Potent action of RFamide-related peptide-3 on pituitary gonadotropes indicative of a hypophysiotropic role in the negative regulation of gonadotropin secretion.

We identified a gene in the ovine hypothalamus encoding for RFamide-related peptide-3 (RFRP-3), and tested the hypothesis that this system produces a hypophysiotropic hormone that inhibits the function of pituitary gonadotropes. The RFRP-3 gene encodes for a peptide that appears identical to human RFRP-3 homolog. Using an antiserum raised against RFRP-3, cells were localized to the dorsomedial hypothalamic nucleus/paraventricular nucleus of the ovine brain and shown to project to the neurosecretory zone of the ovine median eminence, predicating a role for this peptide in the regulation of anterior pituitary gland function. Ovine RFRP-3 peptide was tested for biological activity in vitro and in vivo, and was shown to reduce LH and FSH secretion in a specific manner. RFRP-3 potently inhibited GnRH-stimulated mobilization of intracellular calcium in gonadotropes. These data indicate that RFRP-3 is a specific and potent mammalian gonadotropin-inhibiting hormone, and that it acts upon pituitary gonadotropes to reduce GnRH-stimulated gonadotropin secretion.

[Effect of delayed-action Mirenil (Polfa) on the hypothalamic function in patients with schizophrenia]. / Wpøyw mirenilu prolongatum (Polfa) na czynnosc podwzgórza u chorych z rozpoznaniem schizofrenii.

Hormonal tests--TRH, ITT and L-RH--were performed in 10 cases of paranoid schizophrenia before the paranoid deterioration and after--on average--ten months of treatment with fluphenazine depot injection. The drug did show a clear modulating effect on the mechanisms regulating secretion of gonadotrophins and prolactin --it did not effect the secretion of thyrotropin , thyroxine, triiodothyronine, cortisol, growth hormone or insulin .

Clinical implications of growth hormone feedback mechanisms.

It is now clear that both long-loop and ultrashort-loop feedback mechanisms may operate to autoregulate GH secretion at both central nervous system and pituitary levels. The key components of this control mechanisms involve GH itself, the GH-dependent peptides, somatomedins, and the two specific hypothalamic hypophysiotropic peptides GHRH and somatostatin. Presently, the intimate mechanism(s) and the respective contributions of the different components as well as the possible interactions between them under physiological or pathological conditions await clarification. However, a major role played by the hypothalamic release of somatostatin is clearly emerging. The awareness of GH autoregulation, and that the amounts of GH used for therapy are at or above the total daily GH production suggest caution in the institution of treatment with GH in children with idiopathic short stature, although resilience of the endogenous GH-secretory machinery would be present in these instances. Ideally, a GH replacement therapy should mimic the physiological secretory pattern of GH.

The electrophysiology of the GnRH pulse generator in the rhesus monkey.

The electrophysiological correlates of hypothalamic GnRH pulse generator activity in the rhesus monkey are described. They are characterized by abrupt increases in neuronal multiunit electrical activity (volleys) that are coincident with the initiation of LH pulses in the peripheral circulation. Both the frequency and duration of these volleys can be varied by a variety of endocrinological, pharmacological and environmental perturbations. Their activity throughout the rhesus monkey menstrual cycle as assessed by radiotelemetry is described.

Endocrinological basis of hot flushes.

Hot flushes are frequent among women during natural, surgical, or pharmacological menopause. The available data suggest the involvement of estrogens, progestins, catecholestrogens, catecholamines, dopamine, endorphins, prostaglandins, luteinizing hormone (LH) and luteinizing hormone-releasing hormone (LH-RH) in the pathogenesis of flushes. At present the estrogen withdrawal and pulsatile luteinizing hormone (LH) secretion theories are most commonly accepted for explaining the development of this symptom. The use of LH-RH agonists offers an opportunity to focus on the probable origin and region that regulate the events of this phenomena, since the administration of this drug is associated with hot flushes, in spite of low gonadotropins and normal estrogen levels. Current data may suggest that the origin of this neurovegetative symptom lies in the hypothalamus.

Effects of ovariectomy on GnRH mRNA, proGnRH and GnRH levels in the preoptic hypothalamus of the female rat.

Experiments were carried out to investigate the effects of ovariectomy on gonadotropin-releasing hormone (GnRH) messenger RNA (mRNA), proGnRH and GnRH peptide levels in the hypothalamus of female rats. Intact proestrous female rats and female rats, which had been ovariectomized for 2 weeks, were sacrificed at 9.00 h and the preoptic area (POA) and basal hypothalamus (BH) were dissected out and frozen on dry ice. One group of tissues from proestrous control and ovariectomized females were extracted in acetic acid, centrifuged at 13,000 g and the supernatant purified on a C18 column. The purified extract was then radioimmunoassayed for proGnRH, using a specific antiserum to rat proGnRH (ARK-2), and for GnRH using the E1-14 antiserum. Total cellular RNA was isolated from another group of tissues and prepared as Northern blots. Hybridization with 32P-labeled GnRH cRNA was used to detect GnRH mRNA. A third group of proestrous and ovariectomized female rats were perfused, and 50 microns vibratome sections were cut. These were immunostained with proGnRH or GnRH antiserum, followed by in situ hybridization with 35S-labeled GnRH cRNA to detect GnRH mRNA. Based on the histochemical staining, mRNA was colocalized to the cell soma of neurons containing proGnRH and GnRH throughout the POA and BH. Based on the radioimmunoassay, proGnRH levels were 2 times higher in the POA versus the BH, but GnRH levels were 6-7 times higher in the BH. Ovariectomy significantly decreased proGnRH levels in both the POA and BH, while GnRH decreased in the BH. In contrast, quantitative Northern blot analysis demonstrated that ovariectomy had no effect on mRNA levels in the POA and BH. These data indicate that the effects of ovariectomy on proGnRH and GnRH levels are a result of altered translation, posttranslational processing and/or secretion of GnRH.

Neuroendocrine gene expression in the hypothalamus: in situ hybridization histochemical studies.

1. We have reviewed recent studies in which in situ hybridization histochemistry (ISHH) was used to investigate the regulation of expression of neurohypophysial peptides and hypothalamic releasing hormones. 2. ISHH is a technique in which the presence and quantity of a specific mRNA can be determined in tissue sections with a high degree of resolution and sensitivity. 3. ISHH has been used to measure changes in cellular levels of mRNAs encoding vasopressin, oxytocin, corticotropin-releasing factor, gonadotropin-releasing hormone, thyrotropin-releasing hormone and somatostatin in response to various physiological challenges. 4. A theme emerging from these studies is that changes in levels of mRNA encoding neuroendocrine peptides reflect changes in biosynthesis and secretion.

Accessory olfactory bulb transplants correct hypogonadism in mutant mice.

Transplantation of normal fetal gonadotropin-releasing hormone (GnRH) neurons from the accessory olfactory bulb (AOB) to the third ventricle of GnRH-deficient adult mutant mice reverses the genetically determined reduction in pituitary hormones and poorly developed gonads. The transplanted heterotopic AOB neurons adapt their morphology and secretory functions to what is observed with preoptic GnRH neurons when transplanted into deficient mice and in the normal intact mature animal. This suggests the presence of median eminence trophic factors affecting the growth, terminal sprouting, and functional behavior of the transplanted neurons.

Activity of gonadotropin-releasing hormone neurons during the preovulatory luteinizing hormone surge.

We assessed the frequency of luteinizing hormone (LH) pulsatility (reflecting the activity of gonadotropin-releasing hormone [GnRH] neurons in the hypothalamus) in six women during the periovulatory LH surge, in five women during the early follicular phase, and in seven women in the midfollicular phase (MFP) (calculated as being 3 to 8 days before the LH surge). Collection of blood at 5-minute, versus 15-minute, intervals allowed detection of a larger number of LH pulses in both the MFP (16, versus 27) and periovulatory phase (POP) (11, versus 22) groups of women, but it made no difference in the early follicular phase (EFP) (10 pulses with both methods). During the EFP, the mean number of LH pulses per 4 hours (detected by 5-minute sampling) was 2.0 +/- 0.7 (+/- standard deviation [SD]), and the mean LH amplitude (+/- SD) was 1.3 +/- 0.4 IU/l. There was a significant increase in the number of pulses in the MFP group (3.9 +/- 1.3 pulses/4 hours; P less than 0.05) but no significant change in pulse amplitude (1.1 +/- 0.1 IU/l). During the POP, the mean pulse amplitude was increased (8.5 +/- 1.4 IU/l; P less than 0.001), compared with the MFP and EFP groups, but the mean pulse frequency (3.7 +/- 1.2 pulses/4 hours) was not significantly different from the MFP frequency. We conclude that an acceleration of LH pulsatility occurs several days before the LH surge and does not change thereafter. However, there is an increase in LH pulse amplitude during the LH surge; we attribute this to the increase in pituitary sensitivity at this time.(ABSTRACT TRUNCATED AT 250 WORDS)

Does the seasonal increase in estradiol negative feedback prevent luteinizing hormone surges in anestrous ewes by suppressing hypothalamic gonadotropin-releasing hormone pulse frequency?

To test the hypothesis that the anestrous increase in estradiol negative feedback prevents estrous cycles by suppressing hypothalamic gonadotropin-releasing hormone (GnRH) pulse frequency, a variety of regimens of increasing GnRH pulse frequency were administered to anestrous ewes for 3 days. A luteinizing hormone (LH) surge was induced in 45 of 46 ewes regardless of amplitude or frequency of GnRH pulses, but only 19 had luteal phases. Estradiol administration induced LH surges in 6 of 6 ewes, only 3 having luteal phases. Anestrous luteal phase progesterone profiles were similar in incidence, time course, and amplitude to those of the first luteal phases of the breeding season, which in turn had lower progesterone maxima than late breeding season luteal phases. In the remaining ewes, progesterone increased briefly or not at all, the increases being similar to the transient rises in progesterone occurring in most ewes at the onset of the breeding season. These results demonstrate that increasing GnRH pulse frequency induces LH surges in anestrus and that the subsequent events are similar to those at the beginning of the breeding season. Finally, they support the hypothesis that the negative feedback action of estradiol prevents cycles in anestrus by suppressing the frequency of the hypothalamic pulse generator.

The role of endogenous gonadotropin-releasing hormone in the control of luteinizing hormone and testosterone secretion in the juvenile male monkey, Macaca fascicularis.

To determine what changes occur in the activity of gonadotropin-releasing hormone (GnRH) neurons during pubertal development in primate species we tested the hypotheses that there are morphologic differences between GnRH-containing neurons in juvenile versus adult monkeys, and the low activity of the reproductive axis is governed by hypothalamic GnRH release in monkeys prior to puberty. We removed the brains from 5 juvenile and 5 adult male monkeys (Macaca fascicularis) and blocked, sectioned, and prepared each hypothalamus for light microscopic immunocytochemistry for GnRH-containing cells. The distribution and number of GnRH-containing neurons were similar in adult and juvenile brains; however, GnRH-containing perikarya in adult brains were significantly larger in total cross-sectional area (200 +/- 12 vs. 169 +/- 8 micron 2, P less than 0.05) and in cross-sectional area of the cytoplasm (139 +/- 2 vs. 88 +/- 6 micron 2, P less than 0.05) than in juvenile brains. In another group of 10 juvenile male macaques, we administered an antiserum to GnRH (Fraser #94; 2 ml/kg, i.v.) and monitored the effects on plasma luteinizing hormone (LH) and testosterone concentrations. The percentage of plasma samples with detectable LH levels decreased significantly (from 26.67 +/- 8.3% to 5.3 +/- 3.4%, P less than 0.05) after GnRH antiserum administration; however, plasma testosterone concentrations (0.08 +/- 0.02 ng/ml) remained unchanged. We conclude that during pubertal maturation in primate species there is increased synthesis and release of GnRH from a population of GnRH neurons that are active prior to puberty.

Neuroendocrinology of opioid peptides and their role in the control of gonadotropin and prolactin secretion.

Substantial evidence now exists to indicate that the endogenous hypothalamic opioidergic mechanism(s) represents one of the important controlling systems for release of gonadotropin-releasing hormone. Modulations of frequency and amplitude of the secretory activity of gonadotropin-releasing hormone appears to be mediated through an inhibitory action of endogenous opioids, and the functional coupling of the opioidergic and gonadotropin-releasing hormone systems is an ovarian steroid-dependent event. There is also evidence to implicate suprahypothalamic mechanism(s) that enhance endogenous opioid inhibition of secretion of gonadotropin-releasing hormone. Although exogenous opioid peptides and their synthetic analogs consistently induce the secretion of prolactin, blockade of opioid receptors in humans by naloxone failed to elicit a decrement in the levels of prolactin under a variety of conditions. On the contrary, naloxone induced a remarkable increment in the secretion of prolactin via an increased frequency of pulsatile release which is synchronized with pulses of luteinizing hormone. These observations suggest that a common neuroendocrine mechanism is involved in the opioidergic control of the secretion of both luteinizing hormone and prolactin in women.

[Course and regulation of the menstrual cycle from Robert Schröder to the present]. / Ablauf und Regulation des mensuellen Zyklus seit Robert Schröder bis zur Gegenwart.

The fundamental histological findings about endometrial and ovarian changes during the menstrual cycle by Robert Schröder (1884-1959) are passed in review and some actual conceptions of the regulation of the ovarian cycle are cited. In addition, general findings of molecular processes during gonadotropin actions in follicular cells are described, e.g. activation and desensitization of adenylyl cyclase, the role of 3',5'-adenosinmonophosphate and down-regulation of gonadotropin receptors. In general, the progress in cycle research since the work of R. Schröder was done consists in the transition from the histological description of endometrial changes to recognition of intracellular events which have to be considered by gynaecologists.