Appendicular fracture repair in dogs using the locking compression plate system: 47 cases.

The locking compression plate (LCP) has combination screw holes, making it possible to use the implant in three different ways; as a pure internal fixator using locking head screws, as a conventional compression plate using compression screws, or as a hybrid of the two. The experience with the LCP system in veterinary fracture repair is limited. The objective of this study was to evaluate the outcome of appendicular fractures in dogs, which were repaired with the LCP system combined with less invasive surgical techniques. Medical records and radiographs from 47 dogs were studied retrospectively. Thirty-four percent of the fractures were simple, six percent wedge and 60% comminuted fractures of the humerus (11 %), radius and ulna (30 %), femur (34 %) and of the tibia and fibula (25 %). The fractures were treated using the LCP as an internal fixator; in some cases as a plate and rod construct. Forty-six of 47 fractures reached radiographic union. Mean healing time of the fractures was seven weeks (95% confidence interval from 5.8 to 8.3 weeks). There were statistically significant differences in healing time between juvenile (age under one year) and adults. Complications in the form of implant failures and infections were encountered in approximately 11% of the cases. All implant failures were due to surgical errors. The LCP system in combination with a less invasive surgical approach was found advantageous in comminuted fractures where the LCP was used as a bridging plate, in situations when exact plate contouring was difficult, and when other implants prevented the use of bi-cortical screws.

Bulbospinal neurons implicated in mesopontine-induced anesthesia are substantially collateralized.

Microinjection of pentobarbital or other gamma-aminobutyric acid type A receptor (GABA(A)-R) active anesthetics into a brainstem region in the rat that we have called the mesopontine tegmental anesthesia area (MPTA) induces a general anesthesia-like state that includes suppression of locomotor activity, loss of the righting reflex, atonia, antinociception, and apparent loss of consciousness. The suppression of muscle tone and of nocifensive spinal reflexes suggests a direct or indirect effect at the level of the spinal cord itself, an inference supported by anterograde tracing from the MPTA area. We have now used single and double retrograde tracing to characterize this bulbospinal pathway further. The MPTA contains the majority of all bulbospinal neurons present at mesopontine levels (65.8%). Many of these neurons, although not all, appear to have a highly collateralized projection pattern within the spinal cord. About 40% of the MPTA neurons that project to the lumbar spinal cord also have collaterals at cervical levels, and about 60% of those with projections to the ventral horn also have projections to the dorsal horn (at cervical levels). However, the large majority projects either ipsilaterally or contralaterally. Relatively few ( approximately 13%) send collaterals to both sides of the spinal cord. The pattern of connectivity revealed appears to be consistent with a system designed primarily to modulate motor and sensory functions globally, over the entire neuraxis, rather than regionally or segmentally.

Neural pathways associated with loss of consciousness caused by intracerebral microinjection of GABA A-active anesthetics.

Anesthesia, slow-wave sleep, syncope, concussion and reversible coma are behavioral states characterized by loss of consciousness, slow-wave cortical electroencephalogram, and motor and sensory suppression. We identified a focal area in the rat brainstem, the mesopontine tegmental anesthesia area (MPTA), at which microinjection of pentobarbital and other GABA(A) receptor (GABA(A)-R) agonists reversibly induced an anesthesia-like state. This effect was attenuated by local pre-treatment with the GABA(A)-R antagonist bicuculline. Using neuroanatomical tracing we identified four pathways ascending from the MPTA that are positioned to mediate electroencephalographic synchronization and loss of consciousness: (i) projections to the intralaminar thalamic nuclei that, in turn, project to the cortex; (ii) projections to several pontomesencephalic, diencephalic and basal forebrain nuclei that project cortically and are considered parts of an ascending "arousal system"; (iii) a projection to other parts of the subcortical forebrain, including the septal area, hypothalamus, zona incerta and striato-pallidal system, that may indirectly affect cortical arousal and hippocampal theta rhythm; and (iv) modest projections directly to the frontal cortex. Several of these areas have prominent reciprocal projections back to the MPTA, notably the zona incerta, lateral hypothalamus and frontal cortex. We hypothesize that barbiturate anesthetics and related agents microinjected into the MPTA enhance the inhibitory response of local GABA(A)-R-bearing neurons to endogenous GABA released at baseline during wakefulness. This modulates activity in one or more of the identified ascending neural pathways, ultimately leading to loss of consciousness.

Mesopontine tegmental anesthesia area projects independently to the rostromedial medulla and to the spinal cord.

General anesthetics are presumed to act in a distributed manner throughout the CNS. However, we found that microinjection of GABAA-receptor (GABAA-R) active anesthetics into a restricted locus in the rat brainstem, the mesopontine tegmental anesthesia area (MPTA), rapidly induces a reversible anesthesia-like state characterized by suppressed locomotion, atonia, anti-nociception and loss of consciousness. GABA-sensitive neurons in the MPTA may therefore have powerful control over major aspects of brain and spinal function. Tracer studies have shown that the MPTA projects to the rostromedial medulla, an important reticulospinal relay for pain modulation and motor control. It also projects directly to the spinal cord. But do individual MPTA neurons project to one or to both targets? We microinjected fluorogold into the rostromedial medulla and cholera toxin b-subunit into the spinal cord, or vice versa. Neurons that were double-labeled, and hence project to both targets, were intermingled with single-labeled neurons within the MPTA, and comprised only 11.5% of the total. MPTA neurons that project directly to the spinal cord were larger, on average, than those projecting to the rostromedial medulla, differed in shape, and were much more likely to express GABAA-alpha1Rs as assessed by receptor alpha-1 subunit immunoreactivity (51.4% vs. 18.9%). Thus, for the most part, separate and morphologically distinct populations of MPTA neurons project to the rostromedial medulla and to the spinal cord. Either or both may be involved in the modulation of nociception and the generation of atonia during the MPTA-induced anesthesia-like state.

Projections from the mesopontine tegmental anesthesia area to regions involved in pain modulation.

Pentobarbital microinjected into a restricted locus in the upper brainstem induces a general anesthesia-like state characterized by atonia, loss of consciousness, and pain suppression as assessed by loss of nocifensive response to noxious stimuli. This locus is the mesopontine tegmental anesthesia area (MPTA). Although anesthetic agents directly influence spinal cord nociceptive processing, antinociception during intracerebral microinjection indicates that they can also act supraspinally. Using neuroanatomical tracing methods we show that the MPTA has multiple descending projections to brainstem and spinal areas associated with pain modulation. Most prominent is a massive projection to the rostromedial medulla, a nodal region for descending pain modulation. Together with the periaqueductal gray (PAG), the MPTA is the major mesopontine input to this region. Less dense projections target the PAG, the locus coeruleus and pericoerulear areas, and dorsal and ventral reticular nuclei of the caudal medulla. The MPTA also has modest direct projections to the trigeminal nuclear complex and to superficial layers of the dorsal horn. Double anterograde and retrograde labeling at the light and electron microscopic levels shows that MPTA neurons with descending projections synapse directly on spinally projecting cells of rostromedial medulla. The prominence of the MPTA's projection to the rostromedial medulla suggests that, like the PAG, it may exert antinociceptive actions via this bulbospinal relay.

An integrative approach for the identification of quantitative trait loci.

The genetic dissection of complex traits is one of the most difficult and most important challenges facing science today. We discuss here an integrative approach to quantitative trait loci (QTL) mapping in mice. This approach makes use of the wealth of genetic tools available in mice, as well as the recent advances in genome sequence data already available for a number of inbred mouse strains. We have developed mapping strategies that allow a stepwise narrowing of a QTL mapping interval, prioritizing candidate genes for further analysis with the potential of identifying the most probable candidate gene for the given trait. This approach integrates traditional mapping tools, fine mapping tools, sequence-based analysis, bioinformatics and gene expression.

Comparison of the recurrence rate of gastric dilatation with or without volvulus in dogs after circumcostal gastropexy versus gastrocolopexy.

OBJECTIVE: To compare the recurrence rate of acute gastric dilatation with or without volvulus (GDV) after circumcostal gastropexy (CCGP) or gastrocolopexy (GCP) in dogs. STUDY DESIGN: A prospective, double-blind, multicenter, randomized, controlled, clinical trial with two groups (A and B). ANIMALS: Fifty-four client-owned dogs presented for treatment of GDV. METHODS: Dogs with acute GDV that had not previously had a gastropexy performed were included. The preoperative treatment before gastropexy was standardized. A CCGP was performed on dogs in group A, and a GCP was performed on dogs in group B. Postoperative treatment was standardized, but deviation did occur according to the special needs of particular patients. A minimal follow-up time of 180 days was required for dogs not excluded from the study. The median follow-up time in group A was 700 days; in group B, it was 400 days. The occurrence of abdominal pain and gastrointestinal problems after surgery were recorded by the owners. RESULTS: There was no significant difference in the recurrence rate of GDV between the two groups. At the end of the study, the recurrence rate was 9% and 20% in group A and in group B, respectively. CONCLUSIONS: Both surgical techniques are effective in preventing recurrence of GDV.

Neuropathic pain: what do we do with all these theories?

Only a generation ago there were few ideas as to what might cause neuropathic pain, and even fewer relevant data. In contrast, we can currently point to hundreds of distinct cellular changes that are triggered by nerve injury and that might be relevant to the emergence of pain symptomatology. The number may soon increase to thousands. It is essential, therefore, to redirect efforts towards the development of experimental strategies for testing which of these are essential parts of the pain process and which are tangential. In this paper I point out four such strategies: timing, deletion, prevention and genetic heterogeneity, and summarize how one neuropathic pain theory, the ectopic pacemaker hypothesis, holds up to scrutiny.

Hyperexcitability in sensory neurons of rats selected for high versus low neuropathic pain phenotype.

Selection line rats congenitally high or low for autotomy in the neuroma model of neuropathic pain (HA and LA rats) were found to be correspondingly high and low in a second type of neuropathic pain, the Chung model, which employs an alternative phenotypic endpoint, tactile allodynia. It has been proposed that both phenotypes reflect ectopic hyperexcitability in axotomized primary sensory neurons. To test this hypothesis we made in vitro recordings from sensory neurons in the L4 and 5 dorsal root ganglia. Baseline excitability was similar in HA and LA rats, and axotomy caused an increase in both lines. However, in the one neuronal subclass previously linked to neuropathic pain in these models the increase was significantly greater in HA than LA rats, and only at the time when pain scores in the two lines were diverging. Heritable differences in electrical response to axotomy in a specific afferent cell type appear to be a fundamental determinant of neuropathic pain.

Dye coupling does not explain functional crosstalk within dorsal root ganglia.

Most primary sensory neurons in rat dorsal root ganglia (DRGs) are depolarized during repetitive impulse activity in neighboring neurons that share the same ganglion. We wondered whether this functional crosstalk might be mediated by a network of cytoplasmic bridges (gap junctions) between neighboring neurons and their satellite glia. Neurobiotin was injected intracellularly in whole excised DRGs. Some of the animals were intact, and others underwent transection of the ipsilateral sciatic nerve 7 to 21 days prior to injection. A total of 44 directly injected neurons were recovered histologically. There was little or no evidence of dye spread to neighboring satellite cells or neurons that would have indicated the presence of cytoplasmic bridges, certainly not enough to account for the nearly universal functional coupling that occurs among these neurons. Functional crosstalk within DRGs must therefore employ a different mechanism.

Spinal nerve injury enhances subthreshold membrane potential oscillations in DRG neurons: relation to neuropathic pain.

Primary sensory neurons with myelinated axons were examined in vitro in excised whole lumbar dorsal root ganglia (DRGs) taken from adult rats up to 9 days after tight ligation and transection of the L(5) spinal nerve (Chung model of neuropathic pain). Properties of subthreshold membrane potential oscillations, and of repetitive spike discharge, were examined. About 5% of the DRG neurons sampled in control DRGs exhibited high-frequency, subthreshold sinusoidal oscillations in their membrane potential at rest (V(r)), and an additional 4.4% developed such oscillations on depolarization. Virtually all had noninflected action potentials (A(0) neurons). Amplitude and frequency of subthreshold oscillations were voltage sensitive. A(0) neurons with oscillations at V(r) appear to constitute a population distinct from A(0) neurons that oscillate only on depolarization. Axotomy triggered a significant increase in the proportion of neurons exhibiting subthreshold oscillations both at V(r) and on depolarization. This change occurred within a narrow time window 16-24 h postoperative. Axotomy also shifted the membrane potential at which oscillation amplitude was maximal to more negative (hyperpolarized) values, and lowered oscillation frequency at any given membrane potential. Most neurons that had oscillations at V(r), or that developed them on depolarization, began to fire repetitively when further depolarized. Spikes were triggered by the depolarizing phase of oscillatory sinusoids. Neurons that did not develop subthreshold oscillations never discharged repetitively and rarely fired more than a single spike or a short burst, on step depolarization. The most prominent spike waveform parameters distinguishing neurons capable of generating subthreshold oscillations, and hence repetitive firing, was their brief postspike afterhyperpolarization (AHP) and their low single-spike threshold. Neurons that oscillated at V(r) tended to have a more prolonged spike, with slower rise- and fall-time kinetics, and lower spike threshold, than cells that oscillated only on depolarization. The main effects of axotomy were to increase spike duration, slow rise- and fall-time kinetics, and reduce single-spike threshold. Tactile allodynia following spinal nerve injury is thought to result from central amplification ("central sensitization") of afferent signals entering the spinal cord from residual intact afferents. The central sensitization, in turn, is thought to be triggered and maintained in the Chung model by ectopic firing originating in the axotomized afferent neurons. Axotomy by spinal nerve injury enhances subthreshold membrane potential oscillations in DRG neurons, augments ectopic discharge, and hence precipitates neuropathic pain.

Preoperative open field behavior predicts levels of neuropathic pain-related behavior in mice.

Exploratory open field (OF) activity was assessed in seven different mouse strains and selection lines. We counted the number of beam interruptions made by three cagemate mice at a time. This assay tests reactivity to aversive stimuli, anxiety and emotionality. One hindlimb was then totally denervated by transecting the sciatic and saphenous nerves on one side, and autotomy, a behavior thought to be related to neuropathic pain, was quantified over 35 days. We report that OF activity and autotomy are highly variable across different strains/lines. These results reaffirm the genetic control of these behaviors. We also found that these behaviors are inversely and significantly correlated. We suggest that common genetically-determined neural mechanisms may underlie anxiety, emotionality and neuropathic pain in mice.

Functional cross-excitation between afferent A- and C-neurons in dorsal root ganglia.

Electrophysiological recordings were made in vitro from primary afferent neurons with unmyelinated axons (C-neurons) in excised rat dorsal root ganglia. Spike activity triggered in neurons with myelinated axons (A-neurons) by stimulation of the peripheral nerve or the dorsal root produced a transient depolarization in passive neighboring C-neurons that share the same ganglion. About 90% of neurons sampled responded with this "cross-depolarization". Cross-depolarization was associated with functional excitation as indicated by an increase in firing probability in response to previously subthreshold intracellular test pulses. Furthermore, it yielded a net increase of the input resistance of the affected C-neurons. We suggest that functional coupling among DRG neurons could serve a metabolic role, providing a functionally relevant feedback signal useful for controlling the excitability of nociceptive sensory endings. In addition, the results provide a novel mechanism whereby afferent nociceptors could be stimulated by activity in low-threshold mechanoreceptors, particularly in the event of nerve injury. Hence, the coupling between afferent A- and C-neurons in dorsal root ganglia provides a novel candidate mechanism for neuropathic pain.

Membrane potential oscillations in dorsal root ganglion neurons: role in normal electrogenesis and neuropathic pain.

Abnormal afferent discharge originating at ectopic sites in injured primary sensory neurons is thought to be an important generator of paraesthesias, dysaesthesias, and chronic neuropathic pain. We report here that the ability of these neurons to sustain repetitive discharge depends on intrinsic resonant properties of the cell membrane and that the prevalence of this characteristic increases after nerve injury. Recording from primary sensory neurons in excised rat dorsal root ganglia, we found that some cells show subthreshold oscillations in their membrane potential. The amplitude, frequency, and coherence of these oscillations were voltage sensitive. Oscillations gave rise to action potentials when they reached threshold. Indeed, the presence of oscillations proved to be a necessary condition for sustained spiking both at resting membrane potential and on depolarization; neurons without them were incapable of sustained discharge even on deep depolarization. Previous nerve injury increased the proportion of neurons sampled that had subthreshold oscillations, and hence the proportion that generated ectopic spike discharge. Oscillatory behavior and ectopic spiking were eliminated by [Na(+)](o) substitution or bath application of lidocaine or tetrodotoxin (TTX), under conditions that preserved axonal spike propagation. This suggests that a TTX-sensitive Na(+) conductance contributes to the oscillations. Selective pharmacological suppression of subthreshold oscillations may offer a means of controlling neuropathic paraesthesias and pain without blocking afferent nerve conduction.

Heritability of nociception I: responses of 11 inbred mouse strains on 12 measures of nociception.

It is generally acknowledged that humans display highly variable sensitivity to pain, including variable responses to identical injuries or pathologies. The possible contribution of genetic factors has, however, been largely overlooked. An emerging rodent literature documents the importance of genotype in mediating basal nociceptive sensitivity, in establishing a predisposition to neuropathic pain following neural injury, and in determining sensitivity to pharmacological agents and endogenous antinociception. One clear finding from these studies is that the effect of genotype is at least partially specific to the nociceptive assay being considered. In this report we begin to systematically describe and characterize genetic variability of nociception in a mammalian species, Mus musculus. We tested 11 readily-available inbred mouse strains (129/J, A/J, AKR/J, BALB/cJ, C3H/HeJ, C57BL/6J, C58/J, CBA/J, DBA/2J, RIIIS/J and SM/J) using 12 common measures of nociception. These included assays for thermal nociception (hot plate, Hargreaves' test, tail withdrawal), mechanical nociception (von Frey filaments), chemical nociception (abdominal constriction, carrageenan, formalin), and neuropathic pain (autotomy, Chung model peripheral nerve injury). We demonstrate the existence of clear strain differences in each assay, with 1.2 to 54-fold ranges of sensitivity. All nociceptive assays display moderate-to-high heritability (h2 = 0.30-0.76) and mediation by a limited number of apparent genetic loci. Data comparing inbred strains have considerable utility as a tool for understanding the genetics of nociception, and a particular relevance to transgenic studies.

Heritability of nociception II. 'Types' of nociception revealed by genetic correlation analysis.

Clinical pain syndromes, and experimental assays of nociception, are differentially affected by manipulations such as drug administration and exposure to environmental stress. This suggests that there are different 'types' of pain. We exploited genetic differences among inbred strains of mice in an attempt to define these primary 'types'; that is, to identify the fundamental parameters of pain processing. Eleven randomly-chosen inbred mouse strains were tested for their basal sensitivity on 12 common measures of nociception. These measures provided for a range of different nociceptive dimensions including noxious stimulus modality, location, duration and etiology, among others. Since individual members of inbred strains are identical at all genetic loci, the observation of correlated strain means in any given pair of nociceptive assays is an index of genetic correlation between these assays, and hence an indication of common physiological mediation. Obtained correlation matrices were subjected to multivariate analyses to identify constellations of nociceptive assays with common genetic mediation. This analysis revealed three major clusters of nociception: (1) baseline thermal nociception, (2) spontaneously-emitted responses to chemical stimuli, and (3) baseline mechanical sensitivity and cutaneous hypersensitivity. Many other nociceptive parameters that might a priori have been considered closely related proved to be genetically divergent.

Myelinated afferent fiber types that become spontaneously active and mechanosensitive following nerve transection in the rat.

It is difficult to know which afferent types preferentially develop ectopic firing characteristics following nerve injury because axotomy disconnects the sensory receptor ending from the remainder of the afferent neuron. We compared the prevalence of ectopic firing originating in nerve-end neuromas of nerves serving muscle and skin in the rat. Spontaneous firing was much more prevalent in the injured medial gastrocnemius nerve, a hindlimb muscle nerve, than in the saphenous and sural nerves which primarily innervate hindlimb skin. Ectopic mechanosensitivity, on the other hand, was more prominent in neuromas of the cutaneous nerves. In neuromas of the facial nerve, a cranial nerve which serves striated muscles of the face, there was no spontaneous discharge and very little ectopic mechanosensitivity. We conclude that the development of spontaneous ectopic discharge and ectopic mechanosensitivity depends on the type of myelinated afferent fiber involved.

Effect of age and nerve injury on cross-excitation among sensory neurons in rat dorsal root ganglia.

Spike activity in dorsal root ganglion (DRG) neurons depolarizes passive neighbors that share the same ganglion. We asked whether age or prior nerve injury affect this 'cross-depolarization' signal. Intracellular recordings made from excised DRGs in vitro revealed that the prevalence and duration of cross-depolarization were no greater in adult than in young rats, and that its amplitude was significantly smaller in adults. The amplitude of cross-depolarization was not affected by nerve injury. The decrease in membrane input resistance (R(in)) observed during cross-depolarization was less than that expected from equivalent depolarization alone. This affirms prior evidence that the neural process underlying cross-depolarization causes a net increase in R(in).