Marlena Juszczak

Melatonin, pinealectomy, and release of neurohypophysial hormones : in vitro studies

Abstract: The effect of melatonin as well as pinealectomy on the basal and K+‐evoked release of vasopressin and oxytocin from the neurointermediate lobes in vitro was determined. Pineal removal resulted in a diminution of vasopressin and oxytocin release from the neurointermediate lobes in vitro. Melatonin (10−3 or 10−6 M/1) increased vasopressin and oxytocin release from neurointermediate lobes of sham‐operated rats. Nevertheless, when pinealectomized rats served as donors of the neurointermediate lobes, melatonin (10−3 or 10−6 M/1) increased vasopressin release under basal conditions. For the same tissue, melatonin did not affect the oxytocin release either under basal conditions or during depolarization due to excess potassium. When 10−7 M/l melatonin was used, no changes in either vasopressin or oxytocin release were observed in vitro.

The effect of melatonin on suckling-induced oxytocin and prolactin release in the rat.

There is growing evidence that melatonin (MEL) inhibits oxytocin (OT) release when used in a low dose, while higher doses stimulate the release of the hormone in the rat. In the present study we investigated the effect of exogenous MEL, administered intracerebroventricularly (ICV), on suckling-induced OT and prolactin (PRL) release in the urethane-anesthetized rat. Lactating rats suckled by 8-12 pups were studied on days 8-12 of postpartum, and lactating pups-deprived rats on the same days of postpartum served as a control. Plasma OT and PRL levels as well as hypothalamic and neurohypophyseal OT contents were measured by RIA. Suckling stimulated the secretion of both OT and PRL. The ICV injection of 1 ng/ml MEL produced a significant inhibition of suckling-induced OT as well as PRL secretion. Melatonin in doses of 100 ng/ml or 10 micrograms/ml did not modify the OT release but significantly inhibited PRL release brought about by suckling; 10 pg/ml of MEL was not effective in this regard. Thus, exogenous MEL seems to inhibit suckling-induced OT as well as PRL secretion when applied at doses regarded to be in the range of the physiological level; when applied in higher doses, it was shown not to influence the release of OT following physiological stimulation such as suckling.

The Influence of Melatonin on the Content of Vasopressin and Oxytocin in the Hypothalamus and Neurohypophysis in Euhydrated and Dehydrated Male Rats

Melatonin injected in a single intraperitoneal dose of 100 μg/100 g b. w. to euhydrated rats resulted in a decrease of neurohypophysial oxytocin content but the hypothalamic oxytocin storage as well as the hypothalamo‐neurohypophysial storage of vasopressin were not changed. Following 8 d of once‐daily melatonin treatment the hypothalamic and neurohypophysial oxytocin and vasopressin content was decreased. It might be therefore suggested that melatonin increases the release of neurohypophysial hormones and/or decreases their synthesis.

Melatonin inhibits the substance P-induced secretion of vasopressin and oxytocin from the rat hypothalamo-neurohypophysial system: In vitro studies

The aim of the present investigations was to study the influence of substance P (a member of a family of peptides known as tachykinins) on basal and K(+)-evoked vasopressin (AVP) and oxytocin (OT) release from rat hypothalamo-neurohypophysial system in vitro as well as to determine whether this effect of substance P is sensitive to melatonin. The present results show that substance P stimulates basal AVP and OT release from isolated hypothalamo-neurohypophysial system, when used at the concentrations of 10(-6) and 10(-7)M/l. At the concentration of 10(-9)M/l, however, substance P was found to stimulate the in vitro secretion of AVP, but not that of OT. Melatonin diminished basal release of AVP; it also significantly inhibited the substance P-stimulated secretion of AVP and OT. K(+)-evoked release of the neurohypophysial hormones was not further modified by either substance P or melatonin. The present results show that the stimulatory effect of substance P on basal release of AVP and OT from rat hypothalamo-neurohypophysial system in vitro is sensitive to inhibitory influence of melatonin.

Neurohypophyseal vasopressin in the Syrian hamster: response to short photoperiod, pinealectomy, melatonin treatment, or osmotic stimulation.

In the present study, the effect of photoperiod on vasopressin content in the pituitary neurointermediate lobe (NIL), as well as the ability of pinealectomy to prevent and melatonin to mimic the short photoperiod-induced changes in NIL vasopressin were studied in male Syrian hamsters. The ability of melatonin to modify the hyperosmotically stimulated vasopressin release was also determined. Exposure to short photoperiod (SD) for 4 or 10 weeks increased vasopressin content in the hamster NIL. In long photoperiod (LD)-exposed hamsters, pinealectomy induced a decrease in NIL vasopressin content, whereas no effect of melatonin injections on vasopressin storage in the NIL was detected. In SD-exposed animals, pineal removal failed to alter vasopressin content in the NIL. Hypertonic saline administration led to the expected decrease in vasopressin content in the NIL both in vehicle- and melatonin-treated animals. The hyperosmotically stimulated release of vasopressin was not modified by previous treatment with melatonin. The data from the present study show that, in male Syrian hamsters, exposure of animals to SD increases the vasopressin content in the posterior pituitary, but these changes appear not to be mediated by SD-induced changes in melatonin secretion. Furthermore, the exposure of animals to SD prevents the pinealectomy-induced changes in NIL vasopressin content. Melatonin does not modify the hyperosmotically stimulated vasopressin release in the male Syrian hamster.

Hypothalamic and neurohypophysial vasopressin and oxytocin in melatonin-treated pinealectomized male rats.

The effect of melatonin on hypothalamic and neurohypophysial vasopressin and oxytocin was investigated in normal and pinealectomized rats. Pinealectomy was followed by a decrease of both vasopressin and oxytocin content in the hypothalamus and neurohypophysis. In unpinealectomized rats, melatonin decreased vasopressin and oxytocin storage in the hypothalamo‐neurohypophysial system. Following pineal removal, melatonin did not augment the pinealectomy‐induced decrease of vasopressin and oxytocin in the neurohypophysis; the hypothalamic storage of both neurohormones was even higher when compared with vehicle‐treated animals.

Septal vasopressin induced preservation of social recognition in rats was abolished by pinealectomy.

The role of intraseptal vasopressin (AVP) and the pineal gland in the modulation of social memory was investigated. For social recognition, male pinealectomised (Px) and sham-operated (SO) rats were confronted with juveniles for 4 min, and injected with either 200 pg AVP or 5 ng of its V1 receptor antagonist d(CH2)5Tyr(Me)AVP (AAVP) into the mediolateral septum. Re-exposure to the same and a different juvenile took place after 30 or 120 min. In SO rats, the social memory was facilitated after injection of AVP (120 min) and impaired after AAVP application (30 min). In Px rats, however, neither AVP nor AAVP administration influenced the social memory. Comparison between SO and Px control groups treated with artificial cerebrospinal fluid did not reveal any differences as to social recognition responses. After subcutaneous administration of 250 microg melatonin (substitution for pinealectomy) the social discrimination responses in Px rats after peptide application were influenced in a manner identical to that noted in SO animals. These findings suggest that the modulation of social memory by intraseptal AVP is dependent on an intact pineal function.

Melatonin and the Synthesis of Vasopressin in Pinealectomized Male Rats

The pineal hormone, melatonin, is known to modify, under different experimental conditions, neurohypophysial hormone secretion in the rat. The aim of this study was to investigate the effect of melatonin on the vasopressin biosynthesis rate in the hypothalamus of either pinealectomized or sham-operated rats, using the colchicine method. To estimate whether colchicine affects the function of the neurohypophysis in these animals, the neurohypophysial and plasma vasopressin levels were also measured. The vasopressin synthesis rate was increased after pineal removal, when compared with sham-operated animals, and melatonin strongly inhibited the rise in the hormone synthesis due to pinealectomy. After pineal removal plasma vasopressin concentration was significantly elevated, and melatonin attenuated this effect. On the contrary, the neurohypophysial vasopressin content was significantly decreased after pinealectomy, but it was not further modified by melatonin.Thus, melatonin suppresses the synthesis and secretion of vasopressin in pinealectomized rats. The present results confirm our previous reports as to the inhibitory impact of the pineal on both vasopressin synthesis and release and suggest that melatonin may mediate the effect of the pineal gland on vasopressinergic neuron activity.

Luteinizing hormone-releasing hormone and oxytocin response to hyperosmotic stimulation: in vitro study.

It was shown previously that luteinizing hormone-releasing hormone (LHRH) affects the neurohypophysial oxytocin release in water-deprived rats. However, the detailed mechanisms by which LHRH modifies the oxytocin response to hyperosmotic stimulation have not been explained so far. Using the isolated hypothalamo-neurohypophysial explants obtained from euhydrated rats, the effect of LHRH on the oxytocin secretion was studied under conditions of direct osmotic (i.e., Na(+)- evoked) as well as nonosmotic (i.e., K(+)-evoked) stimulation. Additionally, the oxytocin response to LHRH was investigated using the explants obtained from animals drinking 2% saline for eight days (systemic, i. e., both direct and indirect, osmotic stimulation). LHRH significantly enhanced Na(+)- and K(+)-evoked oxytocin release from explants taken from rats drinking tap water, indicating that LHRH could affect the Na(+)/K(+)-dependent depolarization of perikarya of oxytocin neurones. In contrast, LHRH significantly diminished the K(+)-stimulated hormone release when the neurohypophysial complex was obtained from previously salt-loaded rats, suggesting that peripheral osmotic stimulation somehow modifies the sensitivity of oxytocinergic neurones to LHRH (possible mechanisms are discussed). It is concluded that LHRH may participate in the regulation of oxytocin secretion via both direct and indirect impact on magnocellular oxytocinergic neurones depending on the current functional status of the hypothalamo-neurohypophysial complex.

Hypothalamic gonadotropin-releasing hormone receptor activation stimulates oxytocin release from the rat hypothalamo-neurohypophysial system while melatonin inhibits this process

The present study was undertaken to investigate the influence of gonadotropin-releasing hormone (GnRH) and its agonist and antagonist on oxytocin (OT) release from the rat hypothalamo-neurohypophysial (H-N) system. An additional aim was to determine whether the possible response of oxytocinergic neurons to these peptides could be modified by melatonin through a cAMP-dependent mechanism. The results show that the highly selective GnRH agonist (i.e., [Des-Gly(10),d-His(Bzl)(6),Pro-NHEt(9)]-LHRH; Histrelin) stimulates the secretion of OT from an isolated rat H-N system. Melatonin significantly inhibited basal and histrelin-induced release of OT in vitro, and displayed no significant influence on OT release in the presence of GnRH or its antagonist. Addition of melatonin to a medium containing forskolin resulted in significant reduction of OT secretion from the H-N system. On the other hand, addition of forskolin to a medium containing both histrelin and melatonin did not further alter the inhibitory influence of melatonin on the histrelin-dependent secretion of OT in vitro. Intracerebroventricular (icv) infusion (experiment in vivo) of a GnRH antagonist resulted in substantial inhibition of OT release, thus revealing the stimulatory action of endogenous GnRH. In melatonin-treated animals, blood plasma OT levels were not changed in comparison to the vehicle. Our present data strongly suggests that activation of the GnRH receptor in the hypothalamus is involved in stimulation of OT secretion from the rat H-N system. It has also been shown, under experimental in vitro conditions, that melatonin fully suppresses the response of oxytocinergic neurons to the GnRH agonist - histrelin. The effect of melatonin on OT release is mediated by the cAMP-dependent mechanism, although other mechanisms of action are also possible.