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Induction and inhibition of implantation in lactating rats.

The interrelationship between prolactin and LH in the maintenance of pregnancy during lactation was studied. The reduction of suckled young from eight to two or less, as late as on the morning of Day 4 of pregnancy, resulted in normal implantation. Reintroduction of eight young on Day 4 to lactating pregnant rats deprived of their litters on Day 1 resulted in an inhibition of implantation, but reintroduction on Days 5 or 6 did not inhibit implantation. If oestrogen, HCG or PMSG was given on Day 4 of pregnancy, implantation was induced at the normal time in rats suckling large litters. When LH antiserum was given on the morning of Day 4 or Day 8 to pregnant rats suckling two young each, it blocked implantation and postimplantation survival of blasto-cysts, respectively. When the number of suckling young was increased from two to eight on Day 6, however, LH antiserum blocked pregnancy only to the extent of 70%. Prolactin administered during the preimplantation phase inhibited implantation in pregnant rats suckling a minimum number of young, but had no effect when given during the postimplantation phase. Progesterone failed to block implantation. Prolactin had no inhibitory effect on implantation in the absence of the suckling stimulus or in non-lactating pregnant rats. The inhibition of implantation by prolactin in rats suckling two young could be effectively reversed by the administration of oestrogen, PMSG or HCG on Day 4 of pregnancy.

Animals↗

Sex hormone-dependent brain maturation and sexual behaviour in rats.

In male and female rats the endogenous steroid and gonadotrophin secretion was inhibited by injecting high doses of chlormadinone acetate (CmAc) from day 14 to 24 of life, i. e. during the period of brain maturation. In adulthood the males treated prepubertally with CmAc exhibited reduced sexual activity and fertility, whereas the females did not differ from the controls. More complete sex hormone deficiency during brain maturation was achieved by castration on day 14 of life. Controls were castrated at normal puberty time (40--60 days). Both groups were then substituted with androgens or oestrogens. In the females castrated on day 14 no impairment of sexual behaviour was observed as compared to the later castrated controls. In contrast, the early castrated males showed delayed onset of mounting behaviour. At autopsy, the weights of their sex organs were found to be lower than in the controls despite equal testosterone replacement for several months. These findings speak in favour of a permanently diminished responsiveness to androgens in males having been exposed to more or less severe androgen deficiency during sex specific brain maturation. Hence, the maturation of a male hypothalamus as well as the differentiation appears to depend at least in part on the presence of androgens, whereas in females it runs without hormonal influence.

Animals↗

Use of serum lutenizing hormone in the clinical management of short-term amenorrhea.

Serum luteinizing hormone (LH) values determined by radioimmunoassay can be of aid in evaluating patients with short-term amenorrhea. These values have proved useful in ruling out pregnancy in cases of hypothalamic amenorrhea and amenorrhea occuring during treatment with contraceptive pills and also in diagnosing early pregnancy after clomiphene therapy.

Adult↗

Effects of aging on the hypothalamic-hypophyseal-gonadal axis in female rats.

The failure of reproductive function in aged rats could be due to deficiencies at the level of the ovary, pituitary, hypothalamus, or higher brain centers. The classic explanation that the ovary is depleted of follicles does not receive adequate support on the basis of histologic studies of aged ovaries. Basal serum gonadotropin levels change with increasing age in female rats. Serum follicle-stimulating hormone (FSH) levels rise while serum luteinizing hormone (LH) levels fall. Likewise, the characteristic response to castration is markedly altered in aged female rats with a reduced secretion of FSH and a minimal elevation of LH. However, the pituitaires of these animals are still caapable of responding to exogenous LH-releasing hormone with a delayed LH response whose magnitude simulates that seen in younger female animals. With increasing age there is decreased pituitary and/or hypothalamic sensitivity to the feedback action of estradiol. These data are consistent with the postulation that there is an altered hypothalamic-hypophyseal function in aged rats.

Aging↗

Appearance of hCG in pregnancy plasma following the initiation of implantation of the blastocyst.

Plasma chorionic gonadotropin levels were measured by three different assay methods during early pregnancy in four patients following induction of ovulation with Pergonal and hCG. Radioligand-receptor assay of unextracted samples was subject to non-specific interference by plasma proteins, causing an apparent elevation of gonadotropin levels during the first few days after fertilization. By contrast, the gonadotropin values measured by a highly sensitive LH/hCG bioassay were consistent with those obtained with a sensitive and specific radioimmunoassay for hCG, and showed that the first significant rise in plasma hCG occurred 9 to 13 days after ovulation. These results indicate that hCG does not appear in the maternal circulation until after the initiation of implantation of the blastocyst.

Biological Assay↗

Levels of follicle-stimulating hormone, luteinizing hormone, oestradiol-17 beta and progesterone, and follicular growth in the pseudopregnant rat.

Throughout a period of pseudopregnancy the peripheral blood levels of progesterone, oestradiol-17 beta, follicle-stimulating hormone (FSH) and luteinizing hormone (LH), as well as the size-distribution of ovarian antral follicles were estimated in the rat. The progesterone concentrations, as measured by a competitive protein-binding technique, exceeded metoestrous values (25 ng/ml plasma) from day 3 of pseudopregnancy onwards. The highest levels were found on days 6 and 8 (91 ng/ml). From day 8 onwards the levels decreased gradually but were still above metoestrous values on the day of pro-oestrus after pseudopregnancy. Concentrations of oestradiol-17 beta, as measured by radioimmunoassay, were within the range of those at metoestrus (about 5 pg/ml plasma) until day 10. Thereafter levels increased to a value of 57 pg/ml. Concentrations of FSH, measured by radioimmunoassay, were within the range of metoestrous values until day 10 (about 100 ngNIAMD-rat-FSH RP-1/ml serum), but declined to a level of 33 ng/ml on day 12. Concentrations of LH, measured by radioimmunoassay, were generally within the wide range of metoestrous values (9-60 ng NIAMD-rat-LH RP-1/ml serum), but concentrations found on days 4, 8 and 10 were significantly lower than those found on preceding or subsequent days. Histological determination of the number of follicles present in various volume-classes, showed an increase in antral follicles on days 1 and 2, comparable to the increase observed during metoestrus and dioestrus 1 of the normal cycle. There was no change in the follicles between days 3 and 10 and they resembled those of early dioestrus. Preovulatory growth had occurred by day 12. Injection of human chorionic gonadotrophin (HCG) on days 2, 4 or 6 showed that ovulation could be induced only in some of the larger follicles. On the basis of these results it is suggested that during pseudopregnancy the high progesterone levels present result in a decreased plasma LH level which is insufficient to cause full maturation of the follicles and to stimulate oestrogen secretion to the levels required for induction of an ovulatory surge of LH release.

Animals↗

Induction of lactogenic receptors. II. Studies on the liver of hypophysectomized male rats and on rats bearing a growth hormone secreting tumor.

The role of growth hormone (GH), as well as prolactin and ACTH, in the induction of the PRL receptor was investigated both in hypophysectomized male rat livers and in the livers of male rats bearing a GH secreting tumor. After 7 days of s.c. injections, specific binding (% SB) of PRL in controls and rats treated with oPRL, hGH, ACTH, hCG, estradiol (E2), or testosterone (T) was approximately 1%. Treatment with oPRL plus ACTH increased SB to 4%; adding E2 to this combination produced a further increase to 8%, whereas the addition of T decreased hepatic binding to 1%. Combination of hGH with ACTH was most effective, giving a SB of 33%, which is similar to that observed in the liver of rats bearing a GH secreting tumor (36%). These studies suggest that GH acts synergistically with PRL and/or ACTH to increase lactogenic binding sites in the male rat liver and that sex steroids have a modulating effect on this action.

Adrenocorticotropic Hormone↗

Peripheral plasma progesterone during egg transport in the rabbit.

Peripheral plasma progesterone concentrations were measured in New Zealand rabbits every 6 hr beginning 12 hr before and continuing until 96 hr after either natural mating, hCG injection, or saline injection. The number of ovulation points in naturally mated animals (9.3 +/- 0.6, mean +/- SE) was not significantly different from that in hCG-injected animals (8.6 +/- 1.5). There was a surge in progesterone secretion following both mating and hCG injection. Plasma progesterone concentrations reached a peak prior to ovulation and then fell to basal levels at the time of ovulation. Beginning at approximately 30 hr after the ovulation-inducing stimulus, there was a progressive, significant (P less than 0.001) increase in plasma progesterone concentration, which continued for the duration of the sampling period. The initiation of the postovulatory increase in progesterone secretion corresponds temporally with the movement of eggs from the ampullary-isthmic junction into the isthmus. The progressive increase in plasma progesterone between 30 and 72 hr after the induction of ovulation corresponds with the gradual movement of eggs through the isthmus into the uterus. The data suggest that movement of eggs through the oviductal isthmus is influenced by the postovulatory secretion of progesterone.

Animals↗

Determinantion of testosterone concentration in semen of men with normal or subnormal sperm counts and after vasectomy.

Semen from 58 male subjects, aged 22 to 50, was assayed on an individual basis to determine whether T was present in it. Of the subjects examined 23 were normospermic, 14 oligospermic and 9 azoospermic; 12 men had undergone vasectomy were also included in the study. In 39 of the subjects plasma testosterone was estimated. A competitive protein binding technique was employed for T assays while dried extracts of semen were examined by combined gas-chromatography-mass spectrometry and mass fragmentography. Measurable amounts of T were detected in all seminal specimens assayed. This was confirmed by gas chromatography-mass spectrometry which showed a spectrum suggestive of T. The ratio of unconjugated to conjugated steroid in semen was found to be approximately 1:10. Levels of unconjgated T were similar to those found in plasma of normally menstruating women. The mean seminal concentration of unconjugated T (+/-SD) in the specimens assayed was 0.71 ng/ml+/-0.08 for the normospermic, 0.79+/-0.14 for the ezoospermic, 0.69+/-0.09 for the oligospermic, but only 0.38+/-0.04 for the vasectomized subjects. Plasma levels for this androgen were within the range found in normal men of comparable age. Significant correlation between plasma and seminal T concentration could not be demonstrated and there was no correlation between either of the above parameters and the seminal volume, the number, abnormal form percentage and the motility of spermatozooa in the normo--or oligospermic group. However, when the two groups were pooled into one, significant correlations were found between plasma, (but not seminal T concentration) and the seminal characters examined, perhaps suggesting the number of specimens from the groups should be increased to obtain valid data. Administration of human chorionic gonadotropin produced a marked plasma response as well as a rise of seminal T levels in 3 normospermic subjects whereas cyproterone acetate caused reduction of plasm T levels but had no consistent effect on the seminal concentration of ts steroid although the sensitivity of the seminal method may not have detected smaller changes at this level.

Adult↗

Role of carbohydrate of human chorionic gonadotropin in the mechanism of hormone action.

The role of the carbohydrate part of human chorionic gonadotropin (hCG) was investigated by measuring the ability of hCG derivatives lacking various sugar residues to bind to rat Leydig cells and stimulate them to synthesize testosterone and cyclic adenosine 3':5'-monophosphate (cyclic AMP). Whereas sequential removal of the sialic acid, galactose, N-acetylglucosamine, and mannose residues led to a progressive increase in the effective dose of the hormone required to stimulate steroidogenesis, it resulted in a marked loss in the ability of the hormone to stimulate cyclic AMP accumulation. Low doses of the glycosidase-treated hormone derivatives were additive with hCG when their ability to stimulate testosterone synthesis was analyzed. Nevertheless, the glycosidase-treated derivatives were potent inhibitors of hCG-induced cyclic AMP accumulation, suggesting that removal of the sugars did not influence binding of the hormone to the cell as much as it reduced the ability of the bound hormone to activate adenyl cyclase. This hypothesis was further supported by our finding that the hCG derivatives were highly effective inhibitors of 125I-hGC binding to the intact cells. Removal of sialic acid and galactose enhanced the inhibition, whereas removal of all the sugar residues only decreased the inhibition slightly. The degree of these effects was comparatively small. The possibility that steroidogenesis and cyclic AMP accumulation are altered independently by hCG stimulation is discussed.

Acetylglucosaminidase↗

Pregnant mare's serum gonadotropin. III. Hemispaying and the reversal of the antifertility faculty of pregnant mare's serum gonadotropin.

A single injection of 10 IU of pregnant mare's serum gonadotropin (PMSG) on day 5 of pregancy caused wastage of the fetoplacental unit by day 16 of pregnancy. Semispaying, which seems to subtract approximately 50% of the ovarian steroid contribution, at 24, 48, and 72 hours following the PMSG schedule prevented the antifertility faculty of the hormone preparation. However, hemicastration at 96 hours following the gonadotropin regimen was found to be ineffective, and 100% of the test animals showed complete termination of pregnancy. Experimental data collectively tempt us to propose that an estrogen excess, particularly of follicular origin in both ovaries, is essential for more than 72 hours following gonadotropin sensitization before the antifertility faculty of PMSG can be demonstrated.

Animals↗