Alpha 2-adrenoreceptor subtypes regulate ACTH and beta-endorphin secretions during stress in the rat.

The effect of different alpha 2-adrenoreceptor subtype agonists and antagonists on adrenocorticotrop hormone (ACTH) and beta-endorphin release induced by ether stress was examined. Ether inhalation-induced ACTH and beta-endorphin increase was inhibited by i.c.v. administration of 30 micrograms but not 1 and 10 micrograms clonidine (alpha 2-adrenoreceptor agonist). I.c.v. oxymetazoline (alpha 2A-adrenoreceptor agonist; 1-10-30 micrograms) or the alpha 1-agonist methoxamine (100 micrograms/rat) failed to inhibit the stress-induced rise. Pretreatment with the alpha 1/alpha 2B.C-antagonist prazosin (0.5 mg/kg, i.p.) prevented the effect of clonidine on the ether stress, while the alpha 1/alpha 2A-antagonist WB-4101 (0.5 mg/kg, i.p.) was unable to counteract the inhibitory effect of clonidine. Prazosin alone had no effect on the ether-induced plasma ACTH and beta-endorphin elevation. These results suggest that noradrenaline in the central nervous system may inhibit the stress-induced hypothalamo-pituitary-axis and pituitary beta-endorphin activation via alpha 2B.C-adrenoceptor subtypes and prazosin may antagonize its effect on these receptors.

Domperidone stimulates prolactin secretion in rats with complete destruction of the mediobasal hypothalamus.

The main objective of this study was to further elucidate the functional relationship between endogenous dopamine and the prolactin (PRL)-releasing effect of the dopamine antagonists domperidone and haloperidol. We studied the effect of the above dopamine antagonists on the PRL secretion in control and mediobasal hypothalamus (MBH)-lesioned rats. Significant increase in basal plasma PRL levels was detected 7 days after complete surgical destruction of the MBH. Haloperidol injection (0.5 mg/kg, i.v.) was followed by an increased plasma PRL concentration in the sham-operated animals; however, in the MBH-lesioned rats where the basal PRL levels were high haloperidol failed to produce additional PRL release. In contrast to haloperidol, domperidone (0.1 mg/kg, i.v.) was able to further elevate the MBH-lesion induced high plasma PRL concentration. Moreover, the change in plasma PRL levels of the MBH-lesioned rats was parallel with that in the sham-lesioned animals after domperidone injections. When haloperidol was given prior to the domperidone injection it did not influence the PRL releasing effect of domperidone in MBH-lesioned animals. The PRL stimulatory effect of domperidone (0.3 mg/kg, i.v.) in MBH-lesioned rats was antagonized by dopamine (20 micrograms/kg, i.v.) and bromocryptine (20 micrograms/kg, i.v.). The above results suggest that the stimulatory effect of domperidone on the pituitary PRL secretion is mediated--at least in part--through the pituitary D2 dopamine receptors, but not by the displacement of endogenous dopamine originating from the MBH and reaching the pituitary via portal vessels.

The physiology of social conflict in rats: what is particularly stressful?

The involvement of the opponent's behavior in the aggression-induced stress response was tested. Subjects (male Wistar rats) faced stimulus rats previously submitted either to mediobasal hypothalamic lesion (MBHL) or to sham operation. MBHL intruders were more aggressive but were also more ready to submit compared with sham-operated rats; as residents they were more efficient in inducing submissive behavior in the opponent. The subject's stress response at 15 min was neither dependent on the intensity of aggressive behavior nor on residence status but on dominant-submissive relationships. The submission of the stimulus rat reduced corticosterone plasma levels in the subjects, while the subjects' own submissive behavior elicited a very significant increase in its plasma corticosterone. In contrast, plasma glucose seemed to depend on residence status, whereas plasma free fatty acids reacted to any novel event (including cage switch).

Hypothalamic alpha 2A-adrenoceptors stimulate growth hormone release in the rat.

Oxymetazoline, the relatively selective alpha 2A-adrenoceptor agonist (with more than 60-fold selectivity over the alpha 2B-adrenoceptor subtype), was administered into the lateral ventricle (i.c.v.) of rats and plasma growth hormone (GH) levels were measured. Oxymetazoline was more potent to release GH after i.c.v. administration than was clonidine; 0.01 microgram i.c.v. oxymetazoline already caused a significant release of GH, while at least 0.1 microgram clonidine had to be administered to cause a similar response. The dose-response curve was of an inverted U shape since with 10 micrograms of oxymetazoline the plasma GH did not rise. When oxymetazoline was injected i.c.v. to rats with somatostatin fibres to the median eminence transected by an anterolateral cut in the hypothalamus there was a significant rise in plasma GH, suggesting that oxymetazoline stimulated GHRH rather than inhibited somatostatin release. Pretreatment with CH-38083 (7,8-(methylenedioxy)-14-alpha-hydroxy-alloberban HCl, selective for alpha 2-adrenoceptors but not differentiating between alpha 2A and alpha 2B subtypes), prevented the plasma GH rise normally elicited by 1 microgram i.c.v. oxymetazoline. The alpha 2A- and alpha 1-selective adrenoceptor antagonist, WB-4101 (2-(2,6-dimethoxyphenoxyethyl)-aminomethyl-1,4-benzodioxane hydrochloride), prevented the GH rise normally induced by oxymetazoline while prazosin, the alpha 2B- and alpha 1-selective adrenoceptor antagonist, prolonged the elevation occurring in the control rats between 30 and 60 min after oxymetazoline injection. Since both prazosin and WB-4101 are alpha 1-adrenoceptor antagonists but differ in their action on alpha 2A and alpha 2B subtypes as well as in their action on oxymetazoline-induced GH secretion, the antagonist studies suggest that oxymetazoline stimulates GH release through activation of alpha 2A-adrenoceptors stimulatory to GHRH release, and not by an action through alpha 2B- or alpha 2C- or alpha 1-adrenoceptors. Since WB-4101 also antagonized clonidine action on GH release we also suggest that the major component may be the stimulation of the alpha 2A-adrenoceptors in the clonidine action on GH release.

Do alpha-2 adrenoceptors modify coping strategies in rats?

Previous research has shown that resident rats treated with alpha 2 adrenoceptor blockers display a modified aggressive response towards intruding animals. In the present study we report data on the behavioral changes induced by alpha 2 adrenoceptor blockers in intruder animals. In experiments 1 and 2 intruders smaller in body weight than the residents were treated with 0.0, 0.5, and 1.0 mg/kg CH-38083 and idazoxan, respectively; in experiment 3 weight matched intruders were injected with 1 mg/kg CH-38083 or idazoxan. The treatment of smaller intruders did not change the behavior of residents. In contrast, weight-matched intruders injected with alpha 2 adrenoceptor blockers elicited increased aggression in residents. Social behaviors, exploration and offensive aggression showed insignificant variation in intruders. Defensive behaviors, in contrast, showed major changes: in experiments 1 and 2 a dose-dependent decrease in immobility and a dose-dependent increase in defensive upright was noticed. In experiment 3, high scores of defensive upright were apparent, precluding detection of drug-induced changes. However, when the last 5 min of the encounter were analysed separately, results similar to the first two experiments were observed. Significant negative correlations were found between immobility and defensive upright scores. The results suggest that alpha 2 adrenoceptor blockers induce a shift from a passive (immobility) towards a more active (defensive upright) coping style. These and previous data show that alpha 2 adrenoceptor blockers, other than yohimbine, seem to exert a behavior-activating effect in rats.

Adrenocorticotropin, prolactin and beta-endorphin stimulatory actions of alpha-2-adrenoceptor antagonists.

We studied the effect of glucocorticoid pretreatment, mediobasal hypothalamus lesion (MBHL) and the interaction between clonidine and yohimbine in male Wistar rats to elucidate the sites and/or mechanisms of endocrine actions of alpha 2-antagonists. The pretreatment of 1 mg/kg s.c. dexamethasone for 4 days effectively prevented the stimulatory effect of alpha 2-antagonists yohimbine (5 mg/kg i.p.) and CH-38083 (1 mg/kg i.p.) on adrenocorticotropin (ACTH) secretion, while the action of these antagonists on prolactin (PRL) and beta-endorphin (beta E) remained unchanged. The central (i.c.v.) pretreatment of 5 micrograms/rat clonidine failed to antagonize the prolactin (PRL) and beta E releasing effect of yohimbine. However, it inhibited the yohimbine-induced ACTH secretion. MBHL resulted in a significant enhancement in basal plasma PRL and beta-endorphin (beta E) levels. But basal plasma ACTH levels have not been changed. Yohimbine failed to stimulate ACTH secretion in MBH-lesions rats, while PRL and beta E response to the yohimbine was maintained in these animals. This study confirms that the alpha 2-antagonists stimulate ACTH secretion by a corticosteroid-sensitive mechanism which is located centrally. In contrast, alpha 2-antagonists affect PRL and beta E secretion via a corticosteroid-insensitive mechanism located at the periphery, possible within the pituitary gland.

Diurnal alteration in opiate effects on the hypothalamo-pituitary-adrenal axis: changes in the mechanism of action.

Opioid control of the hypothalamo-pituitary-adrenocortical axis has a characteristic circadian rhythm (Kiem, Kanyicska, Stark and Fekete, 1987). To elucidate the mechanisms leading to circadian alterations of opioid control of the hypothalamo-pituitary-adrenocortical axis, and to look for the receptor type at which the p.m. inhibitory action of opioids occurs, we examined the effect of morphine at different doses and the interaction between naloxone and morphine at different times of day in intact male Wistar rats. In the morning: morphine (10 and 30 mg/kg s.c.) significantly increased corticosterone secretion, while 3 mg/kg s.c. had no effect. Naloxone in a dose of 2.5 mg/kg i.p. significantly antagonized the corticosterone-releasing effect of morphine, suggesting that the secretion of corticosterone induced by morphine is mediated via mu-opioid receptors. In the afternoon: basal plasma corticosterone levels were higher than those in the morning, and morphine caused a significant corticosterone increase only at the dose of 30 mg/kg, and had no effect in the dose of 10 mg/kg. Morphine significantly decreased corticosterone levels in the dose of 3 mg/kg. This inhibitory action lasted approximately 3 h after morphine injection and was able to inhibit the circadian evening rise of corticosterone. The effect of morphine and the interaction between naloxone and morphine on prolactin secretion remained unchanged from a.m. to p.m.; naloxone (2.5 mg/kg i.p.) which inhibited the 30 mg/kg morphine-induced corticosterone rise in the morning failed to antagonize the 3 mg/kg morphine-induced decrease of corticosterone secretion in the afternoon. A high dose of naloxone (10 mg/kg i.p.) effectively prevented the 3 mg/kg morphine-induced p.m. inhibition of corticosterone secretion.(ABSTRACT TRUNCATED AT 250 WORDS)

Prolactin response to morphine in intact and adrenalectomized lactating rats.

To examine the hypothesis that the increased adrenocortical activity during lactation induced the loss of the prolactin (PRL) -releasing effect of morphine, we studied the effect of morphine in adrenalectomized (ADX) and sham-operated primiparous lactating Wistar rats. Animals were adrenalectomized 4 days after delivery. On day 11 of lactation (7 days after ADX), pups were separated from their mother 2 h before morphine or haloperidol injection. Intravenous injection of 5 mg/kg morphine did not change plasma PRL levels in the sham-operated lactating rats, but it resulted in a significant increase of plasma PRL levels in ADX lactating animals, with or without corticosterone replacement. Catalepsy following 10 mg/kg i.v. morphine was also markedly enhanced in ADX lactating animals. The PRL response to 0.5 mg/kg haloperidol was higher in ADX lactating animals than that in the controls. Morphine given 2 h after haloperidol treatment resulted in a further increase of plasma PRL in ADX, but not in the sham-operated lactating animals. These results suggest that adrenal hyperfunction may lead to a loss of sensitivity to morphine during lactation.

Effect of dexamethasone implanted in different brain areas on the morphine-induced PRL, GH and ACTH/corticosterone secretion.

We studied the effect of dexamethasone (DEX) implantation in male Wistar rats to elucidate the site of action of morphine-induced prolactin (PRL), growth hormone (GH), adrenocorticotropic hormone (ACTH) and corticosterone (B) secretion. DEX or cholesterol was implanted in the close vicinity of the paraventricular (PVN), or the arcuate nuclei (ARN) of the hypothalamus or into the hippocampus. Five days after implantation blood samples were taken 30 min after i.p. morphine by decapitation or through an indwelling cannula 15, 30, 60 min after i.v. injection. DEX implanted near the PVN resulted in a blockade of morphine-induced ACTH and B secretion. In contrast, GH response to morphine was enhanced, while that of PRL was unchanged. DEX implanted near the ARN significantly inhibited the PRL-releasing effect of morphine, but was without any influence on the PRL secretion induced by haloperidol. There was a partial reduction in the B response to morphine, and GH secretion was unchanged. Dorsal hippocampal implants were without any effect on the morphine-induced GH, PRL or B secretion. We suggest that the site of glucocorticoid inhibitory action in the hypothalamus is the PVN for the opiate-induced ACTH/B secretion, and the ARN for the morphine-induced PRL release. The enhanced GH response to morphine observed in DEX-PVN implanted rats might be due to a decreased somatostatin tone.

Inhibition of suckling-induced prolactin release by dexamethasone.

The effect of dexamethasone (DEX) treatment (400 and 200 micrograms/kg BW 21 and 2 h before suckling stimulus, respectively) on suckling- and domperidone (DOMP)-induced PRL release was investigated in freely moving, primiparous lactating rats. DEX completely blocked suckling-induced plasma PRL release without affecting DOMP-induced release of the hormone suggesting a central action of DEX. The effect was transient because it could not be detected on the second day of testing. The effect of DEX implanted in three different brain areas on suckling- and DOMP-induced PRL release was also tested. Implants surrounding the hypothalamic paraventricular nuclei and dorsal hippocampus failed to affect PRL release induced by suckling stimulus. Surprisingly, DEX suppressed PRL release induced by suckling stimulus when it was implanted into the medial basal hypothalamus. These findings demonstrate that DEX is a potent inhibitor of the suckling-induced PRL release. They also indicate that the site of action of DEX is not at the anterior pituitary gland or the paraventricular nuclei and hippocampus because DEX treatment and DEX implants had no effect on plasma PRL levels induced by DOMP and suckling stimulus, respectively. Our data suggest that the effect of DEX is mediated through a region of the medial basal hypothalamus. The observed transient block in suckling-induced PRL release may be physiologically relevant during stress in lactating mothers for conserving pituitary stores of the hormone needed for milk production or being able to adapt to a rapid change in osmoregulation.

Reevaluation of the role of alpha 2-adrenoreceptors in morphine-stimulated release of growth hormone.

The relationship between opioidergic and alpha 2-adrenergic system in the regulation of GH secretion was studied using a novel alpha 2-antagonist, CH-38083, and chronic treatment with yohimbine or clonidine. In male Wistar rats morphine (3 mg/kg s.c.), and clonidine (31 micrograms/kg i.p.) induced a significant increase in plasma GH levels. The pretreatment with the alpha 2-antagonist yohimbine (1 and 3 mg/kg) effectively inhibited the GH releasing effect or morphine and clonidine. CH-38083 at the dose of 1 mg/kg did not interfere with the morphine-induced GH secretion, while it fully antagonized the GH-releasing effect of clonidine. Higher doses (3 and 5 mg/kg) of CH-38083 only partly inhibited GH secretion induced by morphine. In rats chronically treated with clonidine (2 micrograms/ml in the drinking water for 14 days) the GH response to an injection of clonidine was blocked, while the effect of morphine on the GH secretion remained unchanged. In long-term castrated rats the effect of clonidine (15, 31 and 250 micrograms/kg i.p.) on the GH secretion was significantly blunted, while the GH-releasing effect of morphine (1, 3 and 5 mg/kg s.c.) remained unchanged. The replacement of testosterone (10 mg/kg for 4 days) in castrates restored the effect of clonidine, whereas it decreased the stimulatory action of morphine on the GH secretion. In rats chronically treated with yohimbine (2 mg/kg i.p. 2-3 times daily for 14 days until sacrifice), the GH response to a high dose of clonidine (0.5 mg/kg i.p.) was blocked, while the effect of morphine (5 mg/kg s.c.) was significantly enhanced.(ABSTRACT TRUNCATED AT 250 WORDS)

Loss of the effects of clonidine on the growth hormone release and hypotensive action in long-term castrated rats: the possible role of testosterone on alpha-2-adrenergic mechanisms of clonidine.

The effect of testosterone on the growth hormone (GH)-releasing and hypotensive actions of clonidine was evaluated in vivo using male Wistar rats. Clonidine at various doses and by various routes (5 micrograms/kg, i.c.v.; 15 micrograms/kg, i.v., and 7, 15, 31, 125 and 250 micrograms/kg, i.p.) induced a significant increase in plasma GH levels in rats. Intracerebroventricular (5 micrograms/kg) injection of clonidine significantly decreased blood pressure. The effects of clonidine on the GH release and blood pressure were lost in long-term castrated rats, but were restored by testosterone replacement in the castrates. In addition, the hypotensive response to clonidine in testosterone-replaced castrated rats lasted longer than in the sham-operated rats. The injection of yohimbine (3 mg/kg, i.p.) strongly inhibited the secretion of GH induced by clonidine treatment in sham-operated animals, and its inhibitory action was significantly attenuated in castrates. In long-term castrated rats, basal blood pressure decreased significantly compared with that of sham-operated rats. This was also restored after testosterone administration to castrates. The results of the present study indicate that the alpha 2-adrenergic receptor-mediated effects of clonidine on the GH release and blood pressure might be dependent on the endogenous testosterone secretion.

The effect of systemically and locally administered steroids on VIP-like immunoreactive cells in the paraventricular nucleus of adrenalectomized rats.

Seven days after adrenalectomy (ADX) the number of vasoactive intestinal peptide (VIP)- and corticotropin-releasing factor (CRF)-immunoreactive cells in the parvocellular part of the rat paraventricular nucleus (PVN) increased markedly. Dexamethasone in the drinking water (started immediately after ADX) or its local implantation around the hypothalamic PVN reduced the ADX-induced increase in the number of VIP- and CRF-like immunoreactive neurones. The present results suggest that PVN might be the site at which glucocorticoids inhibit the increase in VIP-positive cells after ADX.

The possible role of androgenic hormones in maturation of alpha-adrenergic mechanisms in the rat.

Abstract In order to get better characterization of androgenic hormones on the functionality of the alpha(2)-adrenergic system and widen our previous studies, we investigated the effect of clonidine on the growth hormone (GH) secretion in male Wistar rats of various ages: 7-, 14- and 30-day old and adult, adult male castrated rats with and without testosterone treatment. Two different patterns of the GH response to clonidine have been observed in the control and testosterone-treated young animals: clonidine at the dose 15 mug/kg intraperitoneally had no effect on the GH secretion in 7- to 14-day old rat pups. In contrast, its effect appeared following the increase in the plasma testosterone concentration induced by pretreatment with testosterone (5 mg/kg subcutaneously for 4 days) in these animals. In 30-day old rats clonidine affected GH secretion and this influence was more pronounced in the testosterone-treated animals than in the controls. The decrease in the circulating testosterone levels caused by castration in adult male rats caused a decreased GH response to clonidine. Moreover, there was a tendency for the GH response to return in 4-week old animals. The effect of clonidine has been restored by testosterone replacement of castrates. Testosterone administration decreased basal plasma GH levels in the pups. However, it triggered the ultradian surges of GH secretion which were absent in the young animals. Clonidine had no effect on the corticosterone secretion in 7-day old animals. Testosterone treatment induced a response in the 7-day old rat and markedly potentiated its effect on the secretion of this hormone in 14- and 30-day old animals, respectively. Neither progesterone nor hydrocortisone influenced the GH-releasing effect of clonidine. Hydrocortisone markedly inhibited the basal- and clonidine-induced corticosterone secretion. The results of the present study indicate an important role of androgenic hormones in inducing and/or maintaining the effectiveness/sensitivity of the alpha(2)-adrenergic receptor system and suggest a possible role for the androgenic hormones in the maturation of alpha-adrenergic mechanisms in the rat.

Prolactin release induced by opiate agonists, effect of glucocorticoid pretreatment in intact and adrenalectomized rats.

Cortisol administered at a dose of 25 mg/kg 24 h before measurements decreased the prolactin secretion induced by intraventricularly given opioids (dynorphin, beta-endorphin, Met-enkephalin or D-Met-Pro-enkephalinamide). The effect of cortisol was depressed by actinomycin D pretreatment. The cortisol-induced inhibition of the action of morphine was facilitated in adrenalectomized animals; measuring the effects of increasing doses of cortisol a maximal inhibition was obtained at a dose of 5 mg/kg. The opioid-induced corticosterone secretion was not affected 24 h after a single administration of cortisol. The present results show that the cortisol-induced inhibition of opioid-induced prolactin secretion is dependent on protein synthesis and independent of changes in drug metabolism, and of the type of opiate receptor preferentially affected by the opiate agonists employed.

Diurnal variation in prolactin, adrenocorticotropin and corticosterone release induced by opiate agonists in intact and adrenalectomized rats.

Diurnal variations of the effectivity of beta-endorphin (beta-End), dynorphin (DYN), Met-enkephalin (Met-Enk), D-Met2-Pro5-enkephalinamide (D-Met-Pro-Enk) and morphine to induce prolactin (PRL) and adrenocorticotropin (ACTH)/corticosterone (CS) release in intact and adrenalectomized rats have been examined. The response to morphine (10 mg/kg s.c.), Met-Enk (200 micrograms/rat i.c.v.) and D-Met-Pro-Enk (0.5 microgram/rat i.c.v.) did not change with different times of the day, while that to beta-End (0.5 microgram/rat i.c.v.), DYN (1 microgram/rat i.c.v.) and U50-488H, a selective kappa agonist (10 mg/kg s.c.), showed a circadian rhythm in stimulating PRL release, with a higher increase in the afternoon (16.00-17.00 h) than in the morning (08.00-09.00 h). In adrenalectomized rats the loss of this circadian rhythm was shown. The CS release evoked by morphine, D-Met-Pro-Enk, Met-Enk and DYN was demonstrable only in the morning when the basal CS level was significantly lower than in the afternoon. The afternoon release of ACTH by morphine was higher than in the morning in adrenalectomized rats. beta-End and U50-488H were equally active in the morning and in the afternoon in increasing CS secretion. The present results suggest that the diurnal rhythm in the response of CS and PRL release to opioids is in relation with the glucocorticoid secretion.