Enkephalinergic involvement in substantia nigra in the modulation of hypothalamically-induced predatory attack behavior.

The present study was carried out in five cats which did not attack the rats spontaneously. Predatory attack on an anaesthetized rat was elicited by electrical stimulation of lateral hypothalamus at a mean current strength of 650 microA. The attack was accompanied by minimal affective display and culminated in neck biting. Microinfusions of DAME (delta-alanine methionine enkephaline) in 500 ng dose in substantia nigra facilitated the predatory attack and there was a significant reduction in the threshold current strength for affective display as well as somatomotor components. Microinfusions of naloxone, an opioid antagonist in 1.0 microg dose when DAME effect was at its peak reversed the facilitatory effects and the threshold returned to the control levels within 10 minutes of naloxone infusion at the same locus. Microinfusions of naloxone alone in similar dosage completely blocked the predatory attack response as indicated by an increase in the threshold current strength for somatomotor as well as affective display components. The somatomotor were completely inhibited and could not be elicited even when the current strength was increased to 1000 microA. Control injections of saline in similar volumes (0.5 microl) failed to produce any response Microinfusions of naloxone in lower dose (250 ng) failed to produce any blocking effect. These findings indicate that hypothalamically elicited predatory attack is facilitated by enkephalinergic mechanisms operating at the midbrain level.

Differences in opioidergic inhibition of spinal reflexes and Fos expression evoked by mechanical and chemical noxious stimuli in the decerebrated rabbit.

Noxious mechanical and chemical stimuli were applied to the toes of the left hind limb of decerebrated, spinalized rabbits and their effects on a hind limb spinal withdrawal reflex and expression of Fos-like immunoreactivity in the spinal cord were measured. The animals were prepared so as to minimize nociceptive inputs arising from surgery. A single crush stimulus applied with a pair of haemostatic forceps caused long-lasting (c. 20 min) inhibition of reflexes evoked in medial gastrocnemius motoneurons by electrical stimulation of the skin at the heel. Naloxone (0.25 mg/kg i.v.) increased reflexes to more than 1000% of pre-drug controls and reversed crush-evoked inhibition. Mustard oil applied to the toes had no consistent effects on the heel-gastrocnemius reflex before or after naloxone. Both crush and mustard oil stimuli gave rise to unilateral increases in the number of Fos-immunopositive profiles in the superficial dorsal horn of spinal segments L7 and S1. There were significantly more Fos-immunoreactive elements in the central and lateral parts of lamina I of both segments in animals receiving the crush stimulus than there were in animals receiving the mustard oil stimulus. Immunochemical localization of enkephalins in rabbit spinal cord showed a dense network of fibres and terminals in laminae I and II, accompanied by infrequent but distinctly stained neuronal cell bodies. The same pattern, with increased numbers of visible cell bodies, was seen after treatment with colchicine. The present data show that tonic and stimulus-evoked opioidergic inhibition of the heel-gastrocnemius reflex of the rabbit are not epiphenomena of surgical preparation of the hindlimb. Opioid-mediated inhibition of the heel-gastrocnemius withdrawal reflex of the rabbit was evoked by noxious mechanical but not by chemical stimulation of the toes. Of these stimuli, the former gave rise to greater activation of neurons in central and lateral lamina I of segments L7 and S1, the region of termination of afferent fibres from the heel and the location of some enkephalin-positive neuronal cell bodies. Thus, noxious mechanical stimulation of the toes elicits inhibition of the heel-gastrocnemius withdrawal reflex, probably via activation of enkephalinergic neurons in the lateral half of lamina I in the L7 and S1 segments.

Antinociceptive response induced by mixed inhibitors of enkephalin catabolism in peripheral inflammation.

RB101 (N-((R,S)-2-benzyl-3[(S)(2-amino-4-methylthio)butyl dithio]-1-ox-opropyl)-L-phenylalanine benzyl ester) is a recently developed full inhibitor of the enkephalin-catabolizing enzymes able to cross the blood-brain barrier, whereas RB38A ((R)-3-(N-hydroxycarboxamido-2-benzylpropanoyl)-L-phenylalanine) is as potent as RB101 but almost unable to enter the brain. In this study, we have investigated the effects of systemic administration of morphine, RB101 and RB38A on nociception induced by pressure on inflamed peripheral tissues. Antinociceptive test was performed between 4 and 5 days after injection into the rat left hindpaw of Freund's complete adjuvant to produce localized inflammation. Morphine (1, 2 and 4 mg/kg, i.v.) induced antinociception in inflamed paws at all the doses used, and only at the highest dose in non-inflamed paws. RB101 (10 and 20 mg/kg, i.v.) induced an antinociceptive response only in the inflamed paws. RB38A, also induced an antinociceptive effect in the inflamed paws, but only at the highest dose (20 mg/kg, i.v.). The responses induced by morphine and the inhibitors of enkephalin catabolism were antagonized by the systemic administration of naloxone (1 mg/kg) or methylnaloxonium (2 mg/kg) which acts essentially outside the brain. Central injection (i.c.v.) of methylnaloxonium (2 micrograms) blocked the effect of morphine only in non-inflamed paws, and slightly decreased the response induced by RB101 on inflamed paws. These results indicate that the endogenous opioid peptides, probably enkephalins, are important in the peripheral control of nociception from inflamed tissues.

[Effect of synthetic analogs of enkephalins, morphine and their antagonists on the course of experimental traumatic shock]. / Vliianie sinteticheskikh analogov énkefalinov, morfina i ikh antagonistov na techenie éksperimental'nogo travmaticheskogo shoka.

Effects of naloxone, nalorphine, thyroliberin, morphine and two analogues of enkephalins (FK 33-824 and Tyr-D-Ala-Gly-Phe(NO2)-NH2) on the course of traumatic shock were studied in experiments on rabbits. It was found that antagonists of opioid peptides aggravated the course of traumatic shock and morphine and synthetic analogues of enkephalins exerted positive effects during its treatment. Endogenous opioid peptides are suggested to play the protective role in experimental traumatic shock.

Morphine and opioid peptides reduce paraventricular neuronal activity: studies on the rat hypothalamic slice preparation.

Extracellular discharges of neurons in the paraventricular nucleus (PVN) were recorded from slices of rat hypothalamus in vitro. PVN neurons (n = 14) were identified by the criteria of (i) phasic activity patterns and (ii) antidromic invasion from the neurohypophysial tract. Neurons not displaying either of these features were considered unidentified with respect to physiological function (n = 85). The majority of unidentified neurons responded to bath application of morphine (1 microM), [D-Ala2,Met5]enkephalin (1 microM) or beta-endorphin (0.01-1 microM) with a prompt, reversible, dose-related reduction in spike discharge frequency. Naloxone (1 microM) antagonized the opioid-induced depressions in some, but not all, cases. At the concentrations tested, no tachyphylaxis to the effects of the opioids was observed. The opioid effects on putative neurohypophysial neurons were less pronounced; while 2 were depressed, the remaining 12 displayed no change in frequency or pattern of discharge to micromolar concentrations of morphine, [D-Ala2,Met5]enkephalin, or beta-endorphin. Our results indicate that opioids depress neuronal activity in the rat PVN via an interaction with a specific opiate receptor but that this effect is more pronounced on unidentified neurons than on putative neurohypophysial neurons in the slice.

In vitro effects of a synethetic enkephalin analogue (D-Met2,Pro5)-enkephalinamide on the hypothalamus-pituitary--testis function in rats.

The effects of (D-Met2,Pro5)-enkephalinamide, a synthetic analogue of the natural opioid pentapeptides, were investigated on the hypothalamus-pituitary-testis axis using in vitro tissue and isolated cell incubation systems. (D-Met2,Pro5)-enkephalinamide in a concentration range 10(-7)--10(-5) M did not alter the testosterone production and the troph-hormone sensitivity of the isolated Leydig cells, or the gonadotroph hormone release and the luteinizing-hormone--releasing hormone (LH-RH) sensitivity of the isolated anterior pituitary cells. However 10(-5)--10(-5) M synthetic enkephalin analogue induced a dose-dependent inhibition of LH-RH content and release of the mediobasal hypothalamus, which was prevented by equimolar concentrations of naloxone.