Hypothalamo-pituitary Disconnection Research Articles

Mandibular bone loss in ewe induced by hypothalamic–pituitary disconnection

It is supposed that the demographic change will lead to an increase in patients with impaired alveolar bone conditions. Large animal models are of particular interest in this context as they are suitable for developing and testing new dental implants. Recently, we demonstrated that surgical hypothalamo-pituitary disconnection (HPD) causes a pronounced low-turnover situation leading to cortical and trabecular bone loss in sheep. In this study, we aimed to investigate the influence of the HPD procedure on the alveolar bone. Ten adult Merino ewes were randomly assigned to two groups: Control and HPD. After 6months, we analysed the cortical and trabecular bone of all mandibles by histomorphometry and high-resolution peripheral quantitative computed tomography (HR-pQCT). HPD ewes showed a significant decrease in cortical thickness by ~20%, a significant increase in cortical porosity by ~20% and a significant decrease in bone volume by ~30% in comparison with Control ewes. Our results underline the importance of central regulatory mechanisms of bone turnover. However, further studies are needed to understand these central regulatory elements of bone turnover in detail and to judge the value of the HPD sheep for dental research.

Sheep model for osteoporosis: Sustainability and biomechanical relevance of low turnover osteoporosis induced by hypothalamic–pituitary disconnection

Hypothalamo-pituitary disconnection (HPD) leads to low bone turnover and osteoporosis in sheep. To determine the sustainability of bone loss and its biomechanical relevance, we studied HPD-sheep 24 months after surgery (HPD + OVX-24) in comparison to untreated control (Control), ovariectomized sheep (OVX), and sheep 12 months after HPD (HPD + OVX-12). We performed histomorphometric, HR-pQCT, and qBEI analyses, as well as biomechanical testing of all ewes studied. Twenty-four months after HPD, histomorphometric analyses of the iliac crest showed a significant reduction of BV/TV by 60% in comparison to Control. Cortical thickness of the femora measured by HR-pQCT did not change between 12 and 24 months after HPD but remained decreased by 30%. These structural changes were caused by a persisting depression of osteoblast and osteoclast cellular activity. Biomechanical testing of the femora showed a significant reduction of bending strength, whereas calcium content and distribution was found to be unchanged. In conclusion, HPD surgery leads to a persisting low turnover status with negative turnover balance in sheep followed by dramatic cortical and trabecular bone loss with consequent biomechanical impairment.

Low turnover osteoporosis in sheep induced by hypothalamic-pituitary disconnection

The hypothalamus is of critical importance in regulating bone remodeling. This is underscored by the fact that intracerebroventricular-application of leptin in ewe leads to osteopenia. As a large animal model of osteoporosis, this approach has some limitations, such as high technical expenditure and running costs. Therefore we asked if a surgical ablation of the leptin signaling axis would have the same effects and would thereby be a more useful model. We analyzed the bone phenotype of ewe after surgical hypothalamo-pituitary disconnection (HPD + OVX) as compared to control ewe (OVX) after 3 and 12 months. Analyses included histomorphometric characterization, micro-CT and measurement of bone turnover parameters. Already 3 months after HPD we found osteopenic ewe with a significantly decreased bone formation (69%) and osteoclast activity (49%). After a period of 12 months the HPD group additionally developed an (preclinical) osteoporosis with significant reduction (33%) of femoral cortical thickness, as compared to controls (OVX). Taken together, HPD leads after 12 month to osteoporosis with a reduction in both trabecular and cortical bone caused by a low bone turnover situation, with reduced osteoblast and osteoclast activity, as compared to controls (OVX). The HPD-sheep is a suitable large animal model of osteoporosis. Furthermore our results indicate that an intact hypothalamo-pituitary axis is required for activation of bone turnover.

Stimulatory effect of naloxone on plasma cortisol in human: Possible direct stimulatory action at the adrenal cortex

The purpose of the present study was to gain a better insight in the mechanism of naloxone underlying the regulation of adrenal cortisol secretion in humans in vivo; therefore, the stimulatory effect of naloxone on cortisol secretion was assessed in a group of patients with hypothalamo-pituitary disconnection. Patients with hypothalamo-pituitary disconnection because of various pathologies (craniopharingioma, cordoma, suprasellar meningioma, or pituitary macroadenoma) participated in the study. Results Circulating cortisol, but not adrenocorticotropin (ACTH) levels were significantly higher after naloxone administration than after saline. Conclusion Besides the well-known hypothalamo-pituitary stimulatory action on ACTH release in normal humans, the results of the present study suggest that naloxone exerts direct effects on cortisol secretion at the adrenal gland level; another possibility is that naloxone stimulation of cortisol secretion is mediated by other factor than ACTH.

Leptin: Primary Central Site of Action?

The discovery of the fat hormone leptin 16 yr ago (1) heralded an explosion in obesity research that continues to this day. The original premise that this hormone acted simply as an adipostat directing appropriate energy balance regulatory output in the brain has comprehensively unfurled to reveal a far more complex picture. Inevitably perhaps, as the complexity increases so does the controversy. In keeping with this trend, the paper by Qi et al. (2) published in the current issue of Endocrinology calls into question the primacy of the hypothalamic arcuate nucleus in the central leptin signaling pathway. The widely accepted model is that adipocyte-derived leptin acts on first-order neurons in the hypothalamic arcuate nucleus, and the message is relayed to second-order neurons in other hypothalamic regions such as the dorsomedial nucleus, ventromedial nucleus, paraventricularnucleus, and lateral hypothalamic area (3–5). From a multitude of animal experiments, it is unequivocal that the central actions of leptin are anorexigenic. Thus, hypoleptinemia is a powerful orexigenic signal during fasting, and leptin administration decreases appetite and body weight (3–5). Early experiments also reported the pivotal importance of the arcuate nucleus to this response because the anorectic effects of peripherally and centrally administered leptin were abolished in rats following monosodium glutamate lesion of the arcuate nucleus (6, 7). In addition, it has been demonstrated that reactivation of the leptin receptor specifically in the arcuate nucleus leads to decreased food intake and body weight in leptin receptordeficient mice (8) and rats (9). Clearly, receptor distribution provides a good indication of sites of action, and there is fairly universal agreement on the distribution of the signaling (long) form of the leptin receptor in the brain. Thus, it is localized in the arcuate, dorsomedial, ventromedial and paraventricular nuclei, and the lateral hypothalamic area of the adult hypothalamus, regions known to be involved in energy balance and appetite control but with the highest density in the arcuate nucleus (10). Primary targets for leptin in the arcuate nucleus have long been identified as proopiomelanocortin and neuropeptide Y neurons, which coexpress the leptin receptor and in which leptin induces phosphorylation of the signal transducer and activator of transcription 3, a marker of leptin receptor activation (3–5, 10). It follows that deletion of the leptin receptor specifically in arcuate proopiomelanocortin and neuropeptide Y neurons of mice would result in a hyperphagic and obese phenotype; however, the effects on food intake and adiposity are less extreme compared with global leptin receptor deletion, indicating that other leptin-regulated pathways are also involved (11, 12). As mentioned above, the leptin receptor shows a wider hypothalamic distribution besides the arcuate nucleus and leptin receptor deletion specifically in the ventromedial nucleus also results in increased body weight and adiposity in mice (13). Indeed, neuronal activation in many hypothalamic sites is reported in response to leptin administration in rodents (10, 14). However, it remains equivocal whether the responses of different hypothalamic nuclei to leptin are independent or orchestrated by the arcuate nucleus. Now Qi et al. (2) report novel evidence for direct action of leptin on cells in the dorsal, ventromedial, and paraventricular hypothalamic nuclei in sheep, in the presence or absence of an arcuatenucleus.Thus, they found immunostaining for fos, a marker of neuronal activation, in the dorsal, ventromedial and paraventricular hypothalamic nuclei following peripheral leptin administration in their model of hypothalamo-pituitary disconnection in which the arcuate is

The Action of Leptin on Appetite-Regulating Cells in the Ovine Hypothalamus: Demonstration of Direct Action in the Absence of the Arcuate Nucleus

It is widely accepted that leptin acts on first-order neurons in the arcuate nucleus (ARC) with information then relayed to other hypothalamic centers. However, the extent to which leptin mediates its central actions solely, or even primarily, via this route is unclear. We used a model of hypothalamo-pituitary disconnection (HPD) to determine whether leptin action on appetite-regulating systems requires the ARC. This surgical preparation eliminates the ARC. We measured effects of iv leptin to activate hypothalamic neurons (Fos labeling). In ARC-intact animals, leptin increased the percentage of Fos-positive melanocortin neurons and reduced percentages of Fos-positive neuropeptide Y neurons compared with saline-treated animals. HPD itself increased Fos labeling in the lateral hypothalamic area (LHA). Leptin influenced Fos labeling in the dorsomedial nucleus (DMH), ventromedial nucleus, and paraventricular nucleus (PVN) in HPD and normal animals, with effects on particular cell types varying. In the LHA and DMH, leptin decreased orexin cell activation in HPD and ARC-intact sheep. HPD abolished leptin-induced expression of Fos in melanin-concentrating hormone cells in the LHA and in CRH cells in the PVN. In contrast, HPD accentuated activation in oxytocin neurons. Our data from sheep with lesions encompassing the ARC do not suggest a primacy of action of leptin in this nucleus. We demonstrate that first order to second order signaling may not represent the predominant means by which leptin acts in the brain to generate integrated responses. We provide evidence that leptin exerts direct action on cells of the DMH, ventromedial nucleus, and PVN.

Cortisol infusion in late-gestation hypothalamo-pituitary disconnected sheep fetus restores pituitary cell responsiveness to arginine vasopressin

Corticotrophs in the fetal sheep become increasingly responsive to arginine vasopressin (AVP) in late gestation. We previously reported that this may be due in part to corresponding increases in signal transduction (inositol 1,4,5-trisphosphate, IP(3)). These ontogenic changes are prevented by hypothalamo-pituitary disconnection (HPD), which also prevents fetal plasma cortisol concentrations from increasing in late gestation. This led us to hypothesize that cortisol is involved in mediating the changes in pituitary responsiveness. HPD was performed on fetal sheep at 120 days gestational age (dGA). Half of the HPD fetuses were infused with cortisol for 3 days beginning at 135-137 dGA (HPD+C). The remaining HPD fetuses and a group of sham-operated control fetuses were infused with saline. Pituitary cells were isolated and cultured. After 48 h, a subset of cells was stimulated with 100 nM AVP for 2 h, and the medium was collected for ACTH analysis. Another subset of cells was stimulated with 100 nM AVP for 30 min, and the formation of IP(3) was determined. Plasma cortisol concentrations increased rapidly within the first 6 h after infusion (5.2 +/- 1.9 to 29.7 +/- 4.9 ng/ml) but did not increase thereafter. Cells from HPD+C and sham-operated fetuses secreted significantly more ACTH than those from HPD fetuses (% increase from control: 33.0 +/- 8.8%, 47.9 +/- 10.6%, and 11.9 +/- 2.4%, respectively). IP(3) formation was significantly increased in cells from HPD+C and sham-operated compared with HPD fetuses (% increase from control: 17.7 +/- 4.4%, 18.9 +/- 4.3%, and 4.6 +/- 1.5%, respectively). These findings support the idea that cortisol plays a role in mediating the increase in pituitary responsiveness to AVP in the late-gestation fetal sheep.

Hypothalamic input is required for development of normal numbers of thyrotrophs and gonadotrophs, but not other anterior pituitary cells in late gestation sheep

To evaluate the hypothalamic contribution to the development of anterior pituitary (AP) cells we surgically disconnected the hypothalamus from the pituitary (hypothalamo-pituitary disconnection, HPD) in fetal sheep and collected pituitaries 31 days later. Pituitaries (n = 6 per group) were obtained from fetal sheep (term = 147 +/- 3 days) at 110 days (unoperated group) of gestation and at 141 days from animals that had undergone HPD or sham surgery at 110 days. Cells were identified by labelling pituitary sections with antisera against the six AP hormones. Additionally, we investigated the colocalization of glycoprotein hormones. The proportions of somatotrophs and corticotrophs were unchanged by age or HPD. Lactotrophs increased 80% over time, but the proportion was unaffected by HPD. Thyrotrophs, which were unaffected by age, increased 70% following HPD. Gonadotrophs increased with gestational age (LH+ cells 55%; FSH+ cells 19-fold), but this was severely attenuated by HPD. We investigated the possible existence of a reciprocal effect of HPD on multipotential glycoprotein-expressing cells. Co-expression of LH and TSH was extremely rare (< 1%) and unchanged over the last month of gestation or HPD. The increase of gonadotrophs expressing FSH only or LH and FSH was attenuated by HPD. Therefore, the proportions of somatotrophs, lactotrophs and corticotrophs are regulated independently of hypothalamic input in the late gestation fetal pituitary. In marked contrast, the determination of the thyrotroph and gonadotroph lineages over the same time period is subject to complex mechanisms involving hypothalamic factors, which inhibit differentiation and/or proliferation of thyrotrophs, but stimulate gonadotrophs down the FSH lineage. Development of a distinct population of gonadotrophs, expressing only LH, appears to be subject to alternative mechanisms.

Circannual prolactin rhythms: calendar-like timer revealed in the pituitary gland

Although photoperiodic regulation of annual rhythms in reproduction and other functions has been well characterized, the basis for the endogenous generation of circannual rhythms during exposure to constant conditions has not been elucidated. Lincoln and colleagues have recently reported that circannual prolactin rhythms in rams persist after hypothalamo-pituitary disconnection, but not after pinealectomy. Does the pars tuberalis, the site of pituitary gland melatonin receptors, generate circannual rhythms and integrate photoperiodic signals mediated by melatonin?

Ontogeny and Effects of Hypothalamic Pituitary Disconnection on Formation of Inositol Trisphosphate in Fetal Sheep Pituitary Cells

In late gestation fetal sheep, the pituitary becomes increasingly responsive to stimulation by arginine vasopressin (AVP). This change appears to be one important factor mediating the plasma cortisol surge, a critical developmental event. It is not known precisely why pituitary corticotropes become more responsive at this time. In this study we examined the possibility that changes in second messenger generation [inositol trisphosphate (IP(3))] are responsible. Two studies were undertaken. The first was an ontogeny study, where pituitaries were isolated from 100-, 120-, and 140-d gestational age (dGA) fetal sheep. Cells were cultured, stimulated with AVP, and the formation of IP(3) assessed. The amount of IP(3) generated increased with gestational age (percent increases from unstimulated controls were 4.6, 11.5, and 21.5 for 100, 120, and 140 dGA, respectively), with significant differences between the 140-dGA group and both earlier groups apparent. The second study examined the impact of 120-dGA hypothalamo-pituitary disconnection (HPD), which prevents corticotrope maturation, on responsiveness of pituitary cells isolated from 140-dGA fetuses. Cells were stimulated with AVP, and the formation of IP(3) and secretion of ACTH were assessed. Significantly less IP(3) was formed, and ACTH secreted in cells from HPD compared with control fetuses (IP(3) and ACTH levels were 50% and 35% lower, respectively). Results from the HPD study demonstrate that the ontogenic changes in IP(3) after AVP require an intact hypothalamic-pituitary-adrenal axis. These findings suggest that heightened second messenger generation may be a key reason for increased ACTH secretory responsiveness to AVP in the late gestation sheep fetus.

276. Sex difference in the effect of cortisol on the LH response of the pituitary to exogenous GnRH in hypothalamo-pituitary disconnected gonadectomised sheep

We have used the hypothalamo-pituitary disconnected (HPD) sheep model to investigate direct pituitary actions of cortisol to suppress LH secretion in response to exogenous GnRH. We previously observed that, during the non-breeding season, treatment with cortisol did not suppress the LH response to GnRH in HPD gonadectomised rams or ewes.1 In the present experiment, we tested the effect of cortisol on the LH response to exogenous GnRH in gonadectomised HPD sheep during the breeding season. Using a cross-over design, HPD gonadectomised Romney Marsh rams (n = 6) and ewes (n = 5) received a saline or cortisol (250 μg/kg/h) infusion for 30 h on each of two days, one week apart. All animals were treated with 125 ng i.v. injections of GnRH every 2 h during a 6h control period preceding the infusion and during the infusion. Jugular blood samples were taken during the control period and the first 6 h and last 6 h of the infusion (over 3 LH pulses). Mean plasma concentrations of LH and LH pulse amplitudes, driven by programmed GnRH injections, were similar in gonadectomised rams and ewes and there were no significant effects of saline infusion between the control periods or the saline infusion in either sex. The amplitude of LH pulses was significantly (P &lt; 0.05) reduced in rams during the first 6 h of the cortisol infusion compared to the control period, but there were no effects of the cortisol infusion in ewes. These data show that, in the absence of sex steroids, there is a sex difference in the mechanism by which cortisol acts at the pituitary to reduce LH secretion in response to exogenous GnRH in HPD gonadectomized sheep during the breeding season. We conclude that the effect of cortisol to reduce secretion of LH involves an action on the pituitary, at least in gonadectomised rams. (1)Stackpole CA, Turner AI, Clarke IJ and Tilbrook AJ (2003) Biology of Reproduction 36(Supplement 1), 288.

The effect of hypothalamo-pituitary disconnection on the renin-angiotensin system in the late-gestation fetal sheep

The activity of the renin-angiotensin system (RAS) increases significantly in the late-gestation fetal sheep. Fetal cortisol is also increased during this time, and it is thought that the increase in cortisol may modulate the RAS changes. Previous studies have examined the effects of cortisol infusion on RAS activity, but the effects of blocking the peripartum increase in cortisol concentrations on the developmental changes in the RAS are not known. Therefore, we utilized the technique of hypothalamic-pituitary disconnection (HPD), which prevents the cortisol surge from occurring, to investigate the importance of the late-gestation increase in cortisol on the ontogenic changes in RAS activity. HPD of fetal sheep was performed at 120 days of gestational age (dGA), and fetuses were delivered between 135 and 139 dGA. Control fetuses were sham operated. HPD blocked the late-gestation cortisol increase but did not alter renal renin mRNA, renal renin or prorenin protein content, nor plasma renin levels compared with sham operated. However, HPD fetuses had increased ANG II receptor subtype 1 (AT1) mRNA and protein expression in the kidney and lungs. ANG II receptor subtype 2 (AT2) expression was not altered in these tissues at either mRNA or protein level. HPD did not change AT1 or AT2 mRNA in the left ventricle but did result in decreased protein levels for both receptors. These studies demonstrate that blockade of the naturally occurring increase in fetal cortisol concentration in late gestation is associated with tissue-specific alterations in expression of AT1 and AT2 receptors. These changes may impact on fetal tissue maturation and hence have consequences in postnatal life.

Prolactin cycles in sheep under constant photoperiod: evidence that photorefractoriness develops within the pituitary gland independently of the prolactin output signal.

The present study investigated photorefractoriness in the prolactin (PRL) axis in hypothalamopituitary-disconnected (HPD) sheep exposed to prolonged long days. In experiment 1, HPD Soay rams transferred from short (8L:16D) to long (16L:8D) days for 48 wk to induce a cycle of activation, decline (photorefractoriness), and reactivation in PRL secretion were treated chronically with bromocriptine (dopamine-receptor agonist) or vehicle from the onset of photorefractoriness. Bromocriptine (0.01-0.04 mg kg-1 day-1; 12-24 wk of long days) blocked PRL release and caused a rebound response after the treatment, but it had no effect on the long-term PRL cycle (posttreatment PRL minimum, mean +/- SEM, 35.3 +/- 0.6 and 37.0 +/- 0.4 wk for bromocriptine and control groups, respectively; not significant). In experiment 2, HPD rams were treated with sulpiride (dopamine-receptor antagonist) during photorefractoriness. Sulpiride (0.6 mg/kg twice daily; 22-30 wk of long days) induced a marginal increase in blood PRL concentrations, but again, it had no effect on the long-term PRL cycle (PRL minimum, 37.9 +/- 0.4 and 37.6 +/- 0.9 wk for sulpiride and control groups, respectively; not significant). The 24-h blood melatonin profile consistently reflected the long-day photoperiod throughout, and blood FSH concentrations were minimal, confirming the effectiveness of the HPD surgery. The results support the conclusion that photorefractoriness is regulated at the level of the pituitary gland independently of the PRL output signal.

The role of hypothalamic input on corticotroph maturation in fetal sheep.

Corticotropin-releasing hormone receptor type 1 (CRH-R1) expression and vasopressin type 1b (V1b) receptor protein decrease in late-gestation fetal sheep. Because hypothalamo-pituitary disconnection (HPD) has been demonstrated to prevent the morphological maturation of corticotrophs, we hypothesized that hypothalamic input is necessary for the maturational changes in CRH-R1 and V1b receptor levels. We measured CRH-R1 and V1b receptor expression in the anterior pituitaries of fetuses at 140 days gestational age (dGA) that underwent HPD or sham surgery at 120 dGA. CRH-R1 mRNA decreased similarly in HPD and sham-operated fetuses compared with 120 dGA naive fetuses. However, CRH-R1 protein levels were elevated in HPD fetuses compared with sham and were not different from 120 dGA values. V1b protein levels decreased similarly in HPD and sham-operated fetuses compared with 120 dGA naive fetuses. We conclude that hypothalamic input to the pituitary is necessary for the decrease in CRH-R1 receptor protein levels in late-gestation fetal sheep. However, hypothalamic input is not necessary for the decrease in V1b receptor expression seen in late gestation.

Site and mechanism of action of dynorphin A-(1-13) and N-methyl-D-aspartate on ACTH release in fetal sheep.

Dynorphin A (Dyn A) stimulates the release of ACTH in fetal sheep, a response that involves N-methyl-D-aspartate (NMDA) receptors but not the secretogogues corticotropin-releasing hormone or arginine vasopressin. We now find that neither Dyn A-(1-13) (0.5 mg/kg, i.v.) nor NMDA (4 mg/kg, i.v.) elicits ACTH release in postnatal lambs. This led us to hypothesize that Dyn A-(1-13) and NMDA might act to release placental ACTH. However, the ability of Dyn A-(1-13), NMDA, and the kappa-opioid receptor agonist U-50488H (1 mg/kg, i.v.) to release ACTH was lost after either fetal hypophysectomy (n = 4) or hypothalamo-pituitary disconnection (n = 4). These results indicate that neither the placenta nor the fetal pituitary is the site of action for these agonists and suggest a hypothalamic or suprahypothalamic site of action. Furthermore, the release of ACTH by Dyn A-(1-13) and NMDA was abolished after pretreatment with indomethacin, suggesting that they might cause the release of a prostanoid, possibly from the placenta, that subsequently acts at the hypothalamus or serves as a permissive factor in the action of Dyn A-(1-13) and NMDA at the hypothalamus.

Melatonin modulation of prolactin and gonadotrophin secretion. Systems ancient and modern.

Recent studies in sheep indicate that the pineal melatonin signal which transduces effects of photoperiod acts at separate sites in the pituitary gland and brain to regulate seasonality in prolactin (PRL) and gonadotrophin secretion. The pituitary gland is the proposed site for control of PRL based on the observation that hypothalamo-pituitary disconnected (HPD) rams continue to show normal patterns of PRL secretion in response to changes in photoperiod or treatment with melatonin. Lactotrophs do not express melatonin receptors, thus this pituitary effect is assumed to be mediated by cells in the pars tuberalis via "tuberalin". The mediobasal hypothalamus (MBH) is the putative target for gonadotrophin control since: i) gonadotrophin secretion is dependent on pulsatile GnRH secretion from the MBH, ii) local administration of melatonin in the MBH, but not in other areas of the brain and pituitary gland, readily reactivates GnRH-induced LH and FSH secretion in photo-inhibited rams; and iii) treatment of HPD rams with a chronic pulsatile infusion of GnRH stimulates gonadotrophin secretion irrespective of photoperiod. Complementary studies conducted by others in the Syrian hamster, have shown that lesions in the MBH block the action of melatonin on gonadotrophin but not on prolactin secretion; this supports the "dual-site hypothesis". Since all photoperiodic mammals are essentially similar in hyper-secreting PRL under long days, the pituitary control mechanism for PRL is regarded as conserved (ancient) with the pleiotrophic actions of PRL inducing a summer physiology (e.g. growth of summer pelage). In contrast, the variation between species in the timing of the gonadal cycle indicates that evolution has independently modified the melatonin-sensitive neural circuits in the MBH to permit the species-specific timing of the mating season.

Noradrenaline and dopamine regulation of prolactin secretion in sheep: role in prolactin homeostasis but not photoperiodism.

The role of noradrenaline (NA) and dopamine (DA) in the hypothalamic control of prolactin (PRL) secretion was investigated in hypothalamic intact (control) and hypothalamo-pituitary disconnected (HPD) Soay rams. The animals were exposed to alternating 16-weekly periods of short (8 L : 16D) and long days (16 L : 8D) to induce marked cyclical changes in PRL secretion in both groups (as demonstrated previously). Selective NA and DA receptor antagonists (dose: 1.2 micromol/kg) were administered under short days (low endogenous PRL secretion), and agonists (dose: 0.0012-0.12 micromol/kg) were administered under long days (high endogenous PRL secretion). The acute changes in blood PRL concentrations were measured over 4 h as the index of responsiveness. Under short days, treatment with WB4101 (alpha-1 adenoceptor antagonist), and rauwolscine (alpha-2 antagonist), consistently increased PRL secretion in control, but not in HPD rams. The treatments produced similar acute, drug-specific behavioural effects in both groups. Propranolol (beta antagonist) had no effect on PRL secretion, while sulpiride (DA D-2 antagonist) induced a marked increase in blood PRL concentrations in control rams (> 4 h), and a transient effect in HPD rams (15 min). Under long days, when endogenous PRL secretion was increased, phenylephrine (alpha-1 agonist) produced no effects, while bromocriptine (DA D-2 agonist) robustly decreased PRL concentrations in both control and HPD rams, even at the lowest treatment dose. Overall, the positive responses to the antagonists in the control rams, support the view that DA (acting via D-2 receptors), and to a lesser extent NA (acting via alpha-1/alpha-2 receptors), negatively regulate PRL secretion. In contrast, the lack of responses to the antagonists in the HPD rams, support the view that neither DA, nor NA, mediate the photoperiodic control of PRL secretion.

Hypothalamic control of photoperiod-induced cycles in food intake, body weight, and metabolic hormones in rams.

This study used a hypothalamo-pituitary disconnected (HPD) sheep model to investigate the central regulation of long-term cycles in voluntary food intake (VFI) and body weight (BW). VFI, BW, and circulating concentrations of metabolic hormones [alpha-melanocyte-stimulating hormone (alpha-MSH), insulin-like growth factor-1 (IGF-1), insulin, and leptin] were measured in HPD and control Soay rams exposed to alternating 16 weekly periods of long and short days for 80 wk. In the controls, the physiology was cyclical with a 32-wk periodicity corresponding to the lighting regimen. VFI and BW increased under long days to a maximum early into short days, and there were associated increases in blood concentrations of alpha-MSH, insulin, and leptin. In the HPD rams, there were no significant photoperiod-induced changes in any of the parameters. VFI increased after surgery for 8 wk and then gradually declined, although BW increased progressively and the HPD rams became obese. Concentrations of alpha-MSH, insulin, and leptin in peripheral blood were permanently increased (>200%), and levels of IGF-1 decreased (<55%). The HPD lesion effectively destroyed the entire median eminence [no nerve terminals immunostained for tyrosine hydroxylase (TH) and gonadotropin-releasing hormone] and the adjacent arcuate nucleus (no perikarya immunostained for proopiomelanocortin or TH, and no cells expressed neuropeptide Y mRNA). The results support the conclusion that arcuate hypothalamic systems generate long-term rhythms in VFI, BW, and energy balance.

Influence of the degree of stimulation of the pituitary by gonadotropin-releasing hormone on the action of inhibin and testosterone to suppress the secretion of the gonadotropins in rams.

This experiment determined if the degree of stimulation of the pituitary gland by GnRH affects the suppressive actions of inhibin and testosterone on gonadotropin secretion in rams. Two groups (n = 5) of castrated adult rams underwent hypothalamopituitary disconnection and were given two i.v. injections of vehicle or 0.64 microg/kg of recombinant human inhibin A (rh-inhibin) 6 h apart when treated with i.m. injections of oil and testosterone propionate every 12 h for at least 7 days. Each treatment was administered when the rams were infused i.v. with 125 ng of GnRH every 4 h (i.e., slow-pulse frequency) and 125 ng of GnRH every hour (i.e., fast-pulse frequency). The FSH concentrations and LH pulse amplitude were lower and the LH concentrations higher during the fast GnRH pulse frequency. The GnRH pulse frequency did not influence the ability of rh-inhibin and testosterone to suppress FSH secretion. Testosterone did not affect LH secretion. Following rh-inhibin treatment, LH pulse amplitude decreased at the slow, but not at the fast, GnRH pulse frequency, and LH concentrations decreased at both GnRH pulse frequencies. We conclude that the degree of stimulation of the pituitary by GnRH does not influence the ability of inhibin or testosterone to suppress FSH secretion in rams. Inhibin may be capable of suppressing LH secretion under conditions of low GnRH.

The corticotropin-release inhibitory factor hypothesis: a review of the evidence for the existence of inhibitory as well as stimulatory hypophysiotropic regulation of adrenocorticotropin secretion and biosynthesis.

I. Introduction II. Hypothalamic Stimulation of ACTH Release and Biosynthesis A. Corticotropin-releasing factor (CRF) B. Arginine vasopressin (AVP) C. The hypothalamic CRF and AVP neurons and their connections D. Studies of the secretion of CRF and AVP into the hypophysial-portal circulation E. Studies of the neural regulation of CRF and AVP secretion and biosynthesis III. Hypothalamic Inhibition of ACTH Release and Biosynthesis A. Historic studies B. Effect of hypothalamo-pituitary disconnection in adult and fetal sheep C. Effects of the opiate alkaloids and opioid peptides on the HPA axis D. The role of the posterior pituitary in the regulation of corticotropic function E. Hypothalamic ACTH release-inhibitory activity F. Definition of corticotropin release inhibitory factor (CRIF) IV. CRIF: A Consideration of Possible Candidates A. Somatostatin (SST) B. Dopamine C. Atrial natriuretic peptide (ANP) D. Prepro-TRH-(178–199) E. Other substances V. Future Directions