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Background: Assessment of the nociceptive withdrawal reflex (NWR) is frequently applied to probe the excitability level of the spinal nociceptive circuitry. In humans, the NWR threshold (NWR-T) is often estimated by applying electrical stimulation over the sural nerve at the lateral malleolus. Such stimulation may be associated with substantial pain and discomfort rendering completion of the assessment infeasible., New Method: As an alternative to sural nerve stimulation, NWR-Ts were also estimated by electrical stimulation at the arch of the foot. Failure-rates and test-retest reliability of these two procedures were evaluated. A fully-automated interleaved up-down staircase procedure was used to estimate the NWR-T for both stimulation sites. NWRs were detected from EMG measured over the biceps femoris and tibialis anterior muscles, respectively. A total of three repeated measures were performed in two different sessions to evaluate the test-retest reliability of the two methods using Bland-Altman agreement analysis., Results: The failure rate of NWR-T estimation based on electrical stimulation of the sural nerve (29%) was substantially higher than when the NWR was elicited by stimulation at the arch of the foot (5%)., Comparison With Existing Method: The analysis of test-retest reliability indicated that the two methods for NWR-T estimation were equally reliable for within-session comparisons, but stimulation at the arch of the foot enabled NWR-T estimation with superior between-session reliability, Conclusions: These results support a paradigm shift within NWR-T estimation favoring stimulation at the arch of the foot., (Copyright © 2015 Elsevier B.V. All rights reserved.)

This study describes a modified Hargreaves' method for assessing paw withdrawal threshold temperatures for heat (PWT-H) nociception in the hind paws of rats. This method utilises radiant heat to maintain controlled lamp temperatures (CLTs) on a glass floor beneath the rat hind paw. An ascending series of CLTs were applied for 10s, with 5-10min intervals between applications, until characteristic withdrawal behaviour was observed or a cutoff CLT was reached. The average plantar epicutaneous temperatures measured from anaesthetised rats corresponding to CLTs and withdrawal latencies were used for determining PWT-H. The mean PWT-H in 2-month-old (mo) naïve Sprague-Dawley rats (n=38) was 47.6±0.2°C, which is comparable to the noxious threshold temperature for human glabrous skin (46.5±0.5°C). The PWT-H was consistent between trials and daily assessments over four consecutive days. No significant differences were observed between the PWT-H in 2-, 6- to 8-, and >24-mo F344 rats, but the PWT-H in 1-mo rats was significantly reduced. Three hours following plantar incision, the PWT-H decreased to 37.5±0.2°C, consistent with previous observations of C-fibre afferents from incised glabrous skin firing at 36.7±3.6°C. Parallel testing, using the current method and an electronic von Frey device, illustrated similar degrees of incision-induced hyperalgesia, gradual improvements in hyperalgesia, and reversals induced through morphine and gabapentin. In conclusion, the present method facilitates a comparison of PWT-H using electrophysiological and human psychophysical studies involving thermosensation, and as a behavioural assay identical to von Frey testing, this method also measures the threshold for nociception., (Copyright © 2013 Elsevier B.V. All rights reserved.)

The sciatic functional index (SFI) is a remarkable tool to assess dysfunction and functional recovery of the sciatic nerve of rats. Usually measured on hind foot imprints on paper, a new method is now being proposed, by direct analysis of video recorded foot sole images obtained with a treadmill-type walking belt machine functioning with gait speed control (G1). Results were compared with the SFI measured on imprints on paper (G2) and on video recorded foot sole images obtained with a static see-through runway (G3). The right sciatic nerve of 19 adult female Wistar rats was crushed by the application of a controlled load. Impressions/images obtained both preoperatively and at weekly intervals for eight consecutive postoperative weeks were digitized, stored and analyzed in a computer loaded with specific software, the SFI being automatically calculated after measuring the appropriate parameters. SFI differed significantly between G1 and G2 and G1 and G3 (p<0.05), but not between G2 and G3 (p>0.05) during the first and second postoperative weeks, nonsignificant differences (p>0.05) being observed for any comparison between groups during the third through eighth postoperative weeks. We conclude that the three methods yielded equivalent results from the third week onward, but both video recording methods (G2 and G3) permitted a more adequate early evaluation (first and second weeks), since the SFI parameters were more easily identifiable. Images obtained with the walking belt machine are more uniform and sharper, thus contributing to reduce the influence of biases observed with imprints on paper., ((c) 2010 Elsevier B.V. All rights reserved.)

Neurogenic inflammation (NI) is a feature of several inflammatory pain conditions in which females are overrepresented. Therefore, we asked if there are sex differences in the inflammatory response evoked by well known neurogenic stimuli. We compared the amount of plasma extravasation (PE), a measure of inflammation, in the hindpaw skin of male and female rats caused by subcutaneous injection of capsaicin, application of noxious heat (51 degrees C water bath) or electrical stimulation of the saphenous nerve. We also compared the amount of PE in males and females evoked by substance P (SP), the principal neurogenic mediator of PE. PE was quantified using a video camera and digital image analysis to measure changes in reflectance (pixel intensity, PI) of skin due to accumulation of extravasated Evans blue (EB) dye. The increase in PI induced by capsaicin was significantly greater in females compared to males (p<0.001) and in estrus, diestrus, and metestrus females compared to proestrus females. The time to reach maximal capsaicin-induced PE was two times longer in estrus, diestrus, and metestrus females compared to males (p<0.05). PE induced by heat was also significantly greater in females compared to males (p<0.001), however, there was no sex-related difference in PE induced by electrical stimulation or by injection of SP. These findings show that females have a greater inflammatory response when inflammation is induced by capsaicin and noxious heat suggesting possible sex-related changes in TRPV-1 receptor mediated mechanisms. These results add to the growing list of sex difference responses to noxious somatic stimulation.

Objective: To propose an electronic method for sensitivity evaluation in leprosy and to compare it to the Semmes-Weinstein monofilaments., Methods: Thirty patients attending the Dermatology outpatient clinic of HCFMRP-USP were consecutively evaluated by both the electronic aesthesiometer and Semmes-Weinstein monofilaments on hand and foot test points. The intraclass correlation coefficient (ICC) was calculated to determine the variability of the electronic measures and the Kappa coefficient was calculated to determine the agreement between methods according to their categories (altered and non-altered tactile sensitivity)., Results: The ICC was approximately 1, demonstrating repeatability. The Kappa coefficient showed more than 75 and 63% agreement on the hand and foot points, respectively. The mean agreement between the 2 methods for the 7 points of the right and left hand was 77.14 and 75.71%, respectively. The mean agreement for all 10 points was 74.33 and 63.66% on the right and left foot, respectively. In cases of disagreement the detection of altered tactile sensitivity by the electronic esthesiometer on the right and left foot was 90.91 and 84.25%, respectively, with no detection by the monofilaments., Conclusion: The results suggest that the electronic esthesiometer is a reliable and easy application, capable of evaluating alterations of tactile sensitivity in leprosy patients.

A method for quantifying nociceptive withdrawal reflex receptive fields in human volunteers and patients is described. The reflex receptive field (RRF) for a specific muscle denotes the cutaneous area from which a muscle contraction can be evoked by a nociceptive stimulus. The method is based on random stimulations presented in a blinded sequence to 10 stimulation sites. The sensitivity map is derived by interpolating the reflex responses evoked from the 10 sites. A set of features describing the size and location of the RRF is presented based on statistical analysis of the sensitivity map within every subject. The features include RRF area, volume, peak location and center of gravity. The method was applied to 30 healthy volunteers. Electrical stimuli were applied to the sole of the foot evoking reflexes in the ankle flexor tibialis anterior. The RRF area covered a fraction of 0.57+/-0.06 (S.E.M.) of the foot and was located on the medial, distal part of the sole of the foot. An intramuscular injection into flexor digitorum brevis of capsaicin was performed in one spinal cord injured subject to attempt modulation of the reflex receptive field. The RRF area, RRF volume and location of the peak reflex response appear to be the most sensitive measures for detecting modulation of spinal nociceptive processing. This new method has important potential applications for exploring aspects of central plasticity in volunteers and patients. It may be utilized as a new diagnostic tool for central hypersensitivity and quantification of therapeutic interventions.

We describe a method for evaluating the threshold for cutaneous mechanical sensation in rodents, based on a stimulator that drives a probe against the plantar surface of the foot. The stimulator applies loads that can be either constant or linearly increased. We describe withdrawal responses, including forms of movement that precede foot withdrawals. With constant stimuli, response latency declines in a nonlinear fashion as stimulus magnitude is increased. With ramped stimuli the effect of loading rate is complex, reflecting both the rate of change of the stimulus and the animal's reaction time. We demonstrate the utility of using ramped stimuli in experiments that show that thresholds vary spatially across the foot and experiments that show that intradermal capsaicin injections cause allodynia but not hyperalgesia.

In the present study, the activation of p38 mitogen-activated protein kinase (p38 MAPK) in the rostral ventromedial medulla (RVM) following the injection of complete Freund's adjuvant (CFA) into the rat hindpaw was examined in order to clarify the mechanisms underlying the dynamic changes in the descending pain modulatory system after peripheral inflammation. Phospho-p38 MAPK-immunoreactive (p-p38 MAPK-IR) neurons were observed in the nucleus raphe magnus (NRM) and nucleus reticularis gigantocellularis pars alpha (GiA). Inflammation induced the activation of p38 MAPK in the RVM, with a peak at 30 min after the injection of CFA into the hindpaw, which lasted for 1 h. In the RVM, the number of p-p38 MAPK-IR neurons per section in rats killed at 30 min after CFA injection (19.4+/-2.0) was significantly higher than that in the naive group (8.4+/-2.4) [p<0.05]. At 30 min after CFA injection, about 40% of p-p38 MAPK-IR neurons in the RVM were serotonergic neurons (tryptophan hydroxylase, TPH, positive) and about 70% of TPH-IR neurons in the RVM were p-p38 MAPK positive. The number of p-p38 MAPK- and TPH-double-positive RVM neurons in the rats with inflammation was significantly higher than that in naive rats [p<0.05]. These findings suggest that inflammation-induced activation of p38 MAPK in the RVM may be involved in the plasticity in the descending pain modulatory system following inflammation.

Most of functional magnetic resonance imaging (fMRI) experiments have been performed on horizontal bore magnets. Here, we present practical aspects of fMRI based on single-shot, spin-echo echo-planar imaging (EPI) using a widely available, cost effective 89 mm bore vertical 11.7 T microimager. It was demonstrated that reproducible, high-quality fMRI data can be obtained from alpha-chloralose anesthetized adult rat brain. Both coronal and the more extended horizontal EPI images were acquired to measure blood oxygenation level dependent (BOLD) responses to electrical stimulation of fore- and hindpaws. The BOLD patterns observed match the known representations of fore- and hindpaws in the somatosensory cortex in rats. Preliminary results on BOLD signal enhancement using aminophylline are also presented.

Cutaneous reflexes evoked by stimulation of nerves innervating the foot are modulated in a phase-dependent manner during locomotion. The pattern of modulation of these reflexes has been suggested to indicate a functional role of cutaneous reflexes in assisting to maintain stability during walking. We hypothesized that if cutaneous reflexes assist in maintaining stability during gait, then these reflexes should be modulated in a context-dependent manner when subjects are asked to walk in an environment in which stability is challenged. To do this, we asked subjects to walk on a treadmill under five conditions: (1) normally, (2) with the arms crossed, (3) while receiving unpredictable anterior-posterior (AP) perturbations, (4) with the arms crossed while receiving unpredictable AP perturbations, and (5) with the hands holding onto fixed handles. Cutaneous reflexes arising from electrical stimulation of the superficial peroneal (SP; relevant to stumbling) or distal tibial (TIB; relevant to ground contact sensation) nerves were recorded bilaterally, at four points in the step cycle. Reflexes evoked with SP nerve stimulation showed marked facilitation during the most unstable walking condition in 4 of the 7 muscles tested. SP nerve-evoked reflexes in the muscles of the contralateral leg also showed suppression during the most stable walking condition. Reflexes evoked with TIB nerve stimulation were less affected by changes in the walking task. We argue that the specific adaptation of cutaneous reflexes observed with SP nerve stimulation supports the hypothesis that cutaneous reflexes from the foot contribute to the maintenance of stability during walking.

In the present study, a semi-quantitative analysis of Fos expression by mustard oil was performed. For this purpose, mustard oil was applied to the skin of the right hind foot of Wistar rats at various concentrations: 5%, 10%, 30%, 50%, 80% and 100% in liquid paraffin. The distribution and number of activated Fos-positive cells in the stimulated side (ipsilateral) and contralateral side of the spinal cord were investigated following the application. The ED50 of the response was also determined. The number of Fos-labelled cells gradually increased in a dose dependent manner in both sides of superficial layers (laminae I-II) of the spinal cord with increasing concentration of mustard oil. The increase between the doses was found significant in the ipsilateral superficial layers. The increase was significant in the contralateral superficial layers at concentrations above 50%. Very few Fos-labelled cells were observed around the central canal region in all concentrations. Higher doses of the mustard oil did not increase the number of activated cells in the deeper layers. However, the expression in the deeper layers (laminae III-X) does not show a consistent trend. Also, none of the concentrations used produced labelling in neurons of the deep ventral horn neurons or in motor neurons. Forty percent (40%) of mustard oil gave an approximately 1/2 maximum response i.e. an approximate ED 50. This may be important for studies using intrathecal application of antagonist following the mustard oil activation of skin nerve fibres.

Administration of 3 mg/kg 5-hydroxytryptamine2A/2C (5-HT2A/2C) receptor agonist (+/-)-2,5-dimethoxy-4-iodoamphetamine hydrochloride (DOI) induced c-Fos expression in all areas of the rat neocortex. The single footshock 24 h before DOI administration significantly attenuated DOI-induced c-Fos expression in layer IV of the primary somatosensory cortex. These changes in c-Fos expression suggest an adaptational change in the somatosensory cortex by acute stress.

Coupling between functional cortical activity and blood flow is a regulatory principle that adjusts the supply of substrates to the metabolic needs of the tissue. The flow response is usually expressed as the maximum increase over baseline; control system analysis allows the description of the entire time course and the main dynamic features of the regulative principle. In chloralose-anesthetized rats, forepaws were stimulated by trains of electric pulses of 0.3 or 5 ms duration. Blood flow was recorded in the contralateral somatosensory cortex by laser-Doppler flowmetry and correlated with the amplitude of primary somatosensory evoked potentials (SEP). Changes were analyzed by a control system approach. Pulses of 0.3 or 5 ms evoked SEPs of similar amplitude, whereas flow responses differed: 0.3 ms pulses led to a peak and plateau characteristic, 5 ms pulses evoked a plateau characteristic. The flow response evoked by 0.3 ms pulses can be modeled mathematically by an initial feedforward regulative principle followed after some delay by feedback controlled flow stabilization, whereas 5 ms pulses lack the feedforward component. The absence of an electrophysiological difference points to a dissociation between electrophysiological and hemodynamic responses and may be of importance for the understanding of flow coupling.

Aspirin (ASA) is widely used oral analgesics and acts as an inhibitor of cyclo-oxygenase. Also, acetaminophen (APAP) is effective analgesics and may selectively inhibit brain prostaglandin synthetase. However, their mechanisms of action in CNS are poorly defined, although several authors have shown that the antinociceptive effects of ASA and APAP have different underlying mechanisms and play some possible roles on spinal nociceptive processing, such as inhibition of prostaglandin synthesis. To define and characterize antinociceptive profiles of ASA and APAP on various pain models, we performed intraplantar formalin injection test, intrathecal (i.t.) substance P (0.7 microg)-induced nociceptive response test, and i.t. glutamate (20 microg)-induced nociceptive response test after ASA or APAP (from 10 to 300 mg/kg) administered orally to the mouse. In the formalin test, ASA produced an antinociceptive effect during only the 2nd phase (20-40 min), but not the 1st phase (0-5 min), in a dose-dependent manner. However, APAP showed a dose-dependent antinociceptive effect during both phases of the formalin test. In addition, both ASA and APAP reduced nociceptive behavior induced by glutamate administered i.t. in a dose-dependent manner. In substance P-induced nociceptive response, APAP, but not ASA, showed antinociceptive effect in a dose-dependent manner. Our results suggest that ASA and APAP administered orally may be mediated by different nociceptive processing at the spinal cord level.

We have recently observed medio-lateral balance adjustments (BA) preceding reflexive stepping elicited by noxious stimulation. While task specific modulation is evident for BA prior to voluntary leg movement, it is unclear whether rapid BA reactions (prior to 'reflexive' stepping) represent a generic response to evoked limb withdrawal or can be modified to suit task-conditions. This study was designed to establish whether the CNS is able to modify rapid onset latency BAs to match task conditions. Reflexive stepping was evoked by applying a noxious stimulus (50 ms stimulus train, 1 ms pulses, 300 Hz, 4 x perceptual threshold) to the plantar surface of the either the left or right foot. Task conditions were varied prior to stimulation by having subjects maintain one of three different static positions: (1) lean left (70% body weight (BW) on left), (2) neutral (50% BW both sides), (3) lean right (70% BW on right). BAs were denoted by centre-of-pressure (CoP) excursions towards the swing foot after the onset of noxious stimulation (average onset latency of 128 ms). There was a significant increase in frequency of occurrence and a significant increase in magnitude of CoP shift when the stimulation was applied to a loaded limb (leaning with 70% BW on the stimulated foot) as compared to an unloaded limb (30% BW). In addition, 78% of loaded trials featured steps taken with the unstimulated foot, which delayed removal of the stimulated foot. Collectively, the results indicate modifiability of the very rapid onset balance adjustments that precede the onset of limb withdrawal revealing complex control of balance exists even over very brief latencies.

Sciatic nerve section in rats evokes chronic limb edema, pain behavior, and hindpaw hyperalgesia, a syndrome resembling the complex regional pain syndrome type II (CRPS II or causalgia) in man. Glucocorticoids such as methylprednisolone (MP) have been used as analgesic and anti-edematous agents in patients suffering from CRPS, and interestingly these therapeutic effects appear to persist in some patients after stopping the medication. Similar to the CRPS clinical response to glucocorticoids, we now demonstrate that chronic hindpaw edema in the sciatic transection CRPS model is reversed by a continuous infusion of MP (3 mg/kg/day over 21 days), and this anti-edematous effect persists for at least 1 week after discontinuing MP. Furthermore, there is a chronic increase in spontaneous protein extravasation in the hindpaw skin of rats after sciatic transection, similar to the increased protein extravasation observed in the edematous hands of CRPS patients. A 2-week infusion of MP (3 mg/kg/day) reduced spontaneous protein extravasation in the hindpaw skin by 80%. We postulated that increased spontaneous neurogenic extravasation resulted in development of limb edema in both the animal model and the CRPS patient, and that the anti-edematous effects of MP are due to an inhibition of spontaneous extravasation. Additional experiments examined the inhibitory effects of MP infusion on electrically-evoked neurogenic extravasation in the hindpaw skin of normal rats. MP inhibition was dose- and time-dependent, with an ED(50) of 1.2 mg/kg/day for a 14-day continuous infusion of MP, and a maximum inhibitory effect requiring 17 days of MP infusion (3 mg/kg/day). MP (3 mg/kg/day for 14 days) also blocked both capsaicin- and SP-evoked neurogenic extravasation, indicating a post-junctional inhibitory effect. Our interpretation is that increased spontaneous neurogenic extravasation in this CRPS model contributed to the development and maintenance of hindpaw edema, and that chronic MP administration dose- and time-dependently blocked neurogenic extravasation at a post-junctional level, thus reversing spontaneous extravasation and limb edema in this model.

This study employed manipulation of sensory inputs (vision and plantar-surface cutaneous sensation) during gait termination to elicit insight into the roles played by these sensory systems in the control of gait termination. Attenuation of cutaneous sensation was achieved through hypothermic anesthesia. Visual information was occluded using special glasses. The subjects were asked to walk along an 8 m walkway and during randomly selected trials (25% of trials) to terminate their gait in a predetermined area. The centre of mass (COM) was obtained in order to provide an indication of the efficiency and stability during termination when sensory inputs were manipulated. Lack of visual information delayed the initiation of the slowing down of the COM forward progression and increased the step length of the last step of termination. Additionally, lack of vision resulted in the COM moving closer to the base of support (BOS) during double support and more variability, in the COM, when attempting to achieve a final stable position. Insensitivity of the plantar-surface mechanoreceptors led to a longer second step and a more variable foot placement of the first step, and increased the loading rate during the final two steps of termination. Additionally when vision and cutaneous information were absent the resolution of the final stable position was not as effectively controlled. The results demonstrated that visual information about self-motion and object-motion and sensation from the plantar surface of the foot play phase-specific roles in the control of COM during gait termination.

The gain of somatosensory afferent paths from the lower limb to the cerebral cortex was investigated during the acquisition of one target location during plantar flexion. Sensory gain was measured as the magnitude of somatosensory evoked potentials (SEPs) following electrical stimulation of a peripheral nerve in the lower limb, and was recorded from the scalp. We hypothesized gain attenuation of SEPs from sensory paths serving the limb segment responsible for target acquisition. SEP gain was studied as subjects plantar flexed about the anide to a target that was 15 degrees beyond the occurrence of a cutaneous stimulus (cue) to the lateral border of the foot. The "cue" was either fixed in one location or could appear at one of three positions in space. SEP gain was tested during practice and with task acquisition. Electroencephalographic (EEG) recordings were made of primary and secondary complexes of cortical SEPs from sural and tibial nerve stimulation, with 30-40 samples averaged per subject-condition. Electromyographic (EMG) records were made of soleus muscle H-reflexes and M-waves. Target acquisition was recorded as percent correct hits. The results showed significant attenuation in sural and tibial nerve primary SEPs with task acquisition when the cue was fixed or varied in movement space (P<0.05). Secondary SEPs from tibial nerve followed this pattern. Spinal H-reflexes only attenuated with movement per se. We conclude that the CNS preferentially reduces the cerebral inflow of sensory information once such a motor task has been successfully acquired.

The molecular mechanisms underlying mechanoelectrical transduction and the receptors that detect light touch remain uncertain. Studies in Caenorhabditis elegans suggest that members of the DEG/ENaC cation channel family may be mechanoreceptors. Therefore, we tested the hypothesis that subunits of the mammalian epithelial Na(+) channel (ENaC) family are expressed in touch receptors in rat hairless skin. We detected betaENaC and gammaENaC, but not alphaENaC transcripts in cervical and lumbar dorsal root ganglia (DRG). Using immunofluorescence, we found betaENaC and gammaENaC expressed in medium to large lumbar DRG neurons. Moreover, we detected these two subunits in Merkel cell-neurite complexes, Meissner-like corpuscles, and small lamellated corpuscles, specialized mechanosensory structures of the skin. Within these structures, betaENaC and gammaENaC were localized in the nerve fibers believed to contain the sensors responsive to mechanical stress. Thus beta and gammaENaC subunits are good candidates as components of the molecular sensor that detects touch.

The role of plantar pressure sensation in controlling compensatory stepping was explored via hypothermic anesthesia of the foot soles, in 10 healthy young adults. Stepping reactions were evoked by unpredictable platform translation in forward, backward and lateral directions. The findings suggest three specific direction- and phase-dependent roles for the plantar cutaneous afferents: (1) sensing posterior stability limits during initiation of backward steps, (2) sensing and controlling heel-contact and subsequent weight transfer during termination of forward steps, and (3) maintaining stability during the prolonged swing phase of lateral crossover steps.

Inhibition of proinflammatory cytokines reduces hyperalgesia in animal models of painful neuropathy. We set out to investigate the consequences of this treatment for nerve regeneration. Here we examined the sequels of epineurial application of neutralizing antibodies to tumor necrosis factor-alpha (TNF) in chronic constriction injury (CCI) of the sciatic nerve in C57/BL 6 mice. The mice were tested behaviorally for manifestations of thermal hyperalgesia and mechanical allodynia. Nerve regeneration was assessed by morphometry of myelinated nerve fibers in the sciatic nerve and of the epidermal innervation density in the glabrous skin of the hindpaws. Antibodies to TNF reduced thermal hyperalgesia and mechanical allodynia after CCI. Myelinated fiber density in the sciatic nerve was reduced to 30% of normal on day 7 after surgery, and reached 60% on day 45, with no difference between antibody-treated and untreated animals. Epidermal innervation density as shown by PGP 9.5 and CGRP immunohistochemistry was reduced to 25-47% at both time points after CCI, again without differences between antibody treated and untreated mice. Myelinated fiber density but not epidermal innervation density was correlated to thermal and mechanical withdrawal thresholds. We conclude that neutralization of endoneurial TNF attenuates pain related behavior but has no effect on nerve regeneration. Furthermore, the number of epidermal nerve fibers is not relevant to the magnitude of behavioral hyperalgesia in CCI.

The effect of testing surface on the rat hind paw withdrawal threshold in the von Frey hair test is investigated in this study. The data indicate that wire mesh, which is typically used to apply von Frey hairs, may have an effect on the paw withdrawal threshold. For example, in control rats tested on the wire mesh, variability in the withdrawal threshold was observed between the left and the right hind paws (51.04+/-12.29 and 64.31+/-9.37 g, respectively) and on different days of testing (35.24+/-9.54 and 45.83+/-12.97 g for the left and right hind paws, respectively, 7 days later). In an attempt to reduce this variability, a customized platform was used to measure the von Frey hair-induced paw withdrawal in the rat. It consists of an opaque, flat-surfaced plastic platform with holes through which von Frey hairs are inserted and applied to the plantar surface of the paw. In control rats tested with von Frey hairs using this customized platform, variability in the paw withdrawal thresholds between the left and right hind paws in single rats over time as well as between different rats was reduced (49.86+/-6.97 and 49.29+/-6.56 g for the left and right hind paws, respectively, on day 0; 48.29+/-5.82 and 53.00+/-4.59 g for the left and right hind paws, respectively, 7 days later). Furthermore, in rats in which a 2 mm polyethylene cuff was used to constrict the left common sciatic nerve, the ipsilateral as well as the contralateral hind paw withdrawal thresholds were decreased (2.45+/-0.65 and 26.09+/-5.86 g, respectively, 7 days later). In similar rats tested on the wire mesh, the ipsilateral but not the contralateral paw withdrawal threshold decreased (12.80+/-2.21 and 65.00+/-10.28 g, respectively, at 7 days). The data suggest that the flat surface and opaque properties of the customized platform enable accurate, reliable and repeatable measurements of ipsilateral and contralateral paw withdrawal threshold using von Frey hairs in normal and nerve-injured rats.

The pyramidal tract and red nucleus send prominent projections to the spinal cord and are thought to co-operate in producing skilled movements. In the present study, skilled reaching for food located on a shelf and spontaneous grasping, handling and eating pieces of pasta were video-recorded and analyzed in control rats, rats with unilateral ibotenic acid lesions of the red nucleus (RN), unilateral pyramidal tract lesions (PT) and combined lesions. The behavioral results suggest that skilled movements are organized as action patterns, easily recognizable and distinctive for each task. In both skilled and spontaneous reaching, PT lesions reduced success more than RN lesions, suggesting a greater role for the PT in guiding limb movements. Both lesions impaired rotatory movements including limb aiming, pronation and supination. RN lesions additionally abolished the arpeggio movement by which the paw is oriented for searching and grasping. Combined lesions were additive as rats lost both rotatory movements and arpeggio. Nevertheless, even after combined lesions, the rats were able to advance the limb, grasp food and withdraw the limb. The sparing following combined lesions suggests that other neural systems as well as compensatory adjustments assist the impaired limb. The results are discussed in relation to the possible distinctive contributions of the rubrospinal and corticospinal tract to the action patterns that comprise skilled movements in rats.

The effects of wire grates on nerve injury and recovery were examined in rats housed in cages with sawdust-covered solid flooring. For the first 3 weeks of the study, 20 rats were housed on sawdust alone and 20 rats were housed in cages with wire grates placed over the sawdust. For the remaining 9 weeks, 10 animals housed on sawdust had wire grates added to their cages, while grates were removed from the cages of 10 animals. The effects of tactile stimulation on hindpaw plantar skin was measured weekly using the Von Frey filament test. Intraepidermal innervation using PGP 9.5 immunostaining and plantar nerve histology were assessed at the end of the 12-week study. After just 1 week on grates, hindpaw withdrawal thresholds were already markedly decreased and remained low until the grates were removed at 3 weeks. Thresholds returned to normal by 4 weeks after removal of the grates. Wire grates also induced increases in PGP 9.5 immunoreactive intraepidermal fine nerve endings that were normalized after grate removal. Demyelination, Wallerian degeneration and Renaut bodies were induced in the medial plantar nerve in rats housed in cages with wire-grate flooring. Nerve injury was largely resolved after 9 weeks on sawdust flooring. These data demonstrate that wire grates rapidly induce hindpaw tactile hyperesthesia and plantar neuropathy in rats and emphasize a risk of using wire-grate cage flooring in studies assessing hindlimb function and structure., (Copyright 1998 Elsevier Science B.V.)

The antinociceptive effects of morphine (5 micrograms) microinjected into the ventrolateral periaqueductal gray were determined using both the tail flick and the foot withdrawal responses to noxious radiant heating in lightly anesthetized rats. Intrathecal injection of appropriate antagonists was used to determine whether the antinociceptive effects of morphine were mediated by alpha 2-noradrenergic, serotonergic, opioid, or cholinergic muscarinic receptors. The increase in the foot withdrawal response latency produced by microinjection of morphine in the ventrolateral periaqueductal gray was reversed by intrathecal injection of the cholinergic muscarinic receptor antagonist atropine, but was not affected by the alpha 2-adrenoceptor antagonist yohimbine, the serotonergic receptor antagonist methysergide, or the opioid receptor antagonist naloxone. In contrast, the increase in the tail flick response latency produced by morphine was reduced by either yohimbine, methysergide or atropine. These results indicate that microinjection of morphine in the ventrolateral periaqueductal gray inhibits nociceptive responses to noxious heating of the tail by activating descending neuronal systems that are different from those that inhibits the nociceptive responses to noxious heating of the feet. More specifically, serotonergic, muscarinic cholinergic and alpha 2-noradrenergic receptors appear to mediate the antinociception produced by morphine using the tail flick test. In contrast, muscarinic cholinergic, but not monoamine receptors appear to mediate the antinociceptive effects of morphine using the foot withdrawal response.

Behavioral recovery is the major goal of neuroscience research on the peripheral nervous system. However, behavioral methods of assessing peripheral nerve regeneration are still lacking. In the present work we used the median nerve and finger flexors as the experimental model. After resection or crush injury of the median nerve, rats were submitted daily to the grasping test. For this, they were gently lifted by the tail and allowed to grasp a grid connected to an ordinary electronic balance. While grasping the animal continued to be lifted by the tail with increasing firmness until it lost its grip. At this precise moment the value shown by the balance was recorded. In the crushed median nerve, recovery of function was clearly demonstrated by the grasping test. The test also indicated the exact day on which recovery began and its improvement with time. This very simple objective behavioral method provides a sensitive quantitative technique for assessing recovery. The experimental model using the median nerve seems to be better than the sciatic nerve model, as no articular contractures or autotomies were noted in the present experimental animals.

Our understanding of the postural control responses in the event of external perturbation has focused almost exclusively on the early automatic adjustments. The present study addresses another postural reaction that is functionally important: compensatory stepping. The purpose was to identify the relative importance by comparing the prevalence of compensatory stepping with and without instructions constraining the subjects' responses. Subjects stood on two force plates which were mounted on a "moveable" platform. Their posture was perturbed by the translation of the platform either forward or backward at various accelerations. Following a practice period, seven subjects each performed under two different tasks: "constrained" (keep feet in place) and "unconstrained" (no specific instructions given). The primary focus of the analysis was on responses to forward platform translations. Analysis revealed that the frequency of stepping tended to be higher in "unconstrained", as opposed to "constrained", tasks. The frequency of stepping was also related to the interaction between the tasks and the order in which they were given. Specifically, subjects stepped most frequently when they received the "unconstrained" task first. The frequency of stepping also increased as the magnitude of the platform acceleration increased. Time of onset of stepping, as defined from the force plate measures, began as early as 160 ms in one subject and averaged 250 ms across all subjects. These relatively fast response times suggest that step initiation often occurs well before the limits of stability are reached. A novel and unexpected finding was the identification of a third response type, intermediate to stepping and (bilaterally symmetrical) non-stepping responses.(ABSTRACT TRUNCATED AT 250 WORDS)

A comparative analysis of the corticospinal tract nervous propagation and excitability threshold was carried out in young (25 subjects, age range 16-35 years) and in elderly (40 subjects, 51-86 years) populations of healthy volunteers. Motor evoked potentials (MEPs) were recorded from the hand and foot muscles following transcranial magnetic stimulation (TCS) during complete relaxation and active contraction of the target muscles. Threshold intensities corresponded to the stimulator's output eliciting liminal MEPs in about 50% of stimuli during relaxation. It was found that threshold values of magnetic TCS were significantly higher in the elderly (44 +/- 6.4% vs 39 +/- 3.5% for the hand; 66 +/- 10.1% vs 56 +/- 6.7% for the foot; P < 0.001) than in the young subjects. Moreover, this index progressively increased with age (P < 0.001), whilst the propagation time along the central motor tracts did not parallel such an age-related trend.

The incidence of motor nerve and terminal sprouting was quantitatively analyzed in normal unoperated muscles, in homologous muscles contralateral to muscles which have been partially denervated, in partially denervated muscles, and in sham-operated muscles. Muscles were studied by light microscopy after staining motor endplates by a combined silver-cholinesterase stain. In addition, the incidence of endplates containing terminal sprouts, the number of terminal branch points per endplate, and endplate, and endplate size were also assessed in the various groups examined. We observed that following section of the L4 spinal nerve, the incidence of sprouting (preterminal and intranodal) in the contralateral muscle exhibited a 2-fold increase over sham-operated controls. We also found a correlation between nerve terminal sprouting, terminal branch point number and endplate size. All of these parameters were significantly increased in the contralateral muscles as compared to the sham-operated control muscles. These findings suggest that normal muscles undergo sprouting which can be enhanced by contralateral partial denervation. The possible underlying mechanism may be the transneuronal induction of sprouting.

The possibility that morphine might differentially affect spinal neurons responsive to small-diameter thermoreceptive-specific afferents was tested. Systemic morphine enhanced a portion or all of the stimulus-response function of 7 of 9 lamina I spinothalamic tract cells specifically sensitive to cold applied to the glabrous hindpaw in the cat. This result contrasts strongly with the predominant inhibition of nociceptive lamina I neurons by morphine.

Slow-adapting impulses were recorded from the single nerve fiber innervating the Pacinian corpuscle of the cat planta, which was stimulated by constant pressure. Slow-adapting Pacinian corpuscles responded to both low-frequency (0.1-0.01 Hz) and high-frequency (400-800 Hz) vibrations. Simultaneously performed electron microscopy showed that there were no definite ultrastructural differences between slow- and fast-adapting Pacinian corpuscles.

An analysis of response properties of single neurons in the forepaw region of the primary somatosensory neocortex (SI) of the raccoon was undertaken to gain a better understanding of the neural bases of tactile form discrimination. In the course of this investigation a new type of feature detector neuron which responded preferentially to tactile stimulus orientation was discovered. Within a sample of 110 neurons responding to light touch, two categories of cells were noted in terms of the type of stimulation best able to excite them. One category (linear units) was preferentially responsive to indentations of the skin using a narrow elongated stimulus probe applied in a particular orientation. The other category (round field units) gave a maximum response to stimulation with a round stimulus probe applied anywhere within a roughly circular receptive field. The response pattern of some SI neurons to a maintained indentation of the skin consisted of early and late excitatory responses, separated by a depression in responding. The second excitatory response appeared to be more influenced by variations in stimulus parameters than was the first.

Systemic administration of 4-aminopyridine (4-AP) increased the size of the cutaneous receptive fields of 9 of the 15 dorsal horn cells tested. These receptive fields were on the feet and toes of the hind limbs of cats. Receptive field sizes increased with increasing doses of 4-AP. However, 4-AP administration did not change the responses of dorsal horn cells to graded mechanical stimuli administered near the centers of their receptive fields.

The normal recruitment order of EMG spikes of the first deep lumbrical muscle of the cat's hindpaw, usually seen during cortical stimulation, pad pinch and weak plantar nerve stimulation, was temporarily reversed after stimulation of the medial digital nerve of the foot at 50 Hz for 2 min, and normal order was recovered in 2 to 10 min. The longer the period of stimulation of the medial digital nerve was, the longer the time for recovery. Alteration of order is repeatable and reversible after an interval of more than 15 min. After prolonged medial digital nerve stimulation, EMG responses to pad pinch and plantar nerve stimulation were facilitated. Combination of some stimuli (e.g. cortical stimulation and plantar nerve stimulation, pad pinch and plantar nerve stimulation, plantar nerve and medial digital nerve stimulation) also produced reversal of recruitment order during the period of stimulation. A functionally single motor nerve fiber to the first deep lumbrical muscle was isolated from the motor nerve axons in L7 and S1 of the spinal cord of cats, and physiological properties of pairs of motor units whose recruitment order was temporarily altered by plantar nerve stimulation after prolonged stimulation of the medial digital nerve of a foot were examined. Motor units with large action potentials were more facilitated than motor units with small action potentials after prolonged stimulation of the medial digital nerve. The former motor units showed fast contraction time, large twitch tension, low resistance to fatigue and presence of sag-behavior. The latter motor units showed slow contraction time, small twitch tension and high resistance to fatigue. High threshold motor units with middle-sized surface EMG records were recruited after motor units with large potentials had been recruited; motor units with middle-size action potentials were beta motor axons branching to both intrafusal and extrafusal muscle fibers. The beta motor axons also showed fast conduction velocity and the presence of sag-behavior.

Isometric contractions of single motor units were studied in the first deep lumbrical muscle of the cat's hind-foot. Motor units with short twitch contraction times (15-20 msec) generally differed from those with longer ones (23-50 msec; contraction time measured in unpotentiated twitches) in showing (1) a greater maximum tetanic tension, (2) a smaller resistance to fatigue, (3) more post-tetanic potentiation of twitch tension, and (4) no post-tetanic occurrence of repetitive activity in response to single nerve stimuli (such "post-tetanic repetitive activity" was seen in several of the slower units). The ratio between unpotentiated twitch tension and maximum tetanic tension was similar for units with brief and long contraction times. The peak-to-peak amplitude of a single motor unit spike, recorded with gross electrodes, tended to be directly proportional to the maximum tetanic tension of the same motor unit.