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.

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).

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

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.

scholarly journals Does 2-hydroxyflutamide Inhibit Apoptosis in Porcine Granulosa Cells? ^|^mdash; An In Vitro Study

2012 ◽  
Vol 58 (4) ◽  
pp. 438-444 ◽  
Author(s):  
Malgorzata DUDA ◽  
Malgorzata DURLEJ ◽  
Malgorzata KNET ◽  
Katarzyna KNAPCZYK-STWORA ◽  
Zbigniew TABAROWSKI ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
In Vitro Study ◽  
Vitro Study ◽  
Porcine Granulosa Cells

Prolactin, LH, and estradiol-17β in utilization of lipoprotein substrate by porcine granulosa cells in vitro

1988 ◽  
Vol 66 (5) ◽  
pp. 561-566 ◽  
Author(s):  
K. Rajkumar ◽  
P. Klingshorn ◽  
P. J. Chedrese ◽  
B. D. Murphy
Keyword(s):  
Granulosa Cell ◽  
Cell Cultures ◽  
Granulosa Cells ◽  
Low Density Lipoprotein ◽  
Density Lipoprotein ◽  
Low Density ◽  
Porcine Granulosa Cells ◽  
Progesterone Synthesis ◽  
Serum Free Conditions

Porcine granulosa cells cultured under serum free conditions responded by increased progesterone secretion to the addition of the leuteotropic hormones, LH, prolactin, and estradiol. Provision of extracellular substrate for steroidogenesis in the form of porcine high density lipoprotein or low density lipoprotein enhanced progesterone accumulation by granulosa cell cultures. Estradiol, LH, and prolactin all greatly increased progesterone accumulation in the presence of either high or low density lipoproteins. Increases in progesterone accumulation following addition of prolactin or LH in combination with estradiol suggested the presence of a synergistic interaction among leuteotropins. Pre-exposure of granulosa cell cultures to estradiol increased the subsequent stimulatory effect of prolactin on lipoprotein utilization. It is concluded that all three leuteotropins function to enhance and may interact in the utilization of extracellular lipoprotein substrate for progesterone synthesis.

Effect of noradrenaline and dopamine on progesterone and estradiol secretion of human granulosa cells

1993 ◽  
Vol 129 (2) ◽  
pp. 165-168 ◽  
Author(s):  
József Bódis ◽  
Hans R Tinneberg ◽  
Attila Török ◽  
Philippe Cledon ◽  
Volker Hanf ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Direct Action ◽  
Serum Free Medium ◽  
Free Medium ◽  
Vitro Fertilization ◽  
Serum Free ◽  
Human Granulosa Cells ◽  
Estradiol Secretion ◽  
Cells Cultured

The aim of this study was to explore the direct action of noradrenaline and dopamine on progesterone and estradiol secretion of human granulosa cells cultured in serum-free medium. Progesterone and estradiol production was measured in the presence and absence of noradrenaline, dopamine or propranolol using radioimmunoassays; statistical analysis was performed by analysis of variance and Newman-Keul's multiple range test. Twenty-six women aged 31±3 years undergoing in vitro fertilization and embryo transfer for infertility treatment at University Women's Hospital, University of Tübingen, Germany, took part in this study. Noradrenaline significantly inhibited progesterone production by human granulosa cells in a dose-related manner at a concentration of 10−4–10−6 mol/l. Dopamine significantly stimulated estradiol secretion by granulosa cells in an inverse dose-related manner. Both effects were blocked by propranolol. The results suggest that catecholaminergic actions switch over the steroid production of human granulosa cells cultured in serum-free medium from progesterone to estradiol.

scholarly journals Effects of transferrin on aromatase activity in porcine granulosa cells in vitro.

2009 ◽  
Vol 46 (4) ◽  
Author(s):  
Małgorzata Durlej ◽  
Małgorzata Duda ◽  
Katarzyna Knapczyk ◽  
Maria Słomczyńska
Keyword(s):  
Granulosa Cells ◽  
Aromatase Activity ◽  
Porcine Granulosa Cells

scholarly journals Transient Transfection of Porcine Granulosa Cells after 3D Culture in Barium Alginate Capsules

2005 ◽  
Vol 18 (4) ◽  
pp. 677-681 ◽  
Author(s):  
E. Benzoni ◽  
M.L. Torre ◽  
M. Faustini ◽  
S. Stacchezzini ◽  
F. Cremonesi ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Fluorescent Protein ◽  
3D Culture ◽  
Transient Transfection ◽  
Necessary Condition ◽  
In Vitro Toxicity ◽  
Culture Systems ◽  
Porcine Granulosa Cells ◽  
Alginate Capsules

Three-dimensional culture systems in barium alginate capsules can be employed to maintain primary granulosa cells in an undifferentiated state for almost 6 days. This is due to a self-organization of cells in a pseudofollicular structure. The transfection of primary granulosa cells is a necessary condition when employing these culture systems for several purposes, for example as an in vitro toxicity test or the development of oocytes or zygotes. In this work, the feasibility of two transient transfection techniques (liposome-mediated and electroporation) was assessed in primary porcine granulosa cells after a 6-day culture in an artificial extracellular matrix (barium alginate membrane). Human recombinant green fluorescent protein was chosen as a molecular readout, and protein expression was assessed after 48 hours from transfection. Liposome-mediated transfection gave low transfection levels, with increasing yields from 2 to 12 μgDNA/ml of medium; the maximum percentage (85.7%) was reached at 12 μgDNA/ml of medium. Electroporation-mediated transfection yields were higher: the best results (81.7% of transfected cells) were achieved with two 50V pulses and 12 μg/ml DNA. The application of a single or double pulse (50V) at 4 mgDNA/ml gave negligible results. These results indicate that primary granulosa cell cultured in barium alginate capsules can be transfected by electroporation with high transfection yields.

Effects of Ochratoxin A exposure on DNA damage in porcine granulosa cells in vitro

2020 ◽  
Vol 330 ◽  
pp. 167-175
Author(s):  
Tian-Yu Zhang ◽  
Li Kong ◽  
Jia-Xing Hao ◽  
Han Wang ◽  
Zi-Hui Yan ◽  
...  
Keyword(s):  
Dna Damage ◽  
Ochratoxin A ◽  
Granulosa Cells ◽  
Porcine Granulosa Cells
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