Spinal mu, delta and kappa opioids alter chemical, mechanical and thermal sensitivities in amphibians.

Previously we demonstrated the use of chemical (topical acetic acid), thermal (radiant heat) and mechanical (von Frey filament) stimuli as quantifiable behavioral response assays in the northern grass frog, Rana pipiens. Furthermore, response thresholds in all of these sensory modalities are significantly elevated by systemic morphine injections, which can be antagonized by naltrexone. The present study employed these three sensory assays to assess changes in chemical, mechanical and thermal sensitivities following spinal administration of mu, delta and kappa opioids. Significant elevations were observed across all three sensory modalities in each subtype category and these effects were abolished by prior systemic administration of naltrexone. However, naltrexone antagonism of morphine produced hyperalgesia in both the mechanical and thermal modalities. The results support other recent work demonstrating that the spinal site for opioid analgesia is present in amphibians and that the thermal, mechanical and acetic acid assays are measures of true nociceptive activity in the amphibian.

Thermal, mechanical and chemical peripheral sensation in amphibians: opioid and adrenergic effects.

The acetic acid test (AAT) is a quantifiable assay of the response to noxious chemical stimuli on the hindlimb of the northern grass frog, Rana pipiens. AAT is sensitive to both opioid and adrenergic agonist modulation. The present study introduces the novel use of mechanical and thermal stimulus behavioral assays in comparison with the established acetic acid test in studying nociception in frogs. We evaluated mechanical and thermal responses and their sensitivity to systemic morphine (MOR) or dexmedetomidine (DEX) administrations with comparison to AAT. MOR produced dose-related elevations of response thresholds in all three sensory tests, whereas DEX produced elevations in the thermal and AAT assays but had no effect on sensitivity to non-noxious mechanical stimuli. The results suggest a distinct separation of sensory modalities in the frog similar to that observed in mammals and indicate the usefulness of this amphibian model in further studies of nociception.

Sciatic cryoneurolysis in rats: a model of sympathetically independent pain. Part 1: Effects of sympathectomy.

The purpose of this study was to evaluate the effects of preemptive and postlesion sympathectomy in the sciatic cryoneurolysis (SCN) model of neuropathic pain in rats. SCN in rats produces a prolonged significant mechanical allodynia (hypersensitivity to previously non-noxious mechanical stimuli) with no thermal hyperalgesia. In at least two other models, sympathectomy is effective in attenuating existing mechanical allodynia and thermal hyperalgesia or deterring their development after nerve injury. These models appear to mimic the direct sympathetic involvement characteristic of the clinical syndrome termed sympathetically maintained pain (SMP). To investigate these concepts in the SCN model, sympathectomy was performed prior to SCN in animals with established SCN-induced allodynia. Sympathectomy did not alter the pattern of existing allodynia or its development in this model. The results suggest that SCN is a useful and easily reproducible model of sympathetically independent pain (SIP).

Sciatic cryoneurolysis in rats: a model of sympathetically independent pain. Part 2: Adrenergic pharmacology.

The purpose of this study was to investigate the possibility that sympathetic exacerbation of neuropathic pain activity may be mediated in part by the development of abnormal peripheral adrenergic mechanisms at the site of peripheral nerve injury. We evaluated changes in mechanical stimulation withdrawal thresholds before and after the delivery of selected alpha-adrenergic agonists and antagonists to the immediate area of a prior nerve injury in rats that had developed continuous mechanical allodynia subsequent to sciatic cryoneurolysis (SCN). Allodynia was attenuated (thresholds increased) after administration of the alpha 1 antagonist prazosin, but not the alpha 2 antagonist idazoxan. Similar attenuation occurred with infusions of the alpha 2 agonist dexmedetomidine and at the lowest dose (2.0 micrograms) of the mixed alpha agonist clonidine. In contrast, allodynia was exacerbated (thresholds decreased) by infusion of the highest dose of clonidine (20 micrograms). Infusions of the alpha 2 agonist ST-91, a polar analog of clonidine, did not alter withdrawal thresholds. The findings suggest that a minimal peripheral adrenergic modulation of SCN-induced allodynia occurs via mechanisms that are not localized to the prior injury site. However, overall, the SCN model of neuropathic pain appears to be resistant to adrenergic interventions, providing further evidence that SCN in rats is a model of sympathetically independent pain (SIP).

Spinal dynorphin immunoreactivity increases bilaterally in a neuropathic pain model.

Increased spinal levels of dynorphin, an endogenous opioid kappa agonist, are seen in models of both chronic and acute hyperalgesia. This study determined the extent and localization of spinal immunoreactive dynorphin following sciatic cryoneurolysis (SCN), a neuropathic pain model produced by a peripheral nerve freeze lesion. SCN results in behaviors associated with neuropathic pain such as autotomy (the gnawing and scratching of the affected limb), touch-evoked and mechanical allodynia, and spontaneous nociceptive behavior. Following SCN, 4 rats that displayed autotomy and 3 rats that did not were randomly chosen for immunohistochemical staining of dynorphin-like immunoreactivity (DLIR). The area of DLIR above a standardized threshold level was quantified in both dorsal horns of each spinal cord section using a computer-assisted image analyzer to express DLIR in pixels. DLIR was observed both ipsilateral and contralateral to the injured peripheral nerve. In addition, the area of DLIR was significantly greater (P = 0.05) in rats that showed autotomy behavior (mean = 52.6 x 10(3) +/- 25.6) compared to rats with no autotomy (mean = 13.8 x 10(3) +/- 4.78). In sharp contrast to the ipsilateral dynorphin increases observed in other neuropathic pain models, we observe a bilateral increase at 21 days following SCN.

Intrathecal catheterization alone reduces autotomy after sciatic cryoneurolysis in the rat.

There is substantial evidence that sciatic cryoneurolysis (SCN, freeze lesion of the sciatic nerve) is a neuropathic pain model in the rat. During characterization of this model, SCN was performed 4 days after either a sham operation or the insertion of an indwelling intrathecal catheter preparatory to selective spinal drug administration. Body weight and autotomy scores were recorded for the next 22 days until sacrifice. The catheter group experienced significant weight loss (7.5%) by 4 days but rapidly regained to parity with the sham group. Autotomy scores and the frequency of severe autotomy (score > 3) were less at day 22 in the catheter group as compared with the sham-control group (p < 0.005, p < 0.03, respectively). Intrathecal catheterization itself effects the degree of behavioral response to neurogenic pain and thus, should be controlled for in studies using nociceptive animal models.