eGFP and eGFP expression increases as luteinisation progresses.These results demonstrate that in vivo HIFs may play a role in folliculogenesis, but this is restricted to theca cells of antral follicles before hCG stimulation. Following hCG, maximal HIF activity is associated with the time of ovulation. In addition, HIF activity is maintained during luteinisation.

scholarly journals Expression of 11beta-hydroxysteroid dehydrogenases in bovine follicle and corpus luteum

2003 ◽  
Vol 177 (3) ◽  
pp. 445-452 ◽  
Author(s):  
M Tetsuka ◽  
S Yamamoto ◽  
N Hayashida ◽  
KG Hayashi ◽  
M Hayashi ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Follicular Fluid ◽  
Physiological Role ◽  
Stage Iv ◽  
Stage I ◽  
Stage Iii ◽  
Corpora Lutea ◽  
Hydroxysteroid Dehydrogenases ◽  
Target Organs ◽  
Preovulatory Follicles

In glucocorticoid target organs, local concentrations of active glucocorticoid are determined by the relative expression of two 11beta-hydroxysteroid dehydrogenases (HSDs): bi-directional 11beta-HSD type1 (11HSD1) that mainly activates cortisone to cortisol, and dehydrogenase 11beta-HSD type2 (11HSD2) that inactivates cortisol to cortisone. In this study, we examined the expression of mRNA encoding these two 11beta-HSDs in bovine granulosa cells harvested from preovulatory follicles and corpora lutea (CL). Ovaries were obtained from Holstein cows at a local slaughterhouse. Follicles larger than 10 mm in diameter and CL were dissected and follicular fluid and granulosa cells were taken. Corpora lutea were weighed and their stages were morphologically assessed (stage I, days 1-4; stage II, days 5-10; stage III, days 11-17; stage IV, days 8-20). Follicles were classified into four groups according to their hormonal status (oestradiol (E(2)): progesterone (P(4))>1: oestrogen active; E(2):P(4)<1: oestrogen inactive) and stage of the oestrous cycle (luteal or follicular phase). Total RNA was extracted with phenol-chloroform and subjected to a semi-quantitative RT-PCR for 11HSD1, 11HSD2 and beta-actin. Concentrations of steroids in follicular fluid were determined by an enzyme immunoassay. In granulosa cells, only 11HSD1 mRNA was detected. There was a negative correlation between the expression of 11HSD1 and the concentration of cortisol in follicular fluid (P<0.05), indicating 11HSD1 may act as a dehydrogenase in the bovine follicle. Both types of 11beta-HSDs were expressed in CL. The levels of mRNA for both isozymes were high in stage I and II, and were decreased in stage III CL. In stage IV CL, the expression of 11HSD2 but not 11HSD1 mRNA increased. These results indicate that the bovine granulosa cells and CL express 11HSD1 and 11HSD2, and they may play an important physiological role in the bovine ovary through modulating the local glucocorticoid environment.

scholarly journals Cellular heterogeneity of the LH receptor and its significance for cyclic GMP signaling in mouse preovulatory follicles

2020 ◽  
Author(s):  
Valentina Baena ◽  
Corie M. Owen ◽  
Tracy F. Uliasz ◽  
Katie M. Lowther ◽  
Siu-Pok Yee ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Cyclic Gmp ◽  
Cellular Localization ◽  
Cumulus Cells ◽  
Luteinizing Hormone Receptor ◽  
Cellular Heterogeneity ◽  
Signaling Proteins ◽  
Meiotic Arrest ◽  
Lh Receptor ◽  
Preovulatory Follicles

AbstractMeiotic arrest and resumption in mammalian oocytes are regulated by two opposing signaling proteins in the cells of the surrounding follicle: the guanylyl cyclase NPR2, and the luteinizing hormone receptor (LHR). NPR2 maintains a meiosis-inhibitory level of cyclic GMP (cGMP) until LHR signaling causes dephosphorylation of NPR2, reducing NPR2 activity, lowering cGMP to a level that releases meiotic arrest. However, the signaling pathway between LHR activation and NPR2 dephosphorylation remains incompletely understood, due in part to imprecise information about the cellular localization of these two proteins. To investigate their localization, we generated mouse lines in which HA epitope tags were added to the endogenous LHR and NPR2 proteins, and used immunofluorescence and immunogold microscopy to localize these proteins with high resolution. The results showed that the LHR protein is absent from the cumulus cells and inner mural granulosa cells, and is present in only 13-48% of the outer mural granulosa cells. In contrast, NPR2 is present throughout the follicle, and is more concentrated in the cumulus cells. Less than 20% of the NPR2 is in the same cells that express the LHR. These results suggest that to account for the LH-induced inactivation of NPR2, LHR-expressing cells send a signal that inactivates NPR2 in neighboring cells that do not express the LHR. An inhibitor of gap junction permeability attenuates the LH-induced cGMP decrease in the outer mural granulosa cells, consistent with this mechanism contributing to how NPR2 is inactivated in cells that do not express the LHR.

scholarly journals Expression and Localization of Serum/Glucocorticoid-Induced Kinase in the Rat Ovary: Relation to Follicular Growth and Differentiation1

Endocrinology ◽  
2000 ◽  
Vol 141 (1) ◽  
pp. 385-395 ◽  
Author(s):  
Tamara N. Alliston ◽  
Ignacio J. Gonzalez-Robayna ◽  
Patricia Buse ◽  
Gary L. Firestone ◽  
JoAnne S. Richards
Keyword(s):  
In Situ Hybridization ◽  
Subcellular Localization ◽  
Granulosa Cells ◽  
Corpora Lutea ◽  
Pregnant Rats ◽  
Western Blots ◽  
Stromal Compartment ◽  
Luteal Cells ◽  
Preovulatory Follicles

Abstract Expression of serum/glucocorticoid-inducible kinase (Sgk), one member of an inducible serine/threonine kinase family, is induced by FSH/cAMP in rat granulosa cells cultured in defined medium. The FSH-stimulated pattern of sgk expression is biphasic, and transcriptional activation of the sgk gene depends on an intact Sp1/Sp3 binding site within the proximal promoter. To determine whether sgk was expressed in a hormone-dependent and physiologically relevant manner in vivo, the cellular levels of sgk messenger RNA (mRNA) and protein as well as the subcellular localization of this kinase were analyzed in ovaries containing follicles and corpora lutea at specific stages of differentiation. To stimulate follicular development and luteinization, hypophysectomized (H) rats were treated with estradiol (E; HE) and FSH (FSH; HEF) followed by hCG (hCG; HEF/hCG). To analyze Sgk in functional corpora lutea, PRL was administered to HEF/hCG rats, or ovaries of pregnant rats were obtained on day 7, 15, or 22 of gestation. In situ hybridization indicated that sgk mRNA was low/undetectable in granulosa cells of H and HE rats. An acute injection (iv) of FSH to HE rats rapidly increased sgk mRNA at 2 and 8 h. Sgk mRNA was also elevated in granulosa cells of preovulatory follicles of HEF rats and in luteal cells of HEF/hCG and pregnant rats. Northern blots and Western blots confirmed the in situ hybridization data, indicating that the amount and cellular localization Sgk protein were related to that of sgk mRNA. When the subcellular localization of this kinase was analyzed by immunohistochemistry, Sgk protein was nuclear in granulosa cells and some thecal cells of large preovulatory follicles. In contrast, Sgk protein was cytoplasmic in luteal cells as well as some cells within the stromal compartment. Intense immunostaining was also observed in oocytes present in primordial follicles, but not in growing follicles. Collectively, these results show that FSH and LH stimulate marked increases in the cellular content of Sgk, as well as dramatic changes in the subcellular distribution of this kinase. The specific nuclear vs. cytoplasmic compartmentalization of Sgk in granulosa cells and luteal cells, respectively, indicates that Sgk controls distinct functions in proliferative vs. terminally differentiated granulosa cells.

Parathyroid Ca2+-conducting currents are modulated by muscarinic receptor agonists and antagonists

1997 ◽  
Vol 273 (5) ◽  
pp. E880-E890 ◽  
Author(s):  
Wenhan Chang ◽  
Tsui-Hua Chen ◽  
Stacy A. Pratt ◽  
Benedict Yen ◽  
Michael Fu ◽  
...  
Keyword(s):  
Muscarinic Receptors ◽  
Inositol Phosphate ◽  
Receptor Protein ◽  
Dependent Manner ◽  
Rt Pcr ◽  
Pipette Solution ◽  
Pcr Products ◽  
Inositol Phosphate Accumulation ◽  
Receptor Cdna ◽  
Dose Dependent

Parathyroid cells express Ca2+-conducting cation currents, which are activated by raising the extracellular Ca2+ concentration ([Ca2+]o) and blocked by dihydropyridines. We found that acetylcholine (ACh) inhibited these currents in a reversible, dose-dependent manner (50% inhibitory concentration ≈10−8 M). The inhibitory effects could be mimicked by the agonist (+)-muscarine. The effects of ACh were blunted by the antagonist atropine and reversed by removing ATP from the pipette solution. (+)-Muscarine enhanced the adenosine 3′,5′-cyclic monophosphate (cAMP) production by 30% but had no effect on inositol phosphate accumulation in parathyroid cells. Oligonucleotide primers, based on sequences of known muscarinic receptors (M1-M5), were used in reverse transcriptase-polymerase chain reaction (RT-PCR) to amplify receptor cDNA from parathyroid poly (A)+ RNA. RT-PCR products displayed >90% nucleotide sequence identity to human M2- and M4-receptor cDNAs. Expression of M2-receptor protein was further confirmed by immunoblotting and immunocytochemistry. Thus parathyroid cells express muscarinic receptors of M2 and possibly M4 subtypes. These receptors may couple to dihydropyridine-sensitive, cation-selective currents through the activation of adenylate cyclase and ATP-dependent pathways in these cells.

scholarly journals The Differential Metabolomes in Cumulus and Mural Granulosa Cells from Human Preovulatory Follicles

2021 ◽  
Author(s):  
Er-Meng Gao ◽  
Bongkoch Turathum ◽  
Ling Wang ◽  
Di Zhang ◽  
Yu-Bing Liu ◽  
...  
Keyword(s):  
Oocyte Maturation ◽  
Granulosa Cells ◽  
Quantitative Polymerase Chain Reaction ◽  
Cumulus Cells ◽  
Cholesterol Transport ◽  
Vitro Fertilization ◽  
Expression Of Genes ◽  
Preovulatory Follicles ◽  
Morphological Differences

AbstractThis study evaluated the differences in metabolites between cumulus cells (CCs) and mural granulosa cells (MGCs) from human preovulatory follicles to understand the mechanism of oocyte maturation involving CCs and MGCs. CCs and MGCs were collected from women who were undergoing in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) treatment. The differences in morphology were determined by immunofluorescence. The metabolomics of CCs and MGCs was measured by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) followed by quantitative polymerase chain reaction (qPCR) and western blot analysis to further confirm the genes and proteins involved in oocyte maturation. CCs and MGCs were cultured for 48 h in vitro, and the medium was collected for detection of hormone levels. There were minor morphological differences between CCs and MGCs. LC-MS/MS analysis showed that there were differences in 101 metabolites between CCs and MGCs: 7 metabolites were upregulated in CCs, and 94 metabolites were upregulated in MGCs. The metabolites related to cholesterol transport and estradiol production were enriched in CCs, while metabolites related to antiapoptosis were enriched in MGCs. The expression of genes and proteins involved in cholesterol transport (ABCA1, LDLR, and SCARB1) and estradiol production (SULT2B1 and CYP19A1) was significantly higher in CCs, and the expression of genes and proteins involved in antiapoptosis (CRLS1, LPCAT3, and PLA2G4A) was significantly higher in MGCs. The level of estrogen in CCs was significantly higher than that in MGCs, while the progesterone level showed no significant differences. There are differences between the metabolomes of CCs and MGCs. These differences may be involved in the regulation of oocyte maturation.

scholarly journals Effects of Silver Nanoparticles on Proliferation and Apoptosis in Granulosa Cells of Chicken Preovulatory Follicles: An In Vitro Study

Animals ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 1652
Author(s):  
Dorota Katarzyńska-Banasik ◽  
Anna Kozubek ◽  
Małgorzata Grzesiak ◽  
Andrzej Sechman
Keyword(s):  
Silver Nanoparticles ◽  
Granulosa Cells ◽  
Caspase 3 ◽  
In Vitro Study ◽  
Poultry Production ◽  
Cell Death Pathways ◽  
Preovulatory Follicles ◽  
Proliferation And Apoptosis ◽  
Apoptosis Process

The continuous development of poultry production related to the growing demand for eggs and chicken meat makes it necessary to use modern technologies. An answer to this demand may be the use of nanotechnology in poultry farming. One of the promising nanomaterials in this field are silver nanoparticles (AgNPs), which are used as disinfectants, reducing microbial pollution and the amounts of greenhouse gases released. This study aimed to evaluate the effect of AgNPs on the proliferation and apoptosis process in the granulosa cells of chicken preovulatory follicles. The in vitro culture experiment revealed that both 13 nm and 50 nm AgNPs inhibited the proliferation of the granulosa cells. However, a faster action was observed in 50 nm AgNPs than in 13 nm ones. A size-dependent effect of AgNP was also demonstrated for the caspase-3 activity. AgNPs 13 nm in size increased the caspase-3 activity in granulosa cells, while 50 nm AgNPs did not exert an effect, which may indicate the induction of distinct cell death pathways by AgNPs. In conclusion, our study reveals that AgNPs in vitro inhibit granulosa cell proliferation and stimulate their apoptosis. These results suggest that AgNPs may disrupt the final stage of preovulatory follicle maturation and ovulation.

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