scholarly journals FSH, Oxytocin and IGF-I Regulate the Expression of Sirtuin 1 in Porcine Ovarian Granulosa Cells

2020 ◽  
pp. 461-466
Author(s):  
A SIROTKIN ◽  
P DEKANOVÁ ◽  
A HARRATH
Keyword(s):  
Granulosa Cells ◽  
Intracellular Signaling ◽  
Sirtuin 1 ◽  
Signaling System ◽  
Factor I ◽  
Ovarian Granulosa Cells ◽  
Ovarian Cells ◽  
Igf I ◽  
Hormonal Action

The involvement of the mTOR system/enzyme sirtuin 1 (SIRT1) intracellular signaling system in the control of ovarian functions and its role in mediating hormonal action on the ovary has been proposed, but this hypothesis should be supported by a demonstrated influence of hormones on mTOR/SIRT1. Therefore, the aim of our in vitro experiments was to examine the effect of the known hormonal regulators of ovarian functions, such as follicle-stimulating hormone (FSH), oxytocin (OT) and insulin-like growth factor I (IGF-I), on mTOR/SIRT1. The accumulation of SIRT1 in porcine ovarian granulosa cells cultured with and without these hormones (at doses of 1, 10 or 100 ng.ml-1) was evaluated using immunocytochemistry. It was observed that the addition of FSH (at 10 ng.ml-1 but not at 1 or 100 ng/ml) and OT (at all tested doses) increased the expression of SIRT1 in ovarian cells. In addition, 100 ng.ml-1, but not at 1 or 10 ng.ml-1, of IGF-I decreased SIRT1 accumulation. Our observations are the first demonstration that hormones can directly regulate the ovarian mTOR/SIRT1 system and that this system could mediate the action of hormonal regulators on the ovary.

scholarly journals Serum deprivation affects secretory activity of cultured porcine ovarian follicles and granulosa cells and their response to hormones

2009 ◽  
Vol 54 (No. 10) ◽  
pp. 455-460
Author(s):  
A.V. Sirotkin
Keyword(s):  
Granulosa Cells ◽  
Ovarian Follicle ◽  
Secretory Activity ◽  
Ovarian Follicles ◽  
Serum Deprivation ◽  
Ovarian Cells ◽  
Igf I ◽  
Granulose Cells ◽  
Vitro Model

The aim of the present study is to understand the hormonal mechanisms of the effect of malnutrition on ovarian follicle functions. For this purpose, we examined the effect of malnutrition/serum deprivation, addition of metabolic hormones and gonadotropin (IGF-I, leptin and FSH) and their combination on the release of progesterone (P<sub>4</sub>), testosterone (T), estradiol (E<sub>2</sub>) and insulin-like growth factor I (IGF-I) by cultured whole ovarian follicles and on P<sub>4</sub> and IGF-I output by cultured granulosa cells isolated from porcine ovaries. It was observed that in ovarian follicles cultured with nutrients/serum addition of IGF-I reduced release of P<sub>4</sub>, but not of T or E<sub>2</sub>. Exogenous leptin reduced output of E<sub>2</sub>, but not of P<sub>4</sub> or T, and increased IGF-I output. No significant effect of FSH on release of steroid hormones by isolated follicles was found. Serum deprivation did not affect release of P<sub>4</sub>, but reduced output of T and E<sub>2</sub>, and promoted IGF-I release by cultured ovarian follicles. Addition of hormones failed to prevent the effect of malnutrition on the secretory activity of cultured ovarian follicles. In cultured granulose cells, all the tested hormones promoted release of both P<sub>4</sub> and IGF-I. Food restriction/serum deprivation reduced both P<sub>4</sub> and IGF-I output. Additions of either IGF-I, leptin and FSH prevented the inhibitory action of malnutrition on both P<sub>4</sub> and IGF-I release. The present observations (1) confirm the involvement of the hormones IGF-I, leptin and FSH in the control of secretory activity of ovarian cells, (2) demonstrate, that both isolated ovarian granulosa cells and whole follicles cultured in the absence of serum nutrients could be an adequate in-vitro model for studying the effect of malnutrition on ovarian secretory functions, and (3) suggest, that malnutrition could affect ovarian functions through changes in the release of ovarian hormones.

scholarly journals The Effect of Capsaicin on IGF-I and IGF-IR Expression in Ovarian Granulosa Cells

2020 ◽  
Vol 71 (1) ◽  
pp. 1977
Author(s):  
S. GULER ◽  
Β. ZIK
Keyword(s):  
Growth Factor ◽  
Granulosa Cells ◽  
Ovarian Development ◽  
Ovary Development ◽  
Insulin Like Growth Factor ◽  
Sprague Dawley ◽  
Factor I ◽  
Ovarian Granulosa Cells ◽  
Igf I ◽  
Growth Factor I

Capsaicin (trans-8-methyl-N-vanillyl-6-noneadamide) is a pungent ingredient in red peppers from the Capsicum family. Insulin-like growth factor-I (IGF-I) is expressed in granulosa cells and has an important role in ovarian development. However, there are no data about the IGF-I expression in ovarian granulosa cells after capsaicin treatment. The aim of this study was to investigate the expression of IGF-I and its receptor (insulin-like growth factor-I receptor [IGF-IR]) in primary rat ovarian granulosa cells after low and high doses of capsaicin treatment. For this, granulosa cells were isolated and cultured from ovaries of 30-day-old female Sprague-Dawley rats. Granulosa cell plates were divided into four groups as cell control (C), vehicle control (V), and 50 μM and 150 μM capsaicin groups. In experimental groups, granulosa cells were exposed to capsaicin for 24 hours and immunocytochemistry was performed afterwards using anti-IGF-I and anti-IGF-IR antibodies. Both IGF-I and IGF-IR expressions were found to be significantly increased in parallel to the capsaicin doses. Elevated levels of IGF-I may be a risk factor for ovarian development. Because of the crucial role of IGF-I in ovary development, capsaicin treatment can be effective on follicular development and/or disorders characterized by high IGF-I levels.

scholarly journals  The effect of high temperature on swine ovarian function in vitro

2010 ◽  
Vol 55 (No. 8) ◽  
pp. 377-382 ◽  
Author(s):  
AV Sirotkin ◽  
M. Kacaniova
Keyword(s):  
High Temperature ◽  
High Temperatures ◽  
Granulosa Cells ◽  
Ovarian Hormones ◽  
Ovarian Follicles ◽  
Ovarian Granulosa Cells ◽  
Ovarian Cells ◽  
Igf I ◽  
Series Of Experiments ◽  
Cells Cultured

The aim of the present study was to understand the hormonal mechanisms behind the effect of high temperatures on reproductive function. It was proposed that high temperatures can directly alter production of ovarian hormones and/or the response of ovarian cells to hormonal stimulators. To examine this hypothesis, in the 1st series of experiments, we compared the release of progesterone (P<sub>4</sub>), estradiol (E<sub>2</sub>) and expression of the leptin gene in whole ovarian follicles cultured in conditions of normal (37.5&deg;C) and high (41.5&deg;C) temperatures. In the 2<sup>nd</sup> series of experiments, we examined the release of P<sub>4</sub> and insulin-like growth factor I (IGF-I) by ovarian granulosa cells cultured in conditions of normal and high temperatures with and without IGF-I, leptin and FSH. The release of hormones was measured by RIA, while the expression of the leptin gene was evaluated by PCR. It was observed that high temperature significantly increased P<sub>4</sub> and E<sub>2</sub> release and reduced the accumulation of leptin DNA in ovarian follicles. In cultured ovarian granulosa cells, high temperatures promoted the release of both P<sub>4</sub> and IGF-I. The addition of IGF-I, leptin and FSH to granulosa cells cultured at normal temperature promoted the release of both P<sub>4</sub> and IGF-I. High temperature was able to prevent the stimulatory effect of leptin (but not of IGF-I or FSH) on P<sub>4</sub> output and the stimulatory action of both leptin and FSH on IGF-I release by granulosa cells. The present observations (1) demonstrate the possible production of leptin in the porcine ovary, (2) demonstrate for the first time the influence of high temperatures on ovarian P<sub>4</sub>, E<sub>2</sub>, IGF-I and leptin, and (3) suggest, that the negative effect of heat stress on reproductive processes can be due to high temperature-induced malproduction of ovarian hormones and a reduction in the response of ovarian cells to hormonal stimulators.

scholarly journals In Vitro Assessment of Iron Effect on Porcine Ovarian Granulosa Cells: Secretory Activity, Markers of Proliferation and Apoptosis

2011 ◽  
pp. 503-510 ◽  
Author(s):  
A. KOLESAROVA ◽  
M. CAPCAROVA ◽  
M. MEDVEDOVA ◽  
A. V. SIROTKIN ◽  
J. KOVACIK
Keyword(s):  
Growth Factor ◽  
Granulosa Cells ◽  
Caspase 3 ◽  
Secretory Activity ◽  
Cyclin B1 ◽  
Ovarian Granulosa Cells ◽  
Proliferation And Apoptosis ◽  
Igf I ◽  
Iron Sulphate

It would be desirable to expand the existing general knowledge concerning direct action of metals on the ovary. Nevertheless, the results of testing of iron compound on porcine ovarian cells should be interpreted carefully because iron is an essential element which could also induce changes in cellular processes. The aim of this in vitro study was 1) to examine dose-dependent effects of iron on the secretory activity of porcine ovarian granulosa cells, and 2) to outline the potential intracellular mediators mediating these effects. Specifically, we evaluated the effect of iron sulphate on the release of insulin-like growth factor I (IGF-I) and progesterone, as well as the expression of markers of proliferation (cyclin B1) and apoptosis (caspase-3) in porcine ovarian granulosa cells. Concentrations of IGF-I and progesterone were determined by RIA, cyclin B1 and caspase-3 expression by immunocytochemistry (ICC). Our results show a significantly decreased IGF-I secretion by ovarian granulosa cells after iron sulphate addition at the doses 0.5 and 1.0 mg/ml. The iron sulphate additions at doses 0.17 and 1.0 mg/ml had no effect on progesterone secretion. In contrast, iron sulphate addition at doses 0.17-1.0 mg/ml resulted in stimulation of cyclin B1 and caspase-3 expression. In conclusion, the present results indicate a direct effect of iron on 1) secretion of growth factor IGF-I but not steroid hormone progesterone, 2) expression of markers of proliferation (cyclin B1), or 3) apoptosis (caspase-3) of porcine ovarian granulosa cells. These results support an idea that iron could play a regulatory role in porcine ovarian function: hormone release, proliferation and apoptosis.

scholarly journals Do GH, IGF-I and oxytocin interact by regulating the secretory activity of porcine ovarian cells?

2001 ◽  
Vol 171 (3) ◽  
pp. 475-480 ◽  
Author(s):  
AV Sirotkin ◽  
AV Makarevich ◽  
HB Kwon ◽  
J Kotwica ◽  
J Bulla ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Binding Sites ◽  
Secretory Activity ◽  
Prostaglandin E ◽  
Ovarian Granulosa Cells ◽  
Ovarian Cells ◽  
Igf I ◽  
Igf Binding ◽  
Igf Binding Protein ◽  
Igfbp 3

The aims of this study on porcine ovarian granulosa cells were to examine the effect of GH on oxytocin (OT), IGF-I and IGF-I receptors, IGF-binding protein-3 (IGFBP-3), progesterone and prostaglandin E (PGE), as well as to determine whether IGF-I and/or OT may be mediators of GH action. The cells were cultured either with porcine GH (pGH) (1 ng/ml to 10 microg/ml or 100 ng/ml only), antiserum against IGF-I (0.1%), antiserum against OT (0.1%) or a combination of GH (10 ng/ml) with antiserum against IGF-I or antiserum against OT (0.1%). The secretion of IGF-I, OT, IGFBP-3, progesterone and PGE was determined using RIA/IRMA, whilst the IGF-I binding sites were measured using a radioreceptor assay. It was observed that pGH increased the secretion of IGF-I and the abundance of IGF-I binding sites in granulosa cells. Furthermore, GH inhibited OT release, stimulated progesterone and PGE output, but had no significant effect on IGFBP-3 secretion. Immunoneutralization of IGF-I by antiserum against IGF-I inhibited PGE secretion, but it did not influence progesterone or IGFBP-3 secretion. Binding of OT by antiserum suppressed IGFBP-3, PGE, but not progesterone secretion. Neither immunoneutralization of IGF-I nor OT substantially prevented the effects of GH on progesterone, IGFBP and PGE. These observations demonstrate the involvement of GH, IGF-I and OT in the control of porcine ovarian secretory activity and the ability of GH to regulate IGF-I and OT production and IGF-I reception. Nevertheless, lack of correlation between the effects of GH, antiserum against IGF-I and antiserum against OT, as well as the inability of blockade of IGF-I or OT to prevent the effects of GH, suggests that IGF-I and OT, despite their dependence on GH, do not mediate GH action on ovarian cells.

scholarly journals Arachidonic Acid Regulation of Intracellular Signaling Pathways and Target Gene Expression in Bovine Ovarian Granulosa Cells

Animals ◽  
2019 ◽  
Vol 9 (6) ◽  
pp. 374 ◽  
Author(s):  
Nina Zhang ◽  
Liqiang Wang ◽  
Guoya Luo ◽  
Xiaorong Tang ◽  
Lizhu Ma ◽  
...  
Keyword(s):  
Gene Expression ◽  
Signaling Pathways ◽  
Granulosa Cells ◽  
Intracellular Signaling ◽  
Hormone Secretion ◽  
Ovarian Granulosa Cells ◽  
Genes Expression ◽  
Higher Doses ◽  
Gene Expression Levels

In the present study, AA was used to challenge bovine ovarian granulosa cells in vitro and the related parameters of cellular and molecular biology were measured. The results indicated that lower doses of AA increased survival of bovine granulosa cells whereas higher doses of AA suppressed survival. While lower doses of AA induced accumulation of lipid droplet in granulosa cells, the higher dose of AA inhibited lipid accumulation, and AA increased abundance of FABP3, CD36 and SLC27A1 mRNA. Higher doses of AA decreased the secretion of E2 and increased the secretion of P4 accompanied by down-regulation of the mRNA abundance of CYP19A1, FSHR, HSD3B1 and STAR in granulosa cells. The signaling pathways employed by AA in the stimulation of genes expression included both ERK1/2 and Akt. Together, AA specifically affects physiological features, gene expression levels and steroid hormone secretion, and thus altering the functionality of granulosa cells of cattle.

Effects of recombinant bovine somatotrophin, insulin-like growth factor-I and insulin on the proliferation of bovine granulosa cells in vitro

1993 ◽  
Vol 139 (1) ◽  
pp. 67-75 ◽  
Author(s):  
J. G. Gong ◽  
D. McBride ◽  
T. A. Bramley ◽  
R. Webb
Keyword(s):  
Growth Factor ◽  
Granulosa Cells ◽  
Ovarian Follicles ◽  
Dependent Manner ◽  
Insulin Like Growth Factor ◽  
Factor I ◽  
Igf I ◽  
Dose Dependent ◽  
Bovine Somatotrophin

ABSTRACT Treatment of heifers with recombinant bovine somatotrophin (BST) significantly increases the population of small ovarian follicles and peripheral concentrations of somatotrophin, insulin-like growth factor-I (IGF-I) and insulin. To investigate the possible mechanism(s) involved in the action of BST on ovarian follicles, the effects of BST, IGF-I and insulin, given alone or in combination with either FSH or LH, on the proliferation of bovine granulosa cells in vitro were examined using a serum-free culture system. Bovine granulosa cells were obtained from antral follicles classified into three size categories according to diameter: small <5 mm; medium-sized 5–10 mm and large >10 mm. The proliferation of granulosa cells was assessed by the incorporation of [3H]thymidine into the cultured cells. Both FSH and LH (1–1000 ng/ml) inhibited the proliferation of bovine granulosa cells obtained from all three size classes of follicles in a dose-dependent manner. BST, at doses ranging from 1 to 1000 ng/ml, had no effect on the proliferation of granulosa cells from small and medium-sized follicles, but inhibited the division of granulosa cells from large follicles in a dose-dependent manner. Treatment with either IGF-I (10–3000 ng/ml) or insulin (0·5–1000 ng/ml) stimulated, in a dose-dependent manner, the proliferation of granulosa cells obtained from all three size categories of follicles. No synergistic interaction between BST (30 ng/ml) and either FSH (50 ng/ml) or LH (5 ng/ml) was observed in granulosa cells from all three size classes of follicles. In contrast, physiological concentrations of both IGF-I (100 ng/ml) and insulin (1 ng/ml) acted in synergy with both FSH (50 ng/ml) and LH (5 ng/ml) to stimulate the proliferation of granulosa cells from small follicles, whilst no such synergistic interactions were observed in granulosa cells from medium-sized and large follicles. It was concluded that the increase in the number of small ovarian follicles induced by BST treatment in heifers may be mediated by increased peripheral concentrations of IGF-I and/or insulin, possibly acting in synergy with gonadotrophins. Furthermore, insulin probably acts through its own receptor rather than acting via the type-I IGF receptor, as it can stimulate the proliferation of bovine granulosa cells at physiological concentrations. Journal of Endocrinology (1993) 139, 67–75

scholarly journals Leptin affects proliferation-, apoptosis- and protein kinase A-related peptides in human ovarian granulosa cells

2008 ◽  
pp. 437-442
Author(s):  
AV Sirotkin ◽  
M Mlynček ◽  
AV Makarevich ◽  
I Florkovičová ◽  
L Hetényi
Keyword(s):  
Protein Kinase ◽  
Protein Kinase A ◽  
Granulosa Cells ◽  
Intracellular Signaling ◽  
Cyclin B1 ◽  
Dependent Manner ◽  
Cell Functions ◽  
Ovarian Cells ◽  
Kinase A

The aim of our in vitro studies was to understand the role of leptin in controlling proliferation, apoptosis, and protein kinase A (PKA) in human ovarian cells. We analyzed the in vitro effects of leptin (0, 1, 10 or 100 ng/ml) on the accumulation of proliferation-related peptides (PCNA, cyclin B1), apoptosisassociated peptide (Bax) and the intracellular signaling molecule PKA in cultured human granulosa cells using immunocytochemistry and Western immunoblotting. It was observed that leptin stimulated in a dose-dependent manner the accumulation of PCNA (at doses 1-100 ng/ml), cyclin B1 (at doses 10 or 100 ng/ml), Bax (at doses 10 or 100 ng/ml) and PKA (at doses 1-100 ng/ml) in cultured human ovarian cells. These observations suggest the ability of leptin to control directly human ovarian cell functions: proliferation, apoptosis, and intracellular messenger PKA.

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