Baseline Latency Research Articles

A Randomized, Placebo-Controlled Trial of an Amino Acid Preparation on Timing and Quality of Sleep

This study was an outpatient, randomized, double-blind, placebo-controlled trial of a combination amino acid formula (Gabadone) in patients with sleep disorders. Eighteen patients with sleep disorders were randomized to either placebo or active treatment group. Sleep latency and duration of sleep were measured by daily questionnaires. Sleep quality was measured using a visual analog scale. Autonomic nervous system function was measured by heart rate variability analysis using 24-hour electrocardiographic recordings. In the active group, the baseline time to fall asleep was 32.3 minutes, which was reduced to 19.1 after Gabadone administration (P = 0.01, n = 9). In the placebo group, the baseline latency time was 34.8 minutes compared with 33.1 minutes after placebo (P = nonsignificant, n = 9). The difference was statistically significant (P = 0.02). In the active group, the baseline duration of sleep was 5.0 hours (mean), whereas after Gabadone, the duration of sleep increased to 6.83 (P = 0.01, n = 9). In the placebo group, the baseline sleep duration was 7.17 +/- 7.6 compared with 7.11 +/- 3.67 after placebo (P = nonsignificant, n = 9). The difference between the active and placebo groups was significant (P = 0.01). Ease of falling asleep, awakenings, and am grogginess improved. Objective measurement of parasympathetic function as measured by 24-hour heart rate variability improved in the active group compared with placebo. An amino acid preparation containing both GABA and 5-hydroxytryptophan reduced time to fall asleep, decreased sleep latency, increased the duration of sleep, and improved quality of sleep.

Re‐emerging microneurography

Re‐emerging microneurography

Successful Intraoperative Spinal Cord Monitoring During Scoliosis Surgery Using a Total Intravenous Anesthetic Regimen Including Dexmedetomidine

Intraoperative neurophysiological monitoring (IONM) during corrective spinal surgery is widely used. Because of the possible interference with the recording of evoked potentials by inhalational anesthetics, total intravenous anesthetic (TIVA) regimens have been advocated. TIVA regimens may be difficult to use in pediatric populations due to metabolic factors. We report on the results of multimodality IONM during 18 cases in which a TIVA regimen incorporating dexmedetomidine (Precedex, Hespira, Lake Forest, IL) was used. Monitoring techniques included sensory (SSEP) and motor evoked potentials (MEP), as well as pedicle screw stimulation. SSEPs were maintained within an acceptable range of baseline amplitude (50%) and latency (10%), and MEPs remained elicitable throughout each case. We therefore found that the anesthetic regimen did not significantly interfere with any of the monitoring modalities used and conclude that IONM in the presence of dexmedetomidine is feasible under appropriate conditions.

A novel cryptic peptide derived from the rat neuropeptide FF precursor reverses antinociception and conditioned place preference induced by morphine

Neuropeptide FF (NPFF) precursors from different species contain at least three known neuropeptides, i.e. FF (FLFQPQRF-NH 2), AF (AGEGLSSPFWSLAAPQR-NH 2) and SF (SLAAPQRF-NH 2). We demonstrate that the rat NPFF precursor contains another bioactive sequence, NAWGPWSKEQLSPQA, spanning between positions 85 and 99. Synthetic NPFF precursor (85–99) (10 and 20 nmol, i.c.v.) blocked the expression of conditioned place preference induced by morphine (5 mg/kg, s.c.). This peptide alone (10 and 20 nmol, i.c.v.) had no influence on the baseline latency of a nociceptive reaction but reversed the antinociceptive activity of morphine (5 mg/kg, s.c.) in the tail-immersion test in rats. These data suggest the existence of a novel bioactive cryptic peptide within an already known NPFF precursor.

Phenotypic characterization of spatial cognition and social behavior in mice with ‘knockout’ of the schizophrenia risk gene neuregulin 1

Neuregulin-1 (NRG1) has been identified as a candidate susceptibility gene for schizophrenia. In the present study the functional role of the NRG1 gene, as it relates to cognitive and social processes known to be disrupted in schizophrenia, was assessed in mice with heterozygous deletion of transmembrane (TM)-domain NRG1 in comparison with wildtypes (WT). Social affiliative behavior was assessed using the sociability and preference for social novelty paradigm, in terms of time spent in: (i) a chamber containing an unfamiliar conspecific vs. an empty chamber (sociability), or (ii) a chamber containing an unfamiliar conspecific vs. a chamber containing a familiar conspecific (preference for social novelty). Social dominance and aggressive behavior were examined in the resident–intruder paradigm. Spatial learning and memory were assessed using the Barnes maze paradigm, while spatial working memory was measured using the continuous variant of the spontaneous alternation task. Barnes maze data revealed intact spatial learning in NRG1 mutants, with elevated baseline latency to enter the escape hole in male NRG1 mutants reflecting an increase in activity level. Similarly, although a greater number of overall arm entries were found, spontaneous alternation was unaffected in NRG1 mice. Social affiliation data revealed NRG1 mutants to evidence a specific loss of WT preference for spending time with an unfamiliar as opposed to a familiar conspecific. This suggests that NRG1 mutants show a selective impairment in response to social novelty. While spatial learning and working memory processes appear intact, heterozygous deletion of TM-domain NRG1 was associated with disruption to social novelty behavior. These data inform at a novel phenotypic level on the functional role of this gene in the context of its association with risk for schizophrenia.

Characterization of the antinociceptive effects of oxycodone in diabetic mice

We investigated the antinociceptive efficacy of systemic and centrally injected oxycodone on thermal hyperalgesia in streptozotocin-induced diabetic mice. The antinociceptive response was assessed by recording the latency in the tail-flick test using the radiant heat from a 50-W projection bulb on the tail. The tail-flick latency in diabetic mice was significantly shorter than that in non-diabetic mice. Oral (p.o.) and i.t., but not i.c.v., administration of oxycodone prolonged the tail-flick latency in diabetic mice to a level that was considerably longer than the baseline latency in non-diabetic mice. However, morphine did not significantly inhibit the tail-flick response in diabetic mice. The antinociceptive effect of either p.o. or i.t. oxycodone in non-diabetic mice, but not in diabetic mice, was antagonized by pretreatment with a selective μ-opioid receptor antagonist, β-funaltrexamine. In non-diabetic mice, pretreatment with a selective κ-opioid receptor antagonist, nor-binaltorphimine, had no effect on the peak antinociceptive effect of either p.o. or i.t. oxycodone at 30 min after administration, however, it slightly but significantly reduced oxycodone-induced antinociception at 60 and 90 min after administration. On the other hand, pretreatment with nor-binaltorphimine practically abolished the antinociceptive effects of both p.o.- and i.t.-administered oxycodone in diabetic mice. Naltrindole, a selective δ-opioid receptor antagonist, had no effects on the antinociceptive effect of oxycodone in either non-diabetic or diabetic mice. These results suggest that the antinociceptive effects of oxycodone may be mediated by spinal κ-opioid receptors in diabetic mice, whereas it may interact primarily with supraspinal and spinal μ-opioid receptors in non-diabetic mice.

The Role of Adrenergic and Cholinergic Transmission in Volatile Anesthetic-Induced Pain Enhancement

Volatile anesthetic drugs have a biphasic effect on pain transmission. At very small concentrations they enhance pain sensitivity whereas at larger subanesthetic concentrations they have an analgesic effect. Previous work has suggested that nicotinic inhibition could mediate the pronociceptive action of isoflurane. Furthermore, activation of nicotinic receptors facilitates the release of norepinephrine in the spinal cord. We hypothesize that nicotinic modulation of norepinephrine release in the spinal cord mediates isoflurane's pronociceptive action. We used hindpaw withdrawal latency as a measure of pain sensitivity after inhibition of adrenergic activity or treatment with nicotine in mice. Isoflurane's effect on pain is separable by concentration. The 50% effective concentration for pain enhancement is 0.16% isoflurane whereas the 50% effective concentration for the antinociceptive action of isoflurane is 0.8%. Depletion of systemic norepinephrine with the neurotoxin DSP-4 caused a reduction in baseline withdrawal latencies and prevented isoflurane pronociception. Baseline latency was also reduced by intrathecal yohimbine. After treatment with yohimbine, isoflurane had no additional pronociceptive effect. Nicotine administered through intracerebroventricular injection increased baseline latency but did not prevent isoflurane pronociception. Conversely, intrathecal applications of nicotine caused a slight reduction in baseline latency and prevented isoflurane's pronociceptive effect. We conclude that spinal noradrenergic transmission seems to be necessary for isoflurane pronociception to occur. Isoflurane may act by inhibiting tonically active nicotinic receptors that modulate the release of norepinephrine in the spinal cord.

Differential recovery of the electroretinogram, visually evoked cortical potential, and electrically evoked cortical potential following vitrectomy: Implications for acute testing of an implanted retinal prosthesis

To determine the extent to which electrophysiologic tests of the afferent visual pathway are affected by vitrectomy, the procedure was performed in 15 eyes of 11 adult Dutch-belted rabbits. An electroretinogram (ERG), visually evoked cortical potential (VECP), and electrically evoked cortical potential (EECP) were obtained preoperatively and sequentially after surgery. For electrical stimulations, biphasic impulses were delivered to the retina. Post-vitrectomy declines of 49, 25, and 41% from the median baseline amplitudes and increases of 13, 18, and 17% from the median baseline latency values were found for ERG, VECP, and EECP, respectively. At 90 min, 13 to 30% of eyes still had an amplitude more than 10% below baseline on at least one of the three tests, whereas 10 to 47% of eyes had an abnormal latency more than 10% above baseline on at least one of the three tests. Amplitudes were more likely than latencies to return to near baseline, but for eyes that remained subnormal, the decline was greater for amplitudes than latencies. Significant alterations in retinal function, manifested by declines in amplitudes and increases in latencies of the ERG, VECP, and EECP, persist in a large proportion of eyes up to 90 min post-vitrectomy.

Aromatase (Estrogen Synthase) Activity in the Dorsal Horn of the Spinal Cord: Functional Implications

The presence of aromatase (estrogen synthase) in neurons in the dorsal horn of the spinal cord in Japanese quail suggests that estrogens produced locally from androgens could control spinal sensory processes including nociception. We used the hot water nociceptive test (54 degrees C) to appraise the long-term effect of an inhibition of aromatization on the foot withdrawal latency in male quail. Four weeks after the ablation of their main source of testosterone (testes), castrated males displayed a significantly higher foot withdrawal latency than gonadally intact males. A prolonged treatment with subcutaneous capsules filled with testosterone or 17 beta-estradiol restored the baseline latency within 2 weeks. The effect of testosterone in castrated quail was almost completely blocked by systemic injections of Vorozole(TM), a nonsteroidal aromatase inhibitor or tamoxifen, an estrogen receptor antagonist (one injection per day for 10 days). Taken together, these data demonstrate for the first time to our knowledge an effect of estrogens formed by aromatization of androgens on nociception. Because aromatase-immunoreactive neurons and aromatase activity are present in the dorsal horns of the spinal cord, this control of pain thresholds is presumably mediated, at least in part, by estrogens produced at the spinal level that act locally via slow, presumably genomic, mechanisms mediated by the activation of spinal nuclear estrogen receptors.

Use of the elevated T-maze to study anxiety in mice

We have recently suggested that the elevated T-maze (ETM) is not a useful test to study different types of anxiety in mice if a procedure similar to that originally validated for rats is employed. The present study investigated whether procedural (five exposures in the enclosed arm instead of three as originally described for rats) and structural (transparent walls instead of opaque walls) changes to the ETM leads to consistent inhibitory avoidance acquisition (IAA) and low escape latencies in mice. Results showed that five exposures to the ETM provoked consistent IAA, an effect that was independent of the ETM used. However, the ETM with transparent walls (ETMt) seemed to be more suitable for the study of conditioned anxiety (i.e. IAA) and unconditioned fear (escape) in mice, since IAA (low baseline latency with a gradual increase over subsequent exposures) and escape (low latency) profiles rendered it sensitive to the effects of anxiolytic and anxiogenic drugs. In addition to evaluation of drug effects on IAA and escape, the number of line crossings in the apparatus were used to control for locomotor changes. Results showed that whereas diazepam (1.0–2.0 mg/kg) and flumazenil (10–30 mg/kg) impaired IAA, FG 7142 (10–30 mg/kg) did not provoke any behavioral change. Significantly, none of these benzodiazepine (BDZ) receptor ligands modified escape latencies. The 5-HT 1A partial receptor agonist buspirone (1.0–2.0 mg/kg) and the 5-HT releaser fenfluramine (0.15–0.30 mg/kg) impaired IAA and facilitated escape, while the full 5-HT 1A receptor agonist, 8-OH-DPAT (0.05–0.1 mg/kg) and the 5-HT 2B/2C receptor antagonist, SER 082 (0.5–2.0 mg/kg) failed to modify either response. mCPP (0.5–2.0 mg/kg), a 5-HT 2B/2C receptor agonist, facilitated IAA but did not alter escape latency. Neither antidepressant utilized in the current study, imipramine (1.0–5.0 mg/kg) and moclobemide (3.0–10 mg/kg) affected IAA or escape performance in mice. The well-known anxiogenic drugs yohimbine (2.0–8.0 mg/kg) and caffeine (10–30 mg/kg) did not selectively affect IAA, although caffeine did impair escape latencies. Present results suggest the ETMt is useful for the study of conditioned anxiety in mice. However, upon proximal threats (e.g. open arm exposure), mice do not exhibit escape behavior as an immediate defensive strategy, suggesting that latency to leave open arm is not a useful parameter to evaluate unconditioned fear in this species.

Mechanical and thermal hypersensitivity develops following kainate lesion of the ventral posterior lateral thalamus in rats

Damage to the ventral-posterior lateral nucleus (VPL) of the thalamus or its afferent pathways can produce moderate to severe on-going pain and pain in response to normally innocuous stimuli (allodynia) and hypersensitivity to mildly noxious stimuli (hyperalgesia). The present study measured the responses to mechanical and thermal stimuli before and 2, 8, 24 and 48 h after a kainate-induced lesion of the VPL in male rats. Compared with control animals, hypersensitivity to mechanical stimulation of the hindpaw was apparent by 24 h post-lesion. At 48 h, the frequency of mechanical response increased from a baseline response frequency of 17±4.7 to 46±11.6%. Thermal withdrawal latencies 48 h after the lesion decreased from a baseline latency of 9.9±1.8 to 5.3±0.4 s. It is concluded that a neurotoxic lesion of the VPL results in a delayed onset of mechanical and thermal hyperalgesia. This study suggests a potential model for studying the basic mechanisms and potential therapies for central pain syndrome.

Dissociative Tendencies, Attention, and Memory

Two groups of college students were selected on the basis of their scores on the Dissociative Experiences Scale (DES). The high-DES group (score > 20, M = 29.6; n = 54) and low-DES group (score < 10, M = 5.1; n = 54) both completed the standard and a new dualtask version of the Stroop ink-naming task with xs (baseline condition) and color, neutral, and emotionally charged words. Free recall results indicated that high-DES participants remembered fewer emotionally charged words than low-DES participants. We found a crossover interaction for Stroop interference: High-DES participants showed more interference (conflicting color – baseline latency for ink naming) in a selective-attention Stroop task and less interference in the dualtask Stroop task. The interaction between attentional context and dissociation for Stroop interference and the free recall results are consistent with a cognitive-environments view of dissociative tendencies. In this view, dissociative tendencies, which have been otherwise speculated to be largely deleterious, can be advantageous in certain contexts.

PHARMACOLOGICAL CHARACTERIZATION OF A NOVEL CHIMERIC ANALGESIC PEPTIDE AA 501

S361 INTRODUCTION: Many physiological and pharmacological studies have indicated that substance P and opioids are functionally antagonistic in the mediation of nociception and antinociception in vivo. We have previously demonstrated that biphalin, a novel opioid tetrapeptide dimer (Tyr-D-Ala-Gly-Phe-NH-)2, given intrathecally with a peptide substance P antagonist produces potent spinal antinociception in rats (Misterek et al., 1994) [1]. AA 501, a novel compound, binds both to opioid and substance P receptors. Because this novel peptide construct consists of the opioid sequence Tyr-D-Ala-Gly-Phe- covalently linked through a hydrazide bridge to a Z-Trp (N-alpha-Carbobenzoxy-Trp) moiety, and this moiety has putative neurokinin 1 (NK-1) receptor blocking effects, we have now evaluated the activity of AA 501. MATERIAL AND METHODS: After Animal Research Committee approval, adult male Sprague-Dawley rats (225-250 g) were implanted with chronic indwelling IT catheters with tips at the T13-L1 spinal level. Animals had one week to recover from surgery, during which they were habituated daily to the laboratory environment and analgesic testing apparatus. For evaluation of antinociceptive properties of AA 501 we used tail-flick test and formalin test. For measurement of thermal antinociception, a tail-flick apparatus was utilized (baseline latency approximately 3 sec, cutoff 10 sec). Responses were expressed as % maximum possible effect (%MPE): [%MPE=(post-treatment latency-baseline latency)/(cut off time-baseline latency) x100]. In the formalin test 50 [micro sign]l of 5% formalin solution was injected s.c. into the dorsal surface of the rat hind paw. Pain behavior was quantified by periodically counting the occurrence of spontaneous flinching/shaking of the injected paw. AA501 was administered IT 15 minutes before or 9 min after formalin injection. RESULTS: IT administration of AA501 produced dose-dependent antinociception in tail-flick and formalin tests. The time course of antinociceptive responses to lower doses of AA501 (0.25 and 0.5 [micro sign]g) peaked at approximately 50% MPE at 15-30 min and declined to 20-30% MPE at 60 min. Intermediate doses (1 and 4 [micro sign]g) produced an antinociceptive effect that reached 60-80% MPE at 5-45 min. Higher doses (8 and 25 [micro sign]g) produced a maximal antinociceptive response (100% MPE) of long duration (30-75 min). Antinociceptive effects of AA501 were reversed by 10 min pretreatment with naloxone IT 10 [micro sign]g and diminished by SP IT 150 ng. AA501 administered IT in dose 1 [micro sign]g 15 min before formalin injection into the rat hind paw completely blocked the biphasic response to formalin. AA501 administered 9 min after formalin injection also blocked occurrence of the second phase of the response to formalin. DISCUSSION: We have previously shown that a hybrid peptide containing both beta-casomorphin-like and substance P-like structural characteristics possessed an antinociceptive effect in the mouse hot plate test after intrathecal administration (Lipkowski et al., 1994) [2]. AA501 was designed to concurrently activate opioid receptors and block NK-1 receptors. The antinociceptive effects of IT AA501 in the tail-flick and formalin tests are highly significant and dose-dependent, suggesting its value as a potential therapeutic agent or useful tool for study on pain modulation. [Department of Anesthesia funds, NIDA Grant 04128 and The Richard Saltonstall Charitable Foundation.]

Auditory brainstem response forward-masking recovery functions in older humans with normal hearing

We investigated the auditory brainstem response (ABR) recovery from forward masking using toneburst maskers and probes. Two subject groups matched for hearing thresholds were evaluated: normal-hearing young adults (21–40 years) and older subjects (63–77 years) with normal audiometric thresholds. Stimuli consisted of 1, 4 and 8 kHz tonebursts, with 2–4 cycle rise/fall time and no plateau. Forward maskers were tonebursts of the same frequency, with a 5 ms rise/fall time and a 20 ms plateau time. Probes were presented at 40 dB above threshold, and the forward masker was adjusted to a level that just eliminated the ABR to the 40 dB sensation level toneburst when the probe onset occurred at masker offset. Forward-masker intervals varied from 2 to 64 ms. ABR wave V latencies were similar for the young and old age groups regardless of toneburst frequency. Under forward-masking conditions, wave V latency was prolonged for the shorter intervals, and recovered to baseline latency by 64 ms. The forward-masker recovery functions were nearly identical for the two age groups for the 1 kHz toneburst. In contrast, there were clear differences in the recovery functions for the two age groups for the 4 and 8 kHz tonebursts. Specifically, the mean latency shift was greater for the aged group for forward-masker intervals of 16 ms or less. The two age groups showed identical latency shifts for longer forward-masker intervals. These data demonstrate prolonged recovery from forward masking in older human subjects. As these subjects had audiometric thresholds within normal limits, one plausible interpretation of this finding is that the prolonged recovery time is a manifestation of an aging effect on the central auditory nervous system rather than the periphery.

Antinociceptive and morphine modulatory actions of spinal orphanin FQ

Orphanin FQ (nociceptin, OFQ), a heptadecapeptide peptide, has been designated as an endogenous ligand at the orphan receptor ORL1, which lacks affinity for opioid receptor ligands. OFQ-like immunoreactivity has been localized in spinal cord areas that are involved in the processing of nociceptive signals. In this study, the effects of spinally administered OFQ on thermal and mechanical nociceptive stimuli were investigated following intrathecal (i.t.) injection in unanesthetized rats bearing chronic indwelling catheters in the subarachnoid space. Intrathecal OFQ produced two distinct acute actions: an opioid-like antinociceptive effect, and antagonism of morphine-induced antinociception. Chronic administration produces tolerance. Acute actions were as follows. First, injection of OFQ alone in the dose range 10-100 nmol produced sustained antinociceptive effects in the tail-flick (baseline latency 1.5-2 s) and paw-pressure tests. These effects peaked at 45-60 min post-injection, were fully reversible, and were observed in absence of discernable motor impairments. Intrathecal naloxone (5 nmol) significantly attenuated the antinociceptive effects of OFQ (50 nmol i.t.) in tail-flick and paw-pressure tests. A 24-h pretreatment with the irreversible opioid receptor antagonist, beta -flunatrexamine (2 nmol i.t.), also attenuated the antinociceptive effects of OFQ (50, 100 nmol i.t.) and of morphine (7.5 nmol i.t.) in both tests. Low doses (1, 5 nmol) of OFQ, which failed to produce antinociception in the tail-flick test involving a baseline latency of 1.5-2 s, produced a strong antinociceptive response when the baseline latency was increased to 5-6 s. Second, intrathecal OFQ, at doses (0.5, 1.0, and 5.0 nmol) that had no antinociceptive activity in the tail-flick (baseline latency 1.5-2 s) and paw-pressure tests, attenuated the antinociceptive effect of morphine (7.5 nmol i.t.) in these tests. However, a threshold OFQ dose (10 nmol) significantly extended the duration of antinociception induced by morphine (7.5 nmol i.t.) or deltorphin (20 nmol i.t.). Chronic actions were as follows. In rats that were rendered tolerant to spinal morphine, by a continuous intrathecal infusion (7.5 nmol/h) of the agonist for 5 days, the OFQ dose-response curves for its antinociceptive effect in the tail-flick and paw-pressure tests were significantly shifted to the right. In separate experiments, repeated intrathecal injection of OFQ (50 nmol) or morphine (7.5 nmol) produced a significant decline in their antinociceptive effects. Thus, intrathecally administered OFQ produces both development of tolerance to its antinociceptive actions and cross-tolerance to the action of morphine.Key words: orphanin FQ, nociceptin, spinal antinociception, opioid antagonism, tolerance, cross-tolerance.

Antinociceptive and morphine modulatory actions of spinal orphanin FQ

Orphanin FQ (nociceptin, OFQ), a hepatdecapeptide peptide, has been designated as an endogenous ligand at the orphan receptor ORL1, which lacks affinity for opioid receptor ligands. OFQ-like immunoreactivity has been localized in spinal cord areas that are involved in the processing of nociceptive signals. In this study, the effects of spinally administered OFQ on thermal and mechanical nociceptive stimuli were investigated following intrathecal (i.t.) injection in unanesthetized rats bearing chronic indwelling catheters in the subarachnoid space. Intrathecal OFQ produced two distinct acute actions: an opioid-like antinociceptive effect, and antagonism of morphine-induced antinociception. Chronic administration produces tolerance. Acute actions were as follows. First, injection of OFQ alone in the dose range 10-100 nmol produced sustained antinociceptive effects in the tail-flick (baseline latency 1.5-2 s) and paw-pressure tests. These effects peaked at 45-60 min post-injection, were fully reversible, and were observed in absence of discernable motor impairments. Intrathecal naloxone (5 nmol) significantly attenuated the antinociceptive effects of OFQ (50 nmol i.t.) in tail-flick and paw-pressure tests. A 24-h pretreatment with the irreversible opioid receptor antagonist, beta-flunatrexamine (2 nmol i.t.), also attenuated the antinociceptive effects of OFQ (50, 100 nmol i.t.) and of morphine (7.5 nmol i.t.) in both tests. Low doses (1, 5 nmol) of OFQ, which failed to produce antinociception in the tail-flick test involving a baseline latency of 1.5-2 s, produced a strong antinociceptive response when the baseline latency was increased to 5-6 s. Second, intrathecal OFQ, at doses (0.5, 1.0, and 5.0 nmol) that had no antinociceptive activity in the tail-flick (baseline latency 1.5-2 s) and paw-pressure tests, attenuated the antinociceptive effect of morphine (7.5 nmol i.t.) in these tests. However, a threshold OFQ dose (10 nmol) significantly extended the duration of antinociception induced by morphine (7.5 nmol i.t.) or deltorphin (20 nmol i.t.). Chronic actions were as follows. In rats that were rendered tolerant to spinal morphine, by a continuous intrathecal infusion (7.5 nmol/h) of the agonist for 5 days, the OFQ dose-response curves for its antinociceptive effect in the tail-flick and paw-pressure tests were significantly shifted to the right. In separate experiments, repeated intrathecal injection of OFQ (50 nmol) or morphine (7.5 nmol) produced a significant decline in their antinociceptive effects. Thus, intrathecally administered OFQ produces both development of tolerance to its antinociceptive actions and cross-tolerance to the action of morphine.

Involvement of nitric oxide and serotonin in modulation of antinociception and pressor responses evoked by stimulation in the dorsolateral region of the periaqueductal gray matter in the rat

In rats anaesthetized with alphaxalone/alphadolone, electrical stimulation in the periaqueductal gray matter in the region lying lateral and dorsolateral to the aqueduct produced a pressor response and an increase in the latency of the tail flick response to noxious heat applied to the tail. The antinociception and the pressor response were significantly attenuated following microinjection of 15 nmol 5-hydroxytryptamine at the site of stimulation in the periaqueductal gray matter. Microinjection of an equal volume of 165 mM saline had no effect. The inhibitory effects of 5-hydroxytryptamine were blocked by prior intracerebroventricular administration of 100 μg of the nitric oxide synthase inhibitor l-nitroarginine methyl ester. Neither 5-hydroxytryptamine or l-nitroarginine methyl ester had any effect on resting arterial pressure or on the baseline latency of the tail flick reflex. It is suggested that the inhibitory effects of 5-hydroxytryptamine in the dorsolateral periaqueductal gray matter are normally dependent on the functional integrity of local nitric oxide synthase-containing interneurons. Nitric oxide may act in association with 5-hydroxytryptamine to control the excitability of the aversive system in the midbrain.

Effect of transient naloxone antagonism on tolerance development in rats receiving continuous spinal morphine infusion

The magnitude of tolerance and dependence is defined in part by agonist concentration and duration of receptor exposure. Therefore, in the face of continued exposure to an opioid agonist, periodic reduction in opiate receptor occupancy should reduce tolerance. Alternately, we have shown that reversal of opiate agonist action yields increased glutamate release and NMDA-antagonist studies indicated that this release may lead to an exacerbation of tolerance. To address this issue, we observed the effect of transient daily antagonism by naloxone of otherwise continuous opioid receptor exposure on morphine tolerance development. Rats with intrathecal (i.t.) catheters and osmotic minipumps were assigned to one of the following 7-day infusion/treatment groups: group A: i.t. morphine (20 nmol/h) with daily subcutaneous (s.c.) injection of naloxone 0.6 mg/kg, group B: i.t. morphine (20 nmol/h) with daily s.c. saline, group C: i.t. saline (1 μl/h) with daily s.c. injection of naloxone 0.6 mg/kg, or, group D: i.t. saline (1 μl/h) with daily s.c. saline. Hot plate response latency was measured daily before and after the s.c. injection. The infusion was discontinued on day 7 and on day 8 the response of the rat to a probe dose of i.t. morphine (60 nmol) given as a bolus was observed. Elevated hot plate latencies were observed for groups A and B on day 1 of infusion and this declined over the following 3-4-day interval. Group B approached baseline, but by day 5 group A showed a mild hyperalgesia prior to each naloxone injection. Groups C and D showed no change in baseline latency. On day 8, 24 h after termination of morphine infusion, the magnitude of the analgesic response to the probe i.t. morphine was: group D=group C>group B>group A ( P<0.05, 1-way ANOVA). Thus, in contrast to the expectation that tolerance would be reduced by periodic blockade of opiate receptor occupancy, rats that had daily transient receptor antagonism showed a greater tolerance than rats with simple continuous receptor occupancy. These results are, however, consistent with work showing that (i) naloxone will evoke spinal glutamate release in spinal morphine tolerant rats and (ii) spinal NMDA receptor antagonism ameliorates loss of opiate effect in this spinal infusion model.

Behavioral validation of the elevated T-maze, a new animal model of anxiety.

The elevated T-maze test of anxiety has been used to separate in the same rat conditioned from unconditioned responses of fear/anxiety. The test apparatus consists of three elevated arms-one enclosed and two open. Inhibitory avoidance--representing learned fear--is measured by recording the time taken to leave the enclosed arm in three consecutive trials. Unconditioned fear is evaluated by recording the time to escape from the open arm. In this study we investigated procedural questions raised by the use of the elevated T-maze. Experiment 1 showed that restraining the animals at the end of the enclosed arm for 30 s did not change the first (baseline) latency to leave this arm, indicating that aversion for the hands of the experimenter is not a key motivation for this response. The results of Experiment 2 indicated that open-arm experience, but not handling stress is the main cause for inhibitory avoidance acquisition, because rats trained in a T-maze with the three arms enclosed did not show the usual increase in withdrawal latency along the three consecutive trials. The same experiment also showed that the latency to leave the open arm did not undergo habituation over five consecutive trials, evidencing an aversive motivation for this response. The importance of open-arm experience for inhibitory avoidance acquisition was further suggested by the results of Experiment 3, as the removal of a shield that prevents perception of openness tended to increase avoidance latency in the elevated T-maze from the first trial.

Thermal hyperalgesia in rat evoked by intrathecal substance P at multiple stimulus intensities reflects an increase in the gain of nociceptive processing

Substance P (SP) is associated with primary afferent depolarization after a noxious or tissue-damaging peripheral stimulus and is proposed to be a ‘pain transmitter’. However, current work has shown that activation of spinal neurokinin receptors evokes few signs of ‘pain behavior’, but rather decreases nociceptive thresholds, suggesting a modulatory role through additive or multiplicative mechanisms. The current study addressed this question by examining the change in nociceptive threshold under three different radiant thermal stimuli after intrathecal SP administration in unanesthetized rats. With increasing stimulus intensity, the numerical change in withdrawal latency after intrathecal SP decreased, but the fractional change from baseline latency did not change, suggesting that SP increases the gain of spinal nociceptive processing.