334. PROGESTERONE PRODUCTION FROM GRANULOSA CELLS OF SOWS IS ENHANCED EQUALLY BY OMEGA-3 DERIVED PROSTAGLANDIN E3 AND OMEGA-6 DERIVED PROSTAGLANDIN E2

2010 ◽  
Vol 22 (9) ◽  
pp. 134
Author(s):  
R. Smits ◽  
D. T. Armstrong ◽  
L. Ritter ◽  
M. Mitchell ◽  
M. B. Nottle
Keyword(s):  
Prostaglandin E2 ◽  
Granulosa Cells ◽  
Luteinizing Hormone Receptor ◽  
Follicle Cells ◽  
Omega 3 ◽  
Progesterone Production ◽  
Porcine Granulosa Cells ◽  
Significant Difference ◽  
Omega 6

Caughey et al (2005) reported that prostaglandins derived from omega 3 sources eicsapentaenoic acid (EPA C20:5) and docosahexaenoic acid (DHA C22:6) have different properties to those derived from the preferred substrate, arachidonic acid (ARA C20:4; n-6). Armstrong et al (2006) demonstrated that PGE2 increased progesterone when porcine granulosa cells were cultured in vitro with hCG. We hypothesized that PGE3 which is derived from EPA will produce a lower steroidogenic response as progesterone from isolated granulosa cells collected from pre-ovulatory sow ovaries. Ovaries were collected from slaughtered sows and follicles between 3–8 mm were aspirated and through a series of wash steps in HTCM (Hepes TCM 199). Mid-sized granulosa cells were recovered in a solution of BTCM (bicarbonate TCM) containing IGF-1. 0.5 × 106 cells/mL were cultured in 250 µL of Control (BTCM only), PGE2 or PGE3 (320 ng/mL in BTCM, Cayman Chemical Co.) treatments with IGF1 at 25 ng/mL. Cultures were incubated for 22 h at 38oC. Cultures were centrifuged and the supernatant was analysed in duplicate for progesterone. Data was analysed by Univariate GLM ANOVA. There was no significant difference between PGE2 and PGE3 treatments, however the main effect of PGE significantly increased progesterone production relative to the control (P = 0.017). Granulosa cells cultured with omega 3 derived PGE3 did not produce significantly lower progesterone levels than those with PGE2. We conclude that both PGE2 and PGE3 promote a steroidogenic response in cultured porcine granulosa cells. (1) Armstrong DT, Formosa, ER, Amato F, Schultz SJ. 2006. Prostaglandin E2 up-regulates luteinizing hormone receptor (LHR) expression and enhances steroidogenic responses of follicle cells.(2) Caughey GE, James MJ, Cleland LG. 2005. Prostaglandins and leukotrienes. pp. 42–49. In ‘Encyclopaedia of Human Nutrition. Vol. 4’. (Eds B Caballero, L Allen, A Prentice).

Effects of growth hormone and triiodothyronine on insulin-induced progesterone production by granulosa cells from prepubertal gilts

1992 ◽  
Vol 72 (3) ◽  
pp. 589-593 ◽  
Author(s):  
R. N. Kirkwood ◽  
P. A. Thacker ◽  
K. Rajkumar
Keyword(s):  
Growth Hormone ◽  
Granulosa Cells ◽  
Culture Medium ◽  
Low Density Lipoprotein ◽  
Density Lipoprotein ◽  
Bovine Insulin ◽  
Dependent Manner ◽  
Progesterone Production ◽  
Porcine Granulosa Cells

Two experiments were performed using granulosa cells from medium-sized follicles (2–4 mm) derived from prepubertal gilts. Cells were cultured in a serum-free medium at a density of either 1 or 2 × 106 viable cells per well (experiments 1 and 2, respectively). For exp. 1, porcine growth hormone (pGH) (0 or 100 ng mL−1) was included in the culture medium from the time of plating, and low-density lipoprotein (LDL) (100 μg mL−1) was added at 72 h. For exp. 2, granulosa cells were plated in a culture medium containing either pGH (0 or 100 ng mL−1) or triiodothyronine (T3) (0 or 5 ng mL−1) or both pGH T3; LDL was not included. For both experiments, after 24 h of culture, bovine insulin at 0, 10, 100 or 1000 ng mL−1 was included in the medium. Hormones were replaced at 48 and 72 h, and the cultures were terminated at 96 h. Results from exp. 1 indicated that insulin increased (P < 0.01) progesterone production in a dose-dependent manner, both in the presence and absence of LDL. This response was augmented (P < 0.01) by co-culture with pGH. Results from exp. 2 confirmed the augmenting effect of pGH (P < 0.01). It was further observed that T3 increased (P < 0.01) progesterone production when cultured with insulin at 1000 ng mL−1, but at lower insulin-inclusion levels, results were equivocal. The progesterone production response was greatest (P < 0.01) when cells were cultured with both pGH and T3 at insulin levels of 100 or 1000 ng mL−1. There appeared to be little relationship between the media concentrations of insulin-like growth factor 1 and progesterone. The present results suggest that relatively high levels of pGH and T3 will enhance the in vitro steroidogenic capabilities of porcine granulosa cells. Key words: Granulosa cells, GH, T3, insulin

INFLUENCE OF 16-ARYLOXYPROSTAGLANDINS ON THE PRODUCTION OF PROGESTERONE BY HUMAN GRANULOSA CELLS IN VITRO

1976 ◽  
Vol 71 (2) ◽  
pp. 259-263 ◽  
Author(s):  
K. M. HENDERSON
Keyword(s):  
Tissue Culture ◽  
Adenylate Cyclase ◽  
Granulosa Cells ◽  
Farm Animals ◽  
Human Granulosa Cells ◽  
Progesterone Production ◽  
Porcine Granulosa Cells ◽  
Prostaglandin F

SUMMARY 16-Aryloxy analogues of prostaglandin F2α(PGF2α) are potent luteolysins in laboratory and farm animals. When their effect on progesterone production by luteinized human granulosa cells in tissue culture was investigated inhibition of both basal and gonadotrophin-stimulated progesterone production was observed, so revealing characteristics expected of potential human luteolysins. The analogues were, however, unable to inhibit progesterone production stimulated by PGE2, suggesting that like PGF2α these compounds may act by specifically blocking LH-activated adenylate cyclase. The 16-aryloxyprostaglandins similarly inhibited progesterone production by porcine granulosa cells, so that the effects observed with the 16-aryloxyprostaglandins in vitro may be indicative of their potential in vivo.

Effect of insulin on porcine granulosa cells: implications of a possible receptor mediated action

1985 ◽  
Vol 108 (1) ◽  
pp. 104-110 ◽  
Author(s):  
Tetsuo Otani ◽  
Takeshi Maruo ◽  
Nobuyuki Yukimura ◽  
Matsuto Mochizuki
Keyword(s):  
Granulosa Cells ◽  
Glycogen Synthase ◽  
Direct Action ◽  
Insulin Receptors ◽  
Mediated Action ◽  
Porcine Granulosa Cells ◽  
Independent Form ◽  
Significant Difference ◽  
Specific Receptors

Abstract. The mechanism of direct action of insulin on porcine granulosa cells cultured in vitro was investigated. Specific receptors for insulin with two classes of binding sites were present. No significant difference in receptor characteristics was observed between granulosa cells obtained from small (1–2 mm), medium (3–5 mm) or large (6–11 mm) follicles. Insulin (25 mIU/ml) augmented both basal and hCG-stimulated progesterone secretion. Insulin was also essential for hCG-stimulated morphological luteinization of cultured porcine granulosa cells obtained from small follicles. Furthermore, 2 h preincubation with insulin (100 mIU/ml) increased glucose incorportation by cultured porcine granulosa cells as determined by pulsing with d-[14C](U)-glucose. The same dose of insulin also stimulated the glucose-6-phosphate independent form of glycogen synthase. These results suggest that porcine granulosa cells possess insulin receptors and that insulin, mediated by its specific receptors, enhances glucose metabolism by stimulating glycogen synthase. Thus, insulin may play a pivotal role in morphological and funtional differentiation of porcine granulosa cells.

Monoclonal antibodies specific for the binding site on the LH receptor alter progesterone production in cultured granulosa cells

1996 ◽  
Vol 150 (1) ◽  
pp. 9-16 ◽  
Author(s):  
E A Nau ◽  
E M Epsaro ◽  
M S Scherer ◽  
J H Abel
Keyword(s):  
Monoclonal Antibodies ◽  
Binding Site ◽  
Chorionic Gonadotropin ◽  
Granulosa Cells ◽  
Human Chorionic Gonadotropin ◽  
Lh Receptor ◽  
Inhibition Effect ◽  
Progesterone Production ◽  
Porcine Granulosa Cells

Abstract Monoclonal antibodies (mAbs) specific for the LH receptor (LHR) were generated through a modified auto-anti-idiotypic approach in which human chorionic gonadotropin (hCG) was used as the immunogen followed by cyclophosphamide to induce anti-idiotypic antibodies. The purpose of this study was to investigate the effectiveness of these antibodies to alter progesterone production in porcine granulosa cells in vitro. Anti-LHR mAbs were incubated with granulosa cells in the presence or absence of a stimulatory dose of hCG. Progesterone output by treated cells was measured using a RIA procedure. Most of the mAb could inhibit stimulated progesterone production by cultured granulosa cells. It was speculated that two possible mechanisms may cause the inhibition effect observed. Several of the antibodies appeared to block hCG binding thus removing the stimulatory effects of hCG. However, the most potent inhibiting mAbs for progesterone production had little or no effect on hCG binding, suggesting that some other mechanism was responsible for the observed inhibition. In addition, several of the antibodies were found to have a stimulatory effect on progesterone production by granulosa cells even in the absence of a stimulating dose of hCG. It is proposed that these antibodies were able to mimic hCG. Journal of Endocrinology (1996) 150, 9–16

scholarly journals FSH Promotes Progesterone Synthesis by Enhancing Autophagy to Accelerate Lipid Droplet Degradation in Porcine Granulosa Cells

2021 ◽  
Vol 9 ◽  
Author(s):  
Qiang Liu ◽  
Hui Gao ◽  
Feng Yang ◽  
Hanxue Zhang ◽  
Shenming Zeng
Keyword(s):  
Lipid Droplet ◽  
Granulosa Cells ◽  
Akt Inhibitor ◽  
Progesterone Production ◽  
Porcine Granulosa Cells ◽  
Progesterone Synthesis ◽  
Molecular Relationships ◽  
Factor C

Little is known about the molecular relationships among follicle stimulating hormone (FSH), lipid droplet (LD) degradation, and autophagy. In this study, we aimed to investigate the pathway by which FSH regulates autophagy and the potential role of autophagy in progesterone production. Our results revealed that FSH stimulated progesterone production in mammalian follicular granulosa cells (GCs) through a non-canonical pathway. In porcine secondary follicles cultured in vitro, FSH treatment increased the level of the autophagic marker, LC3-II, as well as increased the number of autophagic vacuoles in GCs. The underlying molecular mechanism and biological functions were then investigated in porcine GCs. Our results demonstrated that FSH could upregulate Beclin1 levels in porcine GCs; however, this effect was blocked by LY294002 (a PI3K/AKT inhibitor) and SP600125 (SAPK/JNK inhibitor). Further research confirmed that the transcriptional factor, c-Jun, was phosphorylated by FSH, then translocated into the nucleus from the cytoplasm and bound to the BECLIN1 promoter region, and that LY294002, SP600125, or c-Jun knockdown prevented the increase in Beclin1 levels induced by FSH. Interestingly, inhibition of autophagy using chloroquine or SP600125 decreased progesterone production in porcine GCs treated with FSH, although the expression of StAR and P450scc was not disturbed. Moreover, FSH treatment reduced the average number and size of LDs in porcine GCs, but these effects were eliminated by knocking down the key autophagy genes, ATG5 and BECLIN1; in addition, the effect of FSH on promoting progesterone secretion by the cells was also reduced significantly. Based on the above results, we concluded that FSH promoted progesterone production by enhancing autophagy through upregulation of Beclin1 via the PI3K/JNK/c-Jun pathway to accelerate LD degradation in porcine GCs, independent of the classical steroidogenic pathway.

RGD-mediated adhesion of porcine granulosa cells modulates their differentiation response to FSH in vitro

1995 ◽  
Vol 83 (2-3) ◽  
pp. 169-177 ◽  
Author(s):  
Béatrice Goxe ◽  
Jacques E. Flechon ◽  
Solange Delasalle ◽  
Roland Salesse
Keyword(s):  
Granulosa Cells ◽  
Porcine Granulosa Cells

scholarly journals Time- and Dose-Dependent Effects of 17 Beta-Estradiol on Short-Term, Real-Time Proliferation and Gene Expression in Porcine Granulosa Cells

2017 ◽  
Vol 2017 ◽  
pp. 1-9 ◽  
Author(s):  
Sylwia Ciesiółka ◽  
Joanna Budna ◽  
Karol Jopek ◽  
Artur Bryja ◽  
Wiesława Kranc ◽  
...  
Keyword(s):  
Real Time ◽  
Granulosa Cells ◽  
Hormone Receptor ◽  
Proliferative Activity ◽  
Cell Transformation ◽  
Luteinizing Hormone Receptor ◽  
Luteal Cell ◽  
Ovarian Activity ◽  
Short Term

The key mechanisms responsible for achievement of full reproductive and developmental capability in mammals are the differentiation and transformation of granulosa cells (GCs) during folliculogenesis, oogenesis, and oocyte maturation. Although the role of 17 beta-estradiol (E2) in ovarian activity is widely known, its effect on proliferative capacity, gap junction connection (GJC) formation, and GCs-luteal cells transformation requires further research. Therefore, the goal of this study was to assess the real-time proliferative activity of porcine GCs in vitro in relation to connexin (Cx), luteinizing hormone receptor (LHR), follicle stimulating hormone receptor (FSHR), and aromatase (CYP19A1) expression during short-term (168 h) primary culture. The cultured GCs were exposed to acute (at 96 h of culture) and/or prolonged (between 0 and 168 h of culture) administration of 1.8 and 3.6 μM E2. The relative abundance of Cx36, Cx37, Cx40, Cx43, LHR, FSHR, and CYP19A1 mRNA was measured. We conclude that the proliferation capability of GCs in vitro is substantially associated with expression of Cxs, LHR, FSHR, and CYP19A1. Furthermore, the GC-luteal cell transformation in vitro may be significantly accompanied by the proliferative activity of GCs in pigs.

COMPARISON OF THE STIMULATORY EFFECTS OF OVINE, PORCINE AND HUMAN FOLLICLE-STIMULATING HORMONE AND OF OVINE AND HUMAN LUTEINIZING HORMONE ON THE ACCUMULATION OF CYCLIC AMP BY PORCINE GRANULOSA CELLS

1979 ◽  
Vol 80 (1) ◽  
pp. 9-20 ◽  
Author(s):  
ADA M. LINDSEY ◽  
CORNELIA P. CHANNING
Keyword(s):  
Cyclic Amp ◽  
Granulosa Cells ◽  
Incubation Medium ◽  
Follicle Stimulating Hormone ◽  
Similar Response ◽  
Intracellular Accumulation ◽  
Porcine Granulosa Cells ◽  
Tenfold Increase ◽  
Incubation Periods

The effects of ovine, porcine and human FSH, and ovine and human LH on the accumulation of cyclic AMP by porcine granulosa cells obtained from follicles at various stages of maturation were investigated. During incubation periods of 15 min, 10 μg ovine FSH pretreated with antiserum to LH or 10 μg human FSH resulted in an 11- to 18-fold, five-to ninefold, and less than a twofold increase in intracellular accumulation of cyclic AMP by granulosa cells from small (1–2 mm), medium (3–5 mm) and large (6–12 mm) follicles respectively. Similar patterns of response occurred with addition of porcine FSH. After incubation for 30 and 60 min with ovine, porcine or human FSH, significant accumulation of cyclic AMP in the incubation medium occurred with cells obtained from small and medium-sized follicles. After 60 min of incubation with FSH the accumulation of cyclic AMP in the incubation medium exceeded the intracellular cyclic AMP levels in granulosa cells from small and medium-sized follicles. During incubation periods of 15 min, 1·0 μg ovine LH resulted in less than a twofold, a fourfold and greater than a tenfold increase in intracellular accumulation of cyclic AMP by granulosa cells from small, medium and large follicles respectively. Addition of human LH brought about a similar response. Incubation periods of 30 and 60 min with 1·0 μg ovine or human LH resulted in significant accumulation of cyclic AMP in the incubation medium by granulosa cells from large follicles; cyclic AMP content in the incubation medium was greater after 60 min compared with 30 min of incubation. It was concluded that ovine FSH pretreated with an antiserum to LH had similar effects on cyclic AMP levels as did purified human and porcine FSH, and that the stimulatory effects of the less pure ovine FSH were probably not due to an impurity in the FSH preparation. Porcine granulosa cells obtained from small follicles should be suitable as an in-vitro FSH bioassay while granulosa cells obtained from large follicles should be suitable as an in-vitro LH bioassay.

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