Gonadotropin Output Research Articles

Danazol Treatment for Thrombocytopenia in Myelodysplastic Syndromes: Can an "Old-fashioned" Drug be Effective?

Danazol Treatment for Thrombocytopenia in Myelodysplastic Syndromes: Can an "Old-fashioned" Drug be Effective?

Profound reduction of ovarian estrogen by aromatase inhibition in obese women

It was hypothesized that aromatase inhibitor (AI)-induced interruption of estradiol negative feedback would modulate the reproductive hormone profile of obese women. Regularly cycling women aged 18-40 years with a BMI of 18-25 kg/m(2) (normal weight, n = 10) or >30 kg/m(2) (obese; n = 12) were given AI daily for 7 days. Urinary hormone profiles were compared between groups. Fourteen eumenorrheic, normal weight women not receiving AI stimulation served as historical controls. Urinary metabolites for LH, FSH, estradiol (E1c), and progesterone (Pdg) were measured and normalized to a 28-day cycle. Serum estrone and estradiol were measured in the late follicular phase. Whole-cycle LH, FSH, and luteal Pdg excretion did not differ between obese (BMI = 37.1 + 7 kg/m(2) ) and normal weight women treated with AIs, although LH was greater in stimulated compared with unstimulated normal weight women. Whole cycle mean E1c was lower in AI-stimulated obese and normal weight participants compared with nonstimulated normal weight controls, but obese women treated with AI excreted far less E1c (467.7 ± 217.4 μg/mg Cr) than AI-treated normal weight women (911.4 ± 361.8 μg/mg Cr; P = 0.02). Follicular phase serum estrone and estradiol were also lower in AI-treated obese women versus AI-treated normal weight women (61.7 ± 22.8 and 18.3 ± 3.7 pg/ml versus 99.1 ± 30.5 and 37.7 ± 5.9 pg/ml, respectively; P = 0.034 and 0.005). Normal gonadotropin output and luteal function occur at the expense of reduced E1c excretion in AI-treated women, and this discrepancy is particularly evident in obese women.

Endocrine study of anorexia nervosa.

The main objective of the study was to evaluate the endocrinological picture of anorexia. The sample consisted of 23 anorectic patients (20 females, 3 males) with a control group of 10 normal females and 5 normal males. All participants underwent a work-up which included testing for hypothalamic, hypophyseal, thyroidal, adrenal, gonadal functioning and glucose metabolism. Our results revealed a reduced urinary output and low serum levels of gonadotropins with different responses to LHRH correlating with the stage of the illness. We found reduced urinary estrogens and elevated testosterone levels in females. Males demonstrated a reduction of testosterone. While basal prolactinemia was normal in both sexes, males showed an exaggerated response to TRH. The thyroid function study in anorectic patients revealed a decrease in T3 and in free T3 and an increase in reverse T3. Free T4 was slightly increased with normal T4 levels. Basal TSH was normal with a delayed peak after TRH. We also noticed in the anorectic population reduced basal glucose levels with a flat glucose curve; reduced insulin levels with a slight increase after glucose administration; elevated basal GH with a fair response to L-Dopa; elevated serum cortisol with loss of circadian rhythm and slightly inhibited by dexamethasone. In addition, both noradrenalin and VMA were reduced. We concluded that the multiple endocrine abnormalities found are consistent with hypothalamic dysfunction. The etiology of this dysfunction remains for the endocrinologist highly controversial.

Bone mineral density and depot medroxyprogesterone acetate

Introduction Depot medroxyprogesterone acetate (DMPA) suppresses pituitary gonadotrophin output, thus, suppressing ovulation. Estrogen production from the ovary is also strongly inhibited, and the resulting estrogen deficiency has a detrimental impact on bone. Depot medroxyprogesterone acetate may be particularly detrimental in young women, as it may impede attainment of peak bone mass, and switching to a different contraceptive is recommended. However, the effect of sequential use of DMPA with other contraceptives in this age group has not been investigated. Methods This was a cross-sectional analysis of 218 DMPA users who were 20 years or older (mean, 31 years, ±8.9 SD) at the time of bone mineral density (BMD) estimation. The majority of women had used one or more contraceptive beside DMPA. The most commonly used alternative contraceptive was the oral combined pill (OCP). It was used by 65% of women ( n=143) and for an average duration of 6 years. A logistic regression model was used to estimate the association between potential risk factors and low bone mass. Results The prevalence of low bone mass at either hip or spine ( T≤1) was 41%. The prevalence of a T score below −2.5 was 5%, and 45% of women had already sustained one fracture. Younger age was associated with higher BMD [odds ratios (ORs), 0.054; 95% confidence interval (CI), 0.007–0.431]. However, this protective effect of age was lost once the interaction between the duration of both DMPA and OCP was introduced into the model (OR for low BMD, 1.42; 95% CI, 1.09–1.8). The use of DMPA first before ever use of OCP was particularly detrimental to BMD (OR, 3.94; 95% CI, 1.08–14.0). On the contrary, body mass index was positively associated with BMD (OR, 0.86; 95% CI, 0.8–0.9). No other demographic or anamnestic variables significantly predicted the presence of low BMD in this group of young women. This group of DMPA users appear to be at a very high risk of both low BMD and fractures, possibly independently of DMPA use. This needs to be considered when writing guidelines for risk assessment. Conclusion The use of DMPA before achievement of peak bone mass may be particularly detrimental to bone, but switching DMPA with the OCP in these women does not seem to confer specific benefit in terms of bone density. This needs to be taken into consideration when a change in contraceptive is considered purely for the sake of bone protection.

Age-specific changes in the regulation of LH-dependent testosterone secretion: assessing responsiveness to varying endogenous gonadotropin output in normal men

Pulsatile and thus total testosterone (Te) secretion declines in older men, albeit for unknown reasons. Analytical models forecast that aging may reduce the capability of endogenous luteinizing hormone (LH) pulses to stimulate Leydig cell steroidogenesis. This notion has been difficult to test experimentally. The present study used graded doses of a selective gonadotropin releasing hormone (GnRH)-receptor antagonist to yield four distinct strata of pulsatile LH release in each of 18 healthy men ages 23-72 yr. Deconvolution analysis was applied to frequently sampled LH and Te concentration time series to quantitate pulsatile Te secretion over a 16-h interval. Log-linear regression was used to relate pulsatile LH secretion to attendant pulsatile Te secretion (LH-Te drive) across the four stepwise interventions in each subject. Linear regression of the 18 individual estimates of LH-Te feedforward dose-response slopes on age disclosed a strongly negative relationship (r = -0.721, P < 0.001). Accordingly, the present data support the thesis that aging in healthy men attenuates amplitude-dependent LH drive of burst-like Te secretion. The experimental strategy of graded suppression of neuroglandular outflow may have utility in estimating dose-response adaptations in other endocrine systems.

Chronic cystic ovarian disease in a Holstein cow

Cystic ovarian follicles are commonly found during rectal examination of early postpartum dairy cows, usually presenting with anoestrus and occasionally nymphomania. Most cases self cure with time, or respond to exogenous hormonal treatment. This case report describes a refractory case in a Holstein cow in which a novel treatment approach was used. A gonadotrophin releasing hormone agonist implant was inserted for 180 d in an attempt to suppress pituitary gonadotrophin output, arrest abnormal ovarian follicle growth and prevent steroidogenesis. Frequent serial blood samples were collected before and after implant insertion to monitor changes in pulse release of luteinising hormone. Follow up ultrasound scans and blood samples were done to monitor ovarian structures; progesterone and oestradiol were collected at various times over the 180 d period. A normal, cycling herdmate was enrolled as a control. Prior to implant insertion, high frequency and low amplitude luteinising hormone pulses were detected in the cystic cow. Insertion was followed by a sustained surge in the release of luteinising hormone in both cows, but ovulation was not induced in the cystic cow. Plasma oestradiol levels remained consistently elevated and signs of oestrous behaviour were observed. Long term gonadotrophin releasing hormone agonist treatment failed to suppress either ovarian steroid production or cause regression of the cysts by 180 d.

Hormonal contraception: what is new?

Hormonal contraception has become more effective and more widely used, while the world population has grown from 3000 million in 1960 to 6000 million in 2000. There is a need for improved contraception, because legal abortion is used in a high proportion of pregnancies and illegal abortion continues to be common in some countries. Hormonal contraception now includes different choices of administration and dose regimens. The best selection depends on the benefits and risks of the method and whether there is a medical disability. Medical eligibility for combined oral contraceptives has improved during the past 40 years so that, for most women, all currently available low-dose products are safe. For women with medical conditions, wider eligibility for oral contraceptive use has evolved from better knowledge of the risk factors. The long-term risks of rare cardiovascular and malignant adverse events remain controversial. There are long-term benefits, however, as oral contraceptive use appears to protect against endometrial, ovarian and colorectal cancers. Emergency contraception provides an option that reduces the number of unplanned pregnancies with little or no long-term risk. Endometrial contraception is an option that would ideally have no influence on ovarian function or the bleeding pattern, and cause no significant side-effects. Hormonal male contraception, with indirect suppression of spermatogenesis by decreasing gonadotrophin output, is a further choice. Although hormonal contraception is effective and safe, many research investigations remain to be carried out in order to improve tolerance and achieve wider utilization.

Corticotropin-releasing hormone-immunopositive nerve elements in apposition to chicken gonadotropin-releasing hormone I-containing perikarya in Japanese quail (Coturnix coturnix japonica) brain.

The neuroanatomic basis of how stress inhibits reproduction in birds is not understood. To address this question we used double-label immunofluorescence histochemistry to determine whether corticotropin-releasing hormone (CRH)-immunoreactive (ir) neuronal elements contact chicken gonadotropin-releasing hormone I (cGnRH I)-ir somata in brains of Japanese quail. The double-label system used a sheep anti-cGnRH I primary antibody with a secondary antibody conjugated to dichlorotriazinylaminofluorescein dihydrochloride for green fluorescence and a rabbit anti-CRH antibody with a secondary conjugated to Texas Red for red fluorescence. Immunohistochemical (IHC) distribution of both peptides resembled that in previous reports using single-label IHC. In four areas of the quail brain in which CRH nerve fibers and cGnRH I somata co-occurred (bed nucleus commissural pallii, nucleus preopticus medialis, nucleus septalis lateralis and nucleus accumbens), numerous instances were found of CRH-ir nerve fibers or terminals in apposition to cGnRH I cell bodies. These interactions provide a potential neuroanatomic route by which CRH may directly inhibit the activity of cGnRH-I-containing neurons, thereby inhibiting gonadotropin output and halting or slowing the progression of reproductive cycles. It remains to be demonstrated by electron microscopy whether these interactions, which appear abundant by IHC, represent instances of synaptic contact, as has been demonstrated to occur in analogous areas in mammalian species.

In vitro effects of beta‐endorphin on testicular release of androgens in the lizard podarcis sicula raf

The effects of the proopiomelanocortin-derived opioid peptide, beta-endorphin (β-EP), and of the opioid antagonist, naloxone (NAL), on both basal and pituitary-stimulated androgen secretion from superfused quiescent and active testes were assessed in the adult lizard, Podarcis sicula. In the absence of the homologous pituitary, in vitro treatment with β-EP and/or NAL did not affect basal secretion of androgens from quiescent and active testes. Conversely, in the presence of the homologous pituitary, treatment with β-EP brought about a decrease in androgen secretion in active testes, but no effect on quiescent ones Naloxone counteracted the inhibitory effect of β-EP in active testes, and enhanced maximal pituitary-stimulated secretion of androgens in quiescent but not in active testes. The effects produces by β-endorphin and naloxone were reversible. These results suggest that, in this lizard, opioids might be involved in the control of androgen release. The lack of effect of β-EP and naloxone when added directly to the testes seems to suggest that the opioid agonist and antagonist act on androgen release by modulating pituitary gonadotrophin output. © 1996 Wiley-Liss, Inc.

In vitro effects of beta-endorphin on testicular release of androgens in the lizard Podarcis sicula Raf.

The effects of the proopiomelanocortin-derived opioid peptide, beta-endorphin (beta-EP), and of the opioid antagonist, naloxone (NAL), on both basal and pituitary-stimulated androgen secretion from superfused quiescent and active testes were assessed in the adult lizard, Podarcis sicula. In the absence of the homologous pituitary, in vitro treatment with beta-EP and/or NAL did not affect basal secretion of androgens from quiescent and active testes. Conversely, in the presence of the homologous pituitary, treatment with beta-EP brought about a decrease in androgen secretion in active testes, but no effect on quiescent ones. Naloxone counteracted the inhibitor effect of beta-EP in active testes, and enhanced maximal pituitary-stimulated secretion of androgens in quiescent but not in active testes. The effects produces by beta-endorphin and naloxone were reversible. These results suggest that, in this lizard, opioids might be involved in the control of androgen release. The lack of effect of beta-EP and naloxone when added directly to the testes seems to suggest that the opioid agonist and antagonist act on androgen release by modulating pituitary gonadotrophin output.

Contour of the GnRH pulse independently modulates gonadotropin secretion in the human male.

GnRH pulse frequency, amplitude, and interpulse interval have all been demonstrated to regulate gonadotropin secretion individually. We tested the hypothesis that the contour of the GnRH pulse also modulates gonadotropin output in 10 men with isolated GnRH deficiency in whom a fixed GnRH dose was administered at a constant physiologic frequency by either instantaneous bolus or by 1-, 5-, or 30-min infusions. LH, FSH and free alpha subunit (FAS) responses were also compared to spontaneous gonadotropin secretion in normal adult men. While the LH and FAS pulses following the instantaneous bolus and 1-min infusion of GnRH were indistinguishable, further increases in the duration of gonadotrope stimulation by GnRH were associated with progressive decreases in all parameters of gonadotropin secretion (mean levels, amplitude, peak levels, AUC). FSH secretion was also decreased following variations in the contour of the GnRH pulse, although overall changes were less dramatic than for LH and FAS. The LH pulses following the bolus GnRH stimulation were indistinguishable from spontaneous LH pulses occurring in normal men whereas those stimulated by the 1-, 5-, and 30-min infusions of GnRH became progressively blunted with the lowest levels of secretion occurring after the longest infusion. In sharp contrast, FAS pulse parameters in the GnRH-deficient subjects greatly exceeded those of normal men regardless of the contour of the GnRH stimulus, whereas mean FSH levels were all modestly (although significantly) higher than those of normal adult men. These results demonstrate that the pituitary is sensitive to subtle changes in the contour of the GnRH stimulus, with a more prolonged duration of GnRH stimulation resulting in a diminished pituitary response. Alterations of the contour of endogenous GnRH secretion may represent an additional mechanism for altering gonadotrope function and provide additional evidence for the differential regulation of LH, FAS, and FSH by GnRH. However, the previously reported elevated levels of FAS secretion in GnRH-deficient men undergoing long-term GnRH replacement are not explained by abnormalities of GnRH contour.

Decreased plasma gonadotropin and testosterone levels in arthritic rats: are corticosteroids involved?

Infectious and inflammatory diseases are often accompanied by abnormal reproductive functions, and the present working hypothesis is that proteins (called cytokines or interleukins, ILs) released by activated immune cells are at least in part responsible for these neuroendocrine changes. In order to test this hypothesis, we need paradigms of immune pathologies in which concentrations of cytokines are increased, and those of hormones of the hypothalamic-pituitary-gonadal (HPG) axis are blunted. We chose a rodent model of arthritis, adjuvant-induced arthritis (AIA), in which rats show elevated plasma IL-6 and decreased testosterone (T) concentrations. We describe here the first phase of our studies, in which we determined whether gonadotropin release was also altered, whether this change was responsible for the low T levels, and whether elevated corticosterone participated in the decreased activity of the HPG axis.AIA is induced by the intramuscular injection ofMycobacterium butyricum (MBB) into the tail base of the rat, with swelling of the limbs occurring 11-12 days later. We observed significant decreases in LH and FSH secretion of castrated AIA male rats, suggesting that altered gonadotropin output was independent of the gonads. The absence of significant alterations in GnRH gene expression in the hypothalamus of AIA rats, as well as only modest declines in pituitary responsiveness to GnRH, indicate that these mechanisms are not primarily responsible for the blunted gonadotropin concentrations. Intact AIA rats exhibited a dramatic decline in T levels, but no concimitant rise in LH concentrations. The observation that gonadotropin secretion does not increase despite significantly reduced T levels suggests the presence of an unidentified defect within the GnRH neuronal circuitry that prevents the gonadotrophs to respond to decreased steroid feedback. Testicular responsiveness to hCG was significantly blunted in AIA rats, and this decrease was not reversed by acute blockade of nitric oxide formation or of prostaglandin synthesis. Interestingly, the onset of these hormonal changes preceded the appearance of symptoms (limb swelling), as well as the decrease in body weight that accompanies visible joint enlargement. On the other hand, blunted T secretion coincided with rising levels of ACTH and corticosterone. This suggested that adrenal steroids might be responsible for the decrease in LH and T values, but this hypothesis did not prove valid. Indeed, we observed that adrenalectomized AIA animals implanted with corticosterone pellets retained their low T levels. Furthermore, clamping corticosterone levels was only moderately effective in reversing the inhibitory influence of the arthritic process on LH secretion.In the absence of significant alterations in GnRH gene expression, it is possible that low Gn levels are secondary to an abnormal pattern in GnRH pulse amplitude and/or frequency. While the decrease in plasma LH concentrations may play a role in the dramatically lowered plasma T values, it is more likely that the inability of the testes to respond to gonadotropin is of significance. While we cannot rule out the participation of perceived stress at the onset of the changes in pituitary and testicular function of AIA rats, we hypothesize that cytokines released by the inflamed tissues, an event that may well precede the appearance of overt swelling, are responsible for the activation of the HPA axis and independently, for the decreased activity of the HPG axis. The AIA model may therefore provide an experimental paradigm in which to test hypotheses related to the cross-talk between the immune system and reproductive parameters.

A working hypothesis for the regulation of steroidogenesis and germ cell development in the gonads by glucocorticoids and 11β-hydroxysteroid dehydrogenase (11βHSD)

The relationship between glucocorticoid secretion from the adrenal gland and gonadal function has previously been attributed to central inhibition by the adrenal steroids of pituitary gonadotropin output. This review focuses on the direct actions of glucocorticoids within the gonads, including positive effects on germ cell maturation and both positive and negative effects on the stimulation of gonadal steroidogenesis by LH and FSH. In addition, we address the role in the gonads of 11β-hydroxysteroid dehydrogenase (11βHSD), which interconverts the glucocorticoids with their inactive 11-ketosteroid derivatives. To date, two isoforms of 11βHSD have been described. 11βHSD1, purified and cloned from the liver, has a relatively low affinity for glucocorticoids and acts instead as an 11-oxoreductase, whereas the high affinity 11βHSD2, first identified in the kidney, acts as an efficient 11β-dehydrogenase to inactivate physiological concentrations of glucocorticoid. We propose that in the gonads, 11βHSD1 promotes the positive effects of glucocorticoids on germ cell maturation (by increasing the local concentration of active glucocorticoids), whereas a high affinity 11β-dehydrogenase activity, consistent with that of 11βHSD2, inactivates glucocorticoids and so protects luteal cells from the inhibitory effects of these steroids during the luteal phase of the ovarian cycle.

Developmental expression of the genes encoding transforming growth factor alpha and its receptor in the hypothalamus of female rhesus macaques.

Studies in female rats have shown that transforming growth factor alpha (TGF alpha) stimulates release of luteinizing hormone-releasing hormone (LHRH), the neuropeptide controlling sexual maturation, and that expression of the TGF alpha gene in the hypothalamus increases during both the initiation of normal puberty and after hypothalamic lesions that induce sexual precocity. Since blockade of epidermal growth factor receptors (EGFR), which mediate TGF alpha actions, delayed the normal timing of puberty, it was postulated that TGF alpha/EGFR contributes to the neuroendocrine process that underlies the initiation of normal female puberty. The present study was undertaken to examine the hypothesis that hypothalamic expression of the TGF alpha gene and its receptor changes in relation to the stage of sexual development in nonhuman primates, and to determine whether these changes are accompanied by corresponding alterations in LHRH gene expression. DNA fragments complementary to the coding regions of the rhesus monkey TGF alpha, EGFR and LHRH genes were cloned by reverse transcription-polymerase chain reaction (RT-PCR), sequenced and used to prepare monkey-specific antisense RNA probes. A quantitative RT-PCR was developed in which the cloned sequences were utilized to prepare RNA standards for the quantitation of tissue mRNA levels. Both TGF alpha and EGFR mRNA levels in the medial basal hypothalamus and preoptic area of female monkeys were elevated during neonatal life (1 week to 6 months of age), when FSH secretion is also high, decreased during juvenile development (8-18 months of age), when secretion of both FSH and LH is low, and markedly increased during the expected time of puberty (30-36 months of age). No such changes were observed in either the cerebellum or the cerebral cortex, two brain regions irrelevant to neuroendocrine reproductive control. In contrast to the pronounced alterations in hypothalamic TGF alpha/EGFR gene expression observed during sexual development, LHRH mRNA levels did not vary significantly during this time. Hybridization histochemistry revealed the presence of both TGF alpha and EGFR mRNAs in cells scattered throughout the hypothalamus, but more predominantly in the median eminence, suprachiasmatic nuclei, optic chiasm and cells along the wall of the third ventricle. These results demonstrate that increases in TGF alpha and EGFR gene expression, specific to the neuroendocrine brain, occur during developmental phases in which gonadotropin output is also elevated--most noticeably at the time of puberty.(ABSTRACT TRUNCATED AT 400 WORDS)

Influence of oral contraceptives on integrated secretion of gonadotropins

The mechanism of action of various oral contraceptives has not yet been satisfactorily resolved, as to how gonadotropins affect ovarian function. Alterations of the pulsatile release of LH might be a common denominator. As methodological difficulties for the evaluation of LH pulse pattern with low basal levels exist, we elected to determine the area under the curve (AUC) for LH and FSH for 6 hours before and during treatment with oral contraceptives. LH and FSH were determined every 15 min for 6 hours on day 4 and day 20 of a control cycle and a treatment cycle in 22 women with ovulatory cycles. They received either a combined preparation containing 150 μg desogestrel and 30 μg ethinyl estradiol, a sequential preparation containing 50 μg of ethinyl estradiol and 125 g of desogestrel or only 125 μg desogestrel. There was no difference between the sum of LH pulses on day 4 and day 20 of the control cycle. The AUC for FSH was lower on day 20. When the combined preparation was taken, FSH was suppressed on day 4, and FSH and LH on day 20 of treatment. The degree of suppression was even more pronounced when the sequential OC was taken. Ethinyl estradiol alone was as effective as the combination with desogestrel. Desogestrel alone inhibited ovulation without affecting serum LH and FSH in a comparable manner, suggesting a direct effect on the ovary. The determination of the AUC seems to be a sensitive tool for the evaluation of OC-induced changes in gonadotropin output.

Pituitary Binding Sites for Gonadotrophin‐Releasing Hormone in Booroola Merino Ewes which were Non‐Carriers or Homozygotes of the Fecundity Gene F

Abstract Pituitary binding sites for gonadotrophin-releasing hormone (GnRH) were examined in Booroola ewes on Days 4 and 12 of the oestrous cycle and more than 12 months after ovariectomy. No significant differences were observed in the affinity of the binding sites for [(125) I]D Ala6, des Gly(10) GnRH ethylamide, in non-carriers (++) or homozygotes of the fecundity gene (FF), in intact or ovariectomized animals (overall K(a)= (1.8 +/- 0.1) x 10(9) M(-1); mean +/- SEM, n = 67). In ++ ewes, the number of GnRH binding sites on Day 4 of the oestrous cycle (0.199 +/- 0.025 fmol/mg pituitary, n = 8) was significantly greater (P<0.05) than on Day 12 (0.107 +/- 0.012 fmol/mg pituitary, n = 12). In contrast in FF ewes the number of binding sites on Days 4 and 12 were similar (Day 4: 0.119 +/- 0.030, n = 8; Day 12: 0.092 +/- 0.014, n = 10). On Day 4, but not Day 12, the binding site numbers were significantly higher in ++ compared with FF pituitaries (P<0.05). Likewise on Day 4, but not Day 12, the plasma concentrations of follicle-stimulating hormone were significantly higher (P<0.005) in FF (1.60 +/- 0.15 ng/ml) than ++ ewes (0.96 +/- 0.10 ng/ml). After ovariectomy, no gene-specific differences in binding site numbers were noted despite significantly higher plasma follicle-stimulating hormone concentrations in FF animals. No gene-specific differences in plasma luteinizing hormone concentrations were observed in these animals on Days 4 or 12, or after ovariectomy. Moreover, luteal phase plasma progesterone concentrations were also similar between ++ and FF ewes. We conclude that the gene-specific differences in pituitary responsiveness to GnRH and in pituitary gonadotrophin output are unlikely to be due to differences in GnRH receptor binding affinities.

A serum-free system for culturing human placental trophoblasts

We have compared hormone production by early gestation and term human placental trophoblasts cultured in Ham's F10 medium containing 10% fetal bovine serum with that by cells cultured in serum-free HB102 medium. Mean daily production of progesterone on Days 3 to 7 was approximately 25% less by both early gestation and term cells cultured in HB102 as compared to Ham's F10, but production was maintained at a stable level for at least 7 d longer than the cells in Ham's. Estradiol production from 10(-6) M dehydroepiandrosterone by both early gestation and term cells was comparable in both media. Human placental lactogen production on Days 3 to 7 was 40% less by cells cultured in HB102. Human chorionic gonadotropin (hCG) output by early gestation cells was also 50% less in HB102 but term cells in HB102 produced twice as much hCG as those in Ham's F10. 3B-Hydroxysteroid dehydrogenase (3BHSD) activity in early gestation and term cells and 11B-hydroxysteroid dehydrogenase (11BHSD) activity of early gestation cultures was comparable in the two media. 11BHSD activity was decreased in the term cultures, and this decrease was more marked in Ham's than in HB102. Sulfatase and aromatase activities in the early gestation cultures were comparable in both media; sulfatase activity was comparable and aromatase activity only 20% less in the term cells cultured in HB102. These results indicate that serum-free HB102 supports differentiated function of human trophoblast cells and is useful for studies of placental activity for as long as 14 d in culture.

Short-term use of buserelin in combination with human menopausal gonadotrophins for ovarian stimulation for in-vitro fertilization in endocrinologically normal women

Ten endocrinologically normal women were injected subcutaneously with 500 micrograms D-Ser(TBU)6-EA10-LHRH (buserelin) on days 3,4 and 5 after the start of the menses. Two types of response were observed. Five women (group A) responded promptly and had a mean number of 13.4 oocytes retreived after 11.4 days of stimulation. In the second group (B), two to three times more HMG was needed to obtain a mean number of 7.3 oocytes after 17.2 days of stimulation. The response upon stimulation could be predicted by the serum gonadotrophin output on days 4 and 5 of the cycle. One woman from group B had a premature LH rise on day 16 and luteinization; her cycle was abandoned. In the four other patients of group B, serum and urinary LH concentrations showed that pituitary gonadotrophin secretion had recovered before the ovulatory stimulus, without signs of premature luteinization. Two women in each group became pregnant, one of whom aborted. This short-term GnRH agonist treatment could be an alternative method for ovarian stimulation, although it did not totally prevent the occurrence of an endogenous LH surge.

Studies of the effectiveness of gonadotrophin-releasing hormone, steroids and follicular fluid in modulating ovine gonadotrophin output in vivo and in vitro

Gonadotrophin-releasing hormone (GnRH) readily stimulated LH output by sheep pituitary cells in vitro, and raised plasma LH concentrations in vivo in sheep, in a dose-dependent fashion. However, increases in FSH levels were only marginal by comparison. Dose-dependent decreases in sheep pituitary cell FSH output and in plasma FSH concentrations were caused by sheep follicular fluid and oestradiol-17 beta in vitro, and by bovine follicular fluid and oestradiol benzoate in vivo. In contrast, LH concentrations were only reduced slightly at the higher doses of these reagents. Cumulative suppressive effects of follicular fluid and oestradiol-17 beta (oestradiol benzoate) on FSH levels were observed both in vitro and in vivo. The transient positive feedback effect of oestradiol benzoate on FSH output negated the suppressive effect of bovine follicular fluid on plasma FSH concentrations. Progestagens, androgens and catechol oestrogens also suppressed mean FSH output in vitro, though not as effectively as oestradiol-17 beta. While only 1-5 pg/ml of oestradiol-17 beta was needed to suppress significantly mean FSH output in vitro, greater than 500 pg/ml of the other steroids was required. Seminal plasma inhibin-like peptide failed to suppress mean FSH output by cultured sheep pituitary cells at doses from 1 pg/ml to 500 ng/ml. At higher doses, both FSH and LH output was suppressed and this was accompanied by morphological deterioration of the cells. It is suggested that, to raise plasma FSH concentrations with a view to increasing ovulation rates in sheep, the development of means to reduce the negative feedback effects of steroids, notably oestradiol-17 beta, and inhibin on FSH secretion may be a more appropriate pharmacological strategy than increasing pituitary exposure to GnRH.

Role of zinc in regulating the testicular function. Part 1. Effect of dietary zinc deficiency on serum levels of gonadotropins, prolactin and testosterone in male albino rats

Hypogonadism is a major manifestation of zinc deficiency in both humans and animals. In this study, we try to elucidate the main role of zinc in regulating the testicular function. Ninety male weanling albino rats were divided into three groups: control, zinc-deficient (ZD), and zinc-supplemented (ZS) groups. Six rats from each group were sacrificed every two weeks over a total period of ten weeks. Serum levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), prolactin (PRL) and testosterone (T) were estimated by radioimmunoassay technique. Serum zinc concentrations were also determined. Data revealed that serum zinc concentrations were significantly low in ZD group compared to both control and ZS groups. Normal FSH and LH but low PRL and T levels were demonstrated in sera of ZD group compared to both control and ZS groups. We came to the following conclusions: zinc deficiency has no effect on the pituitary gonadotropin output. Zinc deficiency affects the testicular function either directly through its effect on the testicular steroidogenesis and/or indirectly through its effect on the pituitary synthesis and/or secretion of PRL.