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PGI2-induced activation and sensitization of articular mechanonociceptors.

The effects of PGE2 PGI2 and the stable PGI2 analogue cicaprost on the afferent discharge of ankle joint mechanonociceptors were studied in the anaesthetized rat. Close-arterial injection of PGI2 (0.01-0.1 micrograms) or cicaprost (0.05-5 micrograms) caused both sensitization to mechanical stimulation and excitation of the majority of mechanonociceptors, whereas PGE2 (0.03-3 micrograms) had only weak effects on a small number of nociceptive units. These results suggest the existence of specific PGI2 sensitive receptors (IP receptors) on rat sensory afferent nerves, and support the hypothesis that in the rat endogenous PGI2 plays an important role in the lowering of nociceptive thresholds in inflamed joints.

Animals↗

An intrinsic mechanism to stabilize posture--joint-angle-dependent moment arms of the feline ankle muscles.

The neuromuscular system can use sensory feedback to regulate motion, but these feedback loops involve relatively long delays (50-100 ms) and may produce undesirable oscillations. However, stabilizing changes in muscle force can also be provided intrinsically (i.e. without feedback) by 2 properties of the muscle itself, its force-length and force-velocity relationships. We have discovered another intrinsically stabilizing mechanism in the musculoskeletal architecture of the cat ankle joint. Many of its muscles have their predominant moment arms about the adduction/abduction axis, with smaller moment arms for inversion/eversion and about the principal axis of motion, dorsiflexion/extension. The magnitudes of the moment arms in ab/adduction and in/eversion depend strongly on joint angle, increasing for positions away from neutral that lengthen the muscles. Thus, co-activation of agonist-antagonist pairs, known to occur in these muscles, would provide immediate stabilizing changes in torque when the ankle is perturbed away from neutral position.

Anesthesia↗

Reinnervation of denervated extensor digitorum longus of the rat by the nerve of the soleus does not induce the type I myosin synthesis directly but through a sequential transition of type II myosin isoforms.

The fast-contracting extensor digitorum longus (EDL) muscle of 1-month-old rats was denervated and reinnervated by the nerve innervating the slow-contracting soleus muscle. After variable periods of time, the myosin isoform content of the EDL was analyzed by sensitive electrophoretic techniques, which allowed to discriminate between the slow-type I and the three, IIA, (IID or IIX) and IIB, fast-type II myosin isoforms. Compared to the control EDL, which contains predominantly the IIB isoform, the operated muscles contained variable proportions of all the isoforms. Analysis of the results leads us to conclude that reinnervation of EDL induces a sequential transition of myosin isoforms: IIB----(IID or IIX)----IIA----I.

Animals↗

Prolonged potentiation of transmission through a withdrawal reflex pathway after noxious stimulation of the heel in the rabbit.

The sural-gastrocnemius reflex of the spinalized rabbit was potentiated to an average of 3-6 times control levels after the application of noxious mechanical, thermal or chemical stimuli to the skin of the heel. Facilitation of the reflex was maximal within 1 min of the noxious stimulus, and in many cases persisted for more than 1 h. Prolonged increases in the excitability of the sural-gastrocnemius reflex were not seen after innocuous mechanical or thermal stimulation of the heel. Repetitive electrical stimulation of the sural nerve (100 shocks given at 0.5 Hz) caused persistent facilitation of the reflex when small myelinated A delta fibres or non-myelinated C-fibres were recruited by the conditioning stimulus. Such protracted increases in the excitability of the sural-gastrocnemius pathway would enhance the protective functions of this reflex. The mechanisms described here have probably evolved to provide a high level of reflex protection to the heel after tissue damage has occurred at that site.

Animals↗

Effects on recovery of soleus and extensor digitorum longus muscles of prolonged wheel running during a period of repeated nerve damage.

The right sciatic nerve in NMRI mice was frozen under anaesthesia 13 times at three-week intervals for a total period of 8.5 months. During this period, but not afterwards, one sub-group of these mice had access to running wheels in which the animals ran several kilometres per night, thereby actively or passively training reinnervated or denervated leg muscles, as well as the intact contralateral muscles. A number of distinct effects persisted for as long as 14-18 weeks after the termination of this "endurance training". In reinnervated soleus muscle, tetanic force was significantly higher (37%) in the trained muscles as was muscle weight (36%); in general, negative effects of the nerve damage persisted. In the reinnervated extensor digitorum longus, tetanic force and muscle weight were significantly smaller in the trained animals (by 11 and 16%, respectively) which are considered typical effects of endurance training. The resistance of the soleus neuromuscular junction to block by both curare and Mg2+ was depressed on the damaged side but this property was not influenced by the training; in extensor digitorum longus the pattern was similar. It is concluded that training during the period of repeated cycles of denervation-reinnervation produced significant effects which impressively outlasted the training period. The possible nature of these effects is discussed.

Animals↗

Tachykininergic tone in the spinal cord of the rabbit: dependence on nociceptive input arising from invasive surgery.

In the decerebrated and spinalized rabbit, reflexes evoked in the gastrocnemius medialis muscle nerve by electrical stimulation of teh sural nerve are suppressed after blockade of NK-tachykinin receptors. This observation suggests that endogenous tachykinins tonically enhance transmission between sural nerve afferents and gastrocnemius motoneurons. In the present study we have investigated some possible sources of this tachykininergic tone. Electrical stimulation of the sural nerve at 1 Hz, as used in our previous investigation, leads to increased gastrocnemius reflex responses with successive stimuli. We examined reflexes evoked by pairs of sural stimuli separated by intervals of 10-1000 ms, and found that responses to the second stimuli of such pairs were significantly enhanced at intervals from 50 to 500 ms. Treatment with the NK receptor antagonist CP-99,994 [(+)-(2S,3S)-3-(2-methoxybenzylamino)-2-phenylpiperidine; 1 mg/kg, i.v.] reduced gastrocnemius reflex responses per se, but did not alter the facilitatory effects of pairing sural stimuli. Subsequent treatment with the glutamate N-methyl-D-aspartate receptor blocker dizocilpine (0.5 mg/kg, i.v.) further reduced reflex responses and abolished paired-pulse facilitation. In a second set of experiments, rabbits were prepared so that reflexes could be studied with minimal surgical preparation of the hindlimb. Reflex responses recorded in this way were enhanced by treatment with CP-99,994 (up to 1 mg/kg, i.v.). Subsequent administration of the opioid receptor antagonist naloxone (1-10 microg/kg, i.v.) increased reflexes, as seen previously in surgically-prepared animals. These data show that tachkininergic modulation of spinal reflexes in the rabbit results from the nociceptive input arising from surgical preparation of the leg. In contrast, tonic opioidergic inhibition of reflexes is not substantially dependent on such input.

Animals↗

Noninvasive 3D MR microscopy as a tool in pharmacological research: application to a model of rheumatoid arthritis.

Magnetic resonance microscopy (MRM) was applied to noninvasively image skeletal structures in the hindpaw of the live rat to characterize the progression of a heterologous type II collagen-induced arthritic process. Using a resonator, with optimized filling factor, three-dimensional (3D) gradient-echo images with voxel dimensions of 94 x 81 x 60 micron3 were acquired in 54.6 min. Three-dimensional MRM reduces the slice positioning problem, which is critical in longitudinal studies. Moreover, due to the much smaller slice thickness of images derived from 3D data sets, partial volume effects are less pronounced than in corresponding 2D images. Distinct pathomorphological changes associated with the collagen-induced arthritic process (e.g., increase of metatarsophalangeal joint space, and bone and cartilage erosion) could thus be analyzed under in vivo conditions.

Animals↗

Disrupted bone metabolism in contaminant-exposed white storks (Ciconia ciconia) in southwestern Spain.

In 1998, the Aznalcóllar mine tailings dyke in southwestern Spain broke, flooding the Agrio-Guadiamar river system with acid tailings up to the borders of one of the largest breeding colonies of white storks in the western Palearctic, Dehesa de Abajo. Over the following years, a high proportion of nestlings developed leg defects not seen before the spill, prompting this study. Nestlings with deformed legs had significantly lower plasma phosphorous (P) and higher Ca:P ratios than non-deformed cohorts in the first two years, but in the third year, when more, younger birds were studied, plasma P ranged from much higher to much lower in the affected colony compared with reference birds. Coefficients of variation for phosphorous were 19% and 60%, in reference and contaminated colonies, respectively. Storks from the contaminated colony were unable to control P levels and Ca:P ratios within the narrow limits necessary for normal bone development.

Alkaline Phosphatase↗

Pitx1 determines the morphology of muscle, tendon, and bones of the hindlimb.

The vertebrate forelimb and hindlimb are serially homologous structures; however, their distinctive morphologies suggest that different mechanisms are associated with each limb type to give rise to limb-type identity. Three genes have been implicated in this process; T-box transcription factors Tbx5 and Tbx4, which are expressed in the forelimb and hindlimb, respectively, and a paired-type homeodomain transcription factor Pitx1, expressed in the hindlimb. To explore the roles of Pitx1 and Tbx4 in patterning the hindlimb, we have ectopically misexpressed these genes in the mouse forelimb using transgenic methods. We have developed a novel technique for visualising the structure and organisation of tissues in limbs in 3D using optical projection tomography (OPT). This approach provides unparalleled access to understanding the relationships between connective tissues during development of the limb. Misexpression of Pitx1 in the forelimb results in the transformation and translocation of specific muscles, tendons, and bones of the forelimb so that they acquire a hindlimb-like morphology. Pitx1 also upregulates hindlimb-specific factors in the forelimb, including Hoxc10 and Tbx4. In contrast, misexpression of Tbx4 in the forelimb does not result in a transformation of limb-type morphology. These results demonstrate that Pitx1, but not Tbx4, determines the morphological identity of hindlimb tissues.

Animals↗

The tyrosine kinase inhibitor tyrphostin AG126 reduces the development of acute and chronic inflammation.

Protein tyrosine kinases help to regulate the expression of many genes that play important roles in inflammation. Here we investigate the effects of the tyrosine kinase inhibitor tyrphostin AG126 in two animal models of acute and chronic inflammation, carrageenan-induced pleurisy and collagen-induced arthritis. We report here that tyrphostin AG126 (given at 1, 3, or 10 mg/kg i.p. in the pleurisy model or 5 mg/kg i.p. every 48 hours in the arthritis model) exerts potent anti-inflammatory effects in animal models of acute and chronic inflammation in vivo. These include the inhibition of pleural exudate formation and mononuclear cell infiltration (pleurisy model) and the development of clinical signs and tissue injury (arthritis model). Furthermore, tyrphostin AG126 reduced the staining for nitrotyrosine and poly (ADP-ribose) polymerase (by immunohistochemistry) and the expression of inducible nitric oxide synthase and cyclooxygenase-2 in the lungs of carrageenan-treated rats and in the joints from collagen-treated rats. Thus, we provide the first evidence that prevention of the activation of protein tyrosine kinases reduces the development of acute and chronic inflammation, and that inhibition of the activity of certain tyrosine kinases may represent a novel approach for the therapy of inflammation.

Acute Disease↗

Calpain inhibitor I reduces the development of acute and chronic inflammation.

There is limited evidence that inhibition of the activity of the protease calpain I reduces inflammation. Here we investigate the effects of calpain inhibitor I in animal models of acute and chronic inflammation (carrageenan-induced pleurisy and collagen-induced arthritis). We report here for the first time that calpain inhibitor I (given at 5, 10, or 20 mg/kg i.p. in the pleurisy model or at 5 mg/kg i.p every 48 hours in the arthritis model) exerts potent anti-inflammatory effects (eg, inhibition of pleural exudate formation, mononuclear cell infiltration, delayed the development of the clinical signs and histological injury) in vivo. Furthermore, calpain inhibitor I reduced (1) the staining for nitrotyrosine and poly (ADP-ribose) polymerase (immunohistochemistry) and (2) the expression of inducible nitric oxide synthase and cyclooxygenase-2 in the lungs of carrageenan-treated rats and in joints from collagen-treated rats. Thus, prevention of the activation of calpain I reduces the development of acute and chronic inflammation. Inhibition of calpain I activity may represent a novel therapeutic approach for the therapy of inflammation.

Acute Disease↗

(S)-4C3HPG, a mixed group I mGlu receptor antagonist and a group II agonist, administered intrastriatally, counteracts parkinsonian-like muscle rigidity in rats.

The aim of the present study was to determine whether S-4-carboxy-3-hydroxyphenylglycine (S)-4C3HPG, a mixed group I glutamate metabotropic receptor antagonist and a group II agonist, attenuated parkinsonian-like muscle rigidity in rats. Muscle tone was examined using a combined mechano and electromyographic method, which measured simultaneously the muscle resistance (MMG) of the rat's hind foot to passive extension and flexion in the ankle joint and the electromyographic activity (EMG) of the antagonistic muscles of that joint: gastrocnemius and tibialis anterior. Muscle rigidity was induced by pretreatment with haloperidol (1 mg/kg i.p.). (S)-4C3HPG injected in doses of 5 and 15 microg/0.5 microl bilaterally, into the rostral region of the striatum, decreased both the haloperidol-induced muscle rigidity (MMG) and the enhanced electromyographic activity (EMG). The present results suggest that blockade of mGluR1 receptors and/or activation of mGluR2 ones, localized in the rostral part of the striatum, may be responsible for the anti-parkinsonian effect of (S)-4C3HPG.

Animals↗

An electrophysiological investigation into the monosynaptic H-reflex in the rat.

In order to set up a non-invasive, reliable and reproducible model for investigating alpha-motoneuronal activity, we studied the electrophysiological features of a monosynaptic H-reflex in anaesthetised intact rats, anaesthetised and non-anaesthetised rats transected at the level of the obex. Electrical stimulation of the tibial nerve at the ankle elicited an H-reflex, an F-wave and a direct motor (M) response in the plantaris muscles of all preparations. The H-reflex and F-wave exhibited very similar latencies. The H-reflex had a low threshold and a constant latency. Its amplitude increased as a function of stimulus intensity to reach a maximum value but then decreased when the stimulus intensity was further increased. It could follow high rates of stimulation without any change in shape or latency. The F-wave had a lower amplitude which together with its latency varied from one stimulus to the next. It appeared with intensities of stimulation that elicited an almost maximal M-response and did not decrease when the stimulation was increased. It did not appear systematically from one stimulus to the next. The H-reflex, but neither the F-wave nor the direct motor M-response, was depressed both by vibratory stimuli applied on the Achilles' tendon and following nociceptive stimulation of the flexor reflex afferents. This model could be used for assessing any potential direct effect on motoneurones of a physiological or pharmacological conditioning procedure.

Achilles Tendon↗

Short-latency synaptic patterns among cat peroneal motoneurons.

Motoneurons innervating peroneal muscles in the cat leg (PB, PT and PL, respectively, for peroneus brevis, tertius and longus) were examined for their connections with afferents from these and other leg muscles and with cutaneous afferents. The aim was to investigate (1) whether inputs from nearby muscles and cutaneous areas are likely to assist or oppose the excitation elicited in peroneal motoneurons by PB contractions, and (2) whether reflex connectivity might allow distinction of alpha (i.e. motoneurons innervating skeletal muscle fibres) and beta (i.e. motoneurons innervating both skeletal and intrafusal muscle fibres) subgroups among PB and PT motoneurons. In the three peroneal pools, every motoneuron had excitatory monosynaptic connections with Ia afferents from each of the three peroneal muscles, and nearly every motoneuron received di- or trisynaptic excitation from low-threshold cutaneous afferents in sural or superficial peroneal nerves. Inputs from these sources might facilitate the contraction-induced positive feedback. In contrast, the patterns of short-latency synaptic connections with group I afferents from pretibial flexor and post-tibial extensor muscles were heterogeneous among peroneal motoneurons but did not point to any specific beta pattern.

Animals↗

Arthrosonography--the use of diagnostic ultrasound in septic and traumatic arthritis in cattle--a retrospective study of 25 patients.

The case records, clinical, radiographic, ultrasonographic and arthrocentesis findings from 25 cattle with septic or traumatic arthritis were reviewed. The total of 30 joints involved were: articulatio cubiti (elbow joint)-1, articulatio carpi (carpal joint)-10; articulatio genus (stifle joint)-4, articulatio tarsocruralis (tarsocrural joint)-8; articulatio metacarpo-/metatarsophalangea (metacarpo-/metatarsophalangeal joint -7. Ultrasonographic investigation was performed using 7.5 MHz linear and 5 MHz sector transducers. In eight cattle, a concurrent infection of periarticular tendon sheaths or hygroma was diagnosed. The distension of the joint cavities could be imaged in all cases. Assessing echogenicity, acoustic enhancement, ultrasonographic character of the exudate (liquid or clotted) and findings revealed by aspiration, arthrotomy or at necropsy, the synovial effusion was classified as: serous, fibrinous or fibrino-purulent. Diagnostic ultrasound provided a comprehensive preoperative diagnosis in septic and traumatic arthritis by imaging the accurate localization of soft-tissue swelling and the extent and consistency of joint effusion.

Animals↗

Toe out angle: a functional index for the evaluation of sciatic nerve recovery in the rat model.

In experimental peripheral nerve studies, the rat sciatic nerve model is widely used to examine functional outcome following nerve injury and repair. A variety of evaluation methods exist in the literature, but an adequate selection continues to be a critical point for the researcher. Rats with sciatic nerve injury typically ambulate with an external rotation of the foot. A new functional assessment instrument, the toe out angle (TOA) is quantified using computerized gait analysis. We compared Sciatic Functional Index (SFI) with TOA parameter after peripheral nerve transection and entubulation repair. We found a good correlation between SFI and TOA measurements in terms of predicting functional recovery. Moreover, the TOA provides information on the biomechanical consequences of the external rotation of the foot in the stance phase of walking.

Animals↗

Force output during and following active stretches of rat plantar flexor muscles: effect of velocity of ankle rotation.

During the development of force deficits by repeated stretches, velocity-sensitive changes in the extra force produced during and after subsequent stretching has not been studied. In the present study, repeated dorsiflexion of the foot of rats with maximally contracting plantar flexor muscles was performed at two angular velocities [0.87 (slow muscle stretch) and 10.47rads(-1) (fast muscle stretch)] to examine the active force of the muscles during and following dorsiflexion. Dorsiflexion was performed 30 times with a rest period of 3min between the stretches to minimize muscle fatigue. The ability of rat plantar flexor muscles to produce additional force during the stretch was not velocity sensitive. In contrast, repeated dorsiflexion with fast muscle stretches, but not with slow muscle stretches, resulted in an increase in the force decay with time following the stretches (i.e. increased stress relaxation), as indicated by a change in the time constant of force decay during stress relaxation. Apparently, the stress-relaxation of rat plantar flexor muscles is sensitive to angular velocity of ankle movements; repeated fast, but not slow dorsiflexion, alters the stress relaxation process of active skeletal muscles exposed to stretches which create a force deficit. The change in time constant of force decay during stress relaxation in response to a series of repeated stretches might provide information on the sarcomere length distribution in skeletal muscles.

Animals↗

Forces of individual cat ankle extensor muscles during locomotion predicted using static optimization.

In order to test the principles of the control of synergistic muscles that were proposed in the literature, forces of cat soleus (SO), gastrocnemius (GA), and plantaris (PL) measured during locomotion were compared with the corresponding forces predicted using different optimization criteria. Forces of cat SO, GA, and PL, and the corresponding cat kinematics were recorded simultaneously using E-shaped force transducers and high-speed video, respectively. Measurements were obtained from three cats walking and trotting on a treadmill at five nominal speeds ranging from 0.4 to 1.8 m s-1. Muscle forces were predicted using static optimization and a musculoskeletal model of the cat hindlimb consisting of three segments (foot, shank, and thigh) and three muscles (SO, GA, and PL). Six optimization criteria which had been proposed in the literature were tested. Linear criteria based on the principles of minimum muscle force and stress predicted forces during the stance phase with an average normalized error of 59-322%. Three other criteria--minimization of the sum of the relative muscle forces squared, minimization of the sum of the muscle stresses cubed, and minimization of the upper bound for all of the muscle stresses-showed a better performance: (i) the average error was 43-119% and (ii) the correlation coefficient calculated between the predicted and actual forces exceeded 0.9 for all three muscles. A criterion that was based on the principle of minimum fatigue and accounted for the percentage of slow-twitch fibers in the muscles, had the lowest average error (26-52%). The high correlation (0.97-0.99) between the measured forces and forces predicted by using the minimum fatigue criterion suggested that force sharing among SO, GA, and PL during cat locomotion may be the same for a given set of joint moments and muscle moment arms. It was concluded that static optimization with the appropriate criterion can predict muscle forces adequately for specific movement conditions.

Animals↗