Paw Shaking Research Articles

Science, Medicine, and the Anesthesiologist

Science, Medicine, and the Anesthesiologist

Emergence of Extreme Paw Accelerations During Cat Paw Shaking: Interactions of Spinal Central Pattern Generator, Hindlimb Mechanics and Muscle Length-Depended Feedback.

Cat paw shaking is a spinal reflex for removing an irritating stimulus from paw by developing extremely high paw accelerations. Previous studies of paw shaking revealed a proximal-to-distal gradient of hindlimb segmental velocities/accelerations, as well as complex inter-joint coordination: passive motion-dependent interaction moments acting on distal segments are opposed by distal muscle moments. However, mechanisms of developing extreme paw accelerations during paw shaking remain unknown. We hypothesized that paw-shaking mechanics and muscle activity might correspond to a whip-like mechanism of energy generation and transfer along the hindlimb. We first demonstrated in experiments with five intact, adult, female cats that during paw shaking, energy generated by proximal muscle moments was transmitted to distal segments by joint forces. This energy transfer was mostly responsible for the segmental velocity/acceleration proximal-to-distal gradient. Distal muscle moments mostly absorbed energy of the distal segments. We then developed a neuromechanical model of hindlimb paw shaking comprised a half-center CPG, activating hip flexors and extensors, and passive viscoelastic distal muscles that produced length/velocity-depended force. Simulations reproduced whip-like mechanisms found experimentally: the proximal-to-distal velocity/acceleration gradient, energy transfer by joint forces and energy absorption by distal muscle moments, as well as atypical co-activation of ankle and hip flexors with knee extensors. Manipulating model parameters, including reversal of segmental inertia distal-to-proximal gradient, demonstrated important inertia contribution to developing the segmental velocity/acceleration proximal-to-distal gradient. We concluded that extreme paw accelerations during paw shaking result from interactions between a spinal CPG, hindlimb segmental inertia, and muscle length/velocity-depended feedback that tunes limb viscoelastic properties.

Analgesic Effect of Bangle Rhizome Ethanol Extraction on Thermally Induced Pain in Mus musculus

Introduction: Pain causes deterioration of quality of life. Nonsteroidal anti-inflammatory drug (NSAID) is used to relieve pain with upper gastrointestinal side effects. Zingiber cassumunar Roxb. (known as bangle) is commonly used to relieve pain, but no study has been proved to have analgesic effect. In empirical study, bangle is a potential analgesic due to its bioactive compound named zerumbone. The aim of this study was to investigate the analgesic effect of bangle rhizome ethanol extraction on thermally induced pain.Methods: Five groups of male Mus musculus consisting of 6 mice each were used. A single dose of bangle extract 2 mg/20 g, 4 mg/20 g, 8 mg/20 g, aspirin 1.82 mg/20 g, and aquadest 0.2 ml were administrated. The analgesic effect was evaluated by hot plate test. The test was evaluated at 15th, 30th, 45th minute. Latency period was observed when mice showed pain responses including hind paw shaking, hind paw licking or jumping. The data was analyzed by paired T-test and one-way ANOVA.Results: Results showed that all doses of bangle extract and aspirin 1.82 mg/20 g produced significant latency period (p<0.05) compared to baseline in each group using paired T-test. Latency period of bangle extract 8 mg/20 g was more significant than bangle extract 2 mg/20 g and 4 mg/20 g. One-way ANOVA showed no significant difference in latency period (p=0.1) between bangle extract and aspirin (p>0.05).Conclusion: Bangle extract has analgesic effect on thermally induced pain. The minimum dose for analgesic is 2 mg/20 g or potentially less.

Anti-inflammatory and antinociceptive activities of glucagon-like peptides: evaluation of their actions on serotonergic, nitrergic, and opioidergic systems.

Glucagon-like peptide-1 (GLP-1) and glucagon-like peptide-2 (GLP-2) are gut derived hormones. GLP-1 and GLP-2 were shown to have pleiotropic effects in intestinal and pancreatic diseases. We aimed to investigate the activities of GLP-1 and GLP-2 on nociception and inflammation in mice, involving their actions on serotonergic, nitrergic, and opioidergic systems. Antinociceptive and anti-inflammatory activities of intraperitoneally injected GLPs were evaluated in hotplate latency test, formalin-induced behavioral, and paw edema tests. Ondansetron, a selective 5-HT3 receptor antagonist; L-NAME, a NOS inhibitor; and naloxone, an opioid antagonist were injected to determine the mechanisms of antinociception and anti-inflammation. We also measured blood glucose levels and performed rotarod test in order to evaluate whether the hypoglycemic effect of GLP compounds or alterations in locomotor activity may affect the latency in hotplate test and activity in formalin test. GLP-1 (0.2mg/kg) and GLP-2 (0.05, 0.2mg/kg) significantly increased pain threshold. GLP-1 (0.2mg/kg) and GLP-2 (0.05, 0.1, 0.2mg/kg) significantly decreased formalin-induced licking and shaking behaviors. GLP-1 or GLP-2 showed no significant inhibitory action on formalin-induced swelling in paws of mice. Antinociceptive actions of GLP-1 and GLP-2 were significantly decreased with ondansetron and naloxone, and paw shaking behavior significantly increased with naloxone. GLP-1 and GLP-2 did not impair rotarod performance, and did not cause a significant hypoglycemic effect in our normoglycemic mice after rotarod test. These finding indicated that the antinociceptive and anti-inflammatory effect of GLP-1 was related to opioidergic system. Antinociceptive effect of GLP-2 was partially related to 5-HT3 serotonergic or opioidergic system in hotplate test. However, the anti-inflammatory effect of GLP-2 was not directly related to 5-HT3, NO or opioids.

Control of transitions between locomotor-like and paw shake-like rhythms in a model of a multistable central pattern generator.

The ability of the same neuronal circuit to control different motor functions is an actively debated concept. Previously, we showed in a model that a single multistable central pattern generator (CPG) could produce two different rhythmic motor patterns, slow and fast, corresponding to cat locomotion and paw shaking. A locomotor-like rhythm (~1 Hz) and a paw shake-like rhythm (~10 Hz) did coexist in our model, and, by applying a single pulse of current, we could switch the CPG from one regime to another (Bondy B, Klishko AN, Edwards DH, Prilutsky BI, Cymbalyuk G. In: Neuromechanical Modeling of Posture and Locomotion, 2016). Here we investigated the roles of slow intrinsic ionic currents in this multistability. The CPG is modeled as a half-center oscillator circuit comprising two reciprocally inhibitory neurons. Each neuron is equipped with two slow inward currents, a Na+ current ( INaS) and a Ca2+ current ( ICaS). ICaS inactivates much more slowly and at more hyperpolarized voltages than INaS. We demonstrate that INaS is the primary current driving the paw shake-like bursting. ICaS is crucial for the locomotor-like bursting, and it is inactivated during the paw shake-like activity. We investigate the sensitivity of the bursting regimes to perturbations, using a pulse of current to induce a switch from one regime to the other, and we demonstrate that the transition duration is dependent on pulse amplitude and application phase. We also investigate the modulatory roles of the strength of various currents on characteristics of these rhythms and show that their effects are regime specific. We conclude that a multistable CPG is physiologically plausible and derive testable predictions of the model. NEW & NOTEWORTHY Little is known about how a single central pattern generator could produce multiple rhythms. We describe a novel mechanism for multistability of bursting regimes with strongly distinct periods. The proposed mechanism emphasizes the role of intrinsic cellular dynamics over synaptic dynamics in the production of multistability. We describe how the temporal characteristics of multiple rhythms could be controlled by neuromodulation and how single pulses of current could produce a switch between regimes in a functional fashion.

Task-dependent inhibition of slow-twitch soleus and excitation of fast-twitch gastrocnemius do not require high movement speed and velocity-dependent sensory feedback.

Although individual heads of triceps surae, soleus (SO) and medial gastrocnemius (MG) muscles, are often considered close functional synergists, previous studies have shown distinct activity patterns between them in some motor behaviors. The goal of this study was to test two hypotheses explaining inhibition of slow SO with respect to fast MG: (1) inhibition occurs at high movement velocities and mediated by velocity-dependent sensory feedback and (2) inhibition depends on the ankle-knee joint moment combination and does not require high movement velocities. The hypotheses were tested by comparing the SO EMG/MG EMG ratio during fast and slow motor behaviors (cat paw shake responses vs. back, straight leg load lifting in humans), which had the same ankle extension-knee flexion moment combination; and during fast and slow behaviors with the ankle extension-knee extension moment combination (human vertical jumping and stance phase of walking in cats and leg load lifting in humans). In addition, SO EMG/MG EMG ratio was determined during cat paw shake responses and walking before and after removal of stretch velocity-dependent sensory feedback by self-reinnervating SO and/or gastrocnemius. We found the ratio SO EMG/MG EMG below 1 (p < 0.05) during fast paw shake responses and slow back load lifting, requiring the ankle extension-knee flexion moment combination; whereas the ratio SO EMG/MG EMG was above 1 (p < 0.05) during fast vertical jumping and slow tasks of walking and leg load lifting, requiring ankle extension-knee extension moments. Removal of velocity-dependent sensory feedback did not affect the SO EMG/MG EMG ratio in cats. We concluded that the relative inhibition of SO does not require high muscle velocities, depends on ankle-knee moment combinations, and is mechanically advantageous for allowing a greater MG contribution to ankle extension and knee flexion moments.

Multifunctional central pattern generator controlling walking and paw shaking

Central pattern generators (CPGs) are oscillatory neuronal networks controlling rhythmic motor tasks such as breathing and walking. A multifunctional CPG can produce multiple patterns, e.g. patterns with different periods [1-5]. Here, we investigate whether a pair of cat behaviors -- walking and paw shaking -- could be controlled by a single multifunctional CPG exhibiting multistability of oscillatory regimes. In experiments, both behaviors can be elicited in a spinalized cat, and there is evidence that the same circuitry is used for both rhythms [2,3]. We present a parsimonious model of a half-center oscillator composed of two mutually inhibitory neurons. These cells contains two slowly inactivating inward currents, a persistent Na+ current (INaP) and a low voltage activated Ca++ current (ICaLVA). The dynamics of the multifunctional CPG is based on that the ICaLVA inactivates much slower than INaP and at the more hyperpolarized membrane potentials. Here, we demonstrate the co-existence of two rhythms (Figure ​(Figure1).1). At first, the model demonstrates walking pattern. A switch from a slow, 1-2 Hz walking rhythm to fast, 7-10 Hz paw shake rhythm was elicited by a pulse of conductance of excitatory current delivered to extensor and flexor neurons. Then, a switch back to walking was triggered by a shorter pulse of conductance of inhibitory current delivered to the extensor neuron. Figure 1 Two mutually inhibitory interneurons, IntE (Extensor Interneuron) and IntF (Flexor Interneuron) produce alternating bursting activity at approximately 1.6 Hz representing walking pattern. A switch to paw shaking is executed by a pulse of excitatory conductance ... The CPG model was also incorporated into a neuromechanical model of a cat hindlimb in the AnimatLab environment [6]. The model provides a cellular mechanism of multifunctional CPG operation.

Mid-Lumbar Segments Are Needed for the Expression of Locomotion in Chronic Spinal Cats

In acute experiments performed in decerebrated and spinalized (T13) cats, an intraspinal injection of clonidine, a noradrenergic agonist, restricted to mid-lumbar segments L3-L4, can induce hindlimb locomotion, whereas yohimbine, a noradrenergic antagonist, can block spinal locomotion, and a second spinal lesion at L4 can abolish all locomotor activity. In the present study, we investigated whether the abolition of locomotion after this second spinal lesion was due to an acute spinal shock or to the functional disconnection of the rostral and caudal lumbar segments. In seven cats, first spinalized at T13 and having recovered treadmill locomotion, a second transection was performed at lower lumbar levels. Video and electromyographic recordings were used to evaluate locomotor performance. Results show that after a second transection at L2 or rostral L3 levels, spinal locomotion was maintained; when the second lesion was performed at caudal L3 or L4, all locomotor activity was abolished even after several weeks of attempted locomotor training; vigorous fast paw shakes (FPS) were observed in all cases; and after an intraperitoneal injection of clonidine in cats with a second transection below L4, perineal stimulation induced hyperextension of the hindlimbs but no locomotion. Considering that the main motoneuron pools of the hindlimbs are caudal to L4 and are still functional after the second spinal transection, as evidenced by the presence of FPS, we conclude that the mid-lumbar spinal segments are essential for the specific expression of spinal locomotion but not necessarily for other rhythmic motor patterns.

Motor unit recruitment during vertebrate locomotion

AbstractVertebrate skeletal muscles act across joints and produce segmental accelerations and therefore animal movement when they contract. Different muscles and different motor units vary in their mechanical contractile properties. Early studies on motor unit recruitment demonstrated orderly recruitment of motor units from the slowest to the fastest during a graded contraction. However, many subsequent studies illustrate conditions when alternative recruitment strategies may exist. Motor unit recruitment during locomotion is thus multifactorial and more complex than typically thought.Different types of motor unit vary in their mechanical properties, including rates of force activation and deactivation, maximum unloaded shortening velocities and the shortening velocities at which maximum mechanical power output and maximum mechanical efficiency occur. In short, it would make mechanical sense to perform fast activities with the faster motor units and slow activities with the slower motor units. However, determining patterns of motor unit recruitment during locomotion has presented experimental challenges.Comparisons between distinct muscles have shown that fast fish swimming and the cat paw shake are activities which employ predominantly the fast and not the slower muscle. Glycogen depletion studies have showed that jumping in the bushbaby uses fast without slow motor units within the vastus lateralis and gastrocnemius muscles. Studies in man show that differential recruitment of the different types of muscle fibre occurs at different times within each running stride. It is suggested that vertebrates may have a strategy of recruiting the motor units that are most mechanically suited for the different locomotor demands. However, we have much to learn about motor recruitment patterns during locomotion.

Intraspinal micro stimulation generates locomotor-like and feedback-controlled movements.

Intraspinal microstimulation (ISMS) may provide a means for improving motor function in people suffering from spinal cord injuries, head trauma, or stroke. The goal of this study was to determine whether microstimulation of the mammalian spinal cord could generate locomotor-like stepping and feedback-controlled movements of the hindlimbs. Under pentobarbital anesthesia, 24 insulated microwires were implanted in the lumbosacral cord of three adult cats. The cats were placed in a sling leaving all limbs pendent. Bilateral alternating stepping of the hindlimbs was achieved by stimulating through as few as two electrodes in each side of the spinal cord. Typical stride lengths were 23.5 cm, and ample foot clearance was achieved during swing. Mean ground reaction force during stance was 36.4 N, sufficient for load-bearing. Feedback-controlled movements of the cat's foot were achieved by reciprocally modulating the amplitude of stimuli delivered through two intraspinal electrodes generating ankle flexion and extension such that the distance between a sensor on the cat's foot and a free sensor moved back and forth by the investigators was minimized. The foot tracked the displacements of the target sensor through its normal range of motion. Stimulation through electrodes with tips in or near lamina IX elicited movements most suitable for locomotion. In chronically implanted awake cats, stimulation through dorsally located electrodes generated paw shakes and flexion-withdrawals consistent with sensory perception but no weight-bearing extensor movements. These locations would not be suitable for ISMS in incomplete spinal cord injuries. Despite the complexity of the spinal neuronal networks, our results demonstrate that by stimulating through a few intraspinal microwires, near-normal bipedal locomotor-like stepping and feedback-controlled movements could be achieved.

Weight support and balance during perturbed stance in the chronic spinal cat.

The intact cat maintains balance during unexpected disturbances of stance through automatic postural responses that are stereotyped and rapid. The extent to which the chronic spinal cat can maintain balance during stance is unclear, and there have been no quantitative studies that examined this question directly. This study examined whether the isolated lumbosacral cord of the chronic spinal cat can generate automatic postural responses in the hindlimbs during translation of the support surface. Responses to 16 directions of linear translation in the horizontal plane were quantified before and after spinalization at the T(6) level in terms of forces exerted by each paw against the support, motion of the body segments (kinematics), and electromyographic (EMG) activity. After spinalization, the cats were trained on a daily basis to stand on the force platform, and all four cats were able to support their full body weight. The cats usually required assistance for balance or stability in the horizontal plane, which was provided by an experimenter exerting gentle lateral force at the level of the hips. Three of the four animals could maintain independent stance for a brief period (10 s) after the experimenter stabilized them. The fourth cat maintained weight support but always required assistance with balance. Perturbations were delivered during the periods of independent stance in three cats and during assisted stance in the fourth. A response to translation in the spinal cats was observed only in those muscles that were tonically active to maintain stance and never in the flexors. Moreover, latencies were increased and amplitudes of activation were diminished compared with control. Nevertheless, flexors and extensors were recruited easily during behaviors such as paw shake and stepping. It is concluded that centers above the lumbosacral cord are required for the full elaboration of automatic postural responses. Although the spinal cat can achieve good weight support, it cannot maintain balance during stance except for brief periods and within narrow limits. This limited stability is probably achieved through spinal reflex mechanisms and the stiffness characteristics of the tonically active extensors.

Chapter 16 Orderly Recruitment Tested Across Muscle Boundaries

Publisher Summary The movements and postures of limb and body segments are achieved through partial activation of multiple muscles. In the cat, multiple muscles are coactive during a variety of movements, such as treadmill locomotion, landing, paw shake, head turning, and in humans, during movements such as hand grip and elbow flexion. There are tens to hundreds of heteronymous motor units belonging to different muscles, which are recruited together during discrete motor actions. This chapter discusses whether recruitment is orderly among heteronymous motor units that are activated together. Recruitment order can be assessed in relation to the motoneuron portion of the motor unit. Alpha motoneurons within a motor nucleus (the collection of motoneurons supplying a single muscle) are progressively recruited from slow to fast axonal conduction velocity. Recruitment that is rank-ordered by these properties is commonly referred to as “recruitment in order” by the size principle. The chapter describes the recruitment order with respect to either a muscle and its motor units or a motor nucleus and its motoneurons or motor axons.

Force-sharing among the primary cat ankle muscles.

Force-sharing measurements among the cat ankle extensors have been used for the past 20 years to elucidate the mechanisms underlying normal movement control and to study the force-sharing among synergistic muscles. Despite these efforts, many questions have eluded satisfactory explanation. Specifically, there has been great debate to what degree the soleus muscle is activated during low level locomotion, and whether the force-sharing among the ankle extensors is primarily caused by central or peripheral factors. We have measured the forces and corresponding electromyo-graphical activities in the soleus, gastrocnemius, plantaris, and tibialis anterior in more than 20 cats and for a variety of movements. Based on these measurements, and additional information obtained on the structural and functional properties of the cat ankle muscles, we conclude that the peak soleus forces are virtually maximal for all speeds of locomotion, despite an apparent submaximal level of activation at slow speeds. Furthermore, we found that the force-sharing among the soleus and gastrocnemius encompasses the entire possible range; from large soleus forces and zero gastrocnemius forces during still standing, to large gastrocnemius forces and no soleus forces during a high-frequency paw shake. Finally, we support the idea that shifts in force-sharing from one muscle to the other are primarily caused by central factors; the peripheral factors, although appealing when considering the large differences in size, morphology, and fibre type distribution, are likely only of secondary importance.

Photochemically-induced ischemia of the rat sciatic nerve produces a dose-dependent and highly reproducible mechanical, heat and cold allodynia, and signs of spontaneous pain

Sensory abnormalities and changes in spontaneous behavior were examined after a photochemically induced ischemic lesion of the rat sciatic nerve. Male adult rats were anesthetized and the sciatic nerve was exposed. After the intravenous injection of a photosensitizing dye, erythrosin B, the exposed nerve was irradiated just proximal to the nerve trifurcation with light from an argon laser. Three different irradiation times were used, 30 s, 1 and 2 min. In sham-operated rats, the exposed sciatic nerve was irradiated for 2 min without prior injection of the erythrosin B. Rats were tested for the presence of mechanical, cold and heat allodynia or hyperalgesia. All the animals in the 1- and 2-min irradiation groups developed mechanical, cold and heat allodynia after nerve irradiation. A significant dose-dependent effect of laser exposure time was observed for all modalities tested (2 min>1 min>30 s=sham). The maximum effects were observed at 3 and 7 days postirradiation and remained present for up to 10 weeks. No significant contralateral effects were observed in any of the groups. In three separate groups of rats (1, 2 and 4 min of laser exposure), the presence of possible signs of spontaneous pain (paw shaking, paw elevation and freezing behavior) was tested. A significant and exposure time-dependent increase in spontaneous paw elevation and paw shaking was observed which was maximal at week 1, but resolved at 4 weeks (4 min>2 min>1 min>sham). In addition, animals in all ischemic groups, but not in the sham group, showed a significant increase in freezing behavior up to 4 weeks after nerve irradiation. Light microscopic evaluation of nerves removed 7 days post-irradiation, i.e. when maximal allodynia was observed, showed clear evidence of demyelination of large myelinated fibers. These data indicate that photochemically-induced peripheral nerve ischemia is associated with abnormal pain-related behaviors, including mechanical, thermal and cold allodynia and signs of spontaneous pain. The incidence and severity of the behavioral changes are clearly dependent on the exposure time and are probably due to, at least in part, a demyelinaton. These results partly confirm previous data using a similar technique and suggest that this may represent a new animal model for peripheral neuropathy of ischemic origin. The advantages of the present model are its good reproducibility and the fact that the nerve injury can be easily quantified and graded.

Effect of Adenosine Receptor Agonists and Antagonists on the Expression of Opiate Withdrawal in Rats

The effects of the selective A 1 adenosine receptor agonist N 6-cyclopentyladenosine (CPA) and the selective A 2a agonist 2-[p-(2-carboxethyl)phenylethyl-ethylamino]-5′-ethylcarboxamidoadenosine (CGS 21680) (each at 0.03, 0.1 and 0.3 mg/kg, SC) as well as the selective A 1 adenosine receptor antagonist 8-cyclopentyl-1,3-dipropylxanthine (DPCPX), non-selective antagonists 3-isobutyl-1-methylxanthine (IBMX), aminophylline, 3,7-dimethyl-1-propargyl-xanthine (DMPX) and 8(p-sulfophenyl)-theophylline (8-SPT) were investigated (each at 5, 10 and 30 mg/kg, SC) for their ability to alter the naloxone-precipitated opiate withdrawal syndrome in morphine-dependent rats. Effects of CPA and CGS 21680 on opiate withdrawal in the presence of aminophylline were also investigated. Both CPA and CGS 21680, caused a significant reduction in the incidence of body shakes, teeth chatter and paw shakes and decreased the amount of faecal matter produced. DPCPX, IBMX, DMPX, 8-SPT and aminophylline significantly increased the incidence of jumps and decreased the amount of faecal matter produced. The incidence of body shakes was significantly increased by DMPX, 8-SPT and IBMX. Neither CPA nor CGS 21680 were able to reverse the significant increase in the incidence of jumps caused by aminophylline. These data suggest that there is a role for endogenous adenosine in the modulation of the opiate abstinence syndrome and both A 1 and A 2a adenosine receptors are involved in this phenomenon.

Fictive motor activities in adult chronic spinal rats transplanted with embryonic brainstem neurons.

The present study was designed to examine the effects of an intraspinal transplantation of embryonic brainstem neurons on fictive motor patterns which can develop in hindlimb nerves of adult chronic spinal rats. Seventeen adult rats were spinalized at T8-9 level and, 8 days later, a suspension of embryonic cells obtained either from the raphe region (RR, n = 8) or from the locus coeruleus (LC, n = 9) was injected caudally (T12-13) to the cord transection. Eight control animals (control rats) were spinalized and injected with vehicle under the same conditions. One to three months later, the animals were decorticated and fictive motor patterns were recorded in representative hindlimb nerves. The data revealed that both control and grafted spinal rats could exhibit two distinctly different fictive motor patterns, one which could be associated with stepping and the other with hindlimb paw shaking. They further showed that following transplantation of embryonic RR or LC neurons the excitability of the spinal stepping generator was increased, whereas that of the spinal neural circuits which generate hindlimb paw shaking was not significantly affected. A histological analysis performed on the spinal cord segments below the transection revealed complete absence of serotonin and noradrenaline immunoreactivity in control spinal animals and, in both types of grafted rats, an extensive monoaminergic reinnervation with synaptic contacts between monoaminergic transplanted neurons and host interneurons and/or motoneurons. The possible mechanisms by which grafted monoaminergic neurons can influence the spinal motor networks are discussed.

Lack of physical dependence in mice after repeated systemic administration of the mixed inhibitor prodrug of enkephalin-degrading enzymes, RB101

Development of physical dependence is observed after treatment with opioid agonists, but not after chronic i.c.v. administration of mixed inhibitors of enkephalin-degrading enzymes. The aim of this study was 10 investigate further this promising result of repeated administration of the systemically active mixed inhibitor prodrug RB101, N-[(R,S)-2-benzyl-3[(S)(2-amino-4-methylthio) butyldithio]-1-oxopropyl]-L-phenylalanine benzyl ester. In a comparative study, the naloxone-evoked withdrawal syndrome was quantified in mice chronically treated with i.p. administered morphine or RB101 (6 and 160 mg/kg, respectively) for 5 days, twice daily. After administration of naloxone (5 mg/kg s.c.) on the sixth day, large behavioral changes (jumps, paw shakes. wet-dog shakes, tremor, teeth chattering) and body weight losses occurred in the morphine-treated mice. In contrast, no significant behavioral signs of physical dependence, or body weight changes were observed in the RB101-treated mice. The difference between morphine and RB101 could he partially due to a very low tonic release of enkephalins in the locus coeruleus a brain region critically involved in the development of physical dependence. These results confirm the potential of mixed inhibitors of enkephatin-degiading enzymes as new non-addictive analgesics.

Effect of alpha 2-adrenoceptor agonists on the expression of morphine-withdrawal in rats.

Previous studies using clonidine indicate that alpha 2-adrenoceptors are involved in suppressing opiate-withdrawal symptoms. However, clonidine may act as a partial agonist at alpha 2-adrenoceptors and it also possesses significant alpha 1-receptor agonist activity. The aim of this study was to determine the role of alpha 2-adrenoceptors in the expression of opiate withdrawal signs using morphine-dependent rats. A range of agonists were selected for study on the basis of their differential preferences for alpha-adrenoceptors. Hooded Wistar rats were made physically dependent on morphine (s.c. injection of an emulsion releasing a total of 250 mg/kg of morphine base over 48 h). Test drugs were injected s.c. followed by naloxone (10 mg/kg i.p.) 20 min later. The incidence of 5 selected withdrawal signs was recorded during the following 20 min. The alpha 2-adrenoceptor agonists displayed different profiles of activity. Azepexole (1-10 mg/kg) reduced all signs. Clonidine (80-800 micrograms/kg) reduced all signs except paw shakes while guanfacine (25-250 micrograms/kg) reduced all except jumping and diarrhoea. Talipexole (0.1-1 mg/kg) reduced all signs except diarrhoea which was not affected and jumping which was markedly enhanced. UK 14,304 (80-800 micrograms/kg) reduced jumps, potentiated paw shakes but did not affect body shakes, teeth chattering or diarrhoea. The results suggest that there are subpopulations of alpha 2-adrenoceptors that modulate the expression of opiate withdrawal signs and/or that some of the drugs used affect receptors other than alpha 2-adrenoceptors.

The NMDA antagonist dizocilpine (MK801) attenuates motivational as well as somatic aspects of naloxone precipitated opioid withdrawal

The non-competative N-methyl-D-aspartate (NMDA) receptor antagonist dizocilpine has recently been reported to antagonize certain overt withdrawal signs in morphine dependent rats. The purpose of the present study was to reassess this response and examine the effect of this drug in a model presumably reflective of the motivitional impact of withdrawal using the place conditioning technique. Rats were made opiate dependent by the subcutaneous implantation of a 75 mg morphine pellet. Three-4 days later withdrawal was precipitated by naloxone 0.5 mg/kg. Dizocilpine (0.1–0.5 mg/kg) attenuated many of the subsequent behaviours elicited by naloxone, notably diarrhoea, mouth movements, paw shakes and ptosis. In a separate group of morphine dependent rats, naloxone (0.05 mg/kg) precipitated withdrawal produced a clear place aversion. This place aversion was blocked by dizocilpine (0.02−0.1 mg/kg) pre-treatment prior to conditioning. Therefore dizocilpine may modify both motivational and somatic aspects of opioid withdrawal.

Fictive motor patterns in chronic spinal cats

1. Fictive motor patterns were recorded in hind leg nerves of 10 adult chronic spinal cats (spinalized at T13). Four of these animals had been trained to step with their hind legs on a treadmill (late-spinal animals), whereas the remainder received no training and were examined a short time after spinalization (early-spinal animals). 2. A fictive pattern resembling the locomotor pattern for stepping was evoked in all animals in response to stimulation of the skin of the perineal region. (2-[2,6-Dichloroaniline]-2-imidazoline) hydrochloride (Clonidine) at doses ranging from 100 to 500 micrograms/kg iv facilitated the production of this pattern, particularly in early-spinal animals. 3. The fictive locomotor pattern in late-spinal animals was more complex than that occurring in early-spinal animals. In the latter the pattern consisted of an alternation of activity in flexor and extensor nerves, and changing leg position did not qualitatively alter the pattern, whereas in late-spinal animals the relative durations of the bursts in different flexors were usually not the same, and the pattern of flexor activity was dependent on leg position. 4. Moving the legs from extension to flexion progressively decreased the duration of flexor bursts, increased the cycle period, and decreased the ease with which the pattern could be evoked in both early- and late-spinal animals. 5. 1-beta-3,4-Dihydroxyphenylalanine (DOPA)/Isonocotinic acid 2-[(2-benzylcarbamoyl)ethyl]hydrazide (Nialamide) treatment following Clonidine in early-spinal animals increased the complexity of flexor burst activity. This, and other observations, indicates that DOPA and Clonidine do not have strictly identical actions on the locomotor pattern generator. 6. Stimulation of the paws in late-spinal animals produced two patterns of activity distinctly different from the locomotor pattern. of activity distinctly different from the locomotor pattern. One was a short sequence of high-frequency rhythmic activity (at approximately 8 c/s) in response to gently stimulating one paw with a water jet, and the other was a slow rhythm in flexor nerves in response to squeezing the paw. 7. The main conclusion of this investigation is that three distinctly different fictive motor patterns can be generated in chronic spinal cats depending on the method and site of stimulation. These patterns correspond to three different behaviors (locomotion, paw shake, and rhythmic leg flexion) that can be elicited in behaving chronic spinal cats in response to the same stimuli.(ABSTRACT TRUNCATED AT 400 WORDS)