The Role of Cholesterol Esterification in Ovarian Steroidogenesis: Studies in Cultured Swine Granulosa Cells Using a Novel Inhibitor of Acyl Coenzyme A:Cholesterol Acyltransferase*

The purpose of this study is to evaluate the amphetamine effects on progesterone and estradiol production in rat granulosa cells and the underlying cellular regulatory mechanisms. Freshly dispersed rat granulosa cells were cultured with various test drugs in the presence of amphetamine, and the estradiol/progesterone production and the cytosolic cAMP level were measured. Additionally, the cytosolic-free Ca2+ concentrations ([Ca2+]i) were measured to examine the role of Ca2+ influx in the presence of amphetamine. Amphetamine in vitro inhibited both basal and porcine follicle-stimulating hormone-stimulated estradiol/progesterone release, and amphetamine significantly decreased steroidogenic enzyme activities. Adding 8-Bromo-cAMP did not recover the inhibitory effects of amphetamine on progesterone and estradiol release. H89 significantly decreased progesterone and estradiol basal release but failed to enhance a further amphetamine inhibitory effect. Amphetamine was capable of further suppressing the release of estradiol release under the presence of nifedipine. Pretreatment with the amphetamine for 2 h decreased the basal [Ca2+]i and prostaglandin F2α-stimulated increase of [Ca2+]i. Amphetamine inhibits progesterone and estradiol secretion in rat granulosa cells through a mechanism involving decreased PKA-downstream steroidogenic enzyme activity and L-type Ca2+ channels. Our current findings show that it is necessary to study the possibility of amphetamine perturbing reproduction in females.

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Sirtuin 1 and Sirtuin 3 in Granulosa Cell Tumors

International Journal of Molecular Sciences ◽

10.3390/ijms22042047 ◽

2021 ◽

Vol 22 (4) ◽

pp. 2047

Author(s):

Nina Schmid ◽

Kim-Gwendolyn Dietrich ◽

Ignasi Forne ◽

Alexander Burges ◽

Magdalena Szymanska ◽

...

Keyword(s):

Cell Proliferation ◽

Granulosa Cell ◽

Granulosa Cells ◽

Drug Targets ◽

Growth Regulation ◽

Sirtuin 1 ◽

Ki 67 ◽

Granulosa Cell Tumors ◽

Novel Drug

Sirtuins (SIRTs) are NAD+-dependent deacetylases that regulate proliferation and cell death. In the human ovary, granulosa cells express sirtuin 1 (SIRT1), which has also been detected in human tumors derived from granulosa cells, i.e., granulosa cell tumors (GCTs), and in KGN cells. KGN cells are an established cellular model for the majority of GCTs and were used to explore the role of SIRT1. The SIRT1 activator SRT2104 increased cell proliferation. By contrast, the inhibitor EX527 reduced cell numbers, without inducing apoptosis. These results were supported by the outcome of siRNA-mediated silencing studies. A tissue microarray containing 92 GCTs revealed nuclear and/or cytoplasmic SIRT1 staining in the majority of the samples, and also, SIRT2-7 were detected in most samples. The expression of SIRT1–7 was not correlated with the survival of the patients; however, SIRT3 and SIRT7 expression was significantly correlated with the proliferation marker Ki-67, implying roles in tumor cell proliferation. SIRT3 was identified by a proteomic analysis as the most abundant SIRT in KGN. The results of the siRNA-silencing experiments indicate involvement of SIRT3 in proliferation. Thus, several SIRTs are expressed by GCTs, and SIRT1 and SIRT3 are involved in the growth regulation of KGN. If transferable to GCTs, these SIRTs may represent novel drug targets.

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Role of the plasma membrane in cholesterol esterification in rat hepatoma cells

Journal of Biological Chemistry ◽

10.1016/s0021-9258(19)85179-7 ◽

1993 ◽

Vol 268 (19) ◽

pp. 13838-13843 ◽

Cited By ~ 5

Author(s):

Y. Lange ◽

F. Strebel ◽

T.L. Steck

Keyword(s):

Plasma Membrane ◽

Hepatoma Cells ◽

Cholesterol Esterification ◽

Rat Hepatoma ◽

Rat Hepatoma Cells

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MicroRNA Mediating Networks in Granulosa Cells Associated with Ovarian Follicular Development

BioMed Research International ◽

10.1155/2017/4585213 ◽

2017 ◽

Vol 2017 ◽

pp. 1-18 ◽

Cited By ~ 4

Author(s):

Baoyun Zhang ◽

Long Chen ◽

Guangde Feng ◽

Wei Xiang ◽

Ke Zhang ◽

...

Keyword(s):

Transcription Factors ◽

Granulosa Cells ◽

Messenger Rna ◽

Expression Patterns ◽

Follicular Development ◽

Functional Networks ◽

Signal Pathways ◽

Differentially Expressed Mirnas ◽

Selection Of

Ovaries, which provide a place for follicular development and oocyte maturation, are important organs in female mammals. Follicular development is complicated physiological progress mediated by various regulatory factors including microRNAs (miRNAs). To demonstrate the role of miRNAs in follicular development, this study analyzed the expression patterns of miRNAs in granulosa cells through investigating three previous datasets generated by Illumina miRNA deep sequencing. Furthermore, via bioinformatic analyses, we dissected the associated functional networks of the observed significant miRNAs, in terms of interacting with signal pathways and transcription factors. During the growth and selection of dominant follicles, 15 dysregulated miRNAs and 139 associated pathways were screened out. In comparison of different styles of follicles, 7 commonly abundant miRNAs and 195 pathways, as well as 10 differentially expressed miRNAs and 117 pathways in dominant follicles in comparison with subordinate follicles, were collected. Furthermore, SMAD2 was identified as a hub factor in regulating follicular development. The regulation of miR-26a/b onsmad2messenger RNA has been further testified by real time PCR. In conclusion, we established functional networks which play critical roles in follicular development including pivotal miRNAs, pathways, and transcription factors, which contributed to the further investigation about miRNAs associated with mammalian follicular development.

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The role of IGFs in the regulation of ovarian follicular growth in the brushtail possum (Trichosurus vulpecula)

Reproduction ◽

10.1530/rep-10-0142 ◽

2010 ◽

Vol 140 (2) ◽

pp. 295-303 ◽

Cited By ~ 8

Author(s):

Jennifer L Juengel ◽

Lisa J Haydon ◽

Brigitta Mester ◽

Brian P Thomson ◽

Michael Beaumont ◽

...

Keyword(s):

Granulosa Cell ◽

Granulosa Cells ◽

Cell Function ◽

Antral Follicle ◽

Brushtail Possum ◽

Follicular Growth ◽

Theca Cells ◽

Ovarian Follicular Growth

IGFs are known to be key regulators of ovarian follicular growth in eutherian mammals, but little is known regarding their role in marsupials. To better understand the potential role of IGFs in the regulation of follicular growth in marsupials, expression of mRNAs encoding IGF1, IGF2, IGF1R, IGF-binding protein 2 (IGFBP2), IGFBP4 and IGFBP5 was localized by in situ hybridization in developing ovarian follicles of the brushtail possum. In addition, the effects of IGF1 and IGF2 on granulosa cell function were tested in vitro. Both granulosa and theca cells synthesize IGF mRNAs, with the theca expressing IGF1 mRNA and granulosa cell expressing IGF2 mRNA. Oocytes and granulosa cells express IGF1R. Granulosa and theca cells expressed IGFBP mRNAs, although the pattern of expression differed between the BPs. IGFBP5 mRNA was differentially expressed as the follicles developed with granulosa cells of antral follicles no longer expressing IGFBP5 mRNA, suggesting an increased IGF bioavailability in the antral follicle. The IGFBP protease, PAPPA mRNA, was also expressed in granulosa cells of growing follicles. Both IGF1 and IGF2 stimulated thymidine incorporation but had no effect on progesterone production. Thus, IGF may be an important regulator of ovarian follicular development in marsupials as has been shown in eutherian mammals.

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Effect of Endometriosis on Progesterone Secretion by Human Granulosa Cells: Role of Peritoneal Fluid

Fertility and Sterility ◽

10.1016/j.fertnstert.2005.07.508 ◽

2005 ◽

Vol 84 ◽

pp. S201-S202

Author(s):

P.A. Navarro ◽

F.M. Gomes ◽

C.C. Petean ◽

R.M. Dos Reis ◽

R.A. Ferriani ◽

...

Keyword(s):

Granulosa Cells ◽

Peritoneal Fluid ◽

Human Granulosa Cells ◽

Progesterone Secretion

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Endothelin (ET)-1 and ET-3 inhibit estrogen and cAMP production by rat granulosa cells in vitro

Journal of Endocrinology ◽

10.1677/joe.0.1570209 ◽

1998 ◽

Vol 157 (2) ◽

pp. 209-215 ◽

Cited By ~ 9

Author(s):

AE Calogero ◽

N Burrello ◽

AM Ossino

Keyword(s):

Granulosa Cells ◽

Ovarian Function ◽

Biological Effects ◽

Female Rats ◽

Dependent Manner ◽

Camp Production ◽

Inhibitory Effects ◽

Ovarian Steroidogenesis ◽

Estrogen Production ◽

Etb Receptor

Endothelin (ET)-1 and ET-3, two peptides with a potent vasoconstrictive property, produce a variety of biological effects in different tissues by acting through two different receptors, the ET-1 selective ET(A) receptor and the non-selective ETB receptor. An increasing body of literature suggests that ET-1 acts as a paracrine/autocrine regulator of ovarian function. Indeed, ETB receptors have been identified in rat granulosa cells and ET-1 is a potent inhibitor of progesterone production. In contrast, inconsistent data have been reported about the role of ET-1 on estrogen production and the effects of ET-3 are not known. Therefore, the present study was undertaken to evaluate the effects of ET-1 and ET-3 on estrogen and cAMP production, and the receptor type involved. Given that prostanoids modulate ovarian steroidogenesis and that many actions of ETs are mediated by these compounds, we also evaluated whether the effects of ETs on estrogen and cAMP production might be prostanoid-mediated. ET-1, ET-3, and safarotoxin-S6c (SFX-S6c), a selective ETB receptor agonist, inhibited basal estrogen production by granulosa cells obtained from immature, estrogen-primed female rats, in a concentration-dependent manner. All three peptides were also capable of inhibiting the production of estrogen stimulated by a half-maximal (1 mIU/ml) and a maximally stimulatory (3 mIU/ml) concentration of FSH, ET-1 and ET-3 dose-dependently suppressed basal and FSH (1 mIU/ml)-stimulated cAMP production. ET-3 and SFX-S6c were significantly more potent than ET-1 in suppressing estrogen production, suggesting that this effect was not mediated by the ET(A) receptor. Indeed, BQ-123, a selective ET(A) receptor antagonist, did not influence the inhibitory effects of ET-1 and ET-3 on basal and FSH-stimulated estrogen release. To determine a possible involvement of prostanoids, we evaluated the effects of maximally effective concentrations of ET-1 and ET-3 on estrogen and cAMP production in the presence of indomethacin, a prostanoid synthesis inhibitor. This compound did not have any effect on the suppressive effects of ETs on basal or FSH (1 mIU/ml)-stimulated estrogen or cAMP production. In conclusion, ET-1 and ET-3 were able to inhibit estrogen and cAMP production by rat granulosa cells, indicating that the inhibitory effects of ETs on ovarian steroidogenesis are not limited to progesterone biosynthesis. This effect does not appear to be mediated by prostanoids or by the classical ET(A) and ETB receptors, at least under these experimental conditions.

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Modulation of Cathepsin S (CTSS) Regulates the Secretion of Progesterone and Estradiol, Proliferation, and Apoptosis of Ovarian Granulosa Cells in Rabbits

Animals ◽

10.3390/ani11061770 ◽

2021 ◽

Vol 11 (6) ◽

pp. 1770

Author(s):

Guohua Song ◽

Yixuan Jiang ◽

Yaling Wang ◽

Mingkun Song ◽

Xuanmin Niu ◽

...

Keyword(s):

Cell Proliferation ◽

Granulosa Cells ◽

Hormone Secretion ◽

Vital Role ◽

Regulatory Function ◽

Cathepsin S ◽

Related Gene ◽

Ovarian Granulosa Cells ◽

Proliferation And Apoptosis

Cathepsin S (CTSS) is a member of cysteine protease family. Although many studies have demonstrated the vital role of CTSS in many physiological and pathological processes including tumor growth, angiogenesis and metastasis, the function of CTSS in the development of rabbit granulosa cells (GCS) remains unknown. To address this question, we isolated rabbit GCS and explored the regulatory function of the CTSS gene in cell proliferation and apoptosis. CTSS overexpression significantly promoted the secretion of progesterone (P4) and estrogen (E2) by increasing the expression of STAR and CYP19A1 (p < 0.05). We also found that overexpression of CTSS increased GCS proliferation by up-regulating the expression of proliferation related gene (PCNA) and anti-apoptotic gene (BCL2). Cell apoptosis was markedly decreased by CTSS activation (p < 0.05). In contrast, CTSS knockdown significantly decreased the secretion of P4 and E2 and the proliferation of rabbit GCS, while increasing the apoptosis of rabbit GCS. Taken together, our results highlight the important role of CTSS in regulating hormone secretion, cell proliferation, and apoptosis in rabbit GCS. These results might provide a basis for better understanding the molecular mechanism of rabbit reproduction.

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