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

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.

1972 ◽  
Vol 54 (2) ◽  
pp. 277-284 ◽  
Author(s):  
H. M. A. MEIJS-ROELOFS

SUMMARY Electrical stimulation of the hypothalamus with biphasic pulses was performed in immature female rats. When performed at 27 days of age or later, electrical stimulation in the arcuate nucleus region advanced puberty in all animals, as did stimulation of the anterior hypothalamus at 29 days of age or later. Stimulation in younger rats did not uniformly advance puberty. The responsiveness to electrical stimulation thus seems to develop a few days earlier in the arcuate nucleus region than in the anterior hypothalamus. In a second experiment the possible involvement of follicle-stimulating hormone (FSH) in the advancement of puberty was investigated: the simplified augmented ovarian weight assay for endogenous FSH was performed in rats stimulated in the arcuate nucleus region as well as in controls. A marked increase in ovarian weight, indicating increased FSH levels, was demonstrated in all animals stimulated on day 27 or later; at earlier ages only a percentage of the stimulated animals responded. This percentage paralleled the percentage of animals that showed advancement of puberty. It is concluded that electrical stimulation in both the arcuate nucleus region and the anterior hypothalamus advances the onset of puberty. It is suggested that electrical stimulation causes increased plasma FSH levels and, in consequence, precocious puberty.


1978 ◽  
Vol 89 (1) ◽  
pp. 166-172 ◽  
Author(s):  
T. J. Weiss ◽  
D. T. Armstrong ◽  
J. E. A. McIntosh ◽  
R. F. Seamark

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.


1966 ◽  
Vol 35 (2) ◽  
pp. 199-206 ◽  
Author(s):  
P. S. BROWN ◽  
M. WELLS

SUMMARY The follicle-stimulating hormone (FSH) content of urinary gonadotrophic extracts was assayed by its effect on the ovarian weight of immature mice when given in conjunction with 40 i.u. human chorionic gonadotrophin. About three-quarters of all routine assays gave values of λ between 0·15 and 0·30. Precision was slightly increased when the material was given in three rather than in five injections. Correction of ovarian weight for body weight was either invalid or of no value in reducing variance. Removal of between-litter variance increased precision considerably. Mice of three randomly bred colonies were all satisfactory, and inbred C57BL mice were also suitable for the assay. C3H mice were less sensitive. The efficiency of different methods of extracting FSH from urine was examined. The method of Johnsen (1958) using precipitation with tannic acid was considered the most satisfactory and gave extracts of high potency and low bulk. Limited experiments in which purified human pituitary FSH was assayed with and without added luteinizing hormone, gave results compatible with the assumption that the method is specific for FSH.


1957 ◽  
Vol 16 (1) ◽  
pp. 107-113 ◽  
Author(s):  
W. R. BUTT ◽  
A. C. CROOKE ◽  
JOYCE D. INGRAM ◽  
BRENDA P. ROUND

SUMMARY 1. Follicle stimulating hormone (FSH) has been obtained from the urine of pregnant women. 2. It was prepared by adsorption on kaolin from urine which had been treated with benzoic acid to remove excess human chorionic gonadotrophin (HCG) and was assayed by the procedure which depends on the increase in ovarian weight of immature mice treated simultaneously with HCG. 3. Preliminary results are given for the assay of FSH in normal pregnancy.


1955 ◽  
Vol 13 (1) ◽  
pp. 59-64 ◽  
Author(s):  
P. S. BROWN

SUMMARY Two convenient bioassays of urinary gonadotrophins, using immature mice, are described. The first is based upon the initial doubling of uterine weight. The second, using the ovarian weight response, attempts to increase specificity to follicle stimulating hormone by priming with human chorionic gonadotrophin. The usefulness of both methods is discussed, and the influence of non-specific impurities during the assay of urinary extracts is stressed.


1985 ◽  
Vol 106 (1) ◽  
pp. 61-66 ◽  
Author(s):  
H. M. A. Meijs-Roelofs ◽  
P. Kramer ◽  
P. Osman

ABSTRACT Precocious first ovulation, preceded by an endogenous preovulatory LH surge, could be predictably induced in immature female rats by administering repeated injections of human chorionic gonadotrophin (hCG). Administration of a dose of 0·05–0·075 i.u. hCG, four times a day from day 28 to day 31 of age resulted in a highly constant ovulatory response: at 4·0±0·0 days after the start of treatment 7·7±0·3 (n = 15) ova were found. Use of a higher dose of hCG (0·1 i.u.) resulted in lower numbers of ova (5·6±0·4, n = 7; P<0·005) whereas use of a lower dose of hCG (0·025–0·038 i.u.) resulted in a less constant timing of the induced ovulation at 5·4±0·2 days after the start of treatment (n = 7; P<0·0005). In animals treated with the dose of 0·05–0·075 i.u. hCG, a positive correlation was found between body weight at the start of treatment and the number of ova released (r = 0·75, n = 25; P<0·001). Ovarian follicle dynamics were studied on the various days of hCG treatment (dose 0·05–0·075 i.u.) and compared with the follicle changes that take place after electrical stimulation of the hypothalamus, performed on day 28, a treatment known to result in first ovulation 4–5 days later. In both groups a decrease in the number of the smallest and the middle-sized antral follicles as compared with their respective controls was seen, whereas numbers of follicles in the largest, 'ovulatable' size classes gradually increased. The pattern was more conspicuous in the hCG-treated group, presumably related to greater constancy in timing of the ovulatory response in this group. The present data support the view that endogenous changes in LH secretion during late prepuberty (which have been found to take place) play a significant role in stimulating late-prepubertal follicle growth and the ensuing first ovulation. J. Endocr. (1985) 106, 61–66


1969 ◽  
Vol 45 (1) ◽  
pp. 29-36 ◽  
Author(s):  
R. H. NAQVI ◽  
D. C. JOHNSON

SUMMARY An increase in ovarian weight in immature rats after the injection of chorionic gonadotrophin (HCG) was used to measure variations in endogenous follicle-stimulating hormone (FSH) after steroid treatment. A single injection of several steroids (testosterone, androstenediol, androstenedione, oestradiol benzoate) given 12–96 hr. before treatment with HCG caused a 30–200% increase in ovarian weight. This was not a direct effect of the steroids since hypophysectomy abolished the response, and administration of the compounds concurrently with HCG was ineffective. Within certain limits an increase in the duration of pretreatment enhanced the ovarian response while an increase in the dose of steroid had little effect. Pretreatment with testosterone propionate did not change pituitary FSH activity, indicating that the increase in circulating FSH was due to an increased production of hormone. On the other hand, pituitary FSH in animals treated with oestradiol benzoate was significantly lowered within 72 hr. suggesting an increased release of FSH.


1961 ◽  
Vol 38 (2) ◽  
pp. 181-199 ◽  
Author(s):  
K. Walter ◽  
S. Wysocki

ABSTRACT The activity of chorionic gonadotrophin (HCG) injected in solutions containing different nonspecific substances (polyvinylpyrrolidone, gelatine, tragacanth = »depot-media« is compared with the activity of HCG in distilled water. Intact immature female rats are used for the bioassays, which differ only in injection schedule or the response measured (uterine resp. ovarian weight). Depending on the injection schedule used and the response measured the results of the various bioassays in intact immature rats are different, when one and the same depot-medium is used. Significant indices of discrimination as well as significant slope differences of dose-response curves are observed. The addition of a biologically inactive kaolin extract prepared according to the »crude kaolin-acetone method« to a gonadotrophic extract from the urine of postmenopausal women produced a significant decrease in activity in one type of assay, and a significant slope difference of dose-response curves in another one. In view of these results the difficulties arising for the demonstration of qualitative differences in gonadotrophic preparations and for expressing bioassay-results in terms of the International Reference Preparation are discussed.


1970 ◽  
Vol 63 (2) ◽  
pp. 275-282
Author(s):  
E. T. Bell ◽  
D. W. Christie

ABSTRACT Assays of follicle-stimulating hormone from the National Institutes of Health (NIH-FSH-S4) and the Second International Reference Preparation for Human Menopausal Gonadotrophin (IRP-HMG) have been conducted by the mouse ovarian augmentation test in animals of nine strains from five mouse breeders. Two groups of 50 mice from each colony were used to assay NIH-FSH and the Second IRP-HMG. In the experiment with NIH-FSH dosages of 37.5 to 300.0 μg were given together with 40 IU human chorionic gonadotrophin (HCG) in three or five sc injections over three days. When the Second IRP-HMG was assayed dosages of 0.19 to 1.5 IU were administered in five injections with 20 or 40 IU HCG. Little or no ovarian weight increase occurred following NIH-FSH in six out of nine colonies. In the remaining three the index of precision (λ) was very high. Following administration of the Second IRP-HMG a greater increase in ovarian weight occurred but a satisfactory slope was noted in only three colonies. The λ figures were generally lower than with NIH-FSH. It is concluded, that under the conditions used, six out of the nine colonies were not suitable for the assay of FSH by the ovarian augmentation test. Further work would be required to study the reliability criteria of the assay in the remaining three colonies.


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