Modulation of cardiovascular excitatory responses in rats by transcutaneous magnetic stimulation: role of the spinal cord.

This study investigated the efficacy of magnetic stimulation on the reflex cardiovascular responses induced by gastric distension in anesthetized rats and compared these responses to those influenced by electroacupuncture (EA). Unilateral magnetic stimulation (30% intensity, 2 Hz) at the Jianshi-Neiguan acupoints (pericardial meridian, P 5-6) overlying the median nerve on the forelimb for 24 min significantly decreased the reflex pressor response by 32%. This effect was noticeable by 20 min of magnetic stimulation and continued for 24 min. Median nerve denervation abolished the inhibitory effect of magnetic stimulation, indicating the importance of somatic afferent input. Unilateral EA (0.3-0.5 mA, 2 Hz) at P 5-6 using similar durations of stimulation similarly inhibited the response (35%). The inhibitory effects of EA occurred earlier and were marginally longer (20 min) than magnetic stimulation. Magnetic stimulation at Guangming-Xuanzhong acupoints (gallbladder meridian, GB 37-39) overlying the superficial peroneal nerve on the hindlimb did not attenuate the reflex. Intravenous naloxone immediately after termination of magnetic stimulation reversed inhibition of the cardiovascular reflex, suggesting involvement of the opioid system. Also, intrathecal injection of delta- and kappa-opioid receptors antagonists, ICI174,864 (n=7) and nor-binaltorphimine (n=6) immediately after termination of magnetic stimulation reversed inhibition of the cardiovascular reflex. In contrast, the mu-opioid antagonist CTOP (n=7) failed to alter the cardiovascular reflex. The endogenous neurotransmitters for delta- and kappa-opioid receptors, enkephalins and dynorphin but not beta-endorphin, therefore appear to play significant roles in the spinal cord in mediating magnetic stimulation-induced modulation of cardiovascular reflex responses.     

Nociceptin inhibits rat sympathetic preganglionic neurons in situ and in vitro

In vitro and in situ experiments were conducted to evaluate the hypothesis that the nonclassical opioid peptide nociceptin acting on sympathetic preganglionic neurons (SPNs) inhibits spinal sympathetic outflow. First, whole cell patch recordings were made from antidromically identified SPNs from immature (12-16 day old) rat spinal cord slices. Nociceptin (0.1, 0.3, and 1 microM) concentration dependently suppressed the excitatory postsynaptic potentials (EPSPs) evoked by focal stimulation and hyperpolarized a population of SPNs; these effects were naloxone insensitive. L-Glutamate-induced depolarizations were not significantly changed by nociceptin. Results from this series of experiments indicate that nociceptin inhibits the activity of SPNs by either a presynaptic or postsynaptic site of action, whereby the peptide reduces, respectively, the amplitude of EPSPs or the excitability of SPNs. Second, intrathecal injection of nociceptin (3, 10, and 30 nmol) to urethan-anesthetized rats dose dependently reduced the mean arterial pressure and heart rate; these effects were not prevented by prior intravenous administration of naloxone (1 mg/kg). Physiological saline given intrathecally was without appreciable effects. These results, together with earlier observations of the detection of nociceptin-immunoreactive nerve fibers and nociceptin receptor immunoreactivity in the rat intermediolateral cell column, raise the possibility that the opioid peptide, which may be released endogenously, reduces spinal sympathetic outflow by depressing the activity of SPNs.     

Selective involvement of opioids in mechanisms of synapse-specific plasticity in Helix lucorum snail during sensitization acquisition

Effects of met-enkephalin (opioid peptide) and naloxone (opioid antagonist) on nociceptive sensitization were studied in L-RP11 Helix neurons. In control snails sensitizing stimulation produced reversible membrane depolarization and depression of neural responses evoked by sensory stimuli during the short-term stage of sensitization and facilitation of these responses at the long-term stage. Met-enkephalin (10 but not 0.1 microM) suppressed the neural responses evoked by nociceptive stimuli. Sensitizing stimulation during metenkephalin application prevented the facilitation of neural responses evoked by tactile stimulation of snail head, whereas facilitation of neural responses evoked by chemical stimulation of head or tactile stimulation of foot were similar to that in control sensitized snails. Sensitizing stimulation during met-enkephalin and/or naloxone application prevented the facilitation of neural responses evoked by chemical stimulation of snail head, whereas responses evoked by tactile stimulation of snail head or foot were facilitated (as in neurons of control sensitized snails). Opioids are suggested to be involved in regulation of nociceptive mechanisms and selective induction of long-term plasticity in L-RP11 neural inputs activated by tactile of chemical stimulation of snail head.     

Dynorphin converting enzyme in the rat spinal cord. Decreased activities during acute phase of adjuvant induced arthritis.

This paper describes a study on a dynorphin converting enzyme in spinal cord homogenates from rats with experimental arthritis after adjuvant injection into one hindpaw. The enzyme resembles a neutral cysteine endopeptidase which cleaves the opioid peptide dynorphin B and generates its N-terminal fragment, Leu-enkephalin-Arg6 with opioid activity. It exhibits considerably lower activity against dynorphin A and alpha-neoendorphin, the two other prodynorphin derived peptides. The enzyme showed significantly higher activity in the dorsal part than in the ventral part of the spinal cord. A significant decrease in enzyme activity was observed in the dorsal spinal cord during inflammation as compared to vehicle-injected controls. This decrease paralleled a decrease in the tissue level of Leu-enkephalin-Arg6. These data thus indicate that adjuvant-induced arthritis may generate an important change in a converting enzyme acting on peptide structures, which may be involved in pain modulation. Therefore, a functional role of the present enzyme in the regulation of pain-related peptides is suggested.     

刺激大脑皮层体感Ⅰ区和大脑脚对大鼠脊髓背角神经元伤害感受性反应的影响

在大鼠用玻璃微电极细胞外记录的方法,观察了刺激皮层体感Ⅰ区(SI区)和大脑脚(CP)对皮肤强电刺激诱发的脊髓背角广动力范围(WDR)神经元长潜伏期反应(C-反应)的影响。结果表明刺激SI区对背角WDR神经元C-反应的影响以抑制为主,刺激CP的作用与刺激SI区的作用相似,但刺激CP更为有效。抑制作用的持续时间在不同神经元差别很大,短者在刺激停止后仅持续400ms,长者可达10min以上。静注纳洛酮对抑制作用无明显影响,静注二甲麦角新碱在部分神经元可使抑制作用明显减弱或完全消失,提示5-HT部分参与皮层下行抑制作用的实现,而内鸦片肽则否。     

Effects of the selective kappa opioid antagonist, nor-binaltorphimine, on electrically-elicited feeding in the rat

Lateral ventricular injections of the 'nonspecific' opioid antagonist naloxone (100 micrograms) and the kappa-selective opioid antagonist nor-binaltorphimine (50 micrograms) elevated the electrical brain stimulation frequency threshold for eliciting feeding behavior. Mesopontine aqueductal injections of nor-binaltorphimine, on the other hand, lowered the feeding threshold while naloxone still elevated threshold. These findings suggest the existence of forebrain kappa receptors at which endogenous opioid activity results in a facilitation of feeding while kappa receptors in the brainstem seem to mediate an inhibitory effect.     

Antinociceptive role of galanin in the spinal cord of rats with inflammation, an involvement of opioid systems

The present study investigated the role of galanin in the transmission of nociceptive information in the spinal cord of rats with inflammation. Bilateral decreases in hindpaw withdrawal latencies (HWLs) to thermal and mechanical stimulation were observed after acute inflammation induced by injection of carrageenan into the plantar region of the rat left hindpaw. Intrathecal injection of galanin induced significant increases in the HWLs to thermal and mechanical stimulation in rats with inflammation. The galanin-induced antinociceptive effect was more pronounced in rats with inflammation than that in intact rats. The antinociceptive effect of galanin was partly inhibited by intrathecal injection of naloxone. Furthermore, intrathecal administration of galantide, an antagonist of galanin receptor, could attenuate the antinociceptive effect induced by intraperitoneal injection of morphine, suggesting an involvement of opioid systems in the galanin-induced antinociception. The results indicate that galanin plays an important role in the transmission of nociceptive information in the spinal cord of rats with inflammation, and opioid systems are involved in the galanin-induced antinociception.     

Naloxone reversal of the pentobarbital induced blockade of ovulation in the rat

The effect of naloxone, an opiate antagonist, on the pentobarbital induced suppression of gonadotrophin secretion and subsequent ovulation was studied. Two injections of naloxone, the first at 13.00 h and the second, given simultaneously with an ovulatory blocking dose of pentobarbital at 13.30 h, partially reversed the inhibitory effect of the barbiturate. However, if the first injection of naloxone was given with pentobarbital at 13.30 h and a second at 14.30 h, then the opioid antagonist had no such effect. Naloxone administered alone, enhanced gonadotrophin secretion and increased the number of ova shed. These findings support the view, that endogenous opioids may exert an inhibitory influence on central neural processes involved with ovulation.     

Effect of cerebral ventricles perfusion with naloxone on trigemino-hypoglossal reflex in rats

The goal of this study was to determine whether opioid receptor antagonist naloxone abolishes the influence of periaqueductal central gray (PAG) on nociceptive evoked tongue jerks (ETJ) -- a trigemino-hypoglossal reflex induced by tooth pulp stimulation. In rats under chloralose anesthesia three series of experiments were performed. In the first two groups perfusions of lateral ventricles-cerebellomedullary cistern with McIlwain-Rodnight's solution and naloxone were carried out. In group 3 naloxone was infused through a catheter through the jugular vein. The amplitudes of tongue jerks induced by tooth pulp stimulation were recorded during subsequent 10 min perfusions. Mean amplitude of tongue movements induced by tooth pulp stimulation was regarded as the indicator of the magnitude of trigemino-hypoglossal reflex. We observed that perfusion of the cerebral ventricles with naloxone (100 nmol/ml) increased the trigemino-hypoglossal reflex up to 143%. The amplitude of ETJ was significantly reduced during PAG stimulation with a train of electrical impulses. After obtaining a significant -- 93% -- inhibition of ETJ (7% of the control), naloxone (100 nmol/ml) was added to the perfusion fluid. This led to a significant increase of the reflex up to 68%. Infusion of naloxone through the jugular vein did not affect the reflex. The above results suggest that the inhibition of ETJ due to PAG stimulation is partially reversed by naloxone and mediated via interactions with endogenous opioid systems involved in modulation of nociception.     

Effect of small doses of naloxone on the pulsatile secretion of prolactin in the crossbreed ewe during the non-breeding season

The objective of this work was to study the effect of small doses of naloxone (Nx) on the pulsatile secretion of prolactin (Pr). For this purpose 12 crossbreed ewes were selected and allocated to three groups of four. Group 1 was treated with two injections (at 7 and 19 h) of 40 microg of GnRH. Group 2 was treated with two i.m. injections (at 7 and 19 h) of 0.5mg of naloxone. And the control group 3 was sham treated with injections of 3 ml saline. Blood samples were taken at 20 min intervals during six consecutive hours after injections. When ewes were treated at 7h no significant changes were observed in concentrations of prolactin following treatment with GnRH. Values fluctuated between 200 and 210 ng/ml. In group 2 treated with naloxone there was no change in plasma Pr concentrations during the first 100 min of sampling, however 60 min after Nx treatment Pr decreased significantly (p<0.01) and thereafter Pr plasma levels were consistently less (p<0.001) than control and GnRH treated ewes for the duration of the experiment. The response of the three groups after the second injection (19 h): After the injection of GnRH plasma Pr levels followed much the same pattern observed after the initial treatment, Pr concentrations were similar to those of control ewes. Ewes treated with a second small dose of naloxone (0.5mg i.m.) however, showed a decrease in plasma Pr 60 min after the administration of the endogenous opioid antagonist. Thereafter Nx treated ewes had lesser (p<0.001) plasma Pr levels until the termination of the experiment. It was concluded that Nx an opioid antagonist administered in small intermittent doses can alter Pr plasma concentrations in the ewe, showing that endogenous opioids are important modulators of endocrine function and that the administration of small intermittent doses of opioid antagonists produce significant endocrine changes in ewes.     

The nucleus paragigantocellularis and opioid withdrawal-like behavior

Participation of the nucleus paragigantocellularis (PGi) in mediation of opioid withdrawal was examined in conscious, unrestrained, non-opioid-dependent rats, using electrical stimulation of the PGi. A characteristic series of behaviors, which resembled those seen during naloxone-precipitated withdrawal from dependence on the opioid agonist, butorphanol, was elicited during 30 min of PGi stimulation. Thus, the behavioral syndrome has been termed opioid withdrawal-like. Simultaneous microdialysis measurement of glutamate within the locus ceruleus indicated a positive correlation between extracellular glutamate concentrations and behavioral responses. Behavioral responses were inhibited by 50% during reverse dialysis perfusion of the locus ceruleus with the glutamate receptor antagonist, kynurenic acid, without any effect on glutamate concentrations. Thus, increases in locus ceruleus glutamate partially mediate opioid withdrawal-like behavior. Intracerebroventricular (i.c.v.) injections of the opioid antagonist, naloxone, or of the mu-selective (beta-funaltrexamine) or the delta-selective (naltrindole) opioid antagonists decreased, but did not abolish, stimulation-induced behavioral responses. Similar i.c.v. injections of the kappa-selective antagonist, nor-binaltorphimine, had no effect on behavioral responses to PGi stimulation. Activation of the PGi by electrical stimulation can elicit behaviors similar to those observed during opioid withdrawal. Moreover, additional levels of complexity are evident in the neuropharmacology of PGi stimulation-induced opioid withdrawal-like behavior.     

多巴胺对大鼠背角WDR神经元的抑制不被酚妥拉明...

The inhibitory effects of dopamine (DA) applied spinally on the wide dynamic range (WDR) neurons of dorsal horn in rats were studied with extracellular recording technique. 54 WDR units were tested from 43 rats. With a dosage of DA from 0.26 x 10(-6) to 1.58 x 10(-6) mol/kg, the inhibitory effect of the neurotransmitter on the responses of dorsal horn neurons to noxious transcutaneous electrical stimulation exhibited a gradual increase. After DA (0.52 x 10(-6) mol/kg) administration, the inhibitory effect of DA began to appear in 5 min and reach to maximum in 15 min, whereupon the maximum level could be maintained for about 25 min. This effect of DA could be reversed completely by dopaminergic receptor antagonist, droperidol (0.66 x 10(-6) mol/kg) but not by 2.65 x 10(-6) mol/kg phentolamine or 1.37 x 10(-6) mol/kg naloxone. The results of the present investigation suggest that DA may be involved in the modulation of nociception at the spinal level as an independent neurotransmitter.     

Effect of central naloxone on hormone and blood pressure responses to hemorrhage in conscious sheep

The role of the brain opioid system in the control of hypothalamic-pituitary-adrenal activity was studied in 10 conscious sheep with an indwelling cannula in a cerebral lateral ventricle. On separate days, sheep received infusions of artificial CSF (control) and the opiate antagonist, naloxone (100 micrograms/hr) before and during acute moderate hemorrhage (15 ml/kg over 10 min). Infusion of naloxone before hemorrhage raised plasma ACTH and resulted in a significant increase in cortisol compared to the control infusion. In contrast, ACTH and cortisol responses to hemorrhage tended to be blunted by central naloxone infusion. The responses of vasopressin, aldosterone and the catecholamines remained unaffected by naloxone. The fall in blood pressure and the rise in heart rate accompanying hemorrhage were likewise unaltered. These results suggest that brain opioid peptides have an inhibitory effect on basal ACTH secretion but do not play a major role in modulating the hemodynamic or pituitary-adrenal responses to acute moderate hemorrhage in conscious sheep.     

Nociceptin in rVLM mediates electroacupuncture inhibition of cardiovascular reflex excitatory response in rats.

Electroacupuncture (EA) at Neiguan-Jianshi acupoints through an opioid mechanism inhibits the cardiovascular pressor response induced by mechanical stimulation of the stomach. Because nociceptin also may regulate cardiovascular activity through its action in the brain stem, we hypothesized that this neuromodulator serves a role in the EA-related inhibitory effect. Blood pressure in ventilated male Sprague-Dawley rats (400-600 g) anesthetized by ketamine and alpha-chloralose was measured during balloon inflation of the stomach. Gastric distension with 6-8 ml of air induced consistent pressor reflexes of 26 +/- 1 mmHg that could be repeated every 10 min for 100 min. When nociceptin (10 nM) was microinjected into the rostral ventrolateral medulla (rVLM), the pressor response induced by gastric distension was inhibited by 68 +/- 6%. Thirty minutes of EA also decreased the reflex response by 75 +/- 11%; microinjection of saline into the rVLM did not alter the inhibitory effect of EA. In contrast, microinjection of a nociceptin receptor antagonist into the rVLM promptly reversed the EA response. Pretreatment with the opioid receptor antagonist naloxone did not influence the EA-like inhibitory effect of nociceptin on the distension-induced pressor reflex (22 +/- 1 to 8 +/- 2 mmHg). Furthermore, a mu-opioid receptor agonist microinjected into the rVLM after microinjection of a nociceptin receptor antagonist during EA promptly reversed the nociceptin receptor antagonist-related inhibition of the EA effect. Thus, in addition to the classical opioid system, nociceptin, through opioid receptor-like-1 receptor stimulation in the rVLM, participates in the modulatory influence of EA on reflex-induced increases in blood pressure.     

Responses of units in the magnocellular part of the medial geniculate body to acoustic and somatosensory stimulation

Responses of 150 neurons in the magnocellular part of the medial geniculate body to clicks and to electrodermal stimulation of the contralateral forelimb were investigated in cats immobilized with myorelaxin. Of the total number of neurons 65% were bimodal, 16.6% responded only to clicks, and 15.4% only to electrodermal stimulation. The unitary responses were excitatory (spike potentials) and inhibitory (inhibition of spontaneous activity). Responses beginning with excitation occurred more frequently to stimulation by clicks than to electrodermal stimulation, whereas initial inhibition occurred more often to electrodermal stimulation. The latent period of the initial spike potentials in response to clicks and to electrodermal stimulation was 5–27 and 6–33 (mean 11.6 and 16.2) msec respectively. Positive correlation was found between the latent periods of spike potentials recorded in the same neurons in response to clicks and to electrodermal stimulation, and also to electrodermal stimulation and to stimulation of the dorsal funiculus of the spinal cord. It is concluded that the magnocellular division of the medial genicculate body is a transitional structure between the posterior ventral nucleus and the parvocellular division of the medial geniculate body, and that in addition, it is connected more closely with the auditory than with the somatosensory system. It is suggested that the somatosensory input into the magnocellular division of the medial geniculate body is formed mainly by fibers of the medial lemniscus.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 10, No. 2, pp. 133–141, March–April, 1978.     

猫中缝-脊髓5-羟色胺能和阿片肽能下行抑制系统在抗内脏伤害性反应中作用的比较

在无麻醉的麻痹猫,以伤害性电刺激内脏大神经传入纤维诱发的连续内脏躯体反射放电为指标,分别刺激腓总神经、延髓中缝大核区以及包括导水管周围灰质、中央中核在内的脑区以产生相应的抑制效应。分别全身与脊髓硬膜内给予纳洛酮以及对氯苯丙胺与羟甲丙基甲基麦角酰胺(Methysergide),观察各种抑制效应的变化。发现:(1)静脉注射纳洛酮虽在不少的动物上能暂时地逆转上述效应,但逆转的程度都是不完全的,硬膜内给药也有类似作用;(2)静脉或腹腔内注射对氯苯丙胺一般能较完全而持久地阻断这些抑制效应,但对腓总神经的抑制效应影响较小,硬膜内给药也有相似的效果;(3)羟甲丙基甲基麦角酰胺(静脉或硬膜内注射)主要阻断腓总神经的抑制效应,对中缝大核的抑制效应影响较小。上述结果显示,阿片肽能和5-羟色胺能下行系统均直接参与内脏痛觉传递的延髓中缝-脊髓调节机制,但后者所起的作用可能更大。     

UFP-101 antagonizes the spinal antinociceptive effects of nociceptin/orphanin FQ: behavioral and electrophysiological studies in mice

Nociceptin/orphanin FQ (N/OFQ) modulates various biological functions, including nociception, via selective stimulation of the N/OFQ peptide receptor (NOP). Here we used the NOP selective antagonist UFP-101 to characterize the receptor involved in the spinal antinociceptive effects of N/OFQ evaluated in the mouse tail withdrawal assay and to investigate the mechanism underlying this action by assessing excitatory postsynaptic currents (EPSC) in laminas I and II of the mouse spinal cord dorsal horn with patch-clamp techniques. Intrathecal (i.t.) injection of N/OFQ in the range of 0.1-10 nmol produced a dose dependent antinociceptive effect, which was prevented by UFP-101, but not by naloxone. In contrast the antinociceptive effect of the mu-opioid peptide receptor agonist endomorphin-1 was blocked by naloxone but not by UFP-101. Moreover, N/OFQ and endomorphin-1 induced a significant antinociceptive effect in wild type mice while in mice knockout for the NOP receptor gene only endomorphin-1 was found to be active. In mouse spinal cord slices 1 microM N/OFQ reduced EPSC to 60+/-4% of control values. This inhibitory effect was reversed in a concentration dependent manner by UFP-101 (pA2 value 6.44). The present results demonstrate that N/OFQ-induced spinal antinociception in vivo and inhibition of spinal excitatory transmission in vitro are mediated by receptors of the NOP type.     

刺激中缝背核对大鼠脊髓背角神经元伤害性反应的影响及其传出途径的分析

大量资料表明,中缝背核(DR)在痛觉调节中具有重要作用。本实验用电生理学方法研究DR在痛觉调制中的下行性抑制作用,主要观察刺激DR对清醒制动大鼠脊髓背角神经元伤害性放电的影响。其主要结果是:①刺激DR或电针可以抑制脊髓背角神经元的伤害性反应,吗啡可加强这种抑制效应;②损毁中缝大核(NRM)、纳洛酮、麦角酰二乙胺(LSD)、赛庚啶及对氯苯丙氨酸(PCPA)均能部分阻断DR对脊髓背角神经元伤害性反应的抑制,实验结果表明:刺激DR抑制脊髓背角神经元的伤害性反应,部分是通过NRM间接控制背角神经元的伤害性传入;还有一部分是不通过NRM,可能是DR直接对脊髓背角伤害性信息的调制。在这种下行性抑制通路中有5-HT和阿片样物质的参与。     

The roles of endogenous opioids in the inhibitory action of the hippocampus on preovulatory luteinizing hormone in rats

Whether endogenous opioid peptides were involved in the inhibitory action of the hippocampus (HPC) on luteinizing hormone (LH) release was studied examining the effect of naloxone, an opioid antagonist, on the inhibitory action of electrical stimulation of the HPC and also by examining the effect of metenkephalin administration in the HPC, on preovulatory release of LH in proestrous rats. In rats treated with saline at 13:00 h, either sham stimulation or electrical stimulation of the dorsal HPC, which was performed acutely under ether anesthesia, significantly inhibited the afternoon rise in serum LH. In animals treated with naloxone at a dose of 2.0 mg/kg body weight, the afternoon rise in LH appeared smaller than that in the control group. However, statistical analysis showed no significant difference in LH values compared to the control group. Direct administration of met-enkephalin at a dose of 10 micrograms at 13:00 h through a chronically implanted cannula in the HPC did not induce any significant change in the afternoon rise in LH, regardless of whether it induced behavioral seizures or not. The results suggest that opioid peptides in the HPC do not play a significant role in the inhibitory action of HPC on LH release. Opioids existing in areas other than the HPC may play a certain, but small role.     

Opioids and central baroreflex control: a site of action in the nucleus tractus solitarius

These studies investigated whether the nucleus of the tractus solitarius (NTS) is a central site where opioids modulate baroreceptor reflexes. Microinjections into the NTS of [D-Ala2,MePhe4, Gly-ol5]enkephalin (DAGO) significantly reduced reflex-mediated depressor responses evoked by electrical stimulation of the aortic nerve. Subsequent NTS injections of naloxone restored baroreflexes to control levels. These results demonstrate that the NTS is a central site where exogenously administered opioids can modulate baroreceptor reflexes. NTS injections of naloxone had no effect on baroreflex function, suggesting that tonic activation of opioid receptors at this site plays little or no role in central baroreflex control.