Elevated Plasma Estradiol Research Articles

Vitamin D3 promotes fish oocyte development by directly regulating gonadal steroid hormone synthesis†.

Vitamin D receptors and vitamin D3-metabolizing enzymes have been found to be highly expressed in the ovaries and spermatophores of fish. However, the role of vitamin D3 on fish gonadal development has rarely been reported. In this study, 2-month-old female zebrafish were fed with different concentrations of vitamin D3 diets (0, 700, 1400, and 11200IU/kg) to investigate the effects of vitamin D3 on ovarian development. The diet with 0IU/kg vitamin D3 resulted in elevated interstitial spaces, follicular atresia, and reproductive toxicity in zebrafish ovaries. Supplementation with 700 and 1400IU/kg of vitamin D3 significantly increased the oocyte maturation rate; upregulated ovarian gonadal steroid hormone synthesis capacity; and elevated plasma estradiol, testosterone, and ovarian vitellogenin levels. Furthermore, the current study identified a vitamin D response element in the cyp19a1a promoter and demonstrated that 1.25(OH)2D3-vitamin D response directly activated cyp19a1a production through activating the vitamin D response element. In conclusion, this study shows that an appropriate concentration of vitamin D3 can promote zebrafish ovarian development and affect vitellogenin synthesis through the vdr/cyp19a1a/er/vtg gene axis.

Response of the hen ovary to eCG treatment: Insight into morphology and expression of genes related to steroidogenesis and vitellogenesis

The present study aimed to examine the effect of equine chorionic gonadotropin (eCG) treatment on the chicken ovarian folliculogenesis and steroidogenesis. The expression of vitellogenesis-related genes in the liver was also investigated. Laying hens were injected with 75 I.U./kg of body weight/0.2 mL of eCG, once a day for 7 successive days. On day 7 of the experiment hens, including control hens which were receiving vehicle, were euthanized. The liver and ovarian follicles were harvested. Blood was collected daily through the whole experiment. The eCG treatment resulted in the cessation of egg laying after 3 or 4 days. The eCG-treated hens had heavier ovaries with a higher number of yellowish and yellow follicles arranged in a non-hierarchical way in contrast to ovaries of control hens. Moreover, these birds had elevated plasma estradiol (E2) and testosterone (T) concentrations. The molar ratios of E2:progesterone (P4) and T:P4 were increased in chickens injected with eCG. Real-time polymerase chain reaction revealed changes in mRNA abundances of steroidogenesis-associated genes (StAR, CYP11A1, HSD3β, and CYP19A1) in ovarian follicles: white, yellowish, small yellow, and the largest yellow preovulatory (F3-F1) as well as VTG2, apoVLDL II, and gonadotropin receptors in the liver. In general, the abundances of gene transcripts were higher in eCG-treated hens than in control hens. Western blot analyses showed an elevated abundance of aromatase protein in the prehierarchical and small yellow follicles of eCG-treated hens. Unexpectedly, there was presence of both FSHR and LHCGR mRNA in the liver and the level of expression was shifted in eCG-treated hens. In summary, eCG treatment leads to disruption of the ovarian hierarchy with accompanying changes in circulating steroids and ovarian steroidogenesis.

Role of ovarian sympathetic nerves and cholinergic local system during cold stress.

An increase in the sympathetic tone in the rat ovary induces a polycystic ovary (PCOS-like) phenotype. No information exists about its impact on fertility. In contrast, increased follicular development and improved fertility in rats were found after pharmacological inhibition of acetylcholinesterase, which increased intraovarian acetylcholine (ACh). Now, we studied the impact of sympathetic stress, followed by a recovery period without stress, on the cholinergic and noradrenergic systems of the rat ovary and on fertility. To activate ovarian sympathetic nerves, female Sprague-Dawley rats were exposed to cold stress (4°C/3 h day for 28 days; first period), followed by a 28-day period without cold stress (second period). No changes in estrous cyclicity during the first period was found. At the end of this period, ovarian levels of NA and ACh were increased. Morphometric analysis showed lower numbers of secondary and antral follicles, enhanced follicular atresia and fewer corpora lutea. Plasma progesterone was lower and testosterone was higher than that in controls. At end of the second period, ovarian ACh levels had returned to control levels, but NA levels remained elevated. The second period was also characterized by the presence of cystic follicles in the ovary, by elevated plasma testosterone and estradiol levels, while progesterone levels were decreased. Estrous cyclicity and ovulation during that period were irregular and fertility decreased. Thus, cold stress initially activated both ovarian noradrenergic and cholinergic system. After stress, the ovary did not fully recover and activation of the noradrenergic system persisted and correlated with cystic ovarian morphology and decreased fertility.

Functioning gonadotroph adenoma with severe ovarian hyperstimulation syndrome: A new emergency in pituitary adenoma surgery? Surgical considerations and literature review

The authors reported 2 cases of functioning gonadotroph pituitary adenoma (FGPA) revealed by an ovarian hyperstimulation syndrome (OHSS) in young women. In the first case, OHSS was observed after GnRH analog injection. Pelvic echography revealed multiple voluminous ovarian cysts. Dopamine agonist posology failed in estradiol hypersecretion control, which necessitated endoscopic endonasal transsphenoidal surgery. The patient experienced improvement in pelvic pain as estradiol hypersecretion decreased during the first few postoperative days. Outcome was favorable, and her menstrual cycle was normal after two months. The second case was a young girl with spontaneous pelvic pain and elevated plasma FSH and estradiol levels. FGPA was confirmed on cerebral MRI. Dopamine agonists were introduced, and surgical removal of the pituitary tumor was scheduled for 7 days later. In the meantime, the patient was admitted and underwent surgery for bilateral adnexal torsion related to OHSS. The pituitary tumor was removed one week later. Outcome was favorable, and estradiol and FSH plasma levels were normal after 3 months. The ovarian cysts were no longer visible on echography after 3 months. Given the lack of efficacy of the current standard medical therapy, surgical removal of pituitary adenomas is the reference treatment for FGPA. The authors suggest that severe OHSS related to FGPA should be considered as a relative surgical emergency and that surgery should not be unduly delayed, given the unpredictable risk of adnexal torsion, particularly in case of voluminous ovarian cysts. The authors performed a literature review on this topic.

Elevated seminal plasma estradiol and epigenetic inactivation of ESR1 and ESR2 is associated with CP/CPPS.

Chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) is associated with urinary tract symptoms and hormonal imbalances amongst others. The heterogeneous clinical presentation, unexplored molecular background and lack of prostate biopsies complicate therapy. Here, using liquid biopsies, we performed a comprehensive translational study on men diagnosed with CP/CPPS type III (n = 50; median age 39.8, range 23–65) and age-matched controls (n = 61; median age 36.8, range 20–69), considering biochemical parameters of blood and ejaculates, and epigenetic regulation of the estrogen receptor genes (ESR1 and ESR2) in leukocytes isolated from blood (systemic regulation) and in somatic cells isolated from ejaculates (local regulation). We found elevated 17β-estradiol (E2) levels in seminal plasma, but not in blood plasma, that was significantly associated with CP/CPPS and impaired urinary tract symptoms. In ejaculated somatic cells of CP/CPPS patients we found that ESR1 and ESR2 were both significantly higher methylated in CpG-promoters and expressionally down-regulated in comparison to controls. Mast cells are reported to contribute to CP/CPPS and are estrogen responsive. Consistent with this, we found that E2 –treatment of human mast cell lines (HMC-1 and LAD2) resulted in altered cytokine and chemokine expression. Interestingly, in HMC-1 cells, possessing epigenetically inactivated ESR1 and ESR2, E2 –treatment led to a reduced transcription of a number of inflammatory genes. Overall, these data suggest that elevated local E2 levels associate with an epigenetic down-regulation of the estrogen receptors and have a prominent role in CP/CPPS. Investigating E2 levels in semen could therefore serve as a promising biomarker to select patients for estrogen targeted therapy.

Effect of Ethanolic Stem Bark Extract of Blighia unijugata (Sapindaceae) on Monosodium Glutamate-Induced Uterine Leiomyoma in Sprague-Dawley Rats

Aims: To establish the presence of monosodium glutamate (MSG) as an ingredient of artificial food seasonings on the Ghanaian market, and to evaluate the anti-fibroid property of an ethanolic stem bark extract of Blighia unijugata on MSG-induced uterine leiomyoma in Sprague-Dawley rats and its safety for use. Study Design: Survey and Experimental. Place and Duration of Study: Department of Pharmacology, CHS; September 2012 and May 2013. Methodology: A survey was conducted to ascertain MSG as an ingredient of Food and Drugs Board approved artificial food seasonings on the Ghanaian Market. Phytochemical screening was performed on an ethanolic, aqueous, and petroleum ether extract of B. unijugata. Thin layer and high performance liquid chromatographic analysis were performed on the ethanolic extract of B. unijugata (EBU), selected after phytochemical screening, to obtain fingerprint chromatograms for identification. Preventive and curative studies (measuring total plasma cholesterol and plasma estradiol and uterus weight) using 50 and 100 mg kg-1 EBU, per os, on 600 and 800 mg kg-1 MSG-induced uterine leiomyoma in Sprague-Dawley rats was conducted. Acute and Delayed toxicity on EBU was tested. Results: Of 21 FDB approved artificial food seasonings, 85.7% had MSG as an ingredient. MSG administration to rats elevated significantly (P ≤ .001) cholesterol, estradiol and uterus weight and size (indicating hyperplasia). Curative treatment reduced significantly (P ≤ .01-.001) the elevated plasma cholesterol and estradiol than preventive treatment. Both treatments however significantly decreased (P ≤ .01-.001) elevated uterus weight. The lethal dose was less than 1000 mg kg-1 p.o. Conclusion: MSG is found in almost all artificial food seasoning on the Ghanaian market which could be a risk factor to the development of uterine leiomyoma. The ethanolic extract of Blighia unijugata reversed hyperplasia induced in the uterus by MSG, making it useful as an anti-fibroid drug.

Effect of Ethanolic Stem Bark Extract of Blighia unijugata (Sapindaceae) on Monosodium Glutamate-Induced Uterine Leiomyoma in Sprague-Dawley Rats

Aims: To establish the presence of monosodium glutamate (MSG) as an ingredient of artificial food seasonings on the Ghanaian market, and to evaluate the anti-fibroid property of an ethanolic stem bark extract of Blighia unijugata on MSG-induced uterine leiomyoma in Sprague-Dawley rats and its safety for use. Study Design: Survey and Experimental. Place and Duration of Study: Department of Pharmacology, CHS; September 2012 and May 2013. Methodology: A survey was conducted to ascertain MSG as an ingredient of Food and Drugs Board approved artificial food seasonings on the Ghanaian Market. Phytochemical screening was performed on an ethanolic, aqueous, and petroleum ether extract of B. unijugata. Thin layer and high performance liquid chromatographic analysis were performed on the ethanolic extract of B. unijugata (EBU), selected after phytochemical screening, to obtain fingerprint chromatograms for identification. Preventive and curative studies (measuring total plasma cholesterol and plasma estradiol and uterus weight) using 50 and 100 mg kg-1 EBU, per os, on 600 and 800 mg kg-1 MSG-induced uterine leiomyoma in Sprague-Dawley rats was conducted. Acute and Delayed toxicity on EBU was tested. Results: Of 21 FDB approved artificial food seasonings, 85.7% had MSG as an ingredient. MSG administration to rats elevated significantly (P ≤ .001) cholesterol, estradiol and uterus weight and size (indicating hyperplasia). Curative treatment reduced significantly (P ≤ .01-.001) the elevated plasma cholesterol and estradiol than preventive treatment. Both treatments however significantly decreased (P ≤ .01-.001) elevated uterus weight. The lethal dose was less than 1000 mg kg-1 p.o. Conclusion: MSG is found in almost all artificial food seasoning on the Ghanaian market which could be a risk factor to the development of uterine leiomyoma. The ethanolic extract of Blighia unijugata reversed hyperplasia induced in the uterus by MSG, making it useful as an anti-fibroid drug.

Estrogen and Leptin Regulation of Endocrinological Features of Anorexia Nervosa

Anorexia nervosa (AN) is an eating disorder characterized by profound weight loss, osteoporosis, amenorrhea, and low leptin levels. Leptin is a hormone secreted from adipocytes in direct proportion to body adiposity and low levels of leptin, or hypoleptinemia, is a key endocrinological feature of AN. AN is more prevalent in women, and is characterized by severely reduced estradiol levels. Estrogens are critical regulators of reproduction and metabolism that act by binding to estrogen receptors (ERs). ERs are members of the nuclear receptor superfamily and they regulate target genes by binding to estrogen response elements (EREs). There is an ERE on the long form of the leptin receptor (OB-Rb), which allows estrogen to modulate OB-Rb expression (Machinal et al, 1999). This review will focus on the possible therapeutic benefits of combining estrogens and leptin therapy for AN patients. We propose that combination therapy would decrease the effective dose of each drug and this in turn would reduce the anorexic side effects reported at higher doses. ESR1 and ESR2 are genes that code for estrogen receptors ERα and ERβ, respectively (Osterlund and Hurd, 2001), and are highly expressed in hypothalamic regions that regulate body weight and reproduction. ERs and OB-Rbs are colocalized in specific hypothalamic neurons and as estrogen levels increase, leptin transport across the blood–brain barrier is enhanced. In addition, knockdown of ERα from hypothalamic steroidogenic factor-1 and pro-opiomelanocortin neurons increases food intake, or reduces energy expenditure, or influences reproduction (Xu et al, 2011). In healthy females, leptin regulates the minute-to-minute oscillations in the levels of luteinizing hormone (LH) and estradiol, the most potent estrogen (Licinio et al, 1998). In women with AN, leptin administration increases pulsatile LH, resulting in an enlargement of the ovaries, increased number and sizes of follicles, and elevated plasma estradiol levels (Welt et al, 2004). Therefore, leptin may be necessary for the resolution of amenorrhea in AN. In addition, leptin through OB-Rb reduces food intake by activating transcription factors such as phosphorylated STAT3 (pSTAT3) (Munzberg et al, 2005). Estrogens potentiate leptin-induced pSTAT3 activation in the hypothalamus. Leptin is one of the few treatments for AN patients (Welt et al, 2004); however, at high doses leptin increases weight loss. There are no standardized medications approved for the treatment of AN by the Food and Drug Administration; however, research has focused on the potential therapeutic actions of leptin, as leptin may promote restoration of menstrual cycles and prevent osteoporosis. Estrogen is also given as a treatment for AN; however, sudden introduction of sex steroids to hypo-estrogenic girls can be followed by reductions in food intake and heightened manifestations of AN (Kauli et al, 1982). Therefore, our hypothesis is that combinatorial treatment with very low levels of estrogens and leptin will decrease the effective dose of each hormone because of the enhanced activation of intracellular cascades, and at the same time this would reduce the anorexic side effects reported at higher doses. To our knowledge, estrogenic enhancement of leptin function has not previously been tested with respect to AN.

Elevated levels of the steroidogenic factor 1 are associated with over-expression of CYP19 in an oestrogen-producing testicular Leydig cell tumour

Background and objectivesTesticular Leydig cell tumours (LCTs) are rare, steroid-secreting tumours. Elevated levels of aromatase (CYP19 or CYP19A1) mRNA have been previously described in LCTs; however, little is known about the mechanism(s) causing CYP19 over-expression. We report an LCT in a 29-year-old male with elevated plasma oestradiol caused by enhanced CYP19 transcription.Design and methodsFirst, we measured the intra-tumour expression of CYP19 and determined the use of CYP19 promoters by qPCR. Secondly, we explored CYP19 and promoter II (PII) for gene amplifications and activating mutations in PII by sequencing. Thirdly, we analysed intra-tumour expression of steroidogenic factor 1 (SF-1 (NR5A1)), liver receptor homologue-1 (LRH-1 (NR5A2)) and cyclooxygenase-2 (COX2 (PTGS2)). Finally, we analysed SF-1 for promoter mutations and gene amplifications.ResultsSimilar to what has been recorded in normal Leydig cells, we first found the bulk of tumour CYP19 transcripts to be PII derived, excluding promoter shift as a cause of enhanced transcription. Secondly, we excluded CYP19 and PII gene amplifications, and activating mutations in PII, as causes of elevated CYP19 mRNA. We found SF-1 mRNA to be up-regulated in the tumour, while LRH-1 and COX2 were down-regulated. The finding of elevated SF-1 levels in the tumour was confirmed by immunohistochemistry. The elevated level of SF-1 was not due to promoter mutations or amplifications of the SF-1 gene.ConclusionsOur results strongly suggest that the elevated levels of SF-1 have induced PII-regulated CYP19 transcription in this tumour. These findings are of relevance to the understanding of CYP19 up-regulation in general, which may occur in several tissues, including breast cancer.

An acute stressor alters steroid hormone levels and activity but not sexual behavior in male and female Ocoee salamanders ( Desmognathus ocoee)

Stressors that are chronic have clear suppressive effects on reproductive behaviors in both males and females. Stressors that are acute have effects on reproductive behavior that are less clear. We measured the effects of an acute bout of handling in laboratory-housed male and female Ocoee salamanders ( Desmognathus ocoee), a species with a prolonged mating season. Handling resulted in decreased locomotory activity and elevated plasma corticosterone, a hallmark of the vertebrate stress response. Handling also decreased plasma testosterone in males and elevated plasma estradiol in females. Despite the handling-induced changes in hormone levels, handling had minimal impact on courtship and mating. Other species in which reproduction is insensitive to acute stressors may live in extreme environments with limited reproductive opportunities, whereas Ocoee salamanders live in a relatively temperate environment with multiple reproductive opportunities. Together, these data indicate that an allostatic response to a stressor can alter locomotory activity and elevate corticosterone without suppressing nonessential behaviors like courtship and mating in a species in which reproductive opportunities can occur over a period of multiple months. The lack of reproductive suppression in Ocoee salamanders might be due to the low energetic cost of courtship and mating in this species combined with potentially elevated energetic stores, highlighting the importance of considering energy budgets when making predictions about behavioral effects of acute stressors.

Genetic and physiological determinants of maternal behavior and lamb survival: Implications for low-input sheep management1,2

The relatively intensive supervision afforded many ewes at lambing time is a barrier to the development of low-input sheep management systems. However, in some flocks, reduction in this level of supervision may initially affect lamb mortality and animal welfare. In this review, possibilities for optimizing behavioral interaction between the ewe and lamb are considered, with the goal of improving lamb survival without the need for high levels of human supervision. At birth, ewes show specific behavioral patterns (e.g., licking or grooming, low-pitched bleats, udder acceptance) that facilitate the transition of the lamb from pre- to postnatal life and that accompany the formation of an exclusive olfactory memory for the lamb. The lamb also performs a specific sequence of behaviors directed toward standing, finding the udder, and sucking. The successful accomplishment of these behavior patterns is vital for the formation of a strong attachment between both partners, and for lamb survival. The expression of maternal behavior in the ewe is affected by her previous maternal experience, by nutrition in pregnancy, by breed, by temperament, and, to some extent, by the behavior of her lamb. The maternal care expressed by a ewe at parturition is indicative of her behavior throughout that lactation and in successive pregnancies, suggesting an underlying basis to maternal care intrinsic to that ewe. Studies with Scottish Blackface and Suffolk ewes show that ewes expressing high levels of maternal care have elevated plasma estradiol in late gestation compared with ewes with poorer maternal care, and that circulating estradiol concentration is correlated with maternal behaviors. Although the genetic basis of maternal behaviors has still to be fully determined, there are possibilities of improving maternal behavior by selection, and a better understanding of the neuroendocrine processes underlying individual differences in maternal behavior may help in developing selection strategies. In addition, selection on lamb behaviors, which show some genetic basis, may also be a route to improve lamb survival. Because behavior of both the ewe and lamb is affected by environmental factors, appropriate management, through pregnancy and at parturition, will enhance the expression of maternal behavior and lamb vigor, and so contribute to improving lamb survival.

Physiology of individual late-run Fraser River sockeye salmon (Oncorhynchus nerka) sampled in the ocean correlates with fate during spawning migration

Beginning in 1995, segments of the late-run sockeye salmon (Oncorhynchus nerka) stocks from the Fraser River, British Columbia, have initiated upriver spawning migration up to 6 weeks earlier than historical records; and those fish have experienced high rates of en route mortality. We examined the correlations between physiological and energetic status prior to river entry with subsequent migratory performance of individual salmon using telemetry and noninvasive biopsies for Adams–Thompson–Shuswap (Adams) and Weaver–Harrison (W–H) stocks. Salmon that failed to reach the river were characterized by a tendency to have elevated levels of chronic and acute stress indicators. For one stock of fish (i.e., Adams) that entered the river, those that died before reaching spawning grounds were individuals with low gross somatic energy. Furthermore, females tended to have elevated plasma estradiol levels. When contrasting fish with different behaviours and fates, fish that did not hold in the estuary and subsequently died tended to have less energy than fish that held and reached spawning areas. Females from the former group also had higher 11-ketotestosterone and estradiol levels relative to those from the later group. These data suggest that differences in physiological and energetic status may be associated with high en route mortality in late-run sockeye salmon.

Regulation of Membrane-Associated Prostaglandin E2 Synthase 1 in Pregnant Sheep Intrauterine Tissues by Glucocorticoid and Estradiol

This study was designed to determine the regulatory effect of glucocorticoid and estradiol on expression of ovine intrauterine membrane-associated prostaglandin E(2) synthase 1 (mPTGES1) in late gestation and at labor. For gestational and labor groups, 16 pregnant ewes from 95-147 d gestational age (dGA) and four pregnant ewes at spontaneous term labor were used. The fetal glucocorticoid group, 14 pregnant ewes at 123-125 dGA with fetuses, was divided into the following groups: after sham adrenalectomy (n = 5), adrenalectomy (n = 4), and adrenalectomy with fetal cortisol replacement to late gestation levels (n = 5). For the maternal glucocorticoid group, nine pregnant ewes were treated with saline (n = 4) and three courses of maternal dexamethasone (n = 5). For the estradiol group, 10 pregnant ewes at 119-121 dGA were treated with sesame oil (n = 5) or estradiol (n = 5) to produce labor levels of estradiol in maternal plasma. Endometrial, myometrial, and placental mRNA and proteins were analyzed by Northern and Western blot and immunocytochemistry for mPTGES1. Data were analyzed by Student's t test and ANOVA. There was a significant increase of placental mPTGES1 in late gestation. Glucocorticoids, given to the mother or fetus, significantly stimulated mPTGES1 in placenta. mPTGES1 was elevated only in the endometrium during spontaneous term labor and after estradiol treatment. The mPTGES1 was localized in the myometrial smooth muscle cells, endometrial stromal cells, and placental trophoblast cells. Our study suggested that increased expression of placental mPTGES1 throughout late gestation might result from the increased fetal and maternal circulating glucocorticoids, whereas elevated maternal plasma estradiol concentration might be responsible for the induced mPTGES1 expression in the endometrium during labor.

Possible Relationship between Elevated Plasma ACTH and Tall Stature in Familial Glucocorticoid Deficiency

Familial glucocorticoid deficiency (FGD) is characterized clinically by severe glucocorticoid deficiency associated with failure of adrenal responsiveness to ACTH but not with mineralcorticoid deficiency. Excessive growth was described previously in some patients with FGD, many of whom were shown to have mutations in the ACTH receptor gene. The mechanisms responsible for their excessive growth are unknown. We analyzed the ACTH receptor gene in three patients with FGD and discussed the causes of excessive growth in FGD. No mutations were detected in the coding and promoter regions of the ACTH receptor gene of one female patient who had tall stature (+ 2.41S.D.) and advanced bone age (10 years 9 months) when she was 4 years 9 months old. Her plasma ACTH level had been elevated until then (124-2,684 pg/ml). Moreover, plasma estradiol was elevated for her age (21.3 pg/ml), and it decreased in response to the dexamethasone suppression test (from 25.4 to 6.9 pg/ml). Elevated plasma estradiol was apparently related to the increase in plasma ACTH and played a major role in excessive growth in this patient. On the other hand, the genetic analysis showed that the other two patients who were siblings were homozygous for the R137W mutation. Clinically, they responded well to hydrocortisone replacement therapy with almost normal plasma ACTH levels. Although all patients with the R137W mutation reported previously were tall, our patients were of normal height. We speculate that the major causes of excessive growth in FGD are not only from ACTH receptor mutation, but also from the action of elevated plasma ACTH.

Formation of cystic ovarian follicles associated with elevated luteinizing hormone requires estrogen receptor-beta.

Stringent regulation of LH secretion from the pituitary is vital to ovarian function in mammals. Two rodent models of LH hypersecretion are the transgenic LHbeta-C-terminal peptide (LHbetaCTP) and estrogen receptor-alpha (ERalpha)-null (alphaERKO) mice. Both exhibit ovarian phenotypes of chronic anovulation, cystic and hemorrhagic follicles, lack of corpora lutea, interstitial/stromal hyperplasia, and elevated plasma estradiol and testosterone. Because ERbeta is highly expressed in granulosa cells of the ovary, we hypothesized the intraovarian actions of ERbeta may be necessary for full manifestation of phenotypes associated with LH hyperstimulation. To address this question, we generated female mice that possess elevated LH, but lack ERbeta, by breeding the LHbetaCTP and ERbeta-null (betaERKO) mice. A comparison of LHbetaCTP, alphaERKO, and betaERKO(LHCTP) females has allowed us to elucidate the contribution of each ER form to the pathologies and endocrinopathies that occur during chronic LH stimulation of the ovary. alphaERKO ovaries respond to elevated LH by exhibiting an amplified steroidogenic pathway characteristic of the follicular stage of the ovarian cycle, whereas wild-type(LHCTP) and betaERKO(LHCTP) females exhibit a steroidogenic profile more characteristic of the luteal stage. In addition, the hemorrhagic and cystic follicles of the LHbetaCTP and alphaERKO ovaries require the intraovarian actions of ERbeta for manifestation, because they were lacking in the betaERKO(LHCTP) ovary. In turn, ectopic expression of the Leydig cell-specific enzyme, Hsd17b3, and male-like testosterone synthesis in the alphaERKO ovary are unique to this genotype and are therefore the culmination of elevated LH and the loss of functional ERalpha within the ovary.

Hyperlipidemia and reproductive failure in captive-reared alligators: vitamin E, vitamin A, plasma lipids, fatty acids, and steroid hormones

Blood samples were collected from 26 captive-reared alligators (25 females; one male) and 12 (seven females and five males) wild ‘nuisance’ alligators collected by wildlife personnel in south Louisiana in May 1995. The captive alligators, hatched from artificially incubated eggs in 1972–1973, had received vitamin E supplements during the 3 weeks before the blood sample was collected. Each sample was analyzed for vitamin E (α-tocopherol), vitamin A (retinol), total lipid, triacylglycerol, phospholipid, cholesterol, cholesteryl ester, free fatty acids, steroid hormones and a standard clinical blood panel. The fatty acid composition of the plasma lipid fraction was also analyzed. Results indicated that 18 of the captive females and three of the seven wild females were undergoing vitellogenesis, i.e. had elevated plasma estradiol and elevated plasma calcium. Vitellogenic females had higher vitamin E than non-vitellogenic females (77.4 μg/ml vs. 28.6 μg/ml in captive females; 24.0 μg/ml vs. 21 μg/ml in wild females). Plasma retinol was similar in all groups, ranging from 0.5 to 1.4 μg/ml and close to values reported in birds. All lipid fractions, with the exception of cholesteryl ester, were higher in captive alligators than in wild alligators. There were also significant differences in the fatty acid composition of wild and captive alligators. Plasma eicosapentaenoic and docasahexaenoic acid were higher in wild than in captive alligators, whereas linoleic was higher in captive than in wild.

Effects of estrogenic ( o,p′ -DDT; octylphenol) and anti-androgenic ( p,p′-DDE) chemicals on indicators of endocrine status in juvenile male summer flounder ( Paralichthys dentatus)

Laboratory experiments were conducted with male summer flounder to assess the value of selected measures of endocrine status in fish as indicators of exposure to endocrine-disrupting contaminants. Effects of 1,1,1-trichloro-2-( p-chlorophenyl)-2-( o-chlorophenyl) ethane ( o,p′-DDT), octylphenol and 1,1-dichloro-2,2-bis ( p-chlorophenyl) ethylene ( p,p′-DDE) on hepatosomatic and gonadosomatic indices, plasma steroid hormone levels, vitellogenin production, and gonadal development were evaluated in laboratory-raised, juvenile male summer flounder. Flounder were injected twice with test chemical in a coconut oil carrier. Each chemical was tested at three different concentrations. Estrogenic ( o,p′-DDT; octylphenol) and anti-androgenic ( p,p′-DDE) chemicals were evaluated alone and in combination (octylphenol plus o,p′-DDT or p,p′-DDE). Additionally, some fish were treated with the natural ligand for the estrogen receptor, 17β-estradiol. Blood and tissues from different fish in each treatment were sampled 4, 6 and 8 weeks after the first injection. Fish exposed to a combination of o,p′-DDT plus octylphenol were also sampled after 15 weeks. In all cases, responses of fish exposed to a test chemical were compared to control fish sampled at the same time. The following significant differences, relative to controls, were observed in at least one sampling time or at least one concentration of chemical. 17β-Estradiol-treated flounder exhibited decreased gonadosomatic index (GSI), altered hepatosomatic index (HSI), elevated plasma estradiol, reduced plasma testosterone, and high levels of plasma vitellogenin. Fish treated with o,p′-DDT showed lower GSI, no change in HSI or plasma estradiol, depression of plasma testosterone, and induction of vitellogenesis. Octylphenol treatment resulted in lower GSI, no change in HSI, initially increased plasma estradiol and decreased testosterone, and no vitellogenin production. p,p′-DDE treatment did not significantly alter any indicator relative to controls. In experiments using combinations of chemicals, flounder receiving o,p′-DDT plus octylphenol had lower GSI after 8 weeks and elevated plasma estradiol after 15 weeks exposure. Fish treated with p,p′-DDE plus octylphenol for 8 weeks exhibited a significantly lower GSI. Overall, lower GSI and plasma testosterone levels, relative to controls, were consistent indicators of exposure to estrogenic chemicals in juvenile male flounder. No indicators were found that would identify exposure to the mammalian anti-androgen p,p′-DDE.

Effects of exogenous FSH on follicular recruitment in a viviparous lizard Niveoscincus metallicus (Scincidae)

In the viviparous skink Niveoscincus metallicus clutch size appears to be determined before vitellogenesis, and is not altered later by follicular atresia or embryonic loss. This suggests that the number of follicles recruited is determined by the endocrine environment early in the vitellogenic period. Through a series of experiments in which we manipulated gonadotropin concentrations by administering exogenous FSH, we aimed to investigate this hypothesis. Pre-vitellogenic females showed no response to exogenous ovine FSH. In early vitellogenic females, FSH induced follicular recruitment: follicles were enlarged and clutch size increased by recruitment of a second cohort of follicles; some females also ovulated. Females treated with FSH in mid-vitellogenesis had elevated mean plasma estradiol concentrations compared to controls; no follicular recruitment was observed, but most of these animals ovulated. Females treated with a range of doses of FSH in late vitellogenesis ovulated at least one month before natural ovulation, again without recruitment of extra follicles. It appears therefore that in Niveoscincus metallicus exogenous FSH can induce recruitment of additional follicles only if administered during early vitellogenesis. We conclude that in this species clutch size is determined by proximate environmental factors influencing gonadotropin levels early in follicular recruitment, and cannot be increased even if conditions become more favourable once vitellogenesis is established.

The Acute Effects of Androstenedione Supplementation in Healthy Young Males

We examined the effects of androstenedione supplementation on the hormonal profile of 10 males and its interaction with resistance exercise. Baseline testosterone, luteinizing hormone, estradiol, and androstenedione concentrations were established by venous sampling at 3 hr intervals over 24 hr. Subjects ingested 200 mg of androstenedione daily for 2 days, with second and third day blood samples. Two weeks later, they ingested androstenedione or a placebo for 2 days, in a double-blind, cross-over design. On day 2, they performed heavy resistance exercise with blood sampled before, after, and 90 min post. The supplement elevated plasma androstenedione 2--3-fold and luteinizing hormone approximately 70% but did not alter testosterone concentration. Exercise elevated testosterone, with no difference between conditions. Exercise in the supplemented condition significantly elevated plasma estradiol by approximately 83% for 90 min. Androstenedione supplementation, thus, is unlikely to provide male athletes with any anabolic benefit and, with heavy resistance exercise, elevates estrogen.

Estrogen Secreting Adrenal Adenocarcinoma in an 18-Month-old Boy. Aromatase Activity, Protein Expression, mRNA and Utilization of Gonadal Type Promoter.

We examined clinical, endocrinological and molecular biological aspects of an estrogen-secreting adrenal carcinoma in an 18-month-old male to clarify the pathogenesis of this condition. An 18-month-old boy was referred for evaluation of progressive bilateral gynecomastia and appearance of pubic hair. The patient had elevated plasma estradiol (349 pg/ml) and testosterone (260 ng/dl) levels that completely suppressed FSH and LH levels, and was subsequently diagnosed with an adrenal tumor on the right side. After removal of a 300-g adenocarcinoma, gynecomastia regressed and essentially normal hormone levels were restored. Aromatase activity in the tumor tissue determined by the 3H-water method was 71.0-104.4 pmol/min/mg protein. High levels of aromatase protein and mRNA in the tumor tissue were also demonstrated, while neither aromatase activity nor protein was detected in normal adrenal glands. To investigate the regulation of aromatase expression in the adrenal carcinoma, we examined the usage of alternate promoters responsible for aromatase gene transcription. In the present case, the amounts of aromatase mRNA utilizing gonadal types of exon 1c (1.3) and 1d (II) were significantly higher than those that using other exon 1s. This result suggested that the utilization of a gonadal-type exon 1 might be involved in the over-production of aromatase in estrogen-secreting adrenal carcinoma.