Increase in the expression of thyroid hormone receptors in porcine granulosa cells early in follicular maturation

1992 ◽  
Vol 127 (2) ◽  
pp. 152-160 ◽  
Author(s):  
Takeshi Maruo ◽  
Shinsuke Hiramatsu ◽  
Tetsuo Otani ◽  
Masato Hayashi ◽  
Matsuto Mochizuki
Keyword(s):  
Thyroid Hormone ◽  
Granulosa Cell ◽  
Granulosa Cells ◽  
Northern Blot ◽  
Scatchard Analysis ◽  
Large Follicle ◽  
Follicular Maturation ◽  
Cell Functions ◽  
Porcine Granulosa Cells ◽  
Small Follicle

Thyroid hormone has been demonstrated to synergize with FSH to exert stimulatory effects on the differentiation of porcine granulosa cells. In order to further characterize the nature of thyroid hormone action on granulosa cells, the presence of triiodothyronine (T3) receptors in the nuclei of porcine granulosa cells was examined, and qualitatively and quantitatively compared during follicular maturation. Then, comparative abilities of granulosa cells from varying follicle stages to respond to T3 were assessed in terms of FSH-induced LH/hCG receptor formation and progesterone secretion. Furthermore, the expression of erb-A was analyzed using Northern blot hybridization of porcine granulosa cell RNA with a v-erb-A probe. Binding experiments with [125I] T3 showed that granulosa cell nuclei obtained from small follicles had a greater ability to bind [125I] T3 compared to those from large follicles. Scatchard analysis revealed the presence of nuclear T3 receptors with a single class of binding sites. There was little difference in the affinity of the T3 receptors during follicular maturation. By contrast, the number of the T3 receptors was higher in small follicle granulosa cells compared to that in large follicle granulosa cells. Thus, the increased T3 binding to small follicle granulosa cells relative to large follicle granulosa cells appears to be attributable to the increased number of the nuclear T3 receptors rather than to a change in the affinity. The magnitude of the stimulatory effects of T3 on granulosa cell functions was maximal in small follicle granulosa cells, but negligible in large follicle granulosa cells. Northern blot analysis revealed that granulosa cell RNA contains erb-A transcripts of 5.0 kb and 2.7 kb and that the erb-A transcript abundance is higher in small follicle granulosa cells than that in large follicle granulosa cells. The increased expression of erb-A in immature granulosa cells may be responsible for an increase in the number of T3 receptors in less mature granulosa cells. These results suggest that thyroid hormone acts via its nuclear receptors selectively in immature granulosa cells early in follicular maturation to amplify the FSH actions in the facilitation of granulosa cell differentiation.

Stage-specific expression of c-myc messenger ribonucleic acid in porcine granulosa cells early in follicular growth

1994 ◽  
Vol 131 (3) ◽  
pp. 319-322 ◽  
Author(s):  
Asomi Sato ◽  
Masaki Bo ◽  
Takeshi Maruo ◽  
Shigeki Yoshida ◽  
Matsuto Mochizuki
Keyword(s):  
Granulosa Cell ◽  
Granulosa Cells ◽  
Northern Blot ◽  
Ribonucleic Acid ◽  
Physiological Role ◽  
Northern Blot Hybridization ◽  
Follicular Growth ◽  
Specific Expression ◽  
Messenger Ribonucleic Acid ◽  
Porcine Granulosa Cells

Sato A, Bo M, Maruo T, Yoshida S, Mochizuki M. Stage-specific expression of c-myc messenger ribonucleic acid in porcine granulosa cells early in follicular growth. Eur J Endocrinol 1994;131:319–22. ISSN 0804–4643 In the present study, the expression of c-myc mRNA in porcine granulosa cells harvested from varying follicle stages was analyzed using northern blot hybridization with a c-myc probe, and compared during follicular growth. Northern blot analysis revealed that porcine granulosa cell RNA obtained from small (1–2 mm) follicles contained an abundance of c-myc transcript of 2.4kb, the size of which was identical to the c-myc transcript obtained with human early placental RNA used as the control. In contrast to small-follicle granulosa cell RNA, total RNA preparations from either medium (3–5 mm)-follicle granulosa cells or large (6–11 mm)-follicle granulosa cells exhibited no detectable transcripts with the c-myc probe. These results suggest that expression of c-myc mRNA in porcine granulosa cells is limited only at the early stage of follicular growth. The stage-specific expression of c-myc mRNA in granulosa cells early in follicular growth may play a physiological role in the autonomous growth of immature granulosa cells in early folliculogenesis, seemingly independent of pituitary gonadotropic stimulation. Takeshi Maruo, Department of Obstetrics and Gynecology, Kobe University School of Medicine, Chuo-ku, Kobe 650, Japan

scholarly journals Effect of aromatase inhibitor (fadrozole) on proliferation, estradiol production and telomerase activity in pig granulosa cells in vitro

2009 ◽  
Vol 54 (No. 12) ◽  
pp. 566-574 ◽  
Author(s):  
E. Chronowska ◽  
M. Tománek ◽  
T. Kott
Keyword(s):  
Gene Expression ◽  
Aromatase Inhibitor ◽  
Granulosa Cell ◽  
Granulosa Cells ◽  
Telomerase Activity ◽  
Culture Conditions ◽  
Large Follicle ◽  
Aromatase Gene ◽  
Small Follicle ◽  
Aromatase Gene Expression

The objective of the present work was to study the effect of a nonsteroidal aromatase inhibitor (fadrozole) on proliferation, estradiol production, aromatase expression and telomerase activity (TA) in pig granulosa cells (GC) from small (1–2 mm) and large (5–7 mm) follicles. The cells were treated with fadrozole for 48 h and 72 h in basal and FSH-stimulated conditions. Fadrozole caused a decrease (<i>P</i> < 0.05) of 3H-thymidine incorporation in granulosa cells derived from small (1–2 mm) and large follicles (5–7 mm). The proliferative potential of small-follicle GC was significantly higher (<i>P</i> < 0.01) under all culture conditions. Estradiol production was suppressed (<i>P</i> < 0.01) in both granulosa cell populations cultured in the presence of fadrozole for 48 and 72 h. Fadrozole caused a decrease (<i>P</i> < 0.05) of aromatase gene expression in small-follicle granulosa cell incubated for 72 h and in large-follicle GC after 48 h of culture. Large-follicle GC were characterized by a higher (<i>P</i> < 0.01) level of estradiol production and aromatase gene expression. Telomerase activity decreased (<i>P</i> < 0.05) in large-follicle granulosa cells incubated in the presence of an aromatase inhibitor for 72 h. The TA level in large-follicle granulosa cells was higher (<i>P</i> < 0.01) in comparison to small-follicle GC in all culture conditions after 72 h of incubation. The results of the present study suggest the important role of telomerase in the process of follicular growth and development.

scholarly journals MiRNA-143 mediates the proliferative signaling pathway of FSH and regulates estradiol production

2017 ◽  
Vol 234 (1) ◽  
pp. 1-14 ◽  
Author(s):  
Li Zhang ◽  
XiaoXin Zhang ◽  
Xuejing Zhang ◽  
Yu Lu ◽  
Lei Li ◽  
...  
Keyword(s):  
Signaling Pathway ◽  
Granulosa Cell ◽  
Granulosa Cells ◽  
Loss Of Function ◽  
Cellular Processes ◽  
Cell Functions ◽  
Genes Expression ◽  
Underlying Mechanisms

MicroRNAs (MiRNAs) play important regulatory roles in many cellular processes. MiR-143 is highly enriched in the mouse ovary, but its roles and underlying mechanisms are not well understood. In the current study, we show that miR-143 is located in granulosa cells of primary, secondary and antral follicles. To explore the specific functions of miR-143, we transfected miR-143 inhibitor into primary cultured granulosa cells to study the loss of function of miR-143 and the results showed that miR-143 silencing significantly increased estradiol production and steroidogenesis-related gene expression. Moreover, our in vivo and in vitro studies showed that follicular stimulating hormone (FSH) significantly decreased miR-143 expression. This function of miR-143 is accomplished by its binding to the 3’-UTR of KRAS mRNA. Furthermore, our results demonstrated that miR-143 acts as a negative regulating molecule mediating the signaling pathway of FSH and affecting estradiol production by targeting KRAS. MiR-143 also negatively acts in regulating granulosa cells proliferation and cell cycle-related genes expression. These findings indicate that miR-143 plays vital roles in FSH-induced estradiol production and granulosa cell proliferation, providing a novel mechanism that involves miRNA in regulating granulosa cell functions.

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.

scholarly journals Concanavalin-A Induces Granulosa Cell Death and Inhibits FSH-Mediated Follicular Growth and Ovarian Maturation in Female Rats

Endocrinology ◽  
2013 ◽  
Vol 154 (5) ◽  
pp. 1885-1896 ◽  
Author(s):  
Ethel V. Velasquez ◽  
Mariana Ríos ◽  
María Elena Ortiz ◽  
Carlos Lizama ◽  
Elizabeth Nuñez ◽  
...  
Keyword(s):  
Cell Death ◽  
Granulosa Cell ◽  
Granulosa Cells ◽  
Concanavalin A ◽  
Ex Vivo ◽  
Female Rats ◽  
Carbohydrate Binding ◽  
Con A ◽  
Follicular Maturation

Abstract Reproductive success stems from a finely regulated balance between follicular maturation and atresia, in which the role of carbohydrate structure is poorly understood. Here, we describe for the first time a fraction of purified recombinant human FSH that is capable of bringing about the cell death of granulosa cells and preventing follicular maturation in a rat model. Further analysis by mass spectrometry revealed the presence of the lectin Concanavalin-A (Con-A) within this fraction of recombinant FSH. Using both the fractionated FSH and Con-A, the observed cell death was predominantly located to the granulosa cells. Ex vivo culture of rat follicles demonstrated that follicle degeneration occurred and resulted in the release of a denuded and deteriorated oocyte. Moreover, in vivo experiments confirmed an increase in atresia and a corresponding reduction confined to follicle in early antral stage. As a mechanism of action, Con-A reduces ovarian proliferation, Von Willebrand staining, and angiogenesis. Based on the observation that Con-A may induce granulosa cell death followed by follicle death, our results further demonstrate that follicular carbohydrate moiety is changing under the influence of FSH, which may allow a carbohydrate-binding lectin to increase granulosa cell death. The physiological consequences of circulating lectin-like molecules remain to be determined. However, our results suggest a potential exploitation of carbohydrate binding in fertility and ovarian cancer treatment. This work may shed light on a key role of carbohydrates in the still obscure physiological process of follicular selection and atresia.

Porcine granulosa cell conditioned media as autocrine regulator of progesterone secretion

1993 ◽  
Vol 5 (1) ◽  
pp. 95
Author(s):  
RT Denkova ◽  
IG Ivanov ◽  
LN Kanchev
Keyword(s):  
Granulosa Cell ◽  
Granulosa Cells ◽  
Cultured Cells ◽  
Primary Cultures ◽  
Inhibitory Effect ◽  
Conditioned Media ◽  
Progesterone Secretion ◽  
Porcine Granulosa Cells ◽  
Porcine Granulosa Cell

The ability of porcine granulosa cells to release a progesterone inhibiting substance(s) was examined in vitro. Granulosa cells (SGCs, MGCs and LGCs) were harvested from small, medium or large antral follicles respectively. The effect of granulosa cell conditioned media obtained from small follicles (SGCCM) was studied in the culture of LGCs by estimation of progesterone secretion; the conditioned media evoked the inhibition of progesterone secretion by the LGCs. SGCCM produced by various numbers of cultured granulosa cells showed a dose-related inhibition of progesterone production. A maximum inhibitory effect was noted when a 5-fold concentration of SGCCM was used. The addition of SGCCM had no effect on the growth of the cultured cells. The factor(s) inhibiting progesterone secretion appeared to be a nonsteroidal substance of molecular mass greater than 10 kDa and was heat-stable and trypsin-sensitive. The data presented support the suggestion that the conditioned media generated by primary cultures of SGCs contain nonsteroidal regulators capable of inhibiting progesterone secretion by cultured LGCs; this inhibitory activity can play an important autocrine regulatory role in the process of follicular differentiation.

scholarly journals Binding of LH and FSH to porcine granulosa cells during follicular maturation

Reproduction ◽  
1977 ◽  
Vol 51 (1) ◽  
pp. 23-27 ◽  
Author(s):  
R. Nakano ◽  
T. Akahori ◽  
K. Katayama ◽  
S. Tojo
Keyword(s):  
Granulosa Cells ◽  
Follicular Maturation ◽  
Porcine Granulosa Cells

ACTIVATION INDUCED BY LUTEINIZING HORMONE OF TYPE II PROTEIN KINASE DEPENDENT ON CYCLIC ADENOSINE MONOPHOSPHATE AND PHOSPHORYLATION OF SOLUBLE PROTEINS IN PORCINE GRANULOSA CELLS

1980 ◽  
Vol 84 (1) ◽  
pp. 49-63 ◽  
Author(s):  
D. H. HALPREN-RUDER ◽  
R. A. JUNGMANN ◽  
W. J. GEORGE ◽  
J. R. JETER
Keyword(s):  
Protein Kinase ◽  
Luteinizing Hormone ◽  
Cyclic Amp ◽  
Granulosa Cell ◽  
Granulosa Cells ◽  
Type Ii ◽  
Dependent Protein Kinase ◽  
Porcine Granulosa Cells ◽  
Dependent Protein ◽  
Dose Dependent

The present experiments were designed to study whether exogenous LH could elicit acute cyclic AMP-mediated activation of cyclic AMP-dependent protein kinase and phosphorylation of cellular protein in intact porcine granulosa cells. Incubation of porcine granulosa cells (from 3 to 5 mm diameter follicles) with 2 μg luteinizing hormone/ml (LH) caused a significant rise of cellular cyclic AMP content within 2 min of the addition of LH. The increase was dose-dependent and occurred between doses of 0·2 and 2·0 μg LH/ml. Luteinizing hormone also caused a time- and dose-dependent dissociation of the type II cyclic AMP-dependent protein kinase isozyme in porcine granulosa cells. Luteinizing hormone (0·05–2 μg/ml) significantly dissociated the cyclic AMP-dependent protein kinase between 2 and 30 min after stimulation. The protein kinase dissociation was a specific effect of LH and was not elicited by either adrenocorticotrophic hormone or prolactin. During the period of LH-induced protein kinase activation, several soluble granulosa cell proteins, ranging in molecular weights from about 43 000 to 99 000, became phosphorylated in a time-dependent and hormone-specific manner. The results suggest that cyclic AMP-mediated activation of granulosa cell type II cyclic AMP-dependent protein kinase may be a prerequisite in the short-term molecular action of LH leading to LH-specific phosphorylation of several soluble granulosa cell proteins of an as yet unidentified function.

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