Central neuropeptide B administration activates stress hormone secretion and stimulates feeding in male rats

Neuropeptide B (NPB) was identified to be an endogenous, peptide ligand for the orphan receptors GPR7 and GPR8. Because GPR7 is expressed in rat brain and, in particular, in the hypothalamus, we hypothesized that NPB might interact with neuroendocrine systems that control hormone release from the anterior pituitary gland. No significant effects of NPB were observed on the in vitro releases of prolactin, adrenocorticotropic hormone (ACTH) or growth hormone (GH) when log molar concentrations ranging from 1 pM to 100 nM NPB were incubated with dispersed anterior pituitary cells harvested from male rats. In addition NPB (100 nM) did not alter the concentration response stimulation of prolactin secretion by thyrotropin-releasing hormone, ACTH secretion by corticotropin-releasing factor (CRF) and GH secretion by GH-releasing hormone. However, NPB, when injected into the lateral cerebroventricle (i.c.v.) of conscious, unrestrained male rats, elevated prolactin and corticosterone, and lowered GH levels in circulation. The threshold dose for the effect on corticosterone and prolactin levels was 1.0 nmol, while that for the effect on GH release was 3.0 nmol NPB. Pretreatment with a polyclonal anti-CRF antiserum completely blocked the ability of NPB to stimulate ACTH release and significantly inhibited the effect of NPB on plasma corticosterone levels. NPB administration i.c.v. did not significantly alter plasma vasopressin and oxytocin levels in conscious rats. It did stimulate feeding (minimum effective dose 1.0 nmol) in sated animals in a manner similar to that of the other endogenous ligand for GPR7, neuropeptide W. We conclude that NPB can act in the brain to modulate neuroendocrine signals accessing the anterior pituitary gland, but does not itself act as a releasing or inhibiting factor in the gland, at least with regard to prolactin, ACTH and GH secretion.     

Sex differences in prolactin secretion in parental male and female rats

The present series of experiments report the behavioral and endocrine responses of male and female rats to rat young. In the first experiment, the behavioral responses of intact male and ovariectomized female rats to foster young were measured. While both of these groups exhibited most components of parental behavior after four to six days of exposure to rat young, males failed to build good nests and to crouch over the young. In addition significantly more males killed young than did ovariectomized females. In the second experiment, treatment of male and virgin female rats with a 21-day hormone regimen of estradiol and progesterone stimulated an equally rapid onset of parental behavior in both sexes. Latencies to exhibit parental responses were approx, two days for both males and females. Pup killing was virtually eliminated in males by hormone treatment. Although the behavioral responses of hormone-primed males and females were similar, their prolactin responses to the presentation of rat young differed. Pup presentation stimulated an increase in plasma prolactin content in hormone-primed parental virgin females similar, but not identical, to that found in lactating control rats, while plasma prolactin levels in hormone-primed parental males did not rise in response to the presentation of young. These data indicate that a sex difference in neuroendocrine, but not behavioral, responsiveness to young exists between parental males and females.     

Sex differences in conflict behaviour and in plasma corticosterone levels

Summary The levels of plasma corticosterone and conflict behaviour were followed in male and female water deprived Wistar rats exposed to the punished (0.2 or 0.8 mA) drinking test. The unpunished drinking, performed under familiar or unfamiliar conditions, and plasma corticosterone levels of these male and female rats were determined. Plasma corticosterone was elevated in water-deprived rats compared to rats under normal conditions. In all cases plasma corticosterone levels were considerably higher in water deprived females than in males. The highest levels in both sexes were obtained following drinking punished with 0.8 mA shocks. While the unpunished drinking did not differ between the two sexes, the punished drinking was significantly lower in females than in males. Although female rats displayed less punished licks and had higher plasma corticosterone levels than males, there was no correlation between the two parameters.     

Parallel changes in behavior and plasma levels of hormones in septal rats

(1) Septal lesions in male rats potentiated both behavioral and hormonal responses to stress as measured by plasma levels of corticosterone growth hormone and prolactin. (2) These effects of septal lesions were parallel and transitory. Both behavioral and hormonal abnormalities were maximal 3 days after surgery, decreased at 8 and 13 days after surgery and 2 months after surgery were completely absent. (3) The data are interpreted as suggesting that the septum plays a central role in the mechanisms required for responding to stress because its destruction has parallel effects on behavior, corticosterone, growth hormone and prolactin responses to stress.     

Electroconvulsive therapy (ECT) increases plasma growth hormone, prolactin, luteinising hormone and follicle-stimulating hormone but not thyrotropin or substance P

(1) Using radioimmunoassay, plasma levels of pituitary hormones and substance P were measured during the first 30 min after ECT in 28 sets of peripheral blood samples obtained from 21 patients (14 males and seven females) treated with ECT for endogenous depression. (2) The earliest increase was in plasma growth hormone and follicle-stimulating hormone occurring 5–14 min after ECT, followed by increases in plasma prolactin and luteinising hormone, 15–24 min after ECT. At 25–35 min after ECT only the increase in plasma growth hormone was still significant. (3) ECT had no effect on plasma levels of thyrotrophin or substance P. (4) It is suggested that the observed changes in peripheral levels of pituitary hormones are compatible with non-specific stimulation of the hypothalamo-pituitary system due to the acute stress of ECT or anesthesia.     

Freund's complete adjuvant induces ornithine decarboxylase activity in the central nervous system of male rats and triggers the release of pituitary hormones

In male rats, inoculation of Freund's complete adjuvant (FCA, 0.5 mg/rat of Mycobacterium butyricum in paraffin oil) induced high levels of ornithine decarboxylase (ODC) in the hypothalamus and pituitary gland (285% and 245% of controls, respectively, within 12 h to 2 days). ODC activity also was altered in the cerebellum and left neocortex, but not in the right neocortex. This activity reflected a dynamic equilibrium which is influenced by ODC synthesis, degradation, activation, etc. The circadian rhythms of pituitary ODC activity and plasma prolactin level, 3-4 days after FCA, showed that enhancement of enzymatic activity during the dark phase correlated with a marked release of prolactin (Prl). During this early period after FCA, changes in plasma levels of other pituitary hormones were not significant or were less important. Pretreatment with bromocriptine microcapsules inhibited both basal and FCA-induced pituitary ODC activity, as well as Prl secretion. Further, significant increases in plasma luteinizing hormone and adrenocorticotropic hormone were noted from days 4 and 8, respectively, and onwards. Finally, a phase of reduced corticosterone secretion occurred during the latency period. This study shows that FCA influences central nervous system pathways and supports the idea that endogenous Prl is involved in some early events which lead to the development of adjuvant arthritis.     

Orphanin FQ stimulates prolactin and growth hormone release in male and female rats

Intracerebroventricular administration of Orphanin FQ (5.5, 55 or 550 pmol) caused a dose-related increase in prolactin secretion in both male and female rats and stimulated GH secretion in males. The magnitude of the prolactin secretory response was greater in females than in males. These effects of OFQ on prolactin and growth hormone release are the same as the stimulatory effects of the endogenous opioid peptides.     

Gonadectomy reverses the sexually diergic patterns of circadian and stress-induced hypothalamic-pituitary-adrenal axis activity in male and female rats

Enhanced corticosterone release by female compared to male rats under basal and stress conditions is well documented. The demonstration that gonadectomy enhances stress-induced corticosterone secretion in male rats, but reduces such levels in female rats, suggests a causal association between gonadal steroids and corticosterone release. The present study examined the corticosterone profile of sham gonadectomized and gonadectomized female and male rats under basal and stress conditions. An automated sampling system collected blood from each freely moving, unanaesthetized rat every 10 min (i) over a 24-h period; (ii) following noise stress; and (iii) following an immune-mediated stress (lipopolysaccharide, LPS). Plasma was analysed for corticosterone content using radioimmunoassay. Castration resulted in a significant increase in basal corticosterone release compared to the sham-castrated male rats. Pulsar analysis revealed a significant two-fold increase in the number of corticosterone pulses over 24 h. Corticosterone increases in response to noise stress and to LPS injection were enhanced following castration. Conversely, ovariectomy resulted in a two-fold reduction in the number of corticosterone pulses as well as the stress response compared to sham-ovariectomized female rats. Arginine vasopressin (AVP), corticotrophin-releasing hormone (CRH) and glucocorticoid receptor mRNAs in the paraventricular nucleus and pro-opiomelanocortin (POMC) mRNA in the anterior pituitary were analysed post-LPS administration by in situ hybridization. Significantly higher values were found for AVP, CRH and POMC mRNAs examined for sham females and castrated males compared to sham males and ovariectomized females. This study confirms previous reports concerning the influence of gonadal factors in regulating HPA axis activity and stress responsiveness. The present results extend these observations to the regulation of the dynamic pattern of corticosterone release under basal conditions and suggests that this alteration in pulsatility is important for the differences in stress responsiveness when comparing males and females.     

Hypothalamic-Pituitary-Adrenal Activation Following Endotoxin Administration in the Developing Rat: A CRH-Mediated Effect

The present studies assessed hypothalamic-pituitary-adrenal (HPA) responses following immune activation with endotoxin (ip) in three-day old Long Evans rats. Marked plasma corticosterone (B), adrenocorticotrophic hormone (ACTH) responses and biphasic fluctuations in plasma glucose were maximal at a dose of 0.05 mg/kg. HPA responses peaked between 3–5 h following immune challenge and plasma ACTH and B responses were greater in female than in male rat pups. Plasma levels of corticosterone binding globulin (CBG) were reduced in males and substantially increased in females during the peak HPA response. Changes in plasma glucose were biphasic with slight increases when ACTH and B levels were maximal, but hypoglycemia was evident once plasma B levels returned to resting values. Endotoxin challenge reduced median eminence corticotropin-releasing hormone (CRH) levels at times corresponding with elevated HPA activity, and prior icv injection of the CRH antagonist, a-helical CRH, significantly attenuated elevations in plasma ACTH and B. In addition, α-helical CRH pretreatment completely blocked endotoxin-induced changes in plasma CBG in both males and females. These findings support the view that endotoxin-induced HPA activation in the neonate may occur via CRH.     

Central Nervous System Sites of Action of an Enkephalin Analogue, (D-Met2, Pro5)-Enkephalinamide, and Naloxone on the Secretion of Five Anterior Pituitary Hormones of Male Rats

Central nervous system sites of action of opioid peptides on pituitary hormone secretion were investigated. One nmol of an enkephalin analogue, (D-Met², Pro⁵)-enkephalinamide, and 10 nmol of the opiate antagonist naloxone were injected into ten different regions of the brain of conscious male rats and their effect on the release of five anterior pituitary hormones tested. The injections were made through a special injection cannula which was inserted into the brain through a guide cannula fixed on the skull and implanted into the brain 5 to 7 days earlier. Both compounds injected into the medial septum, medial preoptic area and hypothalamic paraventricular nucleus affected prolactin, growth hormone and luteinizing hormone (LH) secretion. The enkephalin analogue stimulated prolactin and growth hormone and inhibited LH release. Naloxone induced the opposite effect. Drugs given into the hypothalamic ventromedial nucleus caused changes in plasma prolactin and growth hormone levels. Enkephalinamide increased and naloxone decreased plasma concentrations of both hormones. Administration of the compounds into the dorsal raphe area resulted in alterations of prolactin and LH release, the analogue caused elevation of prolactin and inhibition of LH release, whereas the opiate antagonist resulted in opposite changes. Only an LH response was obtained from the hypothalamic dorsomedial nucleus and a growth hormone response from the central amygdala. Also in these cases the enkephalin analogue decreased LH and elevated growth hormone plasma levels, and naloxone brought about a rise in LH and a diminution of growth hormone concentration. None of the regions were effective in inducing a clear-cut adrenocorticotrophin or follicle-stimulating hormone response. The parietal cortex, medial amygdala and the dentate gyrus were entirely ineffective sites. The findings suggest that in the brain there are multiple sites of action of opioids on pituitary trophic hormone secretion and the effective sites are not identical in terms of pituitary hormone response.     

Response of anterior pituitary hormones to chronic stress. The specificity of adaptation

The effect of chronic noise stress on the response of anterior pituitary hormones to the same or to another stressor (forced swimming) was studied in adult male Wistar rats. Both acute stressors increased corticosterone, prolactin, LH and TSH secretion and inhibited GH secretion. Previous chronic exposure to noise reduced corticosterone response to the same stimulus without modifying corticosterone response to a novel acute stress. Neither prolactin nor TSH responses to acute noise were reduced by previous chronic exposure to noise. Since chronic noise increased basal levels of LH and decreased those of GH, the response of these hormones to acute stress was expressed as percent changes of their respective basal values. It was found that chronically stressed rats showed diminished LH response to noise but not to forced swimming. GH showed the same pattern without reaching statistical significance. These data indicate that the response of some anterior pituitary hormones can adapt after repeated exposure to the same stressor. When adaptation occurred, this was specific for the stressor which the animals were repeatedly exposed to. The pituitary-adrenal axis appears to be the most reliable index of adaptation to chronic stress among all the anterior pituitary endocrine axes.     

Somatic and Neuroendocrine Changes in Response to Chronic Corticosterone Exposure During Adolescence in Male and Female Rats

Prolonged stress and repeated activation of the hypothalamic‐pituitary‐adrenal axis can result in many sex‐dependent behavioural and metabolic changes in rats, including alterations in feeding behaviour and reduced body weight. In adults, these effects of stress can be mimicked by corticosterone, a major output of the hypothalamic‐pituitary‐adrenal axis, and recapitulate the stress‐induced sex difference, such that corticosterone‐treated males show greater weight loss than females. Similar to adults, chronic stress during adolescence leads to reduced weight gain, particularly in males. However, it is currently unknown whether corticosterone mediates this somatic change and whether additional measures of neuroendocrine function are affected by chronic corticosterone exposure during adolescence in a sex‐dependent manner. Therefore, we examined the effects of non‐invasively administered corticosterone (150 or 300 μg/ml) in the drinking water of male and female rats throughout adolescent development (30–58 days of age). We found that adolescent animals exposed to chronic corticosterone gain significantly less weight than controls, which may be partly mediated by the effects of corticosterone on food consumption, fluid intake and gonadal hormone function. Our data further show that, despite similar circulating corticosterone levels, males demonstrate a greater sensitivity to these changes than females. We also found that Npy1 and Npy5 receptor mRNA expression, genes implicated in appetite regulation, was significantly reduced in the ventral medial hypothalamus of corticosterone‐treated males and females compared to controls. Finally, parameters of gonadal function, such as plasma sex steroid concentrations and weight of reproductive tissues, were reduced by adolescent corticosterone treatment, although only in males. The data obtained in the present study indicate that chronic corticosterone exposure throughout adolescent development results in significant and sex‐dependent somatic and neuroendocrine changes, and the results also provide an experimental framework for further investigating the impact of corticosterone on metabolic and neuroendocrine function during adolescence.     

Assessment of the effects of a synthetic gonadotropin-releasing hormone associated peptide on hormone release from the in situ and ectopic pituitaries in adult male rats

The in vivo effects of synthetic human gonadotropin-releasing hormone associated peptide (GAP) were evaluated in adult male rats. In normal rats, intracerebroventricular (III ventricle) injection of 5 ng GAP significantly increased plasma LH levels after 60 minutes. Intracerebroventricular administration of 5, 25 or 125 ng of GAP elevated circulating LH levels also at 120 minutes of injection but did not alter plasma FSH, prolactin or testosterone concentrations. In hypophysectomized-pituitary-grafted rats injection of 125 ng GAP directly into the ectopic pituitary induced no changes in peripheral hormone levels. However, intrapituitary graft injection of 25 ng of GnRH significantly elevated circulating levels of LH and testosterone. These results indicate that the ectopic pituitary graft can respond to acute exogenous GnRH stimulation and that the commercially available synthetic GAP fails to inhibit prolactin release in adult male rats.     

Regulation of corticotropin-releasing hormone receptors and hypothalamic pituitary adrenal axis responsiveness during cold stress

The relationship between pituitary corticotropin-releasing hormone (CRH) receptor changes and corticotrope responsiveness was studied during chronic cold stress. Exposure of rats to 4°C caused a gradual increase in plasma adrenocorticotropic hormone (ACTH), reaching 3-fold over the basal levels by 6 h (P< 0.005), followed by a reduction to levels only 1.3-fold over basal by 60 h. Plasma corticosterone was significantly increased after 1 h (P<0.005) and remained elevated for up to the 60 h cold exposure (P < 0.005). No significant changes in plasma thyroid-stimulating hormone, prolactin, growth hormone or vasopressin were observed at 60 h of cold exposure. CRH receptor concentration in the anterior pituitary was unchanged after 18 or 60 h cold stress, whereas in neurointermediate lobe membranes was markedly elevated. Autoradiographic analysis of pituitary CRH receptors confirmed that the increase in CRH binding was confined to the intermediate pituitary. CRH receptor levels in membranes from two brain areas, frontal cortex and amygdala, were unchanged following 60 h cold stress. The areas under the curve for the plasma ACTH and corticosterone levels following an injection of 10μg/kg ovine CRH in conscious rats, were of similar magnitude in control and 60 h cold exposed rats. Plasma ACTH responses to ether stress were significantly higher in rats exposed to cold for 60 h than in controls. In the intermediate pituitary, basal ß-endorphin/lipotropin release from isolated intermediate pituitary cells was significantly lower in cold stressed rats, and despite the increase in CRH receptors CRH-stimulated values were not higher than in controls. Following 60 h cold exposure, immunoreactive CRH content was decreased in neurointermediate pituitary extracts, while it was slightly increased in the median eminence. Exposure to ether for 5 min resulted in a significant decrease in immunoreactive CRH in the median eminence of cold stressed rats, but not in the controls. The data show a lack of correlation between changes in CRH receptor levels and responsiveness of the corticotrope in the anterior and intermediate pituitary lobes. This suggests that postreceptor events and interaction of CRH with other regulators of ACTH secretion are more likely to account for the changes in pituitary responsiveness during chronic stress.     

Differential impact of predator or immobilization stressors on central corticotropin-releasing hormone and bombesin-like peptides in Fast and Slow seizing rat

Lines of rats selectively bred for amygdala excitability, as reflected by kindling rates in response to electrical stimulation, also exhibit differences in tests of anxiety. Inasmuch as corticotropin-releasing hormone (CRH) and bombesin (BN) have been associated with anxiety, regional levels and release of these peptides, as well as plasma adrenocorticotropic hormone (ACTH) and corticosterone, were assessed in 'Slow' and 'Fast' seizing rats following predator exposure (ferret) or immobilization. Ferret exposure elicited a greater increase of plasma ACTH and corticosterone concentrations in the Slow than in the Fast rats. In contrast, immobilization provoked a greater rise of plasma ACTH levels in the Fast rats, paralleling the vigorous struggling observed in this line. In Slow rats, stressor exposure elicited increased levels of ir-BN at the anterior hypothalamus, and increased ir-CRH at the median eminence/arcuate nucleus (Me/Arc), paraventricular hypothalamic nucleus (PVN) and pituitary (Pit), whereas decreased levels of ir-BN were found at the nucleus tractus solitarius (NTS). Fast rats likewise showed decreased ir-BN at the NTS, but unlike the Slow rats, ir-CRH was reduced in the Me/Arc, PVN and Pit in response to both stressors. In vivo microdialysis experiments revealed that in response to ferret exposure, the Slow rats showed a greater CRH release at the central nucleus of the amygdala (CeA) as compared to Fast rats. However, immobilization elicited a more pronounced release of CRH in Fast than in Slow rats. Taken together, the results demonstrate that these two lines of rats show differential endocrinological and neurochemical response patterns to these stressors.     

Lithium and twenty-four hour rhythms of serum corticosterone, prolactin and growth hormone in pigmented eye rats

Lithium, a widely used substance for the treatment of manic-depressive illness, has been reported to alter the phase relationships of a variety of biological rhythms. We have previously found that lithium affects serum melatonin differently in albino compared to pigmented eye rats. The present study was undertaken to investigate the effect of lithium on pituitary rhythms in pigmented eye rats. Six point 24 hour maps were generated throughout a 12 hour light/12 hour dark lighting regime on separate groups of individually housed adult male Long Evans rats (with pigmented eyes), maintained for six weeks on ad lib water and either normal lab chow or lab chow supplemented with 50 mM/Kg of lithium chloride. Animals were sacrificed by rapid decapitation with care to ensure that blood samples were obtained from subjects in the resting, undisturbed state. Plasma lithium levels were 0.57 ± 0.02 mEq/l. In comparison to normal controls, lithium treatment suppressed body weight by 19%, and increased water intake by 100%. Absolute corticosterone levels were not altered by lithium, but the 24-hour pattern was significantly changed. Growth hormone levels were significantly reduced by lithium treatment without alteration of the 24-hour pattern. Prolactin levels were significantly reduced by lithium and the normal 24-hour variation was attenuated. Comparison of these effects of lithium in pigmented eye rats with similar data from albino rats suggests that the effects on growth hormone, body weight and water intake were similar; however, the effects on prolactin and corticosterone differed. Since melatonin has been implicated in the regulation of prolactin and corticosterone rhythms, and this hormone is affected differently in the two types of rat, the observed effects of lithium on these hormone rhythms may be secondary to an alteration of the 24-hour pattern of melatonin by lithium.     

Release of growth hormone, prolactin and somatostatin during perifusion of anterior pituitary and preoptic-medial basal hypothalamus from male and female rats

These experiments were designed to determine whether it is possible using in vitro perifusion to identify a sex difference in anterior pituitary (AP) release of growth hormone (GH) and, if so, to determine whether this difference is correlated with a sex difference in hypothalamic release or content of somatostatin (SRIF). Age-matched rats of both sexes were decapitated at approximately 09.00 h, and blood was collected for determination of non-stress plasma concentrations of GH. Each pituitary was rapidly removed and prepared for perifusion of the AP, and each preoptic-medial basal hypothalamus (PO-MBH) was removed and placed in a separate perifusion chamber. The effluent fractions from perifused APs were assayed for GH and prolactin (Prl), and those from PO-MBH blocks were assayed for SRIF. Non-stress plasma GH concentrations were similar in males and females. During perifusion, baseline GH release was higher (P less than 0.001) from male than from female APs. Release of GH from the APs of both sexes was similarly inhibited (P less than 0.001) by a 1-h administration of SRIF (10(-7) M), and high K+ (50 mM) caused larger (P less than 0.05) GH responses from male than from female APs. In contrast, baseline Prl release was higher (P less than 0.01) from female than from male glands, and Prl release was not affected by SRIF. Male and female PO-MBH tissues showed similar baseline release of SRIF and similar responses to high K+. After perifusion, GH content and concentration were higher in APs from males than from females, but SRIF content in the perifused male and female PO-MBH tissues was similar.(ABSTRACT TRUNCATED AT 250 WORDS)