Inhibition of release of neurohypophysial hormones by endogenous opioid peptides in pregnant and parurient rats

Naloxone, an opiate receptor antagonist, was used to determine whether opioid peptides modulate release of oxytocin (OT) or vasopressin (AVP) in the rat after expulsion of the fetus, i.e. parturition. We measured the concentrations of AVP and OT in plasma and in the neurointermediate lobe of the pituitary of pregnant rats given naloxone (5 mg/kg, s.c.) or saline on day 20 of gestation, and on day 21 either before or during the expulsive stage of labor. Non-pregnant rats in diestrus were giben naloxone for comparison. On days 20 and 21 of gestation, before the onset of parturition, plasma [AVP] but not [OT] was elevated, compared to the non-pregnant controls. After delivery of the first two pups, plasma [OT] approximatelyy doubled, whereas plasma [AVP] remained unchanged. Blocking the action of endogenous opioid peptides with naloxone caused an elevation of plasma [OT] in pregnant animals on days 20 and 21 of gestation and during parturition. Naloxone, however, did not alter plasma [AVP] in either parturient or preparturient animals. In contrast, [AVP], but not [OT], was increased in plasma of non-pregnant rats given naloxone. The content of OT in the neuro-intermediate lobe was similar in pregnant and non-pregnant rats and was unaffected delivery of the first two pups. However, AVP content and the ratio of AVP/OT in the pituitary were lower in pregnant animals before during delivery than in the non-pregnant controls. The content of neither hormone was altered by naloxone. Thus, AVP release apparently increase and pituitary stores of this peptide are decreased by day 20 gestation, when labor has not yet begun. In contrast, OT secretion becomes elevated only during delivery. Inhibition of OT release by opioid peptides may: (1) allow preferential release of AVP during pregnancy; and (2) prevent depletion of pituitary stores of OT and neuronal fatigue during the 1–2 h period of parturition in the rat.     

Differential effects of naloxone on the release of neurohypophysial hormones in normotensive and spontaneously hypertensive rats

The hypothalamo-neurohypophysial system is altered in the spontaneously hypertensive rat (SHR). We hypothesized that an aberrant regulation of vasopressin (VP) and oxytocin (OT) release by endogenous opioid peptides alters this neuroendocrine system in the SHR. Concentrations of the neurohypophysial hormones in plasma and the pituitary were measured in 17-week-old SHRs and two strains of normotensive controls. Wistar Kyoto (WKY) and Sprague-Dawley rats. Animals were decapitated 20 min after s.c. injection of saline (1 ml/kg) or naloxone hydrochloride (1 or 10 mg/kg). In addition, neurohypophysial hormones excreted during the day (08.00-17.30 h) and night (17.30-08.00 h) were determined in urine from 16-week-old animals kept in metabolic cages for 5 days. VP at extrahypothalamic sites was also measured as [VP] in acid extracts of the subfornical organ area, hippocampal commissure-fornix and choroid plexus. Hormones were quantified by radioimmunoassay. The pituitary content, plasma concentration, and urinary excretion of OT were reduced (P less than 0.05) in SHRs, whereas VP content was increased (P less than 0.05) in the pituitary and plasma, but unchanged in urine, of hypertensive animals. In extrahypothalamic tissues, [VP] in the hippocampal commissure-fornix was increased in the SHR. Naloxone elevated (P less than 0.05) the plasma concentration of OT in WKY animals and VP in SHRs. Neither [VP] nor [OT] in plasma was changed by naloxone in Sprague-Dawley rats. Pituitary stores of the neurohypophysial hormones were not altered by naloxone in either hypertensive or normotensive rats. In conclusion, endogenous opioid peptides tonically inhibit OT release in WKY rats, whereas VP release is decreased by opioid peptides in SHRs, 16-17 weeks of age. The neuromodulatory role of opioid peptides in the release of neurohypophysial hormones appears to be altered in the SHR such that VP release is suppressed and OT release is augmented.     

Differential effects on naloxone on the release of neurohypophysial hormones in normotensive and spontaneously hypertensive rats

The hypothalamo-neurohypophysial system is altered in the spontaneously hypertensive rat (SHR). We hypothesized that an aberrant regulation of vasopressin (VP) and oxytocin (OT) release by endogenous opiod peptides alters this neuroendocrine system in the SHR. Concentrations of the neurohypophysial hormones in plasma and the pituitary were measured in 17-week-old SHRs and two strains of normotensive controls, Wistar Kyoto (WKY) and Sprague-Dawley rats. Animals were decapitated 20 min after s.c. injection of saline (1 ml/kg) or naloxone hydrochloride (1 or 10 mg/kg). In addition, neurohypophysial hormones excreted during the day (08.00–17.30 h) and night (17.30–08.00 h) were determined in urine from 16-week-old animals kept in metabolic cages for 5 days. VP at extrahypothalamic sites was also measured as [VP] in acid extracts of the subfornical organ area, hippocampal commissure-fornix and choriod plexus. Hormones were quantified by radioimmunoassay. The pituitary content, plasma concentration, and urinary excretion of OT were reduced (P < 0.05) in SHRs, whereas VP content was increased (P < 0.05) in the pituitary and plasma, but unchanged in urine, of hypertensive animals. In extrahypothalamic tissues, [VP] in the hippocampal commissure-fornix was increased in the SHR. Naloxone elevated (P < 0.05) the plasma concentration of OT in WKY animals and VP in SHRs. Neither [VP] nor [OT] in plasma was changed by naloxone in Sprague-Dawley rats. Pituitary stores of the neurohypophysial hormones were not altered by naloxone in either hypertensive or normotensive rats. In conclusion, endogenous opioid peptides tonically inhibit OT release in WKY rats, whereas VP releas is decreased by opioid peptides in SHRs, 16–17 weeks of age. The neuromodulatory role of opioid peptides in the release of neurohypophysial hormones appears to be altered in the SHR such that VP release is suppressed and OT release is augmented.     

Tetraethylammonium ions and 4-aminopyridine prevent opioid inhibition of neurohypophysial oxytocin release

Naloxone increased the electrically (15 Hz, 1 min) evoked release of oxytocin from isolated neural lobes of rats 3-4-fold. In the presence of 4-aminopyridine or tetraethylammonium ions the evoked release of oxytocin was increased 8-9-fold and remained unaffected by naloxone. Increasing the calcium concentration in the medium from 1.2 to 3 mM caused only a marginal increase of the evoked oxytocin release. In conclusion, blockade of potassium channels and/or the resulting prolongation of the action potential can surmount the opioid inhibition of oxytocin release.     

Serotonin stimulates hypothalamic mRNA expression and local release of neurohypophysial peptides

The neurotransmitter serotonin (5-HT) stimulates the secretion of vasopressin and oxytocin, and 5-HT is involved in the mediation of the vasopressin and oxytocin response to stress. In male Wistar rats, we investigated the 5-HT receptors involved in the 5-HT-induced increase of mRNA expression of vasopressin and oxytocin in the hypothalamic paraventricular nucleus (PVN) and supraoptic nucleus (SON). The 5-HT precursor, 5-hydroxytryptophan, injected in combination with the 5-HT reuptake inhibitor, fluoxetine, increased oxytocin mRNA expression in the PVN, and the concentration of vasopressin and oxytocin in plasma, whereas mRNA in the SON was not affected. Intracerebroventricular infusion of 5-HT agonists selective for the 5-HT1A, 5-HT1B, 5-HT2A and 5-HT2C receptor increased oxytocin mRNA in the SON and PVN. Infusion of agonists selective for the 5-HT2A + 2C receptor increased vasopressin mRNA in the PVN, whereas none of the 5-HT agonists affected vasopressin mRNA in the SON. All the 5-HT agonists infused increased peripheral oxytocin concentration and vasopressin was increased by stimulation of the 5-HT2A, 5-HT2C and 5-HT3 receptor. Intracerebroventricular infusion of 100 nmol 5-HT increased the extracellular hypothalamic concentration of vasopressin as measured by microdialysis in the PVN. To evaluate the involvement of hypothalamic-pituitary system in the 5-hydroxytryptophan and fluoxetine-induced vasopressin secretion, rats were immunoneutralized with a specific anti-corticotropin-releasing hormone antiserum. This treatment reduced plasma vasopressin and oxytocin responses. We conclude that stimulation with 5-hydroxytryptophan or 5-HT agonists increases mRNA expression of oxytocin in the PVN and the SON via stimulation of at least 5-HT1A, 5-HT1B, 5-HT2A and 5-HT2C receptors. Vasopressin mRNA in the PVN was increased only via the 5-HT2 receptor, whereas vasopressin mRNA in the SON does not seem to be affected by 5-HT stimulation. Corticotropin-releasing hormone appears to be partly involved in the mediation of 5-HT induced vasopressin and oxytocin secretion.     

Allopregnanolone and induction of endogenous opioid inhibition of oxytocin responses to immune stress in pregnant rats

In virgin rats, systemic administration of interleukin (IL)-1β (i.e. to mimic infection), increases oxytocin secretion and the firing rate of oxytocin neurones in the supraoptic nucleus (SON). However, in late pregnancy, stimulated oxytocin secretion is inhibited by an endogenous opioid mechanism, preserving the expanded neurohypophysial oxytocin stores for parturition and minimising the risk of preterm labour. Central levels of the neuroactive metabolite of progesterone, allopregnanolone, increase during pregnancy and allopregnanolone acting on GABA(A) receptors on oxytocin neurones enhances inhibitory transmission. In the present study, we tested whether allopregnanolone induces opioid inhibition of the oxytocin system in response to IL-1β in late pregnancy. Inhibition of 5α-reductase (an allopregnanolone-synthesising enzyme) with finasteride potentiated IL-1β-evoked oxytocin secretion in late pregnant rats, whereas allopregnanolone reduced the oxytocin response in virgin rats. IL-1β increased the number of magnocellular neurones in the SON and paraventricular nucleus (PVN) expressing Fos (an indicator of neuronal activation) in virgin but not pregnant rats. In immunoreactive oxytocin neurones in the SON and PVN, finasteride increased IL-1β-induced Fos expression in pregnant rats. Conversely, allopregnanolone reduced the number of magnocellular oxytocin neurones activated by IL-1β in virgin rats. Treatment with naloxone (an opioid antagonist) greatly enhanced the oxytocin response to IL-1β in pregnancy, and finasteride did not enhance this effect, indicating that allopregnanolone and the endogenous opioid mechanisms do not act independently. Indeed, allopregnanolone induced opioid inhibition over oxytocin responses to IL-1β in virgin rats. Thus, in late pregnancy, allopregnanolone induces opioid inhibition over magnocellular oxytocin neurones and hence on oxytocin secretion in response to immune challenge. This mechanism will minimise the risk of preterm labour and prevent the depletion of neurohypophysial oxytocin stores, which are required for parturition.     

Inhibition of striatal acetylcholine release by endogenous serotonin

We have examined the hypothesis that endogenous serotonin (5-HT) exerts an inhibitory influence on the release of acetylcholine (ACh) in striatum. Striatal slices were prepared from adult rats, preincubated with [3H]choline, superfused, and exposed to electrical field stimulation. The stimulation-induced overflow of tritium into the superfusate was used as a measure of ACh release. We observed that fluoxetine, an inhibitor of 5-HT uptake, reduced ACh overflow in slices prepared from caudal striatum, an area of high 5-HT concentration, but not in slices from rostral striatum, an area of low 5-HT concentration. Moreover, methysergide, a 5-HT antagonist, increased ACh efflux in caudal but not rostral striatum. Finally, direct activation of 5-HT receptors with the 5-HT agonist, quipazine, inhibited stimulation-induced ACh overflow in both rostral and caudal striatum. These results suggest that endogenous 5-HT normally is capable of inhibiting striatal ACh release, and that the extent of the modulation is related to the degree of serotonergic innervation. In addition, 5-HT receptors capable of modulating ACh release are present in 5-HT-poor rostral striatum, as well as in 5-HT-rich caudal striatum.     

Diurnal variations of opioid peptides and synenkephalin in vitro release in the amygdala of kindled rats

Pentylenetetrazol (PTZ) kindling was induced in male Wistar rats (250-300 g) by daily intraperitoneal injections of 35 mg/kg of the convulsant agent. Immunoreactive (IR)-Met-enkephalin (IR-ME), IR-Leu-enkephalin (IR-LE), IR-heptapeptide (IR-HE), IR-octapeptide (IR-OC) and IR-synenkephalin (IR-Syn) in vitro release was measured from amygdala slices 24 h after the last stimulus, in groups of eight rats, every 4 h beginning at 08:00 h. Opioid peptides in vitro release displayed diurnal variations. IR-ME and IR-Syn showed maximal levels before the onset of darkness (16:00 h). IR-LE and IR-OC release was enhanced 4 h later (20:00 h), no changes were detected for IR-HE. These results show that endogenous opioid system (EOS) release displays diurnal variations. The peak for the analysed peptides was reached before and during the dark phase. It is suggested that EOS release enhancement in PTZ-kindled rats, seems to be due to a compensatory mechanism against the excitation induced by the blockade of the GABAergic transmission.     

In vitro production and release of opioid peptides in the tooth pulp induced by bradykinin

A possible relationship between met-enkephalin (ME)-like peptides and bradykinin (BK) in the rat incisor pulp was examined in in vitro experiments using whole pulp. ME-like peptide content in the pulp was increased by BK at a concentration of 1 microM, but not in higher concentrations, while the release of ME-like peptides from the pulp into the incubation medium was increased dose-dependently by BK. These effects of BK were inhibited by Des-Arg9-[Leu8]-BK, a potent BK-antagonist, suggesting that the effects of BK were mediated through a specific BK-receptor in the pulp. On the other hand, high K+ did not induce any increased release of ME-like peptides from the pulp and the BK effects were influenced neither in Ca++-free medium nor in the presence of ouabain. These results suggested that ME-like peptide releasing effect of BK was not due to depolarization of the cell membrane and was not active. In addition, kyotorphin, and enkephalin-releaser, could not only elicit a release of ME-like peptides from the pulp, but also a marked increase of the peptide content in the pulp. However, a combination of BK and kyotorphin attenuated the effect of BK or kyotorphin each other. These results suggested that there might be two kinds of mechanisms of ME-like peptide production in the pulp and those mechanisms might interfere mutually.     

Neuroendocrine effects of endogenous opioid peptides in human subjects: A review

In the decade since the discovery of specific opioid receptors in the brain, there have been rapid advances in our understanding of the physiological and pathological roles of the endogenous opioid systems in humans. Endogenous opioid peptides have been demonstrated to play a role as modulators of a number of hormonal functions in humans. In particular they appear to inhibit luteinizing hormone and ACTH release, and the response of arginine vasopressin to osmotic stimuli. They appear to participate in the modulation of carbohydrate homeostasis. In pathophysiological states, they appear to play a role in the decreased pulsatile luteinizing hormone release seen in patients with prolactinomas. Circulating β-endorphin appears to be an important regulator of immune function. Preliminary studies in humans have suggested a role for endogenous opioid peptides in appetite regulation. In the last few years, a few case reports have suggested the possibility of a series of syndromes due to endogenous opioid excess. Within the next decade, we can expect to see the routine use of opioid antagonists in a variety of pathophysiological states.     

Role of endogenous opioid peptides in the acute adaptation to hypoxia.

A non-lethal, hypoxic conditioning stimulus has been shown by Rising and D'Alecy to increase hypoxic survival time in mice. To determine if endogenous opioids alter the hypoxic conditioning-induced increase in hypoxic survival time, we administered naloxone (0.1, 1.0 mg/kg i.p.) or saline (0.3 ml i.p.) 5 min prior to conditioning. Sixty percent of the mice received the hypoxic conditioning stimulus consisting of three sequential hypoxic exposures (4.5% oxygen balance nitrogen for 1.5, 2 and 2.5 min) separated by 5 min of room air. The remaining mice did not receive hypoxic conditioning but instead remained in room air for this time. All mice were tested for hypoxic survival by first exposing them to 20 s of 8.5% oxygen balance nitrogen followed by exposure to 4.5% oxygen balance nitrogen. The hypoxic survival time was recorded as the time from the onset of the 4.5% oxygen to the cessation of spontaneous ventilation. Naloxone (1 mg/kg) completely blocked the adaptation to hypoxia induced by hypoxic conditioning (P = 0.003). Morphine (1, 5, 10 and 20 mg/kg) had no effect on hypoxic adaptation; however, 50 mg/kg morphine decreased the adaptation induced by conditioning (P less than 0.0001) possibly due to high dose toxicity. These data suggest that endogenous opioids are involved in the protective adaptation to hypoxia induced by prior exposure to non-lethal hypoxia.     

Leukocyte opioid receptors mediate analgesia via Ca2+-regulated release of opioid peptides

Opioids are the most powerful analgesics. As pain is driven by sensory transmission and opioid receptors couple to inhibitory G proteins, according to the classical concept, opioids alleviate pain by activating receptors on neurons and blocking the release of excitatory mediators (e.g., substance P). Here we show that analgesia can be mediated by opioid receptors in immune cells. We propose that activation of leukocyte opioid receptors leads to the secretion of opioid peptides Met-enkephalin, β-endorphin and dynorphin A (1–17), which subsequently act at local neuronal receptors, to relieve pain. In a mouse model of neuropathic pain induced by a chronic constriction injury of the sciatic nerve, exogenous agonists of δ-, μ- and κ-opioid receptors injected at the damaged nerve infiltrated by opioid peptide- and receptor-expressing leukocytes, produced analgesia, as assessed with von Frey filaments. The analgesia was attenuated by pharmacological or genetic inactivation of opioid peptides, and by leukocyte depletion. This decrease in analgesia was restored by the transfer of wild-type, but not opioid receptor-lacking leukocytes. Ex vivo, exogenous opioids triggered secretion of opioid peptides from wild-type immune cells isolated from damaged nerves, which was diminished by blockade of Gαi/o or Gβγ (but not Gαs) proteins, by chelator of intracellular (but not extracellular) Ca²⁺, by blockers of phospholipase C (PLC) and inositol 1,4,5-trisphosphate (IP₃) receptors, and was partially attenuated by protein kinase C inhibitor. Similarly, the leukocyte depletion-induced decrease in exogenous opioid analgesia was re-established by transfer of immune cells ex vivo pretreated with extracellular Ca²⁺ chelator, but was unaltered by leukocytes pretreated with intracellular Ca²⁺ chelator or blockers of Gαi/o and Gβγ proteins. Thus, both ex vivo opioid peptide release and in vivo analgesia were mediated by leukocyte opioid receptors coupled to the Gαi/o–Gβγ protein–PLC–IP₃ receptors–intracellular Ca²⁺ pathway. Our findings suggest that opioid receptors in immune cells are important targets for the control of pathological pain.     

Inhibition of shock-elicited ultrasonic vocalization by opioid peptides in the rat: a psychotropic effect

Following three series of electric footshocks (10 shocks/day), one out of three rats in most cages were brought to emit ultrasonic vocalization for several minutes after a single shock. The characteristics of shock-elicited ultrasound were pure tone pulses of a frequency between 22 and 28 kHz, with duration longer than 300 msec. The same type of ultrasound is produced by subordinate male rats during agonistic behavior. The intracerebroventricular injection of beta-endorphin, dynorphin, methionine-enkephalin or leucine-enkephalin attenuated the shock-elicited ultrasonic vocalization. Psychotropic drugs such as diazepam and chlorpromazine also attenuated the shock-elicited ultrasonic vocalization. A test utilizing ultrasonic vocalization in rodents can provide useful data for studying the psychotropic properties of neuropeptides.     

Effect of endogenous opioid peptides on acetylcholine release from the cat superior cervical ganglion: selective effect of a heptapeptide

The present experiments show the presence of both metenkephalin-like and met-enkephalin-Arg6-Phe7-like immunoreactivity in the superior cervical ganglion of the cat; this was determined by radioimmunoassay after high-pressure liquid chromatography separation of tissue extracts. There was measurable efflux of both peptides, as determined by radioimmunoassay of ganglionic perfusates; this measure was increased by thiorphan, an enkephalinase inhibitor. The effect of the 2 peptides on ACh release was determined: The stable analog of methionine-enkephalin, D-Ala2-methionine-enkephalinamide, did not affect ACh release from the ganglion; in contrast, methionine-enkephalin-Arg6-Phe7 significantly depressed evoked ACh release. The effect of met-enkephalin-Arg6-Phe7 to decrease ACh release was antagonized, although only partially, by the opioid antagonist naloxone. Thus, it appears that methionine-enkephalin-Arg6-Phe7 alters ACh release from the superior cervical ganglion by acting, at least in part, on a presynaptic opioid receptor. The results suggest that in the cat superior cervical ganglion, the heptapeptide enkephalin might have a significant role in the regulation of synaptic transmission, which is unrelated to its potential function as a precursor for methionine-enkephalin.     

Involvement of endogenous opioid systems in nociceptin-induced spinal antinociception in rats

The present study investigates the involvement of opioid receptors in the antinociceptive effects of nociceptin in the spinal cord of the rat. Intrathecal administrations of 5 and 10 nmol of nociceptin significantly increase the withdraw response latencies to noxious thermal and mechanical stimulations. This nociceptin-induced antinociceptive effect is significantly attenuated by intrathecal injection of (Nphe(1))nociceptin(1-13)-NH(2), a selective antagonist of the nociceptin receptor (opioid receptor-like receptor ORL1), indicating an ORL1 receptor-mediated mechanism. This antinociceptive effect is also significantly attenuated by intrathecal injections of naloxone (a nonselective opioid receptor antagonist), naltrindole (a selective delta-opioid receptor antagonist), and beta-funaltrexamine (a selective mu-opioid receptor antagonist) in a dose-dependent manner, but not by the selective kappa-opioid receptor antagonist norbinaltorphimine. Since it is unlikely that nociceptin acts by direct binding to opioid receptors, these results suggest a possible interaction between the nociceptin/ORL1 and opioid systems in the dorsal horn of the rat spinal cord.     

The role of opiates and endogenous opioid peptides in the regulation of rat TSH secretion

The administration of morphine to rats at room temperature is reported to suppress serum thyrotropin (TSH) levels by a hypothalamic mechanism. However, it is unknown whether endogenous opioid peptides (EOP) are involved in the control of TSH secretion. The present studies show that naloxone (10 mg/kg, i.p.), an opiate-receptor antagonist, prevented the decline in rat serum TSH which occurs with heat exposure. Morphine sulfate (20 mg/kg, i.p.) treatment prevented the cold-induced elevation in serum TSH, and pretreatment with haloperidol (0.3 mg/kg, i.p.) eliminated morphine's influence. Medial-basal hypothalamic thyrotropin-releasing hormone (TRH) content, measured by RIA, increased in the morphine-treated rats which were exposed to 4 °C. A submaximal intravenous dose of TRH (300 ng/100 g) was given to determine whether morphine suppresses serum TSH through the release of hypothalamic thyrotropic inhibitors. Morphine pretreatment did not alter TSH stimulation by TRH. Morphine alone or combined with TRH did not alter basal or stimulated TSH secretion in vitro.These studies strongly suggest that, in rats, the EOP modulate TSH secretion under conditions such as acute heat exposure which are associated with a decline in serum TSH. Under specific circumstances, the suppression of serum TSH morphine may be dopamine-dependent.     

Role of opioid peptides in pulsatile release of gonadotropins and prolactin in the rat

To determine the role of endogenous opioid peptides in the pulsatile release of gonadotropins and prolactin in the ovariectomized rat, the opiate receptor blocker, naloxone, was administered intravenously, and its effect on plasma FSH, LH and prolactin was determined by multiple sampling prior to and after injection. Naloxone produced a dose-related increase in plasma LH and to a lesser extent FSH and decreased prolactin levels in the experiment in which they were examined. Higher doses of naloxone produced a significant increase in plasma LH pulse amplitude and lengthened the interpulse interval with a consequent decrease in pulse frequency. Minimum values between pulses were also increased. There was no clear effect on FSH pulsations but pulses of prolactin were blocked. Intraventricular (third ventricle) injection of a specific anti beta endorphin antiserum (3 microliter) produced an initial decline followed by an elevation of LH but had no effect on plasma FSH. The normal rabbit serum control injections were without effect. It is hypothesized that initiation of LH pulses in the castrated rat may be related to a periodic removal of tonic beta endorphinergic tone.     

Sympathetic activation triggers endogenous opioid release and analgesia within peripheral inflamed tissue

Stress induces analgesia by mechanisms within and outside the brain. Here we show that the sympathetic nervous system is an essential trigger of intrinsic opioid analgesia within peripheral injured tissue. Noradrenaline, injected directly into inflamed hind paws of male Wistar rats, produced dose-dependent antinociception, reversible by alpha(1)-, alpha(2)- and beta(2)-antagonists. alpha(1)-, alpha(2)- and beta(2)-adrenergic receptors were demonstrated on beta-endorphin-containing immune cells and noradrenaline induced adrenergic receptor-specific release of beta-endorphin from immune cell suspensions. This antinociceptive effect of noradrenaline was reversed by micro - and delta-opioid antagonists as well as by anti-beta-endorphin. Stress-induced peripheral analgesia was abolished by chemical sympathectomy and by adrenergic antagonists. These findings indicate that sympathetic neuron-derived noradrenaline stimulates adrenergic receptors on inflammatory cells to release beta-endorphin, which induces analgesia via activation of peripheral opioid receptors.     

Inhibition of trigemino-hypoglossal reflex in rats by oxytocin is mediated by mu and kappa opioid receptors

Recent studies showed that oxytocin plays an important role in the modulation of pain at different levels of the central nervous system. The present study was undertaken to investigate the effect of oxytocin on trigemino-hypoglossal reflex in rats. With the experimental settings used in this study, we have demonstrated that oxytocin showed significant analgesic effect after intracerebroventricular administration in rats, as assayed by the amplitude of the retractory movements of the tongue after tooth pulp stimulation. Antinociceptive effect of oxytocin was inhibited by subsequent perfusion of cerebral ventricles with oxytocin antagonist, [deamino-Cys1-D-Tyr(OEt)2-Thr4-Orn8]-oxytocin, atosiban. An involvement of opioid system in the oxytocin-induced analgesia was studied after intracerebroventricular administration of different opioid antagonists: non-selective naloxone, mu-selective beta-funaltrexamine, delta-selective naltrindole, and kappa-selective nor-binaltorphimine. It was shown that inhibition of antinociceptive effects was mediated through mu and kappa opioid receptors, indicating that there is a synergy between oxytocin and opioid systems in transmitting and modulating pain stimuli. Co-administration of oxytocin and a mu-selective endogenous opioid ligand endomorphin-2 did not significantly increase the antinociceptive activity of endomorphin-2.     

HPLC separation of vasopressin-like hormones in chicken neurohypophysial extracts

HPLC techniques were used to identify peptides that possess vasopressin-like immunoreactivity in the chicken neurohypophysis. The presence of arginine vasotocin (AVT) was confirmed together with arginine vasopressin (AVP). That the presence of AVP may be confined to the chicken, and not other avian species, was concluded from the absence of the hormone in the neurohypophysis of the duck and turkey. The chicken thus resembles some members of the suiformes and metatheria in possessing two vasopressin-like peptides.