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 The Orphan Nuclear Receptors NURR1 and NGFI-B Modulate Aromatase Gene Expression in Ovarian Granulosa Cells: A Possible Mechanism for Repression of Aromatase Expression upon Luteinizing Hormone Surge

Endocrinology ◽  
2005 ◽  
Vol 146 (1) ◽  
pp. 237-246 ◽  
Author(s):  
Yimin Wu ◽  
Sagar Ghosh ◽  
Yoshihiro Nishi ◽  
Toshihiko Yanase ◽  
Hajime Nawata ◽  
...  
Keyword(s):  
Gene Expression ◽  
Nuclear Receptor ◽  
Granulosa Cells ◽  
Intracellular Camp ◽  
Orphan Nuclear Receptors ◽  
Aromatase Gene ◽  
Ovarian Granulosa Cells ◽  
A Genome ◽  
Responsive Gene ◽  
Aromatase Gene Expression

Ovarian granulosa cells play pivotal roles in many aspects of ovary functions including folliculogenesis and steroidogenesis. In response to FSH and LH, the elevation of intracellular cAMP level in granulosa cells leads to activation of multiple ovarian genes. Here, we report findings from a genome-wide study of the cAMP-responsive gene expression profiles in a human granulosa-like tumor cell line, KGN. The study identified 140 genes that are either activated or repressed by 2-fold or greater after stimulation by the adenylyl cyclase activator forskolin. The induction patterns of some cAMP-responsive genes were further analyzed by quantitative real-time PCR. Consistent with previous observations, the LH-responsive genes, such as the nuclear receptor 4A subfamily (NURR1, NGFI-B, and NOR-1), were rapidly but transiently induced, whereas the FSH-responsive gene CYP19 encoding aromatase was induced in a delayed fashion. Interestingly, ectopic expression of NURR1 or NGFI-B severely attenuated the cAMP-responsive activation of the ovary-specific aromatase promoter. Reduction of the endogenous NURR1 or NGFI-B by small interfering RNA significantly elevated aromatase gene expression. The cis-elements responsible for NURR1/NGFI-B-mediated repression were mapped to the minimal aromatase promoter sequence that confers camp responsiveness. Furthermore, the DNA-binding domain of NURR1 was required for the repression. Taken together, these results strongly suggest a causal relationship between the rapid decline of aromatase mRNA and induction of nuclear receptor subfamily 4A expression, which concomitantly occur upon LH surge at the later stages of ovarian follicular development.

Endometriosis does not Alter Aromatase Gene Expression (CYP19A1) in Mural Lutein-granulosa Cells of Women Undergoing Assisted Reproduction Techniques – a Pilot Study

2011 ◽  
Vol 3 (4) ◽  
pp. 177-182 ◽  
Author(s):  
Lauriane G. De Abreu ◽  
Vanessa S. Silveira ◽  
Carlos A. Scrideli ◽  
Ester S. Ramos ◽  
Rosana M. Dos Reis ◽  
...  
Keyword(s):  
Gene Expression ◽  
Assisted Reproduction ◽  
Granulosa Cells ◽  
Rna Extraction ◽  
Oocyte Retrieval ◽  
Assisted Reproduction Techniques ◽  
Aromatase Gene ◽  
Its Gene ◽  
Ovulatory Follicles ◽  
Aromatase Gene Expression

Purpose The aim of this study was to quantify aromatase gene expression in mural lutein-granulosa cells of women with endometriosis undergoing assisted reproduction techniques (ART, IVF or ICSI). Methods: a case-control study was performed on 11 women with endometriosis (all stages, ASRM criteria) and 11 women with male or tubal causes of infertility undergoing ART. There was no difference between the groups regarding age, amount of gonadotrophins used, days of induction, follicles, and eggs collected. Mural lutein-granulosa cells were harvested from pre-ovulatory follicles during oocyte retrieval and correctly isolated. After cells lyses and storage into Trizol LS Reagent®, RNA extraction and cDNA synthesis were performed. Quantification of relative gene expression for CYP19A1 (aromatase) was performed by real time PCR, using SYBR Green reagents. All experiments were performed in duplicates. Results there was no difference between the groups in the quantitative gene expression of CYP19A1 (aromatase) gene on mural lutein-granulosa cells (P>.05, Mann Whitney). Conclusions These results suggest that this enzyme, aromatase, may have a complex and refined control of its gene expression on this population of granulosa cells, and in spite of previous evidence showing its reduced activity on these cells in endometriosis, gene expression per se may not be affected by the disease.

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.

Quantification of P450 aromatase gene expression in cultured and in vivo grown ovarian follicles in sheep

2014 ◽  
Vol 117 (1) ◽  
pp. 66-72 ◽  
Author(s):  
B.N.V. Lakshminarayana ◽  
V. Praveen Chakravarthi ◽  
K.V. Brahmaiah ◽  
V.H. Rao
Keyword(s):  
Gene Expression ◽  
Ovarian Follicles ◽  
Aromatase Gene ◽  
P450 Aromatase ◽  
Aromatase Gene Expression
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