Induction of luteolysis in the human with a long-acting analog of luteinizing hormone-releasing factor.

Subcutaneous injection of 50 micrograms of a long-acting analog of luteinizing hormone-releasing factor on each of two successive days during mid-luteal phase in normally cycling women induced a short luteal phase and premature menstruation. These events were associated with luteolysis, as evidenced by the consistent and parallel premature decline of progesterone and estradiol levels compared with those in control cycles. This finding may prove to be useful in the prevention or interception of implantation.

Menopausal flushes: a neuroendocrine link with pulsatile luteninizing hormone secreation.

Menopausal flush episodes were found to be invariably associated with the initiation of pulsatile pituitary release of luteinizing hormone. This was not accompanied by a significant change in circulating catecholamine or prolactin concentrations. Since pulsatile luteinizing hormone release results from episodic secretion of luteinizing hormone releasing factor by the hypothalamus, these findings suggest a link between the neuroendocrine mechanisms that initiate such episodic secretion and those responsible for the onset of flush episodes.

Luteal phase defects induced by an agonist of luteinizing hormone-releasing factor: a model for fertility control.

Subcutaneous injection of 50 micrograms of a luteinizing hormone-releasing factor agonist (LRF agonist) for three successive days at the time of menstruation in normal cycling women induces a shortened luteal phase with suboptimal concentrations of circulating estradiol and progesterone. This luteal phase defect follows a reduced concentration of follicle-stimulating hormone during the follicular phase and a resulting inadequate follicular maturation. Since a short luteal phase is associated with an endometrium not conductive to implantation, administration of the LRF agonist at the onset of menstrual cycle may prove to be a practical and novel approach to fertility control.

Direct evidence of estrogen modulation of pituitary sensitivity to luteinizing hormone-releasing factor during the menstrual cycle.

To delineate the role of estradiol in the augmented pituitary gonadotropin responsiveness to synthetic luteinizing hormone releasing factor (LRF) seen during high-estrogen phases of the ovulatory cycles (late follicular and midluteal phases), the anti-estrogenic effect of clomiphene citrate (Clomid) on pituitary response to LRF was evaluated during different phases of the ovulatory cycle. Clomid administration (100 mg/day times 5 days) completely negates the augmented gonadotropin responses to LRF (150 mug) during late follicular and midluteal phases observed during the control studies. Thus, a quantitatively and qualitatively similar pituitary sensitivity to LRF during three distinct phases of the menstrual cycle was induced by Clomid treatment that resembles the LRF responsiveness of themale pituitary. The present study demonstrates the pituitary component of the estrogen-induced changes in the sensitivity to LRF. From this and previous data, we conclude that the increases of estradiol secretion associated with the follicular maturation and corpus luteum formation represent a major component of the feedback signal in the modulation of cyclic gonadotropin release occasioned in a large measure by the augmented pituitary sensitivity to LRF.

Effects of exogenous estrogen and progestin on pituitary responsiveness to synthetic luteinizing hormone-releasing factor.

The effect of estrogen and progestin on pituitary responsiveness to 150 mug synthetic luteinizing hormone-releasing factor (LRF) was assessed in premenopausal women receiving sequential (n=12) and combination (n=7) contraceptive steroids. A marked contrast in the time-course and maximal response to LRF was found; a prompt but quantitatively smaller luteinizing hormone (LH) response was seen during cyclic combination therapy, while a delayed (five times) but enhanced (fivefold) LH response was observed during estrogen segments of cyclic sequential therapy. For follicle-stimulating hormone (FSH), the maximum rise was also higher, and the peak response was similarly delayed in the latter group. The quantitative secretion in response to LRF for LH (area under the curve), but not for FSH, was significantly greater (P < 0.01) in subjects receiving sequential, as compared to subjects receiving combination treatment. In both groups, characteristic gonadotropin responses to LRF were reproducible and were independent of the duration of treatment. Since LRF studies were performed during the estrogen segment of treatment cycle in subjects receiving sequential steroids, our data suggest that estrogen exerts a direct feedback action at the pituitary level and that pituitary responsiveness to LRF is augmented by estrogen.

The effect of ovariectomy on gonadotropin release.

The sequential changes in the concentration and pattern of circulating luteinizing hormone (LH) and follicle-stimulating hormone (FSH)(1) following bilateral ovariectomy were determined in 10 premenopausal women. The initial (1st wk) and delayed (3 wk) secretory responses of serum LH and FSH as related to the phases of the menstrual cycle were examined. Ovariectomy during follicular phase was accompanied by a prompt and much greater rise in both LH and FSH during the 1st wk. This rapid rise was followed by a transient decline between the 7th and 10th day which resulted in a biphasic pattern. In contrast, a slower and progressive rise in serum LH and FSH was observed in subjects ovariectomized during luteal phase of the cycle. The quantitative secretion (area under the curve) during the 1st wk after ovariectomy was significantly greater in patients operated on during the follicular phase than during the luteal phase for both LH (P < 0.05) and FSH (P < 0.01). Thereafter, a similar pattern of gonadotropin rise was observed for patients ovariectonized during either phase of the cycle and reached a plateau by the end of the 3rd wk. At this time, the mean LH concentration increased 6-fold for follicular phase surgery and 8-fold for luteal phase surgery. The mean serum FSH concentration increased 8-fold for follicular phase surgery and 12-fold for luteal phase surgery. The net increase in serum FSH level was higher than that in the serum LH level after surgery in both phases of the cycle and thus a reversal of FSH/LH ratio occurred. These data provide indirect evidence that the phase of ovarian steroid secretion may exert a quantitative influence on the gonadotropin turnover rate within the hypothalamic-pituitary system. The augmented gonadotropin release and the reversal of FSH/LH ratio following ovariectomy presumably could be due to an increased gonadotropin net synthesis which is more pronounced for FSH than for LH.

Augmentation of prolactin secretion by estrogen in hypogonadal women.

The effect of estrogen on prolactin (PRL) release and gonadotropin suppression was assessed in six experiments performed on four hypogonadal women. Ethinyl estradiol at a dose of 1 microgram/kg per day induced a significant elevation of serum PRL levels within the 1st wk of treatment. There was a further rise until a plateau was reached in about 3-4 wk to levels of more than 3 times the initial concentration. This was accompanied by a pattern of increased episodic fluctuation. The corresponding serum luteinizing hormone and follicle-stimulating hormone fell progressively during the study period. These data indicate that a positive feedback relationship between estrogen and PRL release exists in humans.

Characterization of the inappropriate gonadotropin secretion in polycystic ovary syndrome.

To evaluate gonadotropin release in polycystic ovary syndrome (PCO), one or more of the following hypothalamic-pituitary function tests were performed on 24 patients with the syndrome. These tests included (a) the pulsatile pattern and day-to-day fluctuation of gonadotropin release; (b) effects of exogenous estrogen and antiestrogen (clomiphene) administration on gonadotropin release; and (c) pituitary responsiveness to maximal (150 mug) and submaximal (10 mug) luteinizing hormone-releasing factor (LRF) injections. In 10 of the 14 patients sampled frequently (15 min) for 6 h, luteinizing hormone (LH) levels were elevated above the concentration seen in normal cycling women (except the LH surge). These high LH concentrations appeared to be maintained by and temporally related to the presence of exaggerated pulsatile LH release, either in the form of enhanced amplitude or increased frequency. In all subjects, levels of follicle-stimulating hormone (FSH) were low or low normal, and a pulsatile pattern was not discernible. In four patients, daily sampling revealed marked day-to-day fluctuation of LH but not FSH. That the elevated LH levels were not related to a defect in the negative-feedback effect of estrogen was suggested by the appropriate fall of LH in four patients given an acute intravenous infusion of 17beta-estradiol. This infusion had no effect on FSH levels. In addition, clomiphene elicited rises of both LH and FSH that were comparable to the ones observed in normal women given the same treatment. The clomiphene study also suggested that the positive-feed-back mechanism of estrogen on LH release was intact when the preovulatory rises of 17beta-estradiol induced appropriate LH surges. The elevated LH levels appeared to be related to a heightened pituitary responsiveness to the LRF. This was found in the 11 and 2 patients given maximal (150 mug) and submaximal (10 mug) doses of LRF, respectively. The augmented pituitary sensitivity for LH release correlated with the basal levels of both estrone (P less than 0.025) and 17beta-estradiol (P less than 0.02). The net increase in FSH was significantly greater (P less than 0.001) in the PCO patients than the normal women with maximal doses of LRF. With the smaller dose study none of the injections had a discernible effect on FSH concentrations in either subject. The disparity between LH and FSH secretion could be explained by the preferential inhibitory action of estrogen on FSH release, coupled with a relative insensitivity of FSH release. These data indicate that in these PCO patients the abnormalities of the hypothalamic-pituitary regulation of gonadotropin secretion was not an inherent defect but represented a functional derangement consequent to inappropriate estrogen feedback, which led to a vicious cycle of chronic anovulation and inappropriate gonadotropin secretion.

Astrocytes cultured in vitro produce estradiol-17beta and express aromatase cytochrome P-450 (P-450 AROM) mRNA.

Aromatase cytochrome P-450 (P-450AROM) is an enzyme that catalyzes the conversion of androgen to estrogen. Estrogen plays an important role in the neuronal function by promoting the formation of dendrites and may be involved in protecting the neurons in the cerebral cortex against specific pathological conditions such as Alzheimer's disease. However, the cellular origin of estrogen in the brain is not known. The present study demonstrated for the first time the production of estradiol-17beta and expression of P-450AROM mRNA in astrocytes isolated from the cerebral cortex of neonatal rats. Immunocytochemical studies using a monospecific antibody against rat P-450AROM has shown that this enzyme was localized in the cytoplasm of astrocytes. Interleukin-1 (IL-1) has been shown to stimulate the proliferation and differentiation of astrocytes and to affect the aromatase activity in non-neuronal cells such as Sertoli, Leydig, and placental cells. Treatment of astrocytes with IL-1beta induced a dose-dependent inhibition of estradiol production. This inhibitory action of IL-1beta can be reversed by the addition of anti-IL-1beta antibody. Since astrocytes are involved in the synaptic reorganization in the brain by removing cellular debris and by providing the necessary biological factors for neuronal growth, the ability of astrocytes to produce estradiol-17beta and express P-450AROM mRNA in vitro suggests a new role for these cells in protecting and supporting neurons.

Phase I/pharmacokinetic study of high-dose progesterone and doxorubicin.

PURPOSE: We developed a new formulation of progesterone that permits administration of up to 10 g of progesterone as a continuous intravenous infusion over 24 hours and conducted a phase I clinical trial to determine whether progesterone could modulate the in vivo cytotoxicity of the P-glycoprotein substrate doxorubicin. PATIENTS AND METHODS: Thirty-four patients with advanced malignancies were treated with increasing doses of progesterone and a fixed dose of 60 mg/m2 of doxorubicin given as an intravenous bolus 2 hours after starting a 24-hour intravenous infusion of progesterone. RESULTS: Progesterone enhanced doxorubicin-induced myelotoxicity in a dose-dependent fashion without altering the pharmacokinetics of doxorubicin. The steady-state plasma concentration of progesterone at a dose level of 4 g was 4.1 +/- 0.9 mumol/L, which was higher than the minimal concentration required to reverse multidrug resistance (MDR) in vitro. CONCLUSION: Progesterone enhanced the hematologic toxicity of doxorubicin without altering its pharmacokinetics, suggesting that progesterone could modulate P-glycoprotein at the level of pluripotent hematopoietic stem cells. Adequate tissue concentrations of progesterone could be achieved in vivo to modulate doxorubicin toxicity in the bone marrow and thus potentially in tumor tissue as well. Selectivity may potentially be gained by using hematopoietic growth factors to offset the enhanced hematologic toxicity of doxorubicin while leaving the enhancement of toxicity to tumor cells unchanged.

Altered waveform of plasma nocturnal melatonin secretion in premenstrual depression.

The nocturnal secretion of plasma melatonin was determined under dim to dark conditions in eight patients with prospectively confirmed premenstrual syndrome and in eight age- and menstrual cycle phase-matched normal control subjects. Plasma samples for melatonin were collected every 30 minutes from 6 PM to 9 AM during the early follicular, late follicular, midluteal and late luteal phases of the menstrual cycle. Compared with normal controls, patients with premenstrual syndrome had an earlier (phase-advanced) offset of melatonin secretion, which contributed to a shorter secretion duration and a decreased area under the curve. No statistically significant differences were found between women with premenstrual syndrome and normal controls for melatonin onset or peak concentration, or for estradiol or progesterone levels. The data demonstrate that women with premenstrual syndrome have chronobiological abnormalities of melatonin secretion. The fact that these patients respond to treatments that affect circadian physiology, such as sleep deprivation and phototherapy, suggests that circadian abnormalities may contribute to the pathogenesis of premenstrual syndrome.

The effect of an antiserum to luteinizing hormone-releasing hormone on the progesterone-induced luteinizing hormone surge in ovariectomized, estrogen-primed rats.

A rabbit antiserum to synthetic LHRH was shown to be specific to rat hypothalamic LHRH, as revealed by binding studies (RIA). The present study was undertaken to determine the effects of this antiserum (LHRH-AS) on the progesterone-induced LH surge in ovariectomized rats primed with estrogen on the proestrous LH surge in cycling females. An ip injection of 0.2 ml LHRH-AS at 1200 h on proestrus completely inhibited the preovulatory rise in serum LH from 1700-1800 h. In long term ovariectomized rats, a sc injection of estradiol benzoate reduced serum LH levels by 50% at 1200 h on the third day. Subsequent administration of progesterone elicited a LH surge, with peak LH value 4-6 h after progesterone. However, concurrent administration of LHRH-AS effectively prevented the increase in LH release by progesterone, and an iv injection of LHRH-AS was shown to be more effective than ip administration in inhibiting the progesterone stimulation of LH release. Although estrogen and progesterone may induce a change in pituitary responsiveness to hypothalamic LHRH in ovariectomized rats, the present study provides evidence that tonic and/or increased LHRH release is required for the acute increase in LH release caused by progesterone.

Neurosteroidogenesis in astrocytes, oligodendrocytes, and neurons of cerebral cortex of rat brain.

The brain is a steroidogenic organ that expresses steroidogenic enzymes and produces neurosteroids. Although considerable information is now available regarding the steroidogenic capacity of the brain, little is known regarding the steroidogenic pathway and relative contributions of astrocytes, oligodendrocytes, and neurons to neurosteroidogenesis. In the present study, we investigated differential gene expression of the key steroidogenic enzymes using RT-PCR and quantitatively evaluated the production of neurosteroids by highly purified astrocytes, oligodendrocytes, and neurons from the cerebral cortex of neonatal rat brains using specific and sensitive RIAs. Astrocytes appear to be the most active steroidogenic cells in the brain. These cells express cytochrome P450 side-chain cleavage (P450scc), 17alpha-hydroxylase/C17-20-lyase (P450c17), 3beta-hydroxysteroid dehydrogenase (3betaHSD), 17beta-hydroxysteroid dehydrogenase (17betaHSD), and cytochrome P450 aromatase (P450arom) and produce pregnenolone (P5), progesterone (P4), dehydroepiandrosterone (DHEA), androstenedione (A4), testosterone (T), estradiol, and estrone. Oligodendrocytes express only P450scc and 3betaHSD and produce P5, P4, and A4. These cells do not express P450c17, 17betaHSD, or P450arom or produce DHEA, T, or estrogen. Neurons express P450scc, P450c17, 3betaHSD, and P450arom and produce P5, DHEA, A4, and estrogen, but do not express 17betaHSD or produce T. By comparing the ability of each cell type in the production of neurosteroids, astrocytes are the major producer of P4, DHEA, and androgens, whereas oligodendrocytes are predominantly the producer of P5 and neurons of estrogens. These findings serve to define the neurosteroidogenic pathway, with special emphasis on the dominant role of astrocytes and their interaction with oligodendrocytes and neurons in the genesis of DHEA and active sex steroids. Thus, we propose that neurosteroidogenesis is accomplished by a tripartite contribution of the three cell types in the brain.

Dehydroepiandrosterone: biosynthesis and metabolism in the brain.

Dehydroepiandrosterone (DHEA) is abundantly found in brain tissues of several species, including human. However, the cellular origin and pathway by which DHEA is synthesized in brain are not yet known. We have, therefore, initiated pilot experiments to explore gene expression of cytochrome P450 17alpha-hydroxylase (P450c17), the key steroidogenic enzyme for androgen synthesis, and evaluate DHEA production by highly purified astrocytes, oligodendrocytes, and neurons. Using RT-PCR, we have demonstrated for the first time that astrocytes and neurons in the cerebral cortex of neonatal rat brain express P450c17. The presence of P450c17 in astrocytes and neurons was supported by the ability of these cells to metabolize pregnenolone to DHEA in a dose-dependent manner as determined by RIA. These data were further confirmed by production of androstenedione by astrocytes using progesterone as a substrate. However, cortical neurons express a low transcript of P450c17 messenger RNA and produce low levels of DHEA and androstenedione compared with astrocytes. Oligodendrocytes neither express the messenger RNA nor produce DHEA. The production of DHEA by astrocytes is not limited to cerebral cortex, as hypothalamic astrocytes produce DHEA at a level 3 times higher than that produced by cortical astrocytes. Cortical and hypothalamic astrocytes also have the capacity to metabolize DHEA to testosterone and estradiol in a dose-dependent manner. However, hypothalamic astrocytes were 3 times more active than cortical astrocytes in the metabolism of DHEA to estradiol. In conclusion, our data presented evidence that astrocytes and neurons express P450c17 and synthesize DHEA from pregnenolone. Astrocytes also have the capacity to metabolize DHEA into sex steroid hormones. These data suggest that as in gonads and adrenal, DHEA is biosynthesized in the brain by a P450c17-dependent mechanism.

Changes in beta-endorphin-like immunoreactivity in pituitary portal blood during the estrous cycle and after ovariectomy in rats.

beta-Endorphin-like immunoreactivity (beta-EP-LI) was measured by RIA in plasma collected from pituitary portal vessels of rats at various times in the estrous cycle and after ovariectomy. There were no appreciable differences between the mean beta-EP-LI concentration either in the afternoon (1500-1700 h) of estrus (4.1 +/- 1.5 ng/ml) or on diestrous days 1 (4.1 +/- 1.3 ng/ml) or 2 (5.4 +/- 1.5 ng/ml). The concentration increased slightly (6.7 +/- 1.3 ng/ml) but not significantly in the early afternoon (1400-1500 h) of proestrus. The concentration of beta-EP-LI then fell to 0.5 +/- 0.1 ng/ml at 1700-1800 h on proestrus, a level significantly lower than at any other time of the cycle. Portal plasma beta-EP-LI was also low (1.9 +/- 0.5 ng/ml) in animals ovariectomized for 3 weeks. After gel filtration of the portal plasma extracts, most of the beta-EP-LI eluted in the same position as synthetic beta-EP. Dilution of portal plasma produced a displacement curve parallel to that of beta-EP and hypothalamic extract. These results indicate that the secretion of hypothalamic beta-EP into the blood of pituitary portal vessels changes during the estrous cycle, possibly due to cyclic changes in sex steroids.

Hyperprolactinemia decreases the luteinizing hormone-releasing hormone concentration in pituitary portal plasma: a possible role for beta-endorphin as a mediator.

Hyperprolactinemia can reduce the LH secretion in rats, but the mechanism of the effect of PRL is not clear. We have investigated the actions of PRL on the secretion of LHRH and LH and the interaction among PRL, beta-endorphin (beta-EP), and LHRH. The effects of PRL on LHRH and LH secretion were studied in ovariectomized female rats after transplanting four anterior pituitaries to the right kidney capsule of each ovariectomized rat for 2-3 weeks. The level of PRL in rats with pituitary transplants was approximately 5 times higher than that in control rats. The concentration of LHRH in pituitary portal plasma of hyperprolactinemic rats was approximately 4 times lower than that in control rats. Hyperprolactinemic animals also showed lower plasma LH levels than the controls. Since beta-EP inhibits the secretion of LHRH, we have tested whether the reduced secretion of LHRH in hyperprolactinemic ovariectomized rats is associated with an increase in beta-EP activity. This was studied by measuring the concentration of beta-EP in pituitary portal plasma and the response of LHRH and LH to the opiate antagonist naloxone. The level of beta-EP-like immunoreactivity in pituitary portal plasma was significantly higher in hyperprolactinemic rats than in control animals. Naloxone (10 mg/kg, sc) increased both LHRH and LH concentrations in hyperprolactinemic rats, but not in control rats. The present results demonstrate that hyperprolactinemia can reduce LHRH release and suggest a possible involvement of beta-EP in the PRL inhibitory action on LHRH.

Insulin enhancement of luteinizing hormone and follicle-stimulating hormone release by cultured pituitary cells.

The role of insulin in the regulation of basal and gonadotropin-releasing hormone (GnRH)-stimulated release of LH and FSH was investigated in vitro using primary cultures of rat anterior pituitary cells from adult ovariectomized rats. Anterior pituitary cells were incubated for 2 days in the presence or absence of insulin in a serum-free medium. At the end of the insulin treatment, the cells were washed and reincubated in the presence or absence of GnRH, and the LH and FSH released into the medium were measured by RIA. Treatment with insulin (1.0 microgram/ml) for 2 days resulted in significant increases in both the basal and the maximal release of LH and FSH, as well as a 3.2- and 6.3-fold decrease in the ED50 values for GnRH in terms of LH and FSH release, respectively. Treatment with increasing concentrations (0.1-10,000 ng/ml) of insulin, led to a dose-dependent increase in the GnRH (3 X 10(-10) M)-stimulated release of both LH and FSH. This effect of insulin was significant (P less than 0.05) at a physiological concentration of 1 ng/ml (24 microU/ml) with an ED50 value of 40 ng/ml. Increasing duration of exposure to insulin resulted in time-dependent increases in the GnRH (3 X 10(-10) M)-stimulated release of LH, becoming significant at 24 h with maximal enhancement observed by 48 h. The effect of insulin was specific; epidermal or fibroblast growth factor did not enhance LH release. The augmenting effect of insulin was not associated with cellular proliferation or an overall change in protein or LH synthesis. Furthermore, the effect of insulin was independent of the ambient glucose concentration. Insulin was, however, without effect on gonadotrophs cultured in a serum-supplemented medium. Our findings suggest that the gonadotroph constitutes a target cell of insulin and that insulin may act directly on the anterior pituitary in the regulation of gonadotropin release.

Prolactin-releasing action of gonadotropin-releasing hormone in hypogonadal women.

To gain insight into the PRL-releasing effect of GnRH, serum PRL and gonadotropin responses to a 10-microgram iv bolus dose of exogenous GnRH were studied in hypergonadotropic hypogonadal women (HHW) and patients with functional hypothalamic amenorrhea (FHA). The results were compared with those obtained in normal cycling women during the early follicular phase of the cycle. GnRH induced a significant increase in PRL levels (P less than 0.001) in HHW compared to early follicular phase women, in whom no significant response occurred. In HHW, the maximal PRL percent increment was positively correlated with the ratio of the maximal percent increments of FSH and LH (r = 0.93). GnRH induced a significant increase in PRL levels in every FHA patient, but in four of them (high PRL responders), the PRL response was at least 5-fold greater than in the other six (low PRL responders). The clinical profiles, basal hormone concentrations, and LH responses to GnRH were similar in these two groups of FHA patients, but the FSH response to GnRH was greater (P less than 0.05) in the high PRL responders. The maximal percent increment of PRL was also positively correlated with the maximal percent increment of FSH (r = 0.76; P = 0.01). These data demonstrate that in these two hypogonadal models, the PRL response to exogenous GnRH corresponds to the FSH response and suggests that GnRH-stimulated PRL release may be mediated by a paracrine effect between FSH-enriched gonadotrophs and lactotrophs.

Sleep-associated decrease in luteinizing hormone pulse frequency during the early follicular phase of the menstrual cycle: evidence for an opioidergic mechanism.

To investigate the possible role of an opioidergic mechanism(s) in the sleep-associated decrease in LH pulse frequency during the early follicular phase of the menstrual cycle, 10 normal cycling women were studied on days 3 and 4 of their cycles before and after the administration of a specific opiate receptor antagonist, naloxone. Sequential 24-h infusions of naloxone (10-mg iv bolus dose followed by an infusion of 30 micrograms/kg . h) or NaCl were administered randomly. Pulsatile LH activity was assessed for 48 h. Sleep was confirmed by electroencephalogram monitoring during the night hours (2300-0700 h). Significant sleep-associated decreases in LH pulse frequency (P less than 0.01) and mean serum LH levels (P less than 0.01) were found during the NaCl control studies. While naloxone infusion had no effect on LH pulse frequency during the waking hours, it prevented the sleep-associated decrease in pulse frequency and, in fact, significantly (P less than 0.001) increased the LH pulse frequency during the sleeping hours. These observations provide evidence that a diurnal variation of naloxone sensitivity exists in early follicular phase women and that the decrease in LH pulse frequency normally found during sleep is based at least in part on increased opioidergic inhibition.

Nutritional and endocrine-metabolic aberrations in amenorrheic athletes.

Growing evidence suggests that menstrual disturbances in female athletes are related to the metabolic cost of high levels of energy expenditure without compensatory increases in dietary intake. However, the linkage(s) between nutritional deficits and reproductive impairments as a result of slowing of LH pulsatility has not been defined. This study was directed to simultaneously characterize nutritional intake, insulin sensitivity (by rapid iv glucose tolerance test), and 24-h dynamics of insulin/glucose, cortisol, somatotropic [GH/GH-binding protein (GHBP)/insulin-like growth factor I (IGF-I)/IGF-binding proteins (IGFBPs)], and LH axes in highly trained athletes with (cycling athletes; CA) and without (amenorrheic athletes; AA) menstrual cyclicity and in age- and body mass index-matched cycling sedentary controls (CS; n = 8/group). Although daily caloric intake did not differ among the three groups, athletes (CA and AA) consumed less fat and protein than CS. However, the restriction of fat was 50% greater (P < 0.01) in AA than CA and was accompanied by increased carbohydrate (P < 0.05) and fiber (P < 0.01) intake. Athletes, independent of menstrual status, had increased (P < 0.05) insulin sensitivity and reduced insulin levels during the feeding phase of the day. Hypoinsulinemia was more pronounced in AA (P < 0.05) than CA, extending throughout the day, and was accompanied by reduced glucose increments in response to meals (P < 0.05), not seen in CA. Levels of the insulin-dependent IGFBP-1 were markedly elevated (P < 0.001) throughout the diurnal pattern in AA, whereas in CA, a modest elevation (P < 0.001) of IGFBP-1 levels occurred only during the feeding portion of the day. IGFBP-1 levels for the three groups related inversely to 24-h insulin (r = -0.63) and directly to 24-h cortisol (r = 0.69) levels. A 70-80% augmentation (P < 0.001) of 24-h mean GH levels was seen in both groups of athletes, but with distinct pulsatile features. Although pulse amplitude was increased 60% in CA with no change in pulse number, AA displayed more frequent (P < 0.001) pulses, with an elevated (P < 0.01) baseline between pulses. The distorted pattern of GH pulses seen in AA was associated with a 35% decrease in GHBP levels, not seen in CA. Although levels of IGF-I and IGFBP-3 did not differ in either CA or AA, the 2- to 4-fold higher levels of IGFBP-1 in AA than in CA and CS resulted in a 3-fold reduced ratio of IGF-I/IGFBP-1 in AA, which may decrease the bioactivity and hypoglycemic effect of IGF-I. LH pulse frequency was progressively attenuated in the athletes, with a greater (P < 0.001) slowing in AA than CA, unaccompanied by alterations in pulse amplitude or 24-h levels. LH pulse frequency was related positively with insulin (r = 0.65) levels and the ratio of IGF-I/IGFBP-1 (r = 0.69), and negatively with cortisol (r = -0.70) and IGFBP-1 (r = -0.75) concentrations. Stepwise regression analysis suggested that negative influences associated with hypercortisolemia and elevated IGFBP-1 levels predominate in determining GnRH/LH pulsatile activity in these athletes. In sum, although neuroendocrine-metabolic adaptations to the energy cost of exercise training were evident in both groups of athletes, AA displayed alterations distinct from their cycling counterparts, with evidence of a hypometabolic state, including decreased basal body temperature and reduced levels of plasma glucose and serum GHBP, a decrease in the ratio of IGF-I/IGFBP-1, accelerated GH pulse frequency, and elevated interpulse GH levels. Thus, in AA, increased insulin sensitivity, decreased circulating insulin, and a reduced hypoglycemic effect of IGF-I together with elevated GH and cortisol concentrations may comprise a cascade of glucoregulatory adaptations to repartition metabolic fuels for conservation of protein. (ABSTRACT TRUNCATED)