MATURATIONAL CHANGES IN SHEEP OVARIAN FOLLICLES: GONADOTROPHIC STIMULATION OF CYCLIC AMP PRODUCTION BY ISOLATED THECA AND GRANULOSA CELLS

1978 ◽  
Vol 89 (1) ◽  
pp. 166-172 ◽  
Author(s):  
T. J. Weiss ◽  
D. T. Armstrong ◽  
J. E. A. McIntosh ◽  
R. F. Seamark
Keyword(s):  
Cyclic Amp ◽  
Granulosa Cells ◽  
Follicle Stimulating Hormone ◽  
Adenosine Monophosphate ◽  
Ovarian Follicles ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Camp Production ◽  
Stimulation Of ◽  
Stimulating Hormone

ABSTRACT Theca and granulosa tissues isolated from sheep ovarian follicles of different sizes were incubated in the presence of human chorionic gonadotrophin (HCG; 5 IU/ml) or follicle stimulating hormone (FSH; 5 μg NIH-FSH-S11/ml) for 40 min. Changes in the total amounts of cyclic 3′,5′-adenosine monophosphate (cAMP) were used as an index of the responsiveness of these preparations to the hormones. Thecal tissue of both large (4–6 mm in diameter) and small (1–3 mm) follicles responded similarly to gonadotrophins. Granulosa cells from small follicles failed to respond to stimulation by HCG. FSH, however, consistently increased cAMP production in comparison with controls or cells treated with HCG. Granulosa cells of large follicles responded to both HCG and FSH.

FOLLICLE STIMULATING HORMONE-LIKE ACTIVITY IN HUMAN CHORIONIC GONADOTROPHIN PREPARATIONS

1969 ◽  
Vol 60 (1) ◽  
pp. 137-156 ◽  
Author(s):  
C. Robyn ◽  
P. Petrusz ◽  
E. Diczfalusy
Keyword(s):  
Follicle Stimulating Hormone ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Female Rats ◽  
Immature Female ◽  
Ovarian Weight ◽  
Wide Range ◽  
Immature Rats ◽  
Stimulation Of ◽  
Stimulating Hormone

ABSTRACT The follicle stimulating hormone (FSH)-like activity of human chorionic gonadotrophin (HCG) preparations was assayed by the method based on the ovarian weight augmentation in intact immature rats. The potencies ranged from 4.8 to 7.4 IU equivalents of FSH per mg. The FSH-like potency of the Second International Standard Preparation of HCG was 8.5 IU per vial. However, when in intact immature rats the ovarian weight response to HCG preparations was compared at a wide range of doses (40 to 51 200 IU) to that obtained with a human menopausal gonadotrophin (HMG) preparation (0.5 to 128 IU of FSH) in the presence of 40 IU of HCG, significant differences were found. The assays conducted in hypophysectomised immature female rats were invalid, because of lack of parallelism. Antisera were prepared by immunising rabbits with HCG and human hypophysial gonadotrophin (HHG) preparations and the antigonadotrophin profiles (HCG-, FSH- and FSH-like neutralising potencies) of these antisera were established by the use of statistically valid bioassay procedures. The anti-HCG and anti-HHG sera neutralised the FSH activity of HMG preparations as well as the FSH-like activity of HCG preparations. However, 3 to 175 times more antiserum was required to neutralise the equivalent of 1.0 IU of FSH-like activity present in HCG than expected on the basis of the anti-FSH potency of the antisera. On the other hand, there was a high degree of correlation between the neutralising potencies of the antisera when tested against the FSH-like activity and the HCG activity of various HCG preparations. When the FSH-like activity of an HCG preparation was quantitatively neutralised with an anti-HCG serum, some 30 per cent of the HCG activity remained unneutralised, as evidenced by repeated bioassays. Although at least 2000 IU of this »FSH-free« HCG was administered to groups of intact as well as hypophysectomised immature female rats, this high dose of HCG did not induce an increase in ovarian weight beyond that elicited by 40 IU of untreated HCG. Histological examination of the ovaries indicated lack of follicle stimulation in the hypophysectomised, but not in the intact immature animals. There was an excessive stimulation of the interstitial cells in both types of animals. The data indicate that the FSH-like activity of HCG preparations is neither due to a contamination by FSH of pituitary origin, nor is it an evenly distributed intrinsic property of the HCG molecules. It is also concluded that the gonadotrophic activity of biologically pure HCG in immature hypophysectomised female rats consists of a specific stimulation of the interstitial cell apparatus. Such HCG preparations do not induce any follicle stimulation, not even when administered in excessive doses.

Therapeutic use of gonadotrophins and clomiphene

1969 ◽  
Vol 7 (9) ◽  
pp. 33-35
Keyword(s):  
Pregnant Women ◽  
Follicle Stimulating Hormone ◽  
Therapeutic Use ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Human Pituitary ◽  
Synthetic Compound ◽  
Pituitary Glands ◽  
Post Menopausal ◽  
Stimulating Hormone

The three substances now used to stimulate the gonads in infertility are human follicle stimulating hormone (HFSH) obtained mainly from post-menopausal urine, but also from human pituitary glands, human chorionic gonadotrophin (HCG) extracted from the urine of pregnant women, and clomiphene (Clomid - Merrell), a synthetic compound which we reviewed in 1967.1

ANTIGONADOTROPHIC PROFILE OF ANTISERA AGAINST HUMAN GONADOTROPHIN PREPARATIONS

1971 ◽  
Vol 67 (2) ◽  
pp. 262-276 ◽  
Author(s):  
P. Petrusz ◽  
C. Robyn ◽  
E. Diczfalusy
Keyword(s):  
Biological Activity ◽  
Luteinizing Hormone ◽  
Total Dose ◽  
Follicle Stimulating Hormone ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Immunological Activity ◽  
Human Menopausal Gonadotrophin ◽  
Stimulating Hormone

ABSTRACT Forty-two antisera were prepared in rabbits against human chorionic gonadotrophin (HCG), human hypophysial gonadotrophin (HHG), human urinary luteinizing hormone (LH) and human menopausal gonadotrophin (HMG) preparations. The gonadotrophic profiles of the antigens were previously characterized by bioassay, immunoassay and bioimmunoassay methods. The 25 most potent antisera were tested in statistically valid bioassays for their HCG and follicle stimulating hormone (FSH) neutralizing activities as well as for their neutralizing potencies against the FSH-like activity present in HCG preparations. The anti-HCG/anti-FSH ratios of the anti-HCG sera tested varied between 6.2 and > 254, while those of the anti-HHG, anti-LH and anti-HMG sera were close to 2. It was found that the total dose of immunological activity (anti-HCG neutralizing and anti-FSH neutralizing potency) rather than that of the biological activity administered to the rabbits was decisive for obtaining antisera with high anti-HCG and anti-FSH titers. Immunization with a highly purified HCG preparation (> 17 000 IU/mg) resulted in antisera exhibiting lower anti-HCG/anti-FSH ratios than did immunization with partially purified preparations. A highly purified urinary LH preparation which did not contain any detectable FSH activity gave rise to antisera exhibiting anti-HCG/anti-FSH ratios of approximately 2.0. These highly purified HCG and LH preparations were shown previously to possess high anti-FSH neutralizing potencies (Petrusz et al. 1971b). Booster injections did not change significantly the quality or the titer of the antigonadotrophic sera studied. The HCG neutralizing potency of anti-HCG sera was approximately 3 times higher when assayed against a highly purified HCG preparation (> 17 000 IU/mg) as compared to potency estimates obtained against the laboratory standard of HCG (about 2000 IU/mg). It is suggested that consideration should be given to the establishment of standard preparations of antigonadotrophic sera. It is concluded that bioimmunoassays are more suitably than conventional bioassay methods for the assessment of the antigenic purity of human gonadotrophin preparations.

CXADR-like membrane protein (CLMP) in the rat ovary: stimulation by human chorionic gonadotrophin during the periovulatory period

2016 ◽  
Vol 28 (6) ◽  
pp. 742
Author(s):  
Feixue Li ◽  
Xiaoping Miao ◽  
Yonglong Chen ◽  
Thomas E. Curry
Keyword(s):  
Cell Adhesion ◽  
Protein Kinase ◽  
Membrane Protein ◽  
Mrna Expression ◽  
Granulosa Cells ◽  
Receptor Activation ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Rat Ovary ◽  
Stimulation Of

CXADR-like membrane protein (CLMP) is a novel cell–cell adhesion molecule. The present study investigated the spatiotemporal expression pattern of CLMP and its regulation in the rat ovary during the periovulatory period. Real-time polymerase chain reaction analysis revealed that Clmp mRNA was rapidly stimulated in intact ovaries by 4 h after human chorionic gonadotrophin (hCG) treatment. In situ hybridisation analysis demonstrated that Clmp mRNA expression was stimulated in theca cells at 4 h after hCG and remained elevated until 12 h. Clmp mRNA was also upregulated in granulosa cells and was present in forming corpora lutea. Our data indicate that the protein kinase A but not the protein kinase C pathway regulates the expression of Clmp mRNA in granulosa cells. Phosphatidylinositol 3 kinase and p38 kinase are also involved in regulating Clmp mRNA expression. The stimulation of Clmp mRNA by hCG requires new protein synthesis. Furthermore, inhibition of epidermal growth factor receptor activation significantly inhibited Clmp mRNA expression, whereas inhibition of prostaglandin synthesis or progesterone action had no effect. The stimulation of CLMP in the rat ovary may be important in cell adhesion events during ovulation and luteal formation such as maintaining the structure and communication of ovarian follicular and luteal cells.

345 EFFECT OF FOLLICLE STIMULATING HORMONE AND HUMAN CHORIONIC GONADOTROPHIN ON THE IN VITRO MATURATION OF CANINE OOCYTES

2007 ◽  
Vol 19 (1) ◽  
pp. 288
Author(s):  
M.-K. Kim ◽  
H.-J. Oh ◽  
Y. H. Fibrianto ◽  
G. Jang ◽  
H.-J. Kim ◽  
...  
Keyword(s):  
Estrous Cycle ◽  
Uv Light ◽  
Follicle Stimulating Hormone ◽  
Reproductive Hormones ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Nuclear Maturation ◽  
Maturation Rate ◽  
Stimulating Hormone

Growth, maturation, and ovulation of the Graafian follicle depend on appropriate patterns of secretion, sufficient concentrations, and adequate ratios of various reproductive hormones, especially follicle stimulating hormone (FSH) and luteinizing hormone (LH) (Van Tol et al. 1996 Mol. Reprod. Dev. 45, 218–224). The present study investigated the effects of FSH and human chorionic gonadotrophin (hCG) on the nuclear maturation of canine oocytes. In addition, in order to investigate the effect of stage of the estrous cycle on the meiotic competence of canine oocytes matured in vitro, oocytes were collected from various reproductive states and matured in vitro in the presence of the gonadotrophins. Estrous cycle stage was evaluated for each bitch by ovarian morphology, and bitches were categorized according to the stage of the estrous cycle (anestrus, follicular, or diestrus) prior to oocyte collection. Recovered oocytes were cultured in serum-free tissue culture medium (TCM)-199 supplemented with various concentrations of FSH (Exp. 1: 0, 0.1, 1.0, or 10 IU) or hCG (Exp. 2: 0, 0.5, 1.0, or 10 IU) or both (Exp. 3: 1 IU FSH + 1 IU hCG) for 72 h to determine the effective concentration of these hormones, and to examine their combined effect. After maturation culture, oocytes were denuded in PBS containing 0.1% (w/v) hyaluronidase by gentle pipetting. The denuded oocytes were stained with Hoechst 33342 in glycerol and the nuclear state of oocytes was evaluated under UV light. The rates of maturation to the MII stage were significantly higher (P < 0.05) when follicular-stage oocytes were supplemented with 1 IU FSH (6.2%) compared with the other FSH-supplemented groups (0.0 to 3.3%) or to the control (1.8%), or 0.1 or 10.0 IU FSH (0 to 1.2%). Significantly higher (P < 0.05) maturation rate to MII stage was observed in follicular-stage oocytes supplemented with 1.0 IU hCG (7.2%) compared with the control or other hCG-supplemented groups (0 to 1.5%). However, FSH and hCG together did not improve the nuclear maturation rate of canine oocytes (2.4%) compared with FSH (6.2%) and hCG alone (7.2%). In conclusion, FSH or hCG alone significantly increased the maturation of canine oocytes to the MII stage. This work was supported by grant No. M1062503005-06N250300510 from KOSEF, Republic of Korea.

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