Jin Sook Ju

Microinjection of arginine vasopressin into the central nucleus of amygdala suppressed nociceptive jaw opening reflex in freely moving rats.

This study was performed to examine the antinociceptive effect after microinjection of arginine vasopressin (AVP) into the central nucleus of amygdala. We recorded the jaw opening reflex in freely moving rats. After injection of 0.2 or 0.4 nM AVP into the central nucleus of amygdala, digastric electromyogram (dEMG) was suppressed to 55 +/- 5% or 88 +/- 3 of the control. Artificial cerebrospinal fluid had no effects on the basal dEMG activity. V(1) vasopressin receptor antagonist blocked the suppressive effect produced by microinjection of 0.4 nM AVP from 53 +/- 3 to 81 +/- 3% of the control. However, V(2) vasopressin receptor antagonist did not affect changes in dEMG. We observed dEMG activity after intracerebroventricular injection of naloxone, methysergide, or phentolamine. All drugs did not affect the basal dEMG activity at our dose. Naloxone blocked the suppressive effect of 0.4 nM AVP from 42 +/- 4 to 79 +/- 5% of the control. Methysergide also inhibited the suppression of dEMG from 44 +/- 3 to 83 +/- 6% of the control. However, phentolamine, an alpha-adrenergic receptor antagonist, did not affect the suppression of dEMG. These results indicate AVP in the central nucleus of amygdala has potent analgesic effects in the orofacial area. The antinociception of central AVP seems to be mediated by opioid and serotonergic pathways.

Progesterone produces antinociceptive and neuroprotective effects in rats with microinjected lysophosphatidic acid in the trigeminal nerve root

BackgroundIn our present study, we studied the role of demyelination of the trigeminal nerve root in the development of prolonged nociceptive behavior in the trigeminal territory.ResultsUnder anesthesia, the Sprague-Dawley rats were mounted onto a stereotaxic frame and 3 μL of lysophosphatidic acid (LPA, 1 nmol) was injected into the trigeminal nerve root to produce demyelination. This treatment decreased the air-puff thresholds, persisted until postoperative day 130, and then returned to the preoperative levels 160 days after LPA injection. The LPA-treated rats also showed a significant hyper-responsiveness to pin-prick stimulation. We further investigated the antinociceptive and neuroprotective effects of progesterone in rats undergoing demyelination of the trigeminal nerve root. Progesterone (8, 16 mg/kg/day) was administered subcutaneously, beginning on the operative day, for five consecutive days in the LPA-treated rats. Treatment with progesterone produced significant early anti-allodynic effects and delayed prolonged anti-allodynic effects. The expression of protein zero (P0) and peripheral myelin protein 22 (PMP22) were significantly down-regulated in the trigeminal nerve root on postoperative day 5 following LPA injection. This down-regulation of the P0 and PMP22 levels was blocked by progesterone treatment.ConclusionsThese results suggest that progesterone produces antinociceptive effects through neuroprotective action in animals with LPA-induced trigeminal neuropathic pain. Moreover, progesterone has potential utility as a novel therapy for trigeminal neuropathic pain relief at an appropriate managed dose and is therefore a possible future treatment strategy for improving the recovery from injury.

Intracisternal administration of mitogen-activated protein kinase inhibitors reduced IL-1β-induced mirror-image mechanical allodynia in the orofacial area of rats

The present study investigated the role of central mitogen-activated protein kinases (MAPKs) in interleukin-1beta (IL-1beta)-induced mirror-image mechanical allodynia in the orofacial area. Experiments were carried out on Sprague-Dawley rats. Under pentobarbital sodium anesthesia, a polyethylene tube was implanted in the subcutaneous area of one vibrissa pad, which enabled us to inject IL-1beta. For an intracisternal injection, each anesthetized rat was mounted on a stereotaxic frame and a polyethylene tube was implanted. Animals were given a recovery time of at least 72 h from surgery. After a subcutaneous administration of 0.01, 0.1, 1, or 10 pg of IL-1beta, we examined the face withdrawal behavioral responses produced by 10 successive trials of air puffs ipsilateral or contralateral to the IL-1beta injection site. Normal animals did not respond to pressure less than 40 psi. The thresholds of air puffs ipsilateral and contralateral to the IL-1beta injection site were significantly lower in the IL-1beta-treated group, compared with the vehicle-treated group. The decrease in the threshold of air puffs appeared 10 min after an IL-1beta injection and persisted for over 3h. Intracisternal pretreatment with PD98059, a p44/42 MAPK inhibitor, or SB203580, a p38 MAPK inhibitor, significantly reduced the decrease in the threshold of air puffs ipsilateral to the IL-1beta injection site produced by 10 pg of IL-1beta. IL-1beta-induced mirror-image mechanical allodynia was also reduced significantly by intracisternal pretreatment with both PD98059 and SB203580. These results indicate that central MAPK pathways mediate IL-1beta-induced mirror-image mechanical allodynia in the orofacial area.

Interleukin-1β injected intracisternally inhibited NMDA-evoked behavioral response in the orofacial area of freely moving rats

Abstract The brain-derived interleukin-1β (IL-1β) has been involved in the modulation of nociceptive processing. The direction of the effects, however, analgesia or hyperalgesia, is controversial. Here, we report the role of IL-1β injected intracisternally in orofacial pain transmission. Experiments were carried out on 90 male SD rats and surgical procedures were performed under pentobarbital sodium. Intracisternal injection of 0.3 or 0.6 μg of N -methyl- d -aspartic acid (NMDA) produced intense scratching behavioral responses including vocalization, agitation and a desire to escape in a dose-related manner. The intracisternal injection of 1 or 10 ng IL-1β significantly decreased the NMDA-evoked scratching behavioral responses. Pretreatment with an IL-1 receptor antagonist or naloxone, an opioid receptor antagonist, blocked the IL-1β-induced antinociceptive response. These results suggest that cytokine injected intracisternally seems to produce antinociceptive effects in the NMDA-evoked pain model of the orofacial area and the antinociceptive effect seems to be mediated by an opioid pathway.

Effects of Tnf-α injected intracisternally on the nociceptive jaw-opening reflex and orofacial formalin test in freely moving rats

Abstract The present study was performed to investigate the effects of central cytokines on the modulation of nociception in the orofacial area. A nociceptive jaw-opening reflex (JOR) and an orofacial formalin test were monitored after intracisternal administration of tumor necrosis factor (TNF)-α in freely moving rats. Experiments were carried out on 83 male rats weighing 300–350 g and surgical procedures were performed under pentobarbital sodium. After intracisternal injection of Tnf-α, digastric electromyogram (dEMG) and noxious behavioral responses were monitored. In the nociceptive JOR, dEMG was not significantly changed after intracisternal injection of 200 pg and 2 ng Tnf-α. However, 20 ng Tnf-α suppressed dEMG to 72±6% of the control values. The orofacial formalin responses showed two distinct phases separated by a time of relative inactivity with an early short-lasting response (0–9 min, first phase) and a continuous prolonged response (10–45 min, second phase). In the inflammatory orofacial formalin test, intracisternal injection of 20 pg Tnf-α did not change the number of noxious behavioral responses produced by formalin injection. However, 200 pg Tnf-α injected intracisternally significantly increased the number of noxious behavioral responses produced by formalin injection in both the early and late phases, and 2 ng Tnf-α increased formalin induced noxious behavioral responses in only the late phase. A higher dose of 20 ng Tnf-α did not change the number of noxious behavioral responses produced by formalin injection. The hyperalgesic action of Tnf-α injected intracisternally was blocked by pretreatment with the interleukin-1 (IL-1) receptor antagonist. These results suggest that central Tnf-α modulates the transmission of nociceptive information in the orofacial area. However, the hypo/hyperalgesic response of central Tnf-α seems to depend on the orofacial pain model or in a dose-related manner. The hyperalgesic response of central Tnf-α seems to be mediated by the IL-1 receptor.

Intracisternal NMDA produces analgesia in the orofacial formalin test of freely moving rats

The present study was performed to investigate the antinociceptive response to the intracisternal administration of NMDA in the orofacial area. To achieve this purpose, the effects of NMDA injected intracisternally on the orofacial formalin test were monitored in freely moving rats. We also investigated underlying the mechanisms of NMDA-induced antinociceptive response. Experiments were carried out on 80 male SD rats and surgical procedures were performed under pentobarbital sodium (40 mg/kg, i.p.). Fifty microliters of 5% formalin was applied subcutaneously to the vibrissa pad without any restraining of the animals. For each animal, the number of noxious behavioral responses and the time spent grooming, rubbing, and/or scratching the facial region proximal to the injection site were recorded for nine successive 5-min intervals. The orofacial formalin responses showed two distinct phases separated by a time of relative inactivity. Intracisternal administration of NMDA produced intense scratching behavioral responses with dose related manner. NMDA injected intracisternally 30 min prior to formalin injection, however, inhibited noxious behavioral responses produced by a formalin injection significantly. Pretreatment with naloxone 20 min prior to NMDA injection abolished the inhibition of number of scratches and the duration of scratching produced by the intracisternal injection of NMDA in the late phase. Pretreatment with L-NAME, NO synthesis inhibitor, however, did not affect the antinociceptive response produced by NMDA injected intracisternally. These results suggest that NMDA injected intracisternally produces brief pain behavioral responses and also produces delayed antinociceptive effects in the orofacial formalin test. The opioid pathway seems to be involved in the NMDA-induced antinociception in the orofacial area.

Participation of central GABAA receptors in the trigeminal processing of mechanical allodynia in rats

Here we investigated the central processing mechanisms of mechanical allodynia and found a direct excitatory link with low-threshold input to nociceptive neurons. Experiments were performed on male Sprague-Dawley rats weighing 230-280 g. Subcutaneous injection of interleukin 1 beta (IL-1β) (1 ng/10 µL) was used to produce mechanical allodynia and thermal hyperalgesia. Intracisternal administration of bicuculline, a gamma aminobutyric acid A (GABAA) receptor antagonist, produced mechanical allodynia in the orofacial area under normal conditions. However, intracisternal administration of bicuculline (50 ng) produced a paradoxical anti-allodynic effect under inflammatory pain conditions. Pretreatment with resiniferatoxin (RTX), which depletes capsaicin receptor protein in primary afferent fibers, did not alter the paradoxical anti-allodynic effects produced by the intracisternal injection of bicuculline. Intracisternal injection of bumetanide, an Na-K-Cl cotransporter (NKCC 1) inhibitor, reversed the IL-1β-induced mechanical allodynia. In the control group, application of GABA (100 µM) or muscimol (3 µM) led to membrane hyperpolarization in gramicidin perforated current clamp mode. However, in some neurons, application of GABA or muscimol led to membrane depolarization in the IL-1β-treated rats. These results suggest that some large myelinated Aβ fibers gain access to the nociceptive system and elicit pain sensation via GABAA receptors under inflammatory pain conditions.

A role for the purinergic receptor P2X3 in astrocytes in the mechanism of craniofacial neuropathic pain

The purinergic receptor P2X3, expressed in the central terminals of primary nociceptive neurons in the brainstem, plays an important role in pathological pain. However, little is known about expression of P2X3 in the brainstem astrocytes and its involvement in craniofacial pathologic pain. To address this issue, we investigated the expression of P2X3 in astrocytes in the trigeminal caudal nucleus (Vc) in a rat model of craniofacial neuropathic pain, chronic constriction injury of infraorbital nerve (CCI-ION). We found that 1) P2X3-immunoreactivity is observed in the brainstem astrocytes, preferentially in their fine processes, 2) the number of P2X3-positive fine astrocytic processes and the density of P2X3 in these processes were increased significantly in CCI-ION rats, compared to control rats, and 3) administration of MPEP, a specific mGluR5 antagonist, alleviated the mechanical allodynia and abolished the increase in density of P2X3 in fine astrocytic processes caused by CCI-ION. These findings reveal preferential expression of P2X3 in the fine astrocytic processes in the brainstem, propose a novel role of P2X3 in the fine astrocytic process in the mechanism of craniofacial neuropathic pain, and suggest that the expression of astrocytic P2X3 may be regulated by astrocytic mGluR5.