Hong Jia

Inhibitory effects of electrically evoked activation of ventrolateral orbital cortex on the tail-flick reflex are mediated by periaqueductal gray in rats

Abstract The present study found in lightly anesthetized rats that the radiant heat‐evoked tail flick (TF) reflex was markedly inhibited by a unilateral electrical stimulation (a 20 ms train of 0.2 ms, 100 Hz, 30–100 &mgr;A pulses) of the ventrolateral orbital cortex (VLO), with the tail flick latency (TFL) being increased. The mean threshold of VLO stimulation for producing inhibition of the TF reflex was 39.2±8.7 &mgr;A (n=26), and this inhibitory effect increased following increasing stimulation intensity from 40 to 70 &mgr;A. The inhibition developed and remained during the stimulation and disappeared rapidly after termination of the stimulation. When the VLO was stimulated at an intensity of 100 &mgr;A in addition to the inhibition an after‐facilitation of the TF reflex (a decrease in TFL) was observed at 5–10 s after termination of the stimulation. Bilateral electrolytic lesions of the lateral or ventrolateral parts of the periaqueductal gray matter (PAG) dramatically reduced or eliminated the VLO‐evoked inhibition, and the after‐facilitation as well. The difference was significant between the TFL changes produced by VLO stimulation before and after PAG lesion (P<0.01). The results suggest that the antinociception elicited by VLO stimulation is mediated by PAG, leading to activation of the brainstem descending inhibitory system which depresses the nociceptive transmission at the spinal level. The role played by VLO in pain modulation was discussed in association with the proposed endogenous analgesic system consisting of spinal cord‐Sm‐VLO‐PAG‐spinal cord.

Inhibitory effects of electrical stimulation of thalamic nucleus submedius area on the rat tail flick reflex

This study in lightly anesthetized rats found that unilateral electrical stimulation delivered to the ventral part of the thalamic nucleus submedius (Sm), the thalamic reuniens nucleus (Re) and the hypothalamic dorsal area (DA) markedly depressed the TF reflex, and this inhibitory effect increased following increasing stimulation intensity. Stimulation in the dorsal part of Sm did not produce any or only slight depression of the TF reflex. Furthermore, an ipsilateral electrolytic lesion of the ventrolateral orbital cortex (VLO) eliminated the unilateral Sm-evoked inhibition, but not the inhibition elicited by Re and DA and contralateral Sm stimulation. Finally, after bilateral electrolytic lesions of the ventrolateral periaqueductal gray (PAG) the DA and Re and contralateral Sm-evoked inhibitions were also eliminated. The results suggest that the Sm plays an important role in modulation of nociceptive inputs, and this role of Sm is mediated by the VLO and leads to activation of the PAG descending inhibitory system and depression of the nociceptive inputs at the spinal cord level.

Involvement of the frontal ventrolateral orbital cortex in descending inhibition of nociception mediated by the periaqueductal gray in rats

Our previous findings which indicated that electrical stimulation of ventrolateral orbital cortex (VLO) can depress the rat tail flick (TF) reflex and that the VLO-evoked inhibitory effect is blocked by electrolytic lesions of periaqueductal gray (PAG) suggest a role of the VLO in the modulation of nociception. To further investigate the involvement of VLO in this nociceptive modulatory pathway, we tested the effects of microinjections of glutamate (200 mM, 0.7 microliter) into the VLO on the TF reflex. An unilateral microinjection of glutamate into the VLO significantly depressed the TF reflex; and this effect was repeatable. Furthermore, bilateral microinjections of gamma-aminobutyric acid (GABA: 100 mM, 0.5 microliter on each side) into the ventrolateral parts of PAG could eliminate this VLO-evoked inhibition of the TF reflex. These results, along with our previous findings provide further support for a hypothesis that VLO, as a higher center in the frontal cortex, plays an important role in modulation of nociception, and this role is mediated by PAG leading to activation of the brainstem descending inhibitory system which depresses 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 microA (n=22). Increasing stimulation intensities from 30 to 80 microA 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.

Inhibitory effects of glutamate-induced activation of thalamic nucleus submedius are mediated by ventrolateral orbital cortex and periaqueductal gray in 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 mm) 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.

Morphine applied to the ventrolateral orbital cortex produces a naloxone-reversible antinociception in the rat

Our previous findings have indicated that the ventrolateral orbital cortex (VLO) may be involved in modulation of nociception and plays an important role as a higher center of an endogenous analgesic system (a feedback loop) consisting of spinal cord-nucleus submedius (Sm)-VLO-periaqueductal gray (PAG)-spinal cord. To further investigate the neurotransmitter mechanism involved in this nociceptive modulatory pathway, we tested the effects of microinjection of morphine (5 microg, 0.5 microl) into VLO on the tail flick (TF) reflex. The results show that a unilateral microinjection of morphine into VLO dose-dependently suppresses the TF reflex. Furthermore, 6 min after termination of morphine injection, microinjection of opioid receptor antagonist naloxone (1.5 microg, 0.5 microl) into the same VLO site reverses this morphine-evoked inhibition of TF reflex. These results suggest that morphine application to the VLO may directly or indirectly activate VLO neurons projecting to the PAG through the opioid receptor mediation leading to activation of the brainstem descending inhibitory system and depression of the nociceptive inputs at the spinal cord level.

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 microl) into the Sm significantly depressed the TF reflex, whereas a unilateral microinjection of naloxone (5.0 mM, 0.5 microl) 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.

Morphine microinjections into the rat nucleus submedius depress nociceptive behavior in the formalin test

Our previous studies have indicated that the thalamic nucleus submedius (Sm) is involved in modulation of nociception and plays an important role in an endogenous analgesic system (a feedback loop) consisting of spinal cord-Sm-ventrolateral orbital cortex-periaqueductal gray-spinal cord. To investigate whether opioids are involved in this antinociception pathway, the effects of microinjection of morphine and naloxone into the Sm on the nociceptive behavior (agitation) evoked in the formalin test were investigated in the awake rat using an automated movement detection system. The results indicate that a unilateral microinjection of morphine (5 micro g, 0.5 microl) into the Sm suppresses the formalin-induced agitation response, but does not influence spontaneous motor activity, and that the morphine-induced depression can be reversed by microinjection of the opioid receptor antagonist naloxone (1.0 micro g, 0.5 microl) into the same Sm site. The results suggest that opioid receptors in the Sm may be involved in the Sm-mediated depression of persistent inflammatory pain.

Effects of thalamic nucleus submedius lesions on the tail flick reflex inhibition evoked by hindlimb electrical stimulation in the rat.

Bilateral electrolytic lesions of the thalamic nucleus submedius (Sm) facilitated the TF reflex and attenuated the antinociception evoked by hindlimb electrical stimulation with high intensities in lightly anaesthetized rats. However, the antinociception produced by low intensity hindlimb stimulation was unchanged, except that the after-effect was reduced. The results show that the Sm is probably involved in pain modulation and plays an important role in mediation of the antinociception elicited by high intensity peripheral stimulation.

Response of neurons in the thalamic nucleus submedius (Sm) to noxious stimulation and electrophysiological identification of on- and off-cells in rats

&NA; Previous studies have indicated that 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 (Sc)–Sm‐ventrolateral orbital cortex–periaqueductal gray–Sc. However, the function of different types of Sm neurons in nociceptive modulation is unclear. For this reason, on the basis of further studies of properties of the Sm neurons responding to noxious stimuli, the different effects of systemic morphine on the Sm neurons were examined and two classes of nociceptive modulatory neurons, named as off‐ and on‐cells, in this region were identified in lightly anesthetized rats. The results showed that (1) most (84%, 132/157) of the Sm neurons responded to peripheral noxious stimuli. Of these neurons, 66% (n=87) were inhibited, 34% (n=45) excited. All neurons had very large and bilateral, even all body receptive fields. No neuron was found to be responsive to innocuous stimulation; (2) systemic morphine increased the firing rate of neurons inhibited by noxious stimulation, but decreased that of neurons excited by the same stimulation. Furthermore, the effects of morphine could be reversed by systemic naloxone; (3) 45 of Sm neurons examined could be divided into three different classes: off‐cells that decreased the firing rate from tail heating just prior to occurrence of the tail‐flick (TF) reflex (3140±167 ms, n=27), on‐cells that increased the firing rate just before the TF reflex (1720±240 ms, n=8), and neutral‐cells that did not respond to any stimuli and neuronal activities were not related to the TF reflex (n=10). Findings of this study provided electrophysiological evidence for involvement of Sm neurons, as those in the rostral ventromedial medulla, in the opioid‐receptor‐mediated descending nociceptive modulation.