Corpus Luteum Extract Research Articles

Evidence for the presence of oxytocin in the corpus luteum of the goat.

Purified acetic acid extracts of corpora lutea (CL) of non-pregnant goats were found to contain substantial amounts of oxytocin (OT) as measured by radioimmunoassay. OT standard and the CL extracts released prostaglandin F2 alpha (PGF2 alpha) from rat isolated uteri in a quantitatively similar manner. Treatment of both OT standard and CL extract with sodium thioglycolate, oxytocin antiserum or oxytocin antagonist abolished this biological activity. Acid extracts of CL of pregnant goat were found to contain approximately 2% of levels during the cycle by day 21 after fertile mating and this had a reduced ability to release PGF2 alpha from rat uterus. It is concluded that both the immunoreactivity and the biological activity of the CL extracts are due to the presence of an oxytocin-like substance and that tissue levels of oxytocin are low in pregnant compared to non-pregnant goats.

Endocrine control of uterine oxytocin receptors in the ewe.

Specific binding of [3H]oxytocin to high affinity sites (hormone receptors) in membrane preparations from uterine tissues of the ewe has been determined at varying stages of the oestrous cycle and in pregnancy. Mean receptor concentrations in caruncular and intercaruncular endometrium and in myometrium were 14.2, 1.9 and 13.0 fmol/mg protein respectively between days 10 and 13 of the cycle. By the day of oestrus these values had increased to 749, 1085 and 179 fmol/mg protein. These increases in receptor concentrations coincided with luteolysis and falling plasma progesterone levels and followed the preovulatory decline in peripheral oxytocin and rise in ovarian venous oestradiol-17 beta. Receptor concentrations were low in all uterine tissues from pregnant animals between days 14 and 19 after oestrus. Analysis of binding parameters by Scatchard plot suggested a single population of receptor molecules in each of the tissues studied with apparent dissociation constants in the range 1.9-2.2 nmol/l. A number of naturally occurring neurohypophysial peptides inhibited binding of [3H]oxytocin to the receptor from ewes at oestrus; the cross-reactions of arginine vasopressin and vasotocin exceeded that of oxytocin. Use of a receptor binding assay to measure oxytocin in extracts of corpora lutea on days 4 and 10 after oestrus gave values similar to those obtained by radioimmunoassay, suggesting the absence of other receptor-active peptides in the corpus luteum. It is concluded that the oxytocin receptor is present in both components of the endometrium, as well as in the myometrium and that changes in uterine receptor concentrations before oestrus are consistent with receptor activation by steroid hormones.

Luteinizing hormone receptor binding inhibitor in aqueous extract of human corpus luteum.

Aqueous extracts of human luteal tissue significantly inhibited binding of 125I-labelled human luteinizing hormone (LH) to the 2,000-g subcellular fraction of human corpora lutea. In contrast, aqueous extract of nonluteal tissue of the human ovary did not show a comparable activity to inhibit LH binding. Extracts of human corpus luteum had little or no ability to inhibit LH binding to porcine luteal homogenates. The inhibitory effect of the aqueous extract on LH binding to human luteal homogenate was demonstrated to be dose-related. The inhibitory effect of aqueous extracts of human corpora lutea on LH binding increased from the early to mid and late luteal phase of the menstrual cycle. The results of the present study suggest that there is an LH receptor binding inhibitor (LHRBI) in the 30,000-g aqueous extract of the human corpus luteum and the increase in LHRBI in the luteal tissue as the human corpus luteum ages may explain the means whereby the human corpus luteum regulates its own life span.

Inhibitory effect of charcoal-treated aqueous porcine corpus luteum extract upon ovulation in the rabbit.

AbstractAdministration of charcoal-treated aqueous extract of porcine corpus luteum on Days 0, 1, and 2 after mating inhibited coitus-induced ovulation in the rabbit. Inhibition of ovulation was evidenced by a significant (P < 0.01) inhibition of the coitus-induced rise in serum progesterone and lack of formation of corpora lutea. Administration of charcoal-treated heart or lung extract had no significant effect upon the formation of corpora lutea or on the coitus-induced rise in serum progesterone. Administration of the same extract 3 and 4 days after coitus-induced ovulation had no significant effect upon serum progesterone and further formation of corpora lutea. These data are consistent with the existence of an LH binding inhibitor in porcine corpus luteum extract which acts to inhibit the ovulatory response to the coitus-induced LH surge.

DNA synthesis in cultures of calf aortic endothelial cells treated with corpus luteum extract: Its inhibition by 3H-thymidine

In cultures of calf aorta endothelial cells treated with extracts from corpora lutea a paradoxical effect occurred: Cell proliferation was regularly stimulated, but DNA synthesis was more or less inhibited. This inhibition was related to the presence of 3H-thymidine and could be reduced by lowering te 3H-thymidine concentration in the incubation medium. To decide whether the molar concentration or the activity of the 3H-thymidine was responsible for this effect, unlabelled thymidine was added to increase the molar thymidine concentration. A mixture of unlabelled thymidine and 3H-thymidine (ratio 200:1) added to the medium at various concentrations stimulated the DNA synthesis in a dose-dependent was up to 4.1 x 10(-6) M. Furthermore, nonradioactive thymidine slightly promoted the proliferation of calf aorta endothelial cells up to 10(-6) M independently of the presence or absence of corpus luteum extract. It is concluded that the radioactivity of 3H-thymidine is responsible for this effect. Internal 3H-beta radiation may perturb cell cycle progression after incorporation of this radioactive precursor into the cells. This view is supported by the fact that the inhibitory effect can be reduced by adding unlabelled thymidine, i.e. by lowering the specific activity of 3H-thymidine. On the other hand, increasing concentrations of growth promoting factors which stimulate the uptake of 3H-thymidine render the cells more sensitive to this effect.

Demonstration of Luteinizing Hormone/Human Chorionic Gonadotrophin Receptor-Binding Inhibitor in Aqueous Extracts of Frozen Human Corpus Luteum

Aqueous extracts of frozen human corpora lutea were tested for the presence of an inhibitor of luteinizing hormone-receptor site binding (LHRBI) and for the subsequent effect on the stimulatory response of luteinizing hormone (LH) on progesterone synthesis by sheep ovarian cells. In the presence of human corpus luteum extract of normal menstrual cycle (30,000-g supernatant), the binding of 125I human chorionic gonadotrophin (hCG) to granulosa and luteal cells of sheep ovaries was markedly reduced, but the ability of rat testicular LH receptors to bind labelled hCG was less affected. However, extracts of corpora lutea of the first trimester of pregnancy appeared to be less inhibitory on the binding of LH/hCG to ovarian cells and had no effect on the binding of rat testicular cells compared to those of normal menstrual cycle. Addition of both extracts separately inhibited the LH-stimulated in vitro progesterone synthesis by granulosa cell cultures and by incubated sheep corpus luteum slices. These findings provide evidence for the presence of LHRBI in human corpus luteum.

Intraovarian control of progesterone biosynthesis by granulosa cells and corpus luteum

Some of the recent developments on the intraovarian control of follicular development and luteinization have been discussed. Luteinizing hormone (LH) and follicle-stimulating hormone (FSH) regulate the growth and maturation of the follicle. However, the locus of control, by which the responsiveness of follicles to LH and FSH is regulated at different stages of follicular development, resides within the ovary itself. Granulosa cells obtained from small follicles have more FSH receptors and can accumulate more cyclic AMP and progesterone in response to FSH compared to cells obtained from large mature follicles. The situation is reversed in the case of LH responsiveness; larger follicles are more responsive than small ones. The steroidogenic potential of follicular cell types to secrete oestrogen and progesterone is altered with the stage of the follicle. In vivo studies in the rhesus monkey demonstrated that thecal cells of the preovulatory follicle are the principal source of oestrogen and progesterone of the ovarian venous blood. In in vitro cultures, thecal cells alone could secrete more oestrogen than granulosa cells, and mixing of the two cell types augmented oestrogen synthesis. Addition of exogenous testosterone (0.15μg/ml) to granulosa cell cultures resulted in increased secretion of oestrogens. Granulosa cells in culture produced more progesterone than thecal cells. These findings suggest that the thecal layer is the main source of ovarian vein levels of oestrogen and progesterone and the two cell types may be responsible for the high levels of these steroids in the follicular fluid. A luteinization inhibitor, present in the follicular fluid of small and medium-sized follicles may regulate the responsiveness of follicles to gonadotrophins and prevent luteinization of granulosa cells prior to ovulation. Evidence for the presence of another inhibitor, a LH receptor binding inhibitor (LHRBI) in frozen aqueous extracts of porcine corpus luteum and its ability to inhibit progesterone secretion by cultured granulosa cells of mature porcine follicles has also been presented.

Influence of porcine corpus luteum extract on DNA synthesis and proliferation of cultivated fibroblasts and endothelial cells

The angiogenic activity in corpus luteum (corpus luteum angiogenesis factor--CLAP) reported first by us in 1977 was enriched by ammonium sulfate precipitation and further purified by ion exchange chromatography. Extracts produced in this way from corpora lutea of cyclus synchronized pigs have proven effective to stimulate DNA synthesis and proliferation of both calf aorta endothelial cells and primary mouse embryo fibroblasts cultivated either under optimal or suboptimal (depletion) conditions. The mitogenic action of these extracts has been found to be dose-dependent and similar to that of bovine brain extract produced according to the preparation method published by GOSPODAROWICZ.

Characterization of relaxin radioimmunoassay using Bolton-Hunter reagent. First results in plasma during pregnancy, and in placenta, corpora lutea and ovarian cysts in woman.

Application of a porcine relaxin radioimmunoassay system using Bolton and Hunter reagent was characterized in different biological fluids. In pregnant woman, plasma reveals concentrations from undetectable values to 1,800 pg/ml. Maximal concentrations are found between the 8th and the 13th weeks of pregnancy. These peak values are significantly different from those at other stages of pregnancy. Appreciable relaxin concentrations are detected in gestational corpus luteum extract, and in corpora lutea cyst fluid of pregnant and nonpregnant women. No relaxin was detected in a cyclus corpus luteum extract, in a placental extract, or in an ovarian serous cyst. The presence of immunoreactive relaxin in the nonpregnant woman is reported.

Ability of an Extract of Pig Corpus Luteum to Inhibit Binding of 125l-Labeled Human Chorionic Gonadotropin to Porcine Granulosa Cells

SummaryPorcine corpus luteum tissue contains an inhibitor of binding of iodinated human chorionic gonadotropin to porcine granulosa cells. The inhibitor was present in the 1000, 30,000, and 105,000g supernatants of the corpus luteum but not in those of nonluteal ovarian tissue, granulosa cells, lung, or heart tissue. The inhibitor appeared only after storage in the freezer for from 1.5 to 11 weeks. Freezing heart, lung, or nonluteal tissue for a comparable time period did not lead to an increase in inhibitory activity.

The early history of progesterone.

The corpus luteum was first clearly described by Regner de Graaf (1672). That it is an organ of internal secretion was suggested by Louis-Auguste Prenant in 1898 and by Gustav Born about the same time. Clues to its specific action upont the endometrium were discovered through experiments by Ludwig Fraenkel (1903 and 1910), by Leo Loeb (1907) and by Paul Ancel and Paul Bouin (1910). The control of uterine motility by the corpus luteum was demonstrated by Hermann Knaus (1927 ff.). G.W. Corner (1928) demonstrated the necessity of the corpus luteum for survival of the pre-implantation embryo. Using a test based upon Fraenkel and Ancel/ Bouin, Corner and W.M. Allen isolated the hormone progesterone (1929) and first maintained pregnancy with corpus luteum extracts after early ablation of the ovaries (rabbit, 1930). Purification of progesterone was the work of several investigators between 1928 and 1934. The structural formula was worked out by Adolf Butenandt (1933–1934).

Sulfurylation of estradiol-17B by extracts of ovary, corpus luteum, testis and adrenal cortex 1

Soluble enzyme preparations from bovine ovary, corpus luteum and adrenal cortex and from rat testis sulfurylated estradiol-17β in the presence of ATP and Mg ++. Two major products were tentatively identified by chromatographic comparisons as estradiol-3-sulfate and estrone sulfate 2. In addition, two unidentified, highly-polar derivatives were formed from estradiol-17β in relatively small amounts by ovary, corpus luteum and testis. Estradiol-17-sulfate, which was formed readily in similar extracts of rat liver, was not formed in detectable amounts by the extracts of ovary, corpus luteum, testis, and adrenal cortex under the conditions described.

The effect of small doses of progesterone on the uterus of the rat.

It is well known that stimulation, by progesterone, of the formation of a »progestational« endometrium is enhanced by pretreatment with oestrogen. As early as 1928, shortly after the discovery of the corpus luteum hormone, Weichert observed that deciduomata of the traumatized uterus developed only if treatment of the rats with corpus luteum extract had been preceded by the administration of follicular hormone. In 1930, Hisaw & Leonard confirmed these results in rabbits. Shelesnyak (1933) was one of the first to emphasize the importance of an optimal ratio between the doses of both hormones. De Fremery & Geerling (1939) found 0.6 mg. of progesterone daily, preceded by 15 μg. of oestrone or stilboestrol to be the most effective combination. Korenchevsky & Hall (1937) found daily doses of 1 μg. of oestrone followed by 1.5 mg. of progesterone to be even more active than 30 μg. of oestrone followed by 3 mg.

ホルモンと腎機能

M. Ono, a member of our laboratory, has once made public a detailed report on the morphological influences of sex hormones on the kidney of an albino rat, but there is scarcely any systematic study on the functional effect of sex hormones upon the kidney. The present work was attempted to shed light on the effect of sex hormones on the renal function, by measuring GFR and RPF respectively with creatinine clearance and with PAH clearance as to mature male rabbits during 2 weeks of daily administration of various sex hormones in a small dose as well as in a large dose.The results obtained were as follows, compared with the control experiment performed on 9 normal rabbits:(A) Effect of male sex hormones on renal function.(1) Testosterone acetate: GFR showed a considerably great increase (Ca 50-70%) during the administration of this hormone in both small and large doses, while RPF showed a little increase only when given in a small dose.(2) Testosterone propionate: the increase of both GFR and RPF was just as effected as when testosterone acetate was injected, but a degree of the increase was less remarkable in GFR and more conspicuous in RPF than in case of testosterone acetate.(3) Following castration, GFR decreased day by day to as low as -40% on the 28th day, while RPF remained hardly changed throughout the experiment.(B) Effect of female sex hormones on renal function.(1) Follicular hormones: by urinary follicular hormone, in both small and large doses, GFR showed a slight tendency of increasing in one week after the administration of the hormone and returned to a normal value in two weeks. RPF increased in one week after the administration of the hormone in a large dose and in 2 weeks decreased, while, in a small dose administration of the hormone, it showed any significant change neither in one week nor in two weeks. By synthetic follicular hormone, GFR remained scarcely effected in both small and large doses, and RPF decreased slightly more remarkably in cases of a large dose than in the case by urinary follicular hormone, while, in case of a small dose, it showed hardly any change.(2) Corpus luteum hormone: by Oophormin luteum (an extract of corpus luteum of a pig) both GFR and RPF remained unchanged even in a large dose, but the synthetic preparation (progesterone acetate) brought on a slight increase of GFR and RPF in both small and large doses.(C) Effects of male sex hormone on damaged renal function.(1) Testosterone acetate, administered following castration, brought on a remarkable recovery of decreased GFR, while no effect on RPF was observed.(2) GFR and RPF, which had decreased by subcutaneous administration of a sublimate solution, recovered normal by the application of testosterone acetate following.(3) The effects, observed in (1) and (2), occurred in most conspicuous degree on 15th day of daily administration of testosterone acetate, and nothing effective was found already in ca. 15 days after discontinuance of testosterone administration.

性ホルモンと膀胱機能

Embryonically, urinary organ has been closely related to the sexual organ, but no one has reported a systematic study of influences on the bladder function by sexual hormones. With ripe male rabbits, I experimented on the above systematic study by my original cystometric method.The Experiment was tried after or during the injection of different hormones, the dose of which was divided into three doses converted from human's therapeutic dose, and they were graded above normal (large), normal and less normal (small).The obtained results were as follow.(A) The Influences of different steroid hormones:I) Male hormones:(1) Extract of testis (Spermatin) and androgen (Enarmon) were not effective, but testosterone acetate (Enarmon-S) showed that effects of small and normal doses on the normal bladder were hyperactive, but effects of large doses were hypoactive.(2) Castration caused hypofunction and this influence was being made gradually clear during 1 month after the castration. Following 1 month after the castration, this influence was being stabilized.This influence was not recovered by the injection of androgen and extract of testis, but was recovered a little by the small or normal dose of testosterone acetate and much by the large dose of testosterone acetate.II) Female hormones:(1) The large dose of estrogen (Ovahormon benzoat) caused hypofunction a little and the normal or large dose of synthetic estrogenic substance (Estimon, Suron and Rohormon) caused marked hypofunction on the normal Bladder.By the large dose of estrogen, the hypofunctional influence on castrated bladder was not changed and with the normal or large dose of synthetic estrogenic substance, it was improved.(2) Extract of corpus luteum (Oophormon luteum) was not effective, but progestrone acetate (Synthetic Oophormon luteum) showed that its administration caused hyperfunction a little and its normal dose caused so much effect on the normal bladder also.By the extract of corpus luteum, the hypofunctional influence on the castrated bladder was unchanged and with the large dose of progesterone acetate, it was improved as the normal dose of testosteron acetate.(B) The influence of gonadotropic hormones:I) Extract of pituitary anterior lobe (Hypohorin) was not effective on the normall bladder.On the normal bladder, the large dose of the extract caused hyperfunction a little, but it caused hypofunction a little on the castrated bladder.II) Adrenal cortical hormones:Extract of adrenal cortex (Interenin) was not effective, but desoxycorticosterone acetate (Cortate) showed that its normal or large dose caused hyperfunction a little on the normal bladder and recovered the hypofunctional influence a little on the castrated bladder.

Secretory and decidual changes induced in the endometrium of oophorectomized women by administration of estrogen and progesterone.

THERE have been relatively few studies of the histologic response of the human endometrium to progesterone and only one or two in which serial biopsies have been employed to demonstrate the endometrial histology before and during progesterone therapy. Earlier studies of Neustaedter (1) and of Elden (2) using total doses of 12 to 60 rabbit units ofprogestin, produced largely negative results. Kaufmann (3) used corpus luteum extracts indoses of 10 rabbit units daily for eight days and reported the induction of secretory changes in the uterus of a castrate woman. Salmon, Walter and Geist (4) reported 5 patients treated with oral anhydro-hydroxy-progesterone (pregneninolone) in whom, even though there was only one biopsy, a good secretory endometrium seemed to have been induced after eight days. These women had been given estrogen for ten weeks prior to the pregneninolone treatment. Ryden (5) in a comparative study of several estrogens, terminated some of his study intervals by giving a total dose of 25 to 65 mg. of progesterone during the last week. He had serial endometrial biopsies to control his observations but he obtained only incomplete secretory changes. Szarka (6) reported the recovery of a decidual cast after twenty-eight days of progesterone therapy in an ovariectomized woman. She had received 5 mg. of estradiol benzoate daily in the month prior to and during the period of progesterone injections. The progesterone dosage was started at 10 mg. on alternate days and then increased to 20 mg. daily for three weeks. The estrogen dosage was decreased and discontinued during the last week of progesterone; then bleeding started and the endometrium was shed as a decidual cast. This indicated that the estrogen was equally as important as the progesterone in maintaining the decidua.

Prolongation of Pregnancy and Excessive Fetal Development Following Administration of Corpus Luteum Extract in the Treatment of Threatened Abortion

Prolongation of Pregnancy and Excessive Fetal Development Following Administration of Corpus Luteum Extract in the Treatment of Threatened Abortion

Chiari's syndrome

C HIARI in 1855 reported two cases of a syndrome which has occurred very rarely. It consists of amenorrhea with persistent lactation following delivery. In 1882 Frommel found one case in a study of 3?000 gynecological patients in Vienna. Sharpe’ in 1935 reviewed the literature, finding in all, four papers on the subject. AS the condition is so rare and as none of the reported cases had a subsequent pregnancy, the following case seems worth recording. Mrs. F. J., aged 32, was seen July 17, 1934. At the age of 23, an ovarian cyst was removed and an appendectomy done. In 1933, she was delivered by low forceps and episiotomy of a full-term baby girl weighing 8 pounds and 3 ounces, She did not menstruate following the delivery and her breasts continued to lactate. In 1.934, desiring another child, she was sent to me by Dr. Mac Naughton Wilkinson for study and to determine, if possible, the cause of the amenorrhea and lactating breasts. The physical examination was negative except that the breast ducts were full of secretion and the uterus was atrophic. Routine laboratory tests, including a basal metabolism, were negative. Endometrial biopsy revealed an atrophic endometrium. The endocrine preparations of that day were tried without relief. They were thyroid, progynon in oil, and corpus luteum extract. In 1941, the patient returned! stating that in 1940, her breasts had stopped secreting and her periods had returned. Her weight, 142 pounds was the same. The question was, could she become pregnant? Ho%vever, on January 20, 1942, Mrs. F. J. came to the office having missed two periods. Examination showed a pregnant uterus but the breasts were not secreting. On September 7, 1942, she was delivered of a g-pound baby girl by the Bell modification of the Scanzoni maneuver. She was unable to nurse the baby because of insufficient milk. To date, November 2, 1943, she has not menstruated and her breasts are dry. As far as I know, this is the only patient with Chiari’s syndrome who has had a subsequent pregnancy and delivery. An x-ray of the sella turcica and sugar tolerance test would have been interhsting, but there was nothing clinically to suggest difficulty in the pituitary gland.

Falls, Frederick H., Rezek, George H., and Benensohn, S. J.: Clinical and experimental observations on the use of corpus luteum extracts in obstetrics: Surg. Gynec. & Obst. 75: 289, 1942

Falls, Frederick H., Rezek, George H., and Benensohn, S. J.: Clinical and experimental observations on the use of corpus luteum extracts in obstetrics: Surg. Gynec. & Obst. 75: 289, 1942

Progesterone Therapy and Progestin Metabolism in Abortion1

The council on pharmacy and chemistry of the American Medical Association recently stated: ‘It is indeed difficult to conclude from the available data whether or not progesterone or active corpus luteum extracts are of value in the treatment of habitual and threatened abortion’ (1). The accurate evaluation of therapy, of necessity, must remain difficult until we have a better understanding of the etiologic factors in abortion. Diagnosis of threatened abortion has been rendered uncertain by the fact that biologically active prc gestin is identified in human tissues and fluids in only small quantities by the present methods of bioassay. Some of the more recent and more sensitive of these methods have not been applied to clinical studies. Venning's (2) method for the determination of pregnanediol, an excretion product of progestin metabolism, aroused much enthusiasm. It was hoped that, at last, there was a diagnostic implement by which significant alterations in intrinsic progestin levels might be determined.