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The oestrogen-induced surge of LH requires a 'signal' pattern of gonadotrophin-releasing hormone input to the pituitary gland in the ewe.

Two experiments were conducted with ovariectomized and hypothalamo-pituitary disconnected (HPD) ewes to ascertain the pattern of inputs, to the pituitary gland, of gonadotrophin-releasing hormone (GnRH) necessary for the full expression of an oestrogen-induced LH surge. The standard GnRH replacement to these sheep was to give pulses of 250 ng (i.v.) every 2h; at the onset of experimentation, pulses were given hourly. In experiment 1, groups of sheep (n = 7) were given an i.m. injection of 50 micrograms oestradiol benzoate, and after 10 h the GnRH pulse frequency or pulse amplitude was doubled. Monitoring of plasma LH concentrations showed that a doubling of pulse frequency produced a marked increase in baseline values, whereas a doubling of amplitude had little effect on the LH response. In a second experiment, ovariectomized HPD sheep that had received hourly pulses of GnRH for 16 h after an i.m. injection of oil or 50 micrograms oestradiol benzoate were given either a 'bolus' (2.25 micrograms GnRH) or a 'volley' (500 ng GnRH pulses 10 min apart for 30 min, plus a 500 ng pulse 15 min later). Both groups then received GnRH pulses (250 ng) every 30 min for the next 13 h. Oestrogen enhanced the LH responses to the GnRH treatments, and the amount of LH released was similar in ovariectomized HPD ewes given oestrogen plus bolus or volley GnRH treatments and ovariectomized hypothalamo-pituitary intact ewes given oestrogen. These results suggest that the oestrogen-induced LH surge is initiated by a 'signal' pattern of GnRH secretion from the hypothalamus.

Animals↗

Lipocortin 1 mediates an early inhibitory action of glucocorticoids on the secretion of ACTH by the rat anterior pituitary gland in vitro.

Lipocortin 1 (LC1), a mediator of anti-inflammatory steroid action in some peripheral tissues, may contribute to the acute inhibitory effects of these steroids on hypothalamo-pituitary-adrenal (HPA) function. Accordingly, in the present study we have used an in vitro model to examine the potential role of this protein in the regulation of the release of corticotrophin (ACTH) from the anterior pituitary gland. Hypothalamic extracts (0.05-0.4 HE/ml), the 41 amino acid corticotrophin-releasing factor (CRF-41, 1-100 nM), the adenyl cyclase stimulator, forskolin (0.1 microM-10 mM), and the L-Ca2+ channel opener, BAY K8644 (0.01-10 mM), all caused concentration-dependent increases in the release in vitro of immunoreactive (ir)-ACTH from segments of rat anterior pituitary tissue. The secretory responses to submaximal concentrations of these secretagogues were overcome by preincubation of the tissue with dexamethasone (0.1 and 1 microM). LC1 was readily detectable by Western blotting in protein extracts of freshly excised pituitary tissue; a small proportion of the protein was found to be attached to the outer surface of the cell membranes where it was retained by a Ca(2+)-dependent mechanism. Exposure to dexamethasone (0.1 microM) in vitro did not affect the total LC1 content of the pituitary tissue but, over a 2-hour period, it caused a progressive two- to fivefold increase in the amount of LC1 attached to the outer surface of the cell; this response developed in parallel with the inhibitory effects of the steroid on ir-ACTH release. Both the dexamethasone-induced 'externalization' of LC1 by the pituitary tissue and the concomitant steroid-induced inhibition of peptide release were blocked by cycloheximide (1.0 microgram/ml) but not by actinomycin D (0.5 microgram/ml). A stable N-terminal lipocortin 1 fragment, LC1(1-188) (10 pg-10 ng/ml), attenuated (p < 0.01) the release of ir-ACTH evoked by HE (0.1 HE/ml), CRF-41 (1 nM), forskolin (1 mM) and BAY K8644 (1 nM). Conversely, inclusion of the anti-LC1 antibody in the medium substantially overcame the inhibitory effects of dexamethasone (0.1 microM) on the release of ir-ACTH evoked by the secretagogues whilst a control isotype matched antibody was without effect. The results suggest that LC1 plays a key role in effecting the acute inhibitory actions of glucocorticoids on the secretion of ir-ACTH by the rat anterior pituitary gland.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗

A truncated isoform of the thyrotropin-releasing hormone receptor is expressed in the rat central nervous system as well as in the pituitary gland.

Using the reverse transcription-polymerase chain reaction (RT-PCR), a cDNA encoding the entire rat thyrotropin-releasing hormone receptor (TRH-R) was isolated from normal rat pituitary gland mRNA. In addition, a novel truncated isoform of TRH-R which lacks 52 base pairs (bp) in the carboxyl (C-) terminal tail was isolated. This truncation, probably generated by alternate splicing, causes a frame-shift and results in a truncated TRH-R 25 amino acids shorter and with a different C-terminal amino acid sequence than the longer type receptor. This truncated TRH-R mRNA, along with the longer receptor form, was found to be expressed throughout the rat pituitary gland and brain.

Amino Acid Sequence↗

3':5'-cyclic-nucleotide phosphodiesterase in the bovine pituitary gland.

Cyclic-AMP phosphodiesterase activity in the homogenate of the anterior pituitary gland was 2-fold higher than that in the homogenate of the posterior pituitary, whereas cyclic-GMP phosphodiesterase activity was dominant in the posterior homogenate. There were two peaks of cyclic-AMP phosphodiesterase activity with different isoelectric points of 4.3 and 5.2. Fraction I had a molecular weight of 240 000 and a sedimentation coefficient of 6.2 S; fraction II had a molecular weight of 180 000 and a sedimentation coefficient of 3.1 S. Cyclic AMP hydrolytic activity in the supernatant of the posterior lobe corresponded to fraction I in the anterior lobe. Cyclic GMP hydrolytic activity in both the anterior and posterior lobes (activated by Ca2+/calmodulin) had an isoelectric point of 5.2, a molecular weight of 240 000 and a sedimentation coefficient of 6.2 S. Cyclic AMP and GMP hydrolytic activities in both the anterior and posterior lobes appeared in fraction I and did not separate when the preparations were mixed before electric focusing or sucrose density gradient procedures. Cyclic AMP hydrolytic activity in fraction II could be separated from cyclic GMP hydrolytic activity.

3',5'-Cyclic-AMP Phosphodiesterases↗

Phorbol 12-myristate 13-acetate induces prolactin secretion from rat anterior pituitary gland by the activation of protein kinase-C.

The purpose of this study was to characterize the action of phorbol 12-myristate 13-acetate (PMA), a tumor-promoting agent, on rat anterior pituitary gland, focusing the attention on prolactin secretion. PMA elicited a significant increase in prolactin secretion without affecting phosphatidylinositol turnover, considered as an early post-receptor event controlling PRL secretion. However incubation of anterior pituitary glands with PMA caused a loss of protein kinase-C activity in cytoplasm concomitant with an increased enzyme activity in the membrane. The action of PMA on prolactin secretion seems to be mainly dependent from the redistribution and activation of protein kinase-C. In fact, the phorbol ester did not affect pituitary cAMP and cGMP metabolism either in basal conditions or after theophylline.

Animals↗

Effects of tank color on melanin-concentrating hormone levels in the brain, pituitary gland, and plasma of the barfin flounder as revealed by a newly developed time-resolved fluoroimmunoassay.

A pleuronectiform fish, the barfin flounder Verasper moseri, reared in a white tank had a smaller ratio of pigmented area of the skin on non-eyed side, grew faster, and had greater melanin-concentrating hormone (MCH)-immunoreactive cell bodies and MCH gene expression in the brain than in the black tank, indicating that synthesis and release of MCH are higher in fish from a white tank. In the present study, a time-resolved fluoroimmunoassay for MCH was developed. MCH levels were assessed in the brain, pituitary gland, and plasma of barfin flounders reared in a white or black tank. A competitive assay using two antibodies was performed among secondary antibodies in the solid phase, MCH antibodies, samples, and europium-labeled MCH. Displacement curves of serially diluted extracts (brain, pituitary gland, and plasma) of the barfin flounder paralleled that of the MCH standard. MCH levels in the brain and plasma were higher in fish reared in the white tank for 5 months than in the black tank. These results suggest that synthesis and secretion of MCH are enhanced with the white background and that MCH is involved in both somatic growth and the skin pigmentation in the barfin flounder.

Adaptation, Physiological↗

Early development of the pituitary gland in Acipenser naccarii (Chondrostei, Acipenseriformes): an immunocytochemical study.

The distribution and appearance of secretory cells in the pituitary gland were investigated for the first time in a chondrostean species, Acipenser naccarii, from embryos to juveniles, by immunohistochemistry with mammalian and teleost hormone antisera. On 5.5 day post-fertilization (2.5 days pre-hatching), the pituitary of embryos appears as an oval cell mass with a narrow central cavity (hypophysial cleft), close to the ventral border of diencephalon under the third ventricle. At that time no neurohypophysis is observed, the adenohypophysis is not yet structurally divided into pars intermedia (PI) and pars distalis (PD) and only immunoreactive growth hormone cells are detectable. Seven days post-fertilization (1 day pre-hatching) the immunoreactive thyrotropic cells appear in the ventral region and the immunoreactive adrenocorticotropic cells in the posterior dorsal one. At hatching, some immunoreactive melanotropic (ir-MSH) cells are visible in the posterior dorsal region and some immunoreactive prolactin cells in the anterior one. Eight days later the immunoreactive somatolactin cells appear along the posterior dorsal border and the immunoreactive gonadotropic I (ir-GtH I) cells in the ventral region. Here, a few ir-GtH II cells finally appear in 76-86 day old juveniles. The gland elongates after hatching and in 8-day-old larvae two adenohypophysial regions are identified: a posterior (the presumptive PI) and an anterior one (the presumptive PD). In 156-166-day-old juveniles three regions (rostral and proximal pars distalis and pars intermedia) appear and a high number of ir-MSH cells are visible in the rostral region. The first protrusion of neurohypophysis into adenohypophysis is observed in 76-86-day-old juveniles and increases with age, branching into PI. The rostro-caudal distribution of the immunoreactive cells follows the spatial expression of the corresponding hormone gene families observed in zebra fish, suggesting similar differentiating mechanisms in teleosts and chondrosteans.

Adrenocorticotropic Hormone↗

Effects of domperidone on the secretion of prolactin from rat anterior pituitary glands.

The effects of omperidone on prolactin secretion from the anterior pituitary glands of female rats were studied. A perifusion system and radioimmunoassay were used to study secretion; static incubations, and incorporation of [3H]leucine were used to measure biosynthesis. During perifusion, intact anterior lobes showed a constant rate of prolactin secretion for up to 5h (after a 90-min preincubation to stabilize the tissue). Incorporation studies revealed an increase in protein synthesis in perifused hemipituitary glands. When glands were treated with 10nM-domperidone , prolactin secretion began to decline after the first hour, reaching a maximum of 40-50% inhibition after a further 90 min. Growth hormone secretion showed no such decline. Inhibition of prolactin secretion continued for up to 2h after withdrawal of the drug. Treatment with 100nM-dopamine resuled in a more rapid inhibition of secretion, but the effect was reversed on withdrawal of the catecholamine. The prolactin content of perifused hemipituitary glands was measured after treatment with domperidone; the contents of control and treated glands did not differ, but were depleted compared with hemipituitary glands which had not been perifused.

Animals↗

Human chorionic gonadotropin production by the pituitary gland in a premenopausal woman.

OBJECTIVE: Our purpose was to investigate the source of human chorionic gonadotropin production in a nonpregnant, premenopausal woman. STUDY DESIGN: A case of human chorionic gonadotropin production by the pituitary gland in a premenopausal woman is described. RESULTS: Our results confirm that a biologically active human chorionic gonadotropin-like molecule was secreted in a nonpregnant woman. CONCLUSIONS: Our results indicate that the pituitary gland was the most likely source of human chorionic gonadotropin production.

Adult↗

Effects of diethylstilbestrol on the cytogenesis of prolactin cells in the pars distalis of the pituitary gland of the mouse.

This study was carried out to examine the developmental stage when prolactin cells differentiate in mice and to examine the effects of diethylstilbestrol on the development of prolactin cells in the fetal and neonatal pituitary glands. A small number of immunoreactive prolactin cells appeared first on embryonic day 15 in control (injected with oil) pituitary glands, whereas they did not increase in number until postnatal day 2. In diethylstilbestrol-treated mice (5 mg/kg body weight, 24 h before killing), a small number of immunoreactive prolactin cells were detectable as early as embryonic day 14, but not on day 13. They increased in number on embryonic days 15 and 16, and decreased markedly on days 17 and 18, followed by a rapid increase after birth. This transient reduction in the response to diethylstilbestrol was partially restored by treatment with metyrapone, a specific inhibitor of corticosteroid production. These results suggest that in the mouse: (1) differentiation of prolactin cells occurs between embryonic days 13 and 14, (2) prolactin gene expression is suppressed in the nascent prolactin cells presumably due to the presence of high levels of estrogen-binding protein, alpha-fetoprotein, and (3) prolactin gene expression is also suppressed by elevation of circulating glucocorticoids during the perinatal period. The present results suggest that, in the mouse, at least a proportion of prolactin cells are not derived from growth hormone cells, because the diethylstilbestrol-induced prolactin cells appear earlier than growth hormone gene expression.

Animals↗

Dopamine levels in the anterior pituitary gland monitored by in vivo electrochemistry.

The present study was designed to assess the contributions of the long portal vessel and neurointermediate lobe routes of dopamine delivery to the anterior pituitary gland. Dopamine levels were monitored in the anterior pituitary of anesthetized diestrus 1 rats using in vivo differential pulse voltammetry at a carbon paste electrode. Measurements were taken during control periods, following neurointermediate lobectomy and hypophysial stalk section or after inhibition of catecholamine synthesis and dopamine infusion. Neurointermediate lobectomy resulted in a slight rise in plasma prolactin and a significant fall in voltammetric current. Subsequent stalk section elevated prolactin significantly and further reduced the voltammetric output. Inhibiting catecholamine synthesis significantly elevated plasma prolactin and reduced the voltammetric current. Stepped infusions of dopamine then suppressed plasma prolactin and elevated the voltammetric output in a dose related manner. The final values of both parameters were not significantly different from pre-inhibition levels. These data provide direct estimates of the relative contributions of various vascular routes to dopamine in the anterior pituitary and support the sufficiency of dopamine as the physiological inhibitor of prolactin secretion.

Animals↗

Size and shape of the pituitary gland during pregnancy and post partum: measurement with MR imaging.

Cranial magnetic resonance (MR) imaging was performed in 38 pregnant and postpartum women and 30 nonpregnant age-matched control subjects to establish standards for pituitary gland size and shape during this period. Gland height and infundibulum width were measured on midline T1-weighted sagittal images. Gland convexity or concavity was graded qualitatively. Throughout pregnancy, gland height increased linearly by approximately 0.08 mm/wk. No gland exceeded 10 mm in height during pregnancy. Increases in gland convexity also correlated with progression of pregnancy. The largest glands were seen in the immediate postpartum period; during this period, five of 12 glands measured 10.0-11.8 mm. Beyond the first week post partum, glands rapidly returned to normal size, apparently regardless of the status of breast-feeding. The mean diameter of the infundibulum was 2.2 mm (range, 0.8-4.0 mm). The pituitary gland enlarges throughout pregnancy but should probably not exceed 10 mm during most of this period. Size of up to 12 mm may be acceptable immediately post partum.

Adolescent↗

[Physiological significance of the self-priming effect of LH-RH on the anterior pituitary glands].

To investigate the intracellular mechanisms of the self-priming effect of LH-RH on the anterior pituitary gland, the responses of pituitary and serum gonadotropins to two administrations of this decapeptide at a 60 min, interval were studied in male rats. A clearly recognized self-priming effect in male rats was significantly suppressed by the administration of RNA synthesis inhibitor. Although the pituitary LH content decreased significantly after the first administration, the content increased to the control level immediately before the second administration. This recovery was prevented by the administration of RNA synthesis inhibitor. In spite of the large second response, the rate of the second LH depletion was low. The number of secretory granules in gonadotrophs showed similar changes to the pituitary LH content. The dilatation of the Golgi apparatus was observable at 60 min. after the first administration. These results of rat experiments indicate that the second response includes the newly synthesized gonadotropins, and a clinical LH-RH two step test could be employed to evaluate both pituitary gonadotropin synthesizing and secreting functions.

Animals↗

Thyroid hormone regulation of messenger ribonucleic acid encoding thyrotropin (TSH)-releasing hormone is independent of the pituitary gland and TSH.

Cellular levels of mRNA encoding pro TRH in the rostral paraventricular nucleus are reduced by thyroid hormones. To determine whether this regulatory effect of thyroid hormones requires a functional pituitary gland or, specifically, TSH, we examined the effect of T3 on proTRH mRNA in hypophysectomized, thyro-parathyroidectomized male rats with or without bovine TSH replacement. Hypophysectomy plus thyro-parathyroidectomy reduced serum T4 and TSH to undetectable levels in all animals and elevated TRH mRNA in the paraventricular nucleus over that of sham-operated animals. Eleven consecutive daily injections of T3 significantly reduced TRH mRNA levels in both sham controls and thyro-parathyroidectomized rats. However, 11 daily injections of bovine TSH (1 U/day) failed to alter the effect of T3 on TRH mRNA levels. These results demonstrate that the regulatory influence of thyroid hormones on the biosynthesis of TRH within the thyrotropic center of the brain is independent of the pituitary gland and of TSH.

Animals↗

Effect of serotonin on the basal and gonadotrophin-releasing hormone-induced release of luteinizing hormone from rat pituitary glands in vitro.

The effect of serotonin (5-HT) on the basal and gonadotrophin-releasing hormone (GnRH)-stimulated release of luteinizing hormone (LH) was studied in rat adenohypophysis in vitro. Anterior pituitary glands from ovariectomized rats were incubated for 1h in the presence of different doses of 5-HT (0.01 to 3 mumol/l). Serotonin added to the culture medium slightly dimished the basal release of LH and markedly inhibited the release of LH induced by GnRH. Responsiveness to GnRH (3 nmol/l) was significantly reduced, in a dose-dependent manner, by the simultaneous treatment of glands with 5-HT. Maximal inhibition to 65% of the response obtained with GnRH alone, was attained with 1 mumol/l 5-HT. The EC50 value was estimated to be about 1.9 X 10(-7) M. The inhibitory effect of 5-HT was evident within 30 min of incubation. Furthermore, 5-HT appear to exert a short-lasting action, since the rate of basal and GnRH-induced release of LH was reduced during the first hour of incubation, but after 2h the suppressive effects of 5-HT were no longer apparent. Methysergide, a serotonin receptor blocking agent, partially antagonized the inhibitory effect of 5-HT on LH release, either basal or GnRH-stimulated. This suggests that a receptor-mediated component may be involved in the mechanism of 5-HT action. The present results indicate that 5-HT can affect the release of LH by acting directly at the pituitary gland level.

Animals↗