BIOTRANSFORMATION OF PREGNENOLONE AND PROGESTERONE BY RABBIT OVARIAN FOLLICLES AND CORPORA LUTEA

1973 ◽  
Vol 74 (4) ◽  
pp. 775-782 ◽  
Author(s):  
Edward V. YoungLai
Keyword(s):  
Granulosa Cells ◽  
Salt Solution ◽  
Hydroxysteroid Dehydrogenase ◽  
Ovarian Follicles ◽  
Rabbit Serum ◽  
Corpora Lutea ◽  
Normal Rabbit Serum ◽  
Theca Cells ◽  
Theca Interna

ABSTRACT Rabbit ovarian follicles obtained prior to and after mating and corpora lutea (24 h and 48 h post-coitus) were incubated with radioactive pregnenolone and progesterone to determine whether these substrates could serve as precursors of androgens and oestrogens. Incubations were carried out for 3 h in Hanks balanced salt solution: medium 199: normal rabbit serum: 55:30:15. Granulosa cells and corpora lutea converted pregnenolone to progesterone but no labelled oestrogens could be detected. Trace amounts of androgens were synthesized by the granulosa cells. Whole sliced follicles and theca formed progesterone from pregnenolone and androgens from both substrates. Oestradiol-17β was only synthesized by the whole sliced follicles and in one experiment by theca cells. Mating caused an increase in the 3β-hydroxysteroid dehydrogenase for pregnenolone and formation of testosterone but no oestrogens by the theca cells. These results indicate that both theca interna and granulosa cells are needed for oestrogen biosynthesis by rabbit follicles.

TESTOSTERONE PRODUCTION BY ISOLATED RABBIT FOLLICLES IN THE PRESENCE OF LUTEINIZING HORMONE AND INHIBITORS OF STEROIDOGENIC ENZYMES

1976 ◽  
Vol 82 (3) ◽  
pp. 637-643 ◽  
Author(s):  
E. V. YoungLai
Keyword(s):  
Protein Synthesis ◽  
Luteinizing Hormone ◽  
Hydroxysteroid Dehydrogenase ◽  
Ovarian Follicles ◽  
Endocrine Function ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Steroidogenic Enzymes ◽  
Partial Inhibition ◽  
Testosterone Production

ABSTRACT Rabbit ovarian follicles were incubated with luteinizing hormone (LH) and various inhibitors of steroidogenesis in order to determine what enzymes were necessary for testosterone production. Incubations were carried out in minimum Eagle's medium: normal rabbit serum: 95: 5 with medium being changed every 15 min and stored at −15° C until assayed for testosterone by radioimmunoassay. Inhibitors and LH were added at different times after the start of the incubations. Of the inhibitors tested only SU 10603, an inhibitor of the 17α-hydroxylase enzyme completely prevented the testosterone response to LH while aminoglutethimide (inhibitor of 20α-hydroxycholesterol dehydrogenase) and U 30870 (inhibitor of 3β-hydroxysteroid dehydrogenase) only showed partial inhibition. These results suggest that cholesterol is an obligatory intermediate for the production of testosterone by rabbit ovarian follicles and that normal LH stimulated endocrine function can resume after treatment with inhibitors. The results are also in agreement with previous data using inhibitors of protein synthesis in the presence of LH.

Localization of oxytocin and neurophysin in baboon (Papio anubis) corpus luteum by immunocytochemistry

1986 ◽  
Vol 113 (4) ◽  
pp. 570-575 ◽  
Author(s):  
Firyal S. Khan-Dawood
Keyword(s):  
Corpus Luteum ◽  
Rabbit Serum ◽  
Corpora Lutea ◽  
Positive Staining ◽  
Normal Rabbit Serum ◽  
Fallopian Tubes ◽  
Papio Anubis ◽  
Luteal Cells ◽  
Luteal Tissue ◽  
Immunocytochemical Reaction

Abstract. Immunoreactive oxytocin is detectable in the corpora lutea of women and cynomolgus monkeys by radioimmunoassay. To localize the presence of oxytocin and neurophysin I in ovarian tissues of subhuman primates, three corpora lutea and ovarian stromal tissues and two Fallopian tubes obtained during the menstrual cycle of the baboon and decidua from two pregnant baboons were examined using highly specific antisera against either oxytocin or neurophysin I and preoxidase-antiperoxidase light microscopy immunohistochemistry. Oxytocin-like as well as neurophysin I-like immunoreactivities were found in some cells of all the corpora lutea only, but could not be demonstrated in ovarian stromal tissues, Fallopian tubes and decidua. Specificity of the immunocytochemical reaction was further confirmed by immunoabsorption of the antiserum with excess oxytocin or neurophysin, after which the immunoreactivities for both oxytocin and neurophysin in the luteal tissue were negative. Similar controls using normal rabbit serum gave no positive staining for either oxytocin or neurophysin. Counterstaining of the positive immunoreactivities for oxytocin and neurophysin I with Mayer's haematoxylin and eosin demonstrated clearly that the oxytocin and neurophysin I appeared as granular material mainly within the cytoplasm of the luteal cells. The localization of immunoreactive oxytocin and neurophysin I in the corpus luteum of the baboon demonstrates directly the presence of these two neurohypophysial peptides within primate luteal cells and suggests their local production.

scholarly journals Transcriptome Comparisons Identify New Cell Markers for Theca Interna and Granulosa Cells from Small and Large Antral Ovarian Follicles

PLoS ONE ◽  
2015 ◽  
Vol 10 (3) ◽  
pp. e0119800 ◽  
Author(s):  
Nicholas Hatzirodos ◽  
Katja Hummitzsch ◽  
Helen F. Irving-Rodgers ◽  
Raymond J. Rodgers
Keyword(s):  
Granulosa Cells ◽  
Ovarian Follicles ◽  
Cell Markers ◽  
Theca Interna

scholarly journals Implication of cortisol and 11β-hydroxysteroid dehydrogenase enzymes in the development of porcine (Sus scrofa domestica) ovarian follicles and cysts

Reproduction ◽  
2007 ◽  
Vol 133 (6) ◽  
pp. 1149-1158 ◽  
Author(s):  
Neera Sunak ◽  
Daphne F Green ◽  
Lalantha R Abeydeera ◽  
Lisa M Thurston ◽  
Anthony E Michael
Keyword(s):  
Granulosa Cells ◽  
Hydroxysteroid Dehydrogenase ◽  
Ovarian Follicles ◽  
Cyst Fluid ◽  
Ovarian Cysts ◽  
Follicle Growth ◽  
Antral Follicles ◽  
Cortisol Metabolism ◽  
Porcine Granulosa Cells ◽  
Follicle Diameter

This study investigated cortisol inactivation by 11β-hydroxysteroid dehydrogenase (11β HSD) enzymes in porcine granulosa cells from antral follicles at different developmental stages and in ovarian cysts. In granulosa cells, cortisol oxidation increased threefold with antral follicle diameter (P < 0.001). This trend was paralleled by a threefold increase in NADP+-dependent 11β-dehydrogenase activity in granulosa cell homogenates with follicle diameter. Intact granulosa cells from ovarian cysts exhibited significantly lower enzyme activities than cells from large antral follicles. Neither intact cells norcell homogenates displayed net 11-ketosteroid reductase activities. Since porcine follicular fluid (FF) from large antral follicles and ovarian cysts contains hydrophobic inhibitors of glucocorticoid metabolism by type 1 11β HSD, this studyalso investigated whether levels of 11β HSD inhibitors changed during follicle growth and could affect cortisol metabolism in granulosa cells. The extent of inhibition of 11β HSD1 activity in rat kidney homogenates decreased progressively from 50 ± 8% inhibition by FF from small antral follicles (P < 0.001) to 23 ± 6% by large antral FF (P < 0.05). Cyst fluid inhibited 11β HSD1 activity by 59 ± 4% (P < 0.001). Likewise, net cortisol oxidation in granulosa cells was significantly decreased by large antral FF (35–48% inhibition, P < 0.05) and cyst fluid (45–75% inhibition, P < 0.01). We conclude that inactivation of cortisol by 11β HSD enzymes in porcine granulosa cells increases with follicle development but is significantly decreased in ovarian cysts. Moreover, changes in ovarian cortisol metabolism are accompanied by corresponding changes in the levels of paracrine inhibitors of 11β HSD1 within growing ovarian follicles and cysts, implicating cortisol in follicle growth and cyst development.

scholarly journals 11β-Hydroxysteroid dehydrogenase expression and activities in bovine granulosa cells and corpora lutea implicate corticosteroids in bovine ovarian physiology

2007 ◽  
Vol 193 (2) ◽  
pp. 299-310 ◽  
Author(s):  
L M Thurston ◽  
D R E Abayasekara ◽  
A E Michael
Keyword(s):  
Molecular Mass ◽  
Granulosa Cells ◽  
Luteal Phase ◽  
Hydroxysteroid Dehydrogenase ◽  
Follicular Phase ◽  
Corpora Lutea ◽  
Luteal Cells ◽  
Antral Follicles ◽  
Follicle Diameter ◽  
Dependent Type

Cortisol–cortisone metabolism is catalysed by the bi-directional NADP(H)-dependent type 1 11β-hydroxysteroid dehydrogenase (11βHSD1) enzyme and the oxidative NAD+-dependent type 2 11βHSD (11βHSD2). This study related the expression of 11βHSD1 and 11βHSD2 enzymes (mRNA and protein) to net 11-ketosteroid reductase and 11β-dehydrogenase (11β-DH) activities in bovine follicular granulosa and luteal cells. Granulosa cells were isolated from follicles of < 4, 4–8, > 8 and > 12 mm in diameter in either the follicular or luteal phase of the ovarian cycle. Luteal cells were obtained from corpora lutea (CL) in the early non-pregnant luteal phase. Enzyme expression was assessed by reverse transcription-PCR and western blotting, while enzyme activities were measured over 1 h in cell homogenates using radiometric conversion assays with 100 nM [3H]cortisone or [3H]cortisol and pyridine dinucleotide cofactors. Irrespective of follicle diameter, the expression of 11βHSD2 and NAD+-dependent oxidation of cortisol predominated in granulosa cells harvested in the follicular phase. In contrast, in granulosa cells obtained from luteal phase follicles and in bovine luteal cells, expression of 11βHSD1 exceeded that of 11βHSD2 and the major enzyme activity was NADP+-dependent cortisol oxidation. Increasing follicular diameter was associated with progressive increases in expression and activities of 11βHSD2 and 11βHSD1 in follicular and luteal phase granulosa cells respectively. In follicular phase granulosa cells from antral follicles < 12 mm, 11βHSD1 migrated with a molecular mass of 34 kDa, whereas in the dominant follicle, CL and all luteal phase granulosa cells, a second protein band of 68 kDa was consistently detected. In all samples, 11βHSD2 had a molecular mass of 48 kDa, but in large antral follicles (> 8 mm), there was an additional immunoreactive band at 50 kDa. We conclude that 11βHSD2 is the predominant functional 11βHSD enzyme expressed in follicular phase granulosa cells from growing bovine antral follicles. In contrast, in bovine granulosa cells from dominant or luteal phase follicles, and in bovine luteal cells from early non-pregnant CL, 11βHSD1 is the major glucocorticoid-metabolising enzyme. The increasing levels of cortisol inactivation by the combined NADP+- and NAD+-dependent 11β-DH activities suggest a need to restrict cortisol access to corticosteroid receptors in the final stages of follicle development.

3β-HYDROXYSTEROID DEHYDROGENASE ACTIVITY IN THE MOUSE OVARY

1965 ◽  
Vol 32 (3) ◽  
pp. 365-371 ◽  
Author(s):  
M. M. FERGUSON
Keyword(s):  
Dehydrogenase Activity ◽  
Hydroxysteroid Dehydrogenase ◽  
The Other ◽  
Interstitial Tissue ◽  
Corpora Lutea ◽  
Mouse Ovary ◽  
Colour Reaction ◽  
Theca Interna

SUMMARY Sections of ovaries from 30 Swiss white mice were incubated with ten steroid substrates to demonstrate 3β-hydroxysteroid dehydrogenase activity histochemically. The substrates were: (1) 3β-hydroxypregn-5-en-20-one (pregnenolone), (2) 3β,17α-dihydroxypregn-5-en-20-one (17α-hydroxypregnenolone), (3) 3β-hydroxyandrost-5-en-17-one (DHA), (4) 3β,17β-dihydroxyandrost-5-ene (androstenediol), (5) 3β-sulphoxypregn-5-en-20-one (pregnenolone sulphate), (6) 3β-sulphoxy-17α-hydroxypregn-5-en-20-one (17α-hydroxypregnenolone sulphate), (7) 3β-sulphoxyandrost-5-en-17-one (DHA sulphate), (8) 3β-acetoxypregn-5-en-20-one (pregnenolone acetate), (9) 3β-acetoxyandrost-5-en-17-one (DHA acetate), and (10) 3β-acetoxy-17β-hydroxyandrost-5-ene (androstenediol acetate). Pregnenolone, 17α-hydroxypregnenolone, DHA and androstenediol gave a colour reaction in the corpora lutea, interstitial tissue, theca interna and stroma of all ovaries examined. The granulosa of many follicles, some thought to be atretic, also contained diformazan granules. 17α-Hydroxypregnenolone did not give as intense a reaction as the other free steroids. No diformazan was deposited with DHA sulphate as substrate. Pregnenolone sulphate and 17α-hydroxypregnenolone sulphate were used by the same tissues as were the free steroids, although they were much less well utilized. Utilization of 3β-acetoxy derivatives was similar to that of the free steroids.

scholarly journals Expression of FOXL2 and RSPO1 in Hen Ovarian Follicles and Implication of Exogenous Leptin in Modulating Their mRNA Expression in In Vitro Cultured Granulosa Cells

Animals ◽  
2019 ◽  
Vol 9 (12) ◽  
pp. 1083 ◽  
Author(s):  
Weihe Niu ◽  
Izhar Hyder Qazi ◽  
Sichen Li ◽  
Xiaoling Zhao ◽  
Huadong Yin ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Granulosa Cells ◽  
Ovarian Follicles ◽  
Control Group ◽  
Follicle Development ◽  
Theca Cells ◽  
Increasing Trend ◽  
Group A ◽  
Ovulatory Follicles

In this study, using a laying hen model, we determined the expression of FOXL2 and RSPO1 in different central and peripheral tissue and ovarian follicles at different stages of development. At the same time, mRNA expression of both genes in granulosa and theca cells harvested from follicles at different stages of folliculogenesis was also evaluated. Finally, we assessed the effect of leptin treatment on expression of FOXL2 and RSPO1 in in vitro cultured granulosa cells harvested from 1–5 mm to F3–F1 follicles. Our RT-qPCR results revealed that a comparatively higher expression of FOXL2 and RSPO1 was observed in ovary, hypothalamus, and pituitary. Abundant mRNA expression of FOXL2 was observed in small prehierarchical follicles (1–1.9 and 2–2.9 mm follicles; p < 0.05), whereas mRNA expression of RSPO1 showed an increasing trend in large hierarchical follicles (F5–F1), and its abundant expression was observed in post-ovulatory follicles. FOXL2 mRNA expression was stable in granulosa cells harvested from 3–5 mm to F4 follicles, and exhibited a significantly higher expression in large hierarchical follicles. Conversely, relatively low mRNA expression of FOXL2 was observed in theca cells. RSPO1 mRNA expression was relatively lower in granulosa cells; however, theca cells exhibited a significantly higher mRNA expression of RSPO1 in F4 to F1 follicles. In the next experiment, we treated the in vitro cultured granulosa cells with different concentrations (1, 10, 100, and 1000 ng/mL) of exogenous leptin. Compared to the control group, a significant increase in the expression of FOXL2 was observed in groups treated with 1, 10, and 100 ng/mL leptin, whereas expression of RSPO1 was increased in all leptin-treated groups. When treated with 100 ng/mL leptin, FOXL2 and RSPO1 expression was upregulated in cultured granulosa cells harvested from both large hierarchical (F3–F1) and small prehierarchical follicles (1–5 mm). Based on these findings and evidence from mainstream literature, we envisage that FOXL2 and RSPO1 genes (in connection with hypothalamic-hypophysis axis) and leptin (via modulation of FOXL2 and RSPO1 expression) might have significant physiological roles, at least in part, in modulating the ovarian mechanisms, such as follicle development, selection, and steroidogenesis in laying hens.

Immunohistochemical localization of 17α-hydroxylase/C17–20 lyase and aromatase cytochrome P-450 in the human ovary during the menstrual cycle

1992 ◽  
Vol 135 (3) ◽  
pp. 589-NP ◽  
Author(s):  
T. Tamura ◽  
J. Kitawaki ◽  
T. Yamamoto ◽  
Y. Osawa ◽  
S. Kominami ◽  
...  
Keyword(s):  
Menstrual Cycle ◽  
Corpus Luteum ◽  
Granulosa Cells ◽  
Polyclonal Antibodies ◽  
Immunohistochemical Localization ◽  
Corpora Lutea ◽  
Human Ovary ◽  
Normal Human ◽  
Theca Interna ◽  
Cytochrome P 450

ABSTRACT Immunohistochemical localization of 17α-hydroxylase/C17–20 lyase (P-45017α,lyase) and aromatase cytochrome P-450 (P-450arom) in normal human ovaries during the menstrual cycle was studied using specific polyclonal antibodies which were raised against corresponding enzymes. In the follicular phase of matured follicles, P-45017α,lyase was localized in theca interna cells and P-450arom in granulosa cells. P-45017α,lyase was expressed in theca interna cells before P-450arom was expressed in granulosa cells. The corpus luteum showed immunoreactivity to both enzymes and, after menstruation, immunoreactivity decreased gradually until it could not be detected in the corpus albicans. In corpus luteum graviditatis the immunoreactivity continued to be expressed strongly. In some atretic follicles, P-45017α,lyase and/or P-450arom continued to be expressed. In the stromal layer, P-45017α,lyase was detected in secondary interstitial cells, which originated from the theca interna of atretic follicles, and P-450arom was detected in hilar cells. Immunoreactivity to both enzymes was also detected in oocytes of developing follicles. These results are consistent with the two cell theory in the human ovary. They also suggest that androgens and oestrogens are produced not only by follicles and corpora lutea but also by stroma and oocytes. Journal of Endocrinology (1992) 135, 589–595

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