Tail-flick Reflex Research Articles

Descending facilitatory modulation of a behavioral nociceptive response by stimulation in the adult rat anterior cingulate cortex

It is well documented that the descending endogenous analgesia system, including the periaqueductal gray (PAG) and the rostral ventral medulla (RVM), play an important role in modulation of nociceptive transmission and morphine- and cannabinoid-produced analgesia. Neurons in the PAG receive inputs from different nuclei of higher structures, including the anterior cingulate cortex (ACC). However, it is unclear if stimulation of neurons in the ACC modulates spinal nociceptive transmission. The present study has examined the effects of electrical stimulation and chemical activation of metabotropic glutamate receptors (mGluRs) in the ACC on a spinal nociceptive tail-flick (TF) reflex induced by noxious heating. Activation of the ACC at high intensities (up to 500 μA) of electrical stimulation did not produce any antinociceptive effect. Instead, at most sites within the ACC (n = 36 of 41 sites), electrical stimulation produced significant facilitation of the TF reflex (i.e. decreases in TF latency). Chemical activation of mGluRs within the ACC also produced a facilitatory effect. Descending facilitation from the ACC apparently relays at the RVM. Electrical stimulation in the RVM produces a biphasic modulatory effect, showing facilitation at low intensities and inhibition at higher intensities. The present study provides evidence that activation of mGluRs within the ACC can facilitate spinal nociception. Copyright 2000 European Federation of Chapters of the International Association for the Study of Pain

Activation of cGMP-dependent protein kinase Iα is required for N-methyl- d-aspartate- or nitric oxide-produced spinal thermal hyperalgesia

The effect of a selective cyclic guanocine 3′,5′-monophosphate (cGMP)-dependent protein kinase Iα inhibitor, Rp-8-[(4-chlorophenyl)thio]-cGMPS triethylamine (Rp-8-p-CPT-CGMPS), on either N-methyl- d-aspartate (NMDA)- or N-ethyl-2-(1-ethyl-2-hydroxy-2-nitrosohydrazino)ethanamine (NOC-12, a nitric oxide (NO) donor)-produced thermal hyperalgesia was examined in the rat. Intrathecal administration of NMDA (15 pg/10 μl) or NOC-12 (10, 20 and 30 μg/10 μl) produced a marked curtailment of the tail-flick latency. Maximal NMDA- or NOC-12-produced facilitation of the tail-flick reflex was significantly and dose-dependently blocked by intrathecal pretreatment with Rp-8-p-CPT-CGMPS (7.5, 15 and 30 μg/10 μl). Rp-8-p-CPT-CGMPS given alone did not markedly alter baseline tail-flick latency. These results suggest that the activation of cGMP-dependent protein kinase Iα is required for NMDA- or NO-produced facilitation of thermal hyperalgesia at the spinal cord level.

Delta opioid receptor mediated actions in the rostral ventromedial medulla on tail flick latency and nociceptive modulatory neurons

The rostral ventromedial medulla (RVM) is critical for the modulation of dorsal horn nociceptive transmission. Three classes of RVM neurons (ON, OFF, and NEUTRAL) have been described that have distinct responses to noxious stimuli and mu opioid receptor (MOR) agonists. The present study in barbiturate anesthetized rats investigated the effects of the delta 2 opioid receptor (DOR2) agonist, [D-Ala2]deltorphin II (DELT), microinfused into the RVM on the tail flick reflex and activity of RVM neurons. Tail flick latencies increased dose-dependently after administration of DELT (0.6 nmol and 1.2 nmol). Furthermore, DELT inhibited the tail flick related increase in ON cell activity and shortened the tail flick related pause in OFF cell activity. The activity of NEUTRAL cells was not affected. The antinociceptive effects and corresponding changes in ON and OFF cell activity produced by DELT were antagonized by the DOR2 antagonist, naltriben methanesulfonate, administered at the same site. These DOR2 mediated effects on noxious stimulation-evoked changes in RVM neuronal activity are similar to those reported for MOR agonists and suggest that both DOR2 and MOR produce analgesia through activation of OFF cells.

Does selective destruction of the vagal afferent inflow facilitate or reduce the development of fever?

Secondary afferent neurons in the (i) rostral commissural nucleus tractus solitarii (NTS) or (ii) caudal NTS in rats were destroyed with kainate. Lipopolysaccharide (3 μg/kg i.p.) in (i) led to a rise in colonic temperature ( T c) and random changes in the tail-flick reflex (TFR) latency. In (ii), lipopolysaccharide was not accompanied by fever, but induced a sustained hypoalgesia. In subdiaphragmatically vagotomized rats (iii) lipopolysaccharide failed to change T c and TFR. Structural alterations in NTS and other brainstem nuclei in (iii) were electron microscopically visualized. The changes in the responses to lipopolysaccharide in the three groups may be related to degenerative events in the brainstem nuclei.

Role of Vagal Afferents and the Rostral Ventral Medulla in Intravenous Serotonin-Induced Changes in Nociception and Arterial Blood Pressure

THURSTON–STANFIELD, C. L., J. T. RANIERI, R. VALLABHAPURAPU AND D. BARNES–NOBLE. Role of vagal afferents and the rostral ventral medulla in intravenous serotonin-induced changes in nociception and arterial blood pressure. PHYSIOL BEHAV 67(5) 753–767, 1999.—Intravenous administration of serotonin inhibits the nociceptive tail-flick (TF) reflex, partially through activation of vagal afferents. The present study examined the role of the rostral ventral medulla (RVM) in i.v. serotonin-produced inhibition of the TF reflex. In Experiment 1, the effects of anesthetic blockade of the RVM on serotonin-produced inhibition of the TF were determined. Lidocaine attenuated the serotonin-produced inhibition of the TF reflex, but had no effect on the cardiovascular effects of serotonin. In Experiment 2, the effects of i.v. serotonin on neural activity in the RVM in intact and cardiopulmonary deafferented rats were determined. Neurons in the RVM were classified as ON and OFF cells, where ON cells were excited by noxious heat, and OFF cells were inhibited. The effects of i.v. serotonin on TF latency, blood pressure, and ON or OFF cell activity were then determined. In intact rats, serotonin produced a dose-dependent increase in TF latency, triphasic changes in blood pressure, and bi- or triphasic changes in ON or OFF cell activity. The changes in blood pressure included an initial sharp decrease in blood pressure (Bezold–Jarisch reflex), followed by a brief pressor response, followed by a delay depressor response. ON cells were generally excited, although there was a period during which the excitation decreased. OFF cells were initially excited, followed by a period of inhibition, followed by a second period of excitation. Bilateral cervical vagotomy attenuated the increase in TF latency, the Bezold–Jarisch reflex, and the excitation of OFF cells, and potentiated the excitation of ON cells and the pressor response. Bilateral sinoaortic deafferentation attenuated the Bezold–Jarisch reflex and potentiated the pressor response. These findings indicate that i.v. serotonin inhibits the TF reflex through at least two distinct mechansims, one of which requires the RVM. In addition, serotonin produces a vagally mediated excitation of OFF cells and inhibition of ON cells that may mediate some of the antinociception.

Interactions between social stress and morphine in the periaqueductal gray: effects on affective vocal and reflexive pain responses in rats.

Endogenous opioid systems within the mesencephalic periaqueductal gray matter (PAG) appear to be intricately involved in many affective, defensive, submissive, and reflexive responses, and these systems are activated by aversive stimuli. The present experiments evaluated the influence of opioid receptors within the PAG on affective vocal and reflexive responses to aversive stimuli in socially inexperienced, as well as defensive and submissive responses in defeated, adult male Long-Evans rats. Defeat stress consisted of: (1) an aggressive confrontation with a "resident" stimulus rat in which the experimental "intruder" rat exhibited escape, defensive and submissive behaviors [i.e. upright, supine postures and ultrasonic vocalizations (USV)], and subsequently, (2) protection from the resident rat with a wire mesh screen for ca. 25 min. Defeat stress was immediately followed by an experimental session with thermal antinociceptive and tactile startle stimuli (20 psi airpuffs). The mu opioid receptor agonist morphine (0.3, 1, 3 microgram IC) attenuated startle-induced USV and the tail-flick reflex in socially inexperienced and defeated rats, with both groups of rats demonstrating equal sensitivity to morphine. Morphine decreased defeat-induced USV and increased the display of the crouch posture in defeated rats; these morphine effects in socially inexperienced and defeated rats were re- versed with the opioid receptor antagonist naltrexone (0.1 mg/kg IP). These results reveal that the ventrolateral PAG is an important site in which mu opioid receptor agonists such as morphine mediate affective vocal and submissive responses, yet this structure is not critical in the display of defeat stress-augmented effects of morphine. Endogenous opioid mechanisms appear to participate in the organization of defensive behavior, namely, to facilitate a shift from active to passive forms of coping.

Morphine applied to the thalamic nucleus submedius produces a naloxone reversible antinociceptive effect in the rat

Our previous studies have indicated that the thalamic nucleus submedius (Sm) is involved in nociceptive modulation and plays an important role in an endogenous analgesic system (a feedback loop) consisting of spinal cord – Sm – ventrolateral orbital cortex (VLO) – periaqueductal gray (PAG) – spinal cord. To further investigate the neurotransmitter and receptor mechanisms in this nociceptive modulatory pathway, we tested the effects of microinjection of morphine and naloxone into the Sm on the rat tail flick (TF) reflex. A unilateral microinjection of morphine (8.0 mM, 0.5 μl) into the Sm significantly depressed the TF reflex, whereas a unilateral microinjection of naloxone (5.0 mM, 0.5 μl) into the Sm facilitated the TF reflex. Five minutes after morphine application into Sm, injection of naloxone in this nucleus markedly reversed the inhibition evoked by applying morphine in Sm. These findings suggest that the endogenous opioid peptides may be involved in the antinociceptive effects evoked by activation of the Sm–VLO–PAG pathway which depressed the nociceptive inputs at the spinal level via the brainstem descending inhibitory system, and exert a tonic descending influence.

Electrically-evoked inhibitory effects of the nucleus submedius on the jaw-opening reflex are mediated by ventrolateral orbital cortex and periaqueductal gray matter in the rat

In previous studies we have shown that electrical stimulation of the nucleus submedius inhibits the rat radiant heat-induced tail flick reflex, and that this antinociceptive effect is mediated by the ventrolateral orbital cortex and periaqueductal gray. The aim of the present study was to examine whether electrical stimulation of the nucleus submedius could inhibit the rat jaw-opening reflex, and to determine whether electrolytic lesions of the ventrolateral orbital cortex or the periaqueductal gray could attenuate the nucleus submedius-evoked inhibition. Experiments were performed on pentobarbital-anesthetized rats. The jaw-opening reflex elicited by electrical stimulation of the tooth pulp or the facial skin was monitored by recording the evoked digastric electromyogram. Conditioning stimulation was delivered unilaterally to the nucleus submedius 90 ms prior to each test stimulus to the tooth pulp. After that, electrolytic lesions were made in ventrolateral orbital cortex or periaqueductal gray, and the effect of nucleus submedius stimulation on the jaw-opening reflex was re-examined. Unilateral electrical stimulation of nucleus submedius was found to significantly depress the jaw-opening reflex (mean threshold of 28.0±1.4 μA, n=48), and the magnitude of inhibition increased linearly when the stimulus intensity was increased from 20 to 70 μA, resulting in depression of the digastric electromyogram amplitude from 18.4±5.4% to 74.0±4.9% of the control ( P<0.01, n=37). The onset of inhibition occured 60 ms after the beginning of nucleus submedius stimulation and lasted about 100 ms, as determined by varying the conditioning-test time interval. Furthermore, ipsilateral lesions of the ventrolateral orbital cortex or bilateral lesions of the lateral or ventrolateral parts of periaqueductal gray eliminated the nucleus submedius-evoked inhibition of the jaw-opening reflex. These data suggest that the nucleus submedius plays an important role in modulation of orofacial nociception, and provide further support for a hypothesis that the antinociceptive effect of nucleus submedius stimulation is mediated by ventrolateral orbital cortex and activation of a descending inhibitory system in the periaqueductal gray.

Involvement of excitatory amino acid receptors and nitric oxide in the rostral ventromedial medulla in modulating secondary hyperalgesia produced by mustard oil

We have recently reported a model of secondary hyperalgesia in which facilitation of the thermal nociceptive tail-flick reflex following topical mustard oil is largely dependent on descending influences from the rostral ventromedial medulla (RVM). The current study was designed to examine a potential role for excitatory amino acid receptors and nitric oxide in the RVM in modulating this hyperalgesia. Topical application of mustard oil (100%) to the lateral surface of the hind leg of awake rats produced a short-lived (60 min) facilitation of the tail-flick reflex that was dose-dependently attenuated by microinjection of the selective N-methyl- d-aspartate (NMDA) receptor antagonist APV (1–100 fmol) into the RVM. Microinjection of a greater dose of APV (1000 fmol) into the RVM produced a significant inhibition of the tail-flick reflex in the presence, but not absence, of mustard oil. In contrast, microinjection of the non-NMDA receptor antagonist DNQX (10 nmol) into the RVM further enhanced the magnitude and duration of the hyperalgesic response, and produced a facilitation of the tail-flick reflex following injection into the RVM of naive animals. Similar to APV, microinjection of the nitric oxide synthase inhibitor l-NAME (100–1000 nmol) into the RVM attenuated mustard oil hyperalgesia, while the greatest dose (1000 nmol) produced a significant inhibition of the tail-flick reflex in the presence, but not absence, of mustard oil. A role for nitric oxide synthase in the RVM in mustard oil hyperalgesia was further demonstrated by a significant increase in the number of NADPH-d labeled cells in the RVM at the time of maximal hyperalgesia. Involvement of NMDA receptors and nitric oxide in the RVM in descending nociceptive facilitation was supported by the observation that microinjection of either NMDA or the NO ⋅ donor GEA 5024 into the RVM of naive animals dose-dependently facilitated the tail-flick reflex. The hyperalgesia produced by NMDA injection into the RVM was blocked by prior intra-RVM injection of either APV or l-NAME. These results support the notion that secondary hyperalgesia produced by mustard oil involves concurrent activation of dominant descending facilitatory, as well as masked inhibitory systems from the RVM. Additionally, the data suggest that descending facilitation involves activation of NMDA receptors and production NO ⋅ in the RVM, whereas inhibition involves activation of non-NMDA receptors in the RVM.

Pharmacological evidence for a periaqueductal gray–nucleus raphe magnus connection mediating the antinociception induced by microinjecting carbachol into the dorsal periaqueductal gray of rats

A previous study demonstrated that microinjection of carbachol (CCh) into the dorsal periaqueductal gray matter (dPAG) of rats increases the latency for the tail flick reflex. Several other studies have implicated the raphe magnus (NRM) and the reticularis paragigantocellularis (NRPG) nuclei as relay stations through which descending pathways from the PAG project to the spinal cord via the dorsolateral funiculus (DLF). In the present study, the effects of microinjecting CCh into the dPAG on the tail flick test were examined in rats in which the ipsilateral DLF was previously lesioned, or saline or lidocaine (2%) was microinjected into the NRM or ipsilateral NRPG. The DLF lesion did not change the baseline threshold of the animals in the test, but abolished the CCh-induced increase in the tail flick latency from the dPAG. The neural block of the NRM or NRPG with lidocaine also did not change significantly the latency for the tail flick reflex. The increase in the tail flick latency produced by CCh from the dPAG was not changed by the neural block of the NRPG, but was significantly reduced by the neural block of the NRM. These results are interpreted as indicative that the central antinociceptive mechanisms activated by CCh from the dPAG depend on a descending pathway that projects to the spinal cord via DLF utilizing at least the NRM, but not the NRPG, as an intermediary relay station.

Inhibitory effects of 5-hydroxytryptamine microinjection into thalamic nucleus submedius on rat tail flick reflex are mediated by 5-HT 2 receptors

Our previous findings indicated that electrical or chemical activation of the thalamic nucleus submedius (Sm) produced significant antinociceptive effects and that these effect were blocked by lesion or depression of the ventrolateral orbital cortex (VLO) or the periaqueductal gray (PAG) suggesting a role of the Sm in modulation of nociception. To further investigate the neurotransmitter mechanism involved in this nociceptive modulatory pathway, we tested the effects of microinjection of 5-hydroxytryptamine (5-HT, 50 mM, 0.5 μl) into Sm on the tail flick (TF) reflex. The results show that a unilateral microinjection of 5-HT into Sm significantly depresses the TF reflex; and that this effect is repeatable and dose-dependent. Furthermore, microinjection of 5-HT 2 receptor antagonist cyproheptadine (CPT, 0.3 mM, 0.5 μl) into the same Sm site reverses this 5-HT-evoked inhibition of TF reflex. These results suggest that 5-HT application to the Sm may activate Sm neurons through the 5-HT 2 receptors leading to activation of the brainstem descending inhibitory system via the VLO and depression of the nociceptive information at the spinal level.

Inhibitory effects of electrical stimulation of ventrolateral orbital cortex on the rat jaw-opening reflex

In previous studies, we have shown that electrically or chemically evoked activation of the ventrolateral orbital cortex (VLO) depresses the rat tail-flick (TF) reflex, and this antinociceptive effect is mediated by the periaqueductal gray (PAG). The aim of the present study was to examine whether electrical stimulation of the VLO could inhibit the rat jaw-opening reflex (JOR), and to determine whether electrolytic lesions of the PAG could attenuate this VLO-evoked inhibition. Unilateral electrical stimulation of the VLO significantly depressed the JOR elicited by tooth pulp or facial skin stimuli, with a mean threshold of 30.5±2.3 μA ( n=22). Increasing stimulation intensities from 30 to 80 μA resulted in greater reduction of the dEMG amplitude from 22.9±5.0% to 69.7±3.7% of the baseline value ( P<0.01, n=22). The inhibitory effect appeared 50 ms after the beginning of VLO stimulation and lasted about 150 ms, as determined by varying the conditioning-test (C-T) time interval. Unilateral lateral or ventrolateral lesions of the PAG produced only a small attenuation of the VLO-evoked inhibition of the JOR, but bilateral lesions eliminated this inhibition. These findings suggest that the VLO plays an important role in modulation of orofacial nociceptive inputs, and provide further support for the hypothesis that the antinociceptive effect of VLO is mediated by PAG leading to activation of a brainstem descending inhibitory system and depression of nociceptive inputs at the trigeminal level. The role played by VLO in pain modulation is discussed in association with the proposed endogenous analgesic system consisting of medullary cord–Sm–VLO–PAG–medullary cord.

Development of mustard oil-induced hyperalgesia in rats

Age-dependent changes in nociceptive responses were investigated using either the electromyogram (EMG) recorded from the hamstring muscle in response to electrical stimulation of the hind foot in spinal transected rats or measurement of the tail-flick (TF) reflex latency in intact rats. The development of hyperalgesia produced by topical application of mustard oil was subsequently studied. In experiments involving EMG recordings, rats were tested from day 2 to day 34 after birth (4-day interval) and as adults. In experiments involving measurement of the TF reflex, rats were tested from day 5 to day 30 after birth (5-day interval) and as adults. It was found that the latency and the duration of an early component of the EMG decreased with an increase in animal age, and was similar to adult animals at approximately 18 days after birth. The thermal tail withdraw threshold was lower in pups in comparison with older rats, and took more than 30 postnatal days to become similar to that of adult rats. Although nociceptive behaviors such as biting, body movement, and vocalization could be produced in intact rats by mustard oil in rats as young as 5 days old, the intensities of these responses were subjectively less than those of adult rats. Mustard oil application enhanced significantly the EMG response to electrical stimulation and the effect increased with increasing age. Similarly, mustard oil applied to a hind leg facilitated the TF reflex (decreased response latency). In both experiments, it took approximately 34–40 postnatal days for mustard oil-produced hyperalgesia to become similar to that of adult rats. These data confirm that nociceptive processing is not mature in the young animal and that a developmental period after birth is required for hyperalgesia-related mechanisms to mature.

Spinal serotonergic receptors mediate facilitation of a nociceptive reflex by subcutaneous formalin injection into the hindpaw in rats

It is documented that spinal nociceptive transmission receives descending facilitatory and inhibitory modulation from supraspinal structures. The rostral ventral medulla (RVM), including the nucleus raphe magnus (NRM), nuclei reticularis gigantocellularis (NGC) and gigantocellularis pars alpha (NGC α), is the major bulbar relay of descending modulatory influences. Pharmacological studies show that facilitation of a spinal nociceptive tail-flick (TF) reflex induced by stimulation in the NGC and NGC α is mediated by spinal serotonergic receptors. The present series of experiments provide evidence that activation of spinal serotonergic systems are critical for both induction and maintenance of secondary hyperalgesia induced by subcutaneous injection of formalin into one hindpaw. Subcutaneous injection of formalin produced facilitation of tail withdrawal (mechanical) and the TF reflex (thermal). Facilitatory effects persisted for at least 30 min. Peripheral blockade of the activity by local injection of a hydrophilic lidocaine derivative (QX-314, 5%) into the injected hindpaw abolished both mechanical and thermal facilitation, indicating that peripheral input is important to maintain long-lasting facilitation. Intrathecal application of a serotonergic receptor antagonist methysergide at a dose (64 nmol) which completely blocked descending facilitation produced by electrical- or chemical-stimulation in the NGC and NGC α also significantly attenuated or completely abolished facilitation of tail withdrawal and the TF reflex induced by formalin. Methysergide was effective whether the injection was performed before or after the formalin injection. These results suggest that activation of descending facilitatory serotonergic influences by a prolonged noxious stimulation could contribute to secondary hyperalgesia observed at the tail.

Inhibitory effects of glutamate-induced activation of thalamic nucleus submedius are mediated by ventrolateral orbital cortex and periaqueductal gray i n rats

This study found that in lightly-anesthetized rats a unilateral micro-injection of glutamate (200 mm, 0.5 μ1) into the thalamic nucleus submedius (Sm) markedly depressed the radiant heat-evoked tail flick (TF) reflex. After injection, the mean TFL increased 25.6 ± 6.5% ( n = 24) of the baseline at 5 min, up to a peak value (48.4 ± 7.2%) at 20 min, and recovered to the baseline level at 60 min. This inhibitory effect was dose-related and repeatable over a time interval of 1.0–1.5 h in the same animal. Furthermore, micro-injections of γ-aminobutyric acid (GABA) (100 m M) into the ipsilateral ventrolateral orbital cortex (VLO) (0.7μl), or bilaterally into the lateral or ventrolateral parts of the periaqueductal gray (PAG) (0.5 μ1 on each side), eliminated the Sm-evoked inhibition. After GABA was injected into VLO or PAG, the Sm applications of glutamate failed to produce any significant changes in TFL, with the TFL changes being similar to the saline control ( p>0.05). These results confirmed our previous findings that electrical stimulation of Sm depressed the rat TF reflex and that this inhibitory effect was blocked by electrolytic lesion of the VLO or PAG. Therefore, the present study provides further support for the hypothesis that Sm plays an important role in modulation of nociception, and that its effects are mediated by the VLO-PAG pathway, leading to activation of the brainstem descending inhibitory system and depression of the nociceptive inputs at the spinal cord level.

Cannabinoid receptor-mediated inhibition of the rat tail-flick reflex after microinjection into the rostral ventromedial medulla

Systemic administration of cannabinoids produce profound antinociception in rodents. The purpose of this study was to examine the contribution of the rostral ventromedial medulla (RVM) to cannabinoid-mediated inhibition of the tail-flick reflex. Rats received direct injections of two selective cannabinoid agonists, WIN55,212-2 and HU210, into the RVM. Both compounds significantly elevated tail-flick latencies by over 50%. WIN55,212-3, the inactive enantiomer, was without effect. Furthermore, co-administration of the selective cannabinoid receptor antagonist, SR141716A greatly attenuated the antinociception produced by HU210. Finally, injections of WIN55,212-2 outside the region of the RVM did not affect tail-flick latencies. These results demonstrate that the cannabinoid receptor system participates in the descending control of nociception and raise the possibility that actions of endogenous cannabinoids in the RVM may modulate nociceptive responsiveness.

Antinociception following opioid stimulation of the basolateral amygdala is expressed through the periaqueductal gray and rostral ventromedial medulla

The amygdala, periaqueductal gray (PAG), and rostral ventromedial medulla (RVM) are critical for the expression of some forms of stress-related changes in pain sensitivity. In barbiturate anesthetized rats, microinjection of agonists for the μ opioid receptor into the amygdala results in inhibition of the tail flick (TF) reflex evoked by radiant heat. We tested the idea that TF inhibition following opioid stimulation of the amygdala is expressed through a serial circuit which includes the PAG and RVM. Rats were anesthetized and prepared for microinjection of DAMGO (0.5 μg/0.25 μl) into the basolateral amygdala (BLA) and lidocaine HCl (2.5%/0.4–0.5 μl) into either the ventrolateral PAG or RVM. Lidocaine did not significantly alter baseline values for TF latency or TF amplitude. When injected into the PAG prior to DAMGO application in the BLA, lidocaine significantly attenuated DAMGO-induced antinociception for the entire 40 min testing session. Similar treatment in the RVM also resulted in an attenuation of antinociception although rats showed significant recovery of TF inhibition by 40 min after lidocaine injection. Since acute injection of lidocaine into the RVM also affected baseline heart rate, separate animals were prepared with small electrolytic lesions placed in the RVM. Chronic RVM lesions also blocked TF inhibition produced by amygdala stimulation but did not affect heart rate. These results, when taken together with similar findings in awake behaving animals, suggest that a neural circuit which includes the amygdala, PAG, and RVM is responsible for the expression of several forms of hypoalgesia in the rat.

Pharmacological and neuroanatomical evidence for the involvement of the anterior pretectal nucleus in the antinociception induced by stimulation of the dorsal raphe nucleus in rats

Several studies have shown that the anterior pretectal nucleus (APtN) is involved in descending inhibitory pathways that control noxious inputs to the spinal cord and that it may participate in the normal physiological response to noxious stimulation. Among other brain regions known to send inputs to the APtN, the dorsal column nuclei (DCN), pedunculopontine tegmental nucleus (PPTg), deep mesencephalon (DpMe), and dorsal raphe nucleus (DRN) are structures also known to be involved in antinociception. In the present study, the effects of stimulating these structures on the latency of the tail withdrawal reflex from noxious heating of the skin (tail flick test) were examined in rats in which saline or hyperbaric lidocaine (5%) was previously microinjected into the APtN. Brief stimulation of the PPTg, DpMe or DRN, but not the DCN, strongly depressed the tail flick reflex. The antinociceptive effect of stimulating the DRN, but not the PPTg or DpMe was significantly reduced, but not abolished, by the prior administration of the local anaesthetic into the APtN. The antinociception induced by stimulation of the PPTg or DpMe, therefore, is unlikely to depend on connections between these structures and the APtN. Similar inhibition of the effect of stimulating the DRN was obtained from rats previously microinjected with naloxone (2.7 nmol) or methysergide (2 nmol) into the APtN. Strongly labelled cells were identified in the DRN following microinjection of the fluorescent tracer Fast Blue into the APtN. These results indicate that the APtN may participate as a relay station through which the DRN partly modulates spinal nociceptive messages. In addition, they also indicate that endogenous opioid and serotonin can participate as neuromodulators of the DRN-APtN connection.

Circuitry underlying antiopioid actions of orphanin FQ in the rostral ventromedial medulla.

Several laboratories recently identified a 17 amino-acid peptide, termed "nociceptin" or "orphanin FQ (OFQ)", as the endogenous ligand for the LC132 (or "opioid receptor-like1") receptor. Taken together with the fact that the cellular effects of OFQ are to a large extent opioid-like, the close relationship between the LC132 receptor and known opioid receptors raised expectations that the behavioral effects of this peptide would resemble those of opioids. However studies of the role of OFQ in nociception have not provided a unified view. The aim of the present study was to use a combination of electrophysiological and pharmacological techniques to characterize the actions of OFQ in a brain region in which the circuitry mediating the analgesic actions of opioids has been relatively well characterized, the rostral ventromedial medulla (RVM). Single-cell recording was combined with opioid administration and local infusion of OFQ in the RVM of rats lightlyanesthetized with barbiturates. The tail flick reflex was used as a behavioral index of nociceptive responsiveness. Two classes of physiologically identifiable RVM neurons with distinct responses to opioids have been characterized. -cells are activated, although indirectly, by opioids, and there is strong evidence that this activation is crucial to opioid antinociception. -cells, thought to enable nociception, are directly inhibited by opioids. Cells of a third class, cells, do not respond to opioids and whether or not they have any role in nociceptive modulation remains an open question. OFQ infused within the RVM profoundly suppressed the firing of all classes of RVM neurons, blocking opioid-induced activation of -cells. The antinociceptive effects of a micro-opioid agonist infused at the same site were significantly attenuated in these animals. Those of systemically administered morphine, which can produce its antinociceptive effects by acting at a number of CNS sites, were not blocked by RVM OFQ. Inasmuch as activation of -cells can account for the antinociceptive action of opioids within the RVM, these results demonstrate that, at least within the medulla, OFQ can exert a functional "antiopioid" effect by suppressing firing of this cell class. However to the extent that antinociceptive and pronociceptive outflows from various brain regions involved in both transmission and modulation of nociception are active under different conditions, focal application of OFQ in different regions could potentially produce either hypalgesia or hyperalgesia.

Morphine and Dextrorphan Lose Antinociceptive Activity but Exhibit an Antispastic Action in Chronic Spinal Rats

Within 3–4 weeks after spinal transection, morphine-induced antinociception, assessed with the tail flick reflex in rats, is profoundly reduced. The cause of this decrement is unknown. The present studies were conducted to determine whether this phenomenon reflects a general loss in opiate activity or a selective decline in opiate antinociception. This was accomplished by assessing the effect of morphine on two different responses, the tail flick reflex and the hindlimb spasticity that develops in chronic spinal rats. Because excitatory amino acid antagonists are also antinociceptive in acute spinal rats, the effect of one such drug, dextrorphan, on these two behaviors was also evaluated in chronic spinal animals. The antinociceptive and antispastic effect of subcutaneous (6 mg/kg) and intrathecal (5 μg) morphine injections were assessed in intact and chronic (21–28 days) spinal rats, whereas the effect of subcutaneous (25 and 40 mg/kg) and intrathecal (350 μg) dextrorphan was assessed in acute (1 day) and chronic spinal rats. The antinociceptive effect of both drugs was significantly reduced in chronic spinal animals, relative to saline controls. However, each drug treatment produced a significant antispastic effect in the same animals, indicating a selective decline in opiate action. This outcome also suggests that excitatory amino acid antagonists may be useful as adjunct antispastic agents.