Locus coeruleus modulation of dorsal horn unit responses to cutaneous stimulation

A single injection of N,N′-bis(β-chloroethyl)-4,4′-bipiperidine (BPM) into 30-day old rats produced brain lesions in the ventromedial hypothalamus, the dorsal vagal complex (dorsal motor nucleus of the vagus and nucleus of the solitary tract), and the medial septum. The brain lesions were associated with a chronic increase in the rate of body weight gain and the development of obesity. The lesions are similar to those seen in mice following BPM or gold thioglucose administration, however, the development of the obesity in the rat follows a more protracted time course.     

Three transverse dipolar generators in the human cervical and lumbo-sacral dorsal horn: evidence from direct intraoperative recordings on the spinal cord surface

In the context of the intraoperative study of spinal cord surface evoked potentials in patients operated upon for chronic pain and spasticity, we have undertaken an analysis of the dipolar dorso-ventral organization of surface spinal cord evoked potentials in man. Averaged evoked potentials to peripheral nerve electrical stimulations were obtained from the dorsal and ventral pial surface of the cervical and lumbo-sacral spinal cord (7 pairs from 5 patients), using a small silver ball macroelectrode, positioned during open neurosurgical approaches. We found that the dorsally recorded N13 and N24 waves reversed into ventral P13 and P24 waves respectively. A second negative potential, N2, and a late prolonged positivity, P, similarly reversed into a P2 and an N wave respectively. Our data add up to a collection of skin, oesophageal, epidural, pial and intramedullary recordings in man and animals to provide the evidence for a transverse dipolar organization of the human postsynaptic N13, N24 and N2 potentials, originating from deep layers of the cord dorsal horn, and for a similar organization of the P wave, which has been shown to correlate with presynaptic inhibition on primary afferent fibres.     

Delimiting subterritories of the human subthalamic nucleus by means of microelectrode recordings and a Hidden Markov Model

Positive therapeutic response without adverse side effects to subthalamic nucleus deep brain stimulation (STN DBS) for Parkinson's disease (PD) depends to a large extent on electrode location within the STN. The sensorimotor region of the STN (seemingly the preferred location for STN DBS) lies dorsolaterally, in a region also marked by distinct beta (13–30 Hz) oscillations in the parkinsonian state. In this study, we present a real‐time method to accurately demarcate subterritories of the STN during surgery, based on microelectrode recordings (MERs) and a Hidden Markov Model (HMM). Fifty‐six MER trajectories were used, obtained from 21 PD patients who underwent bilateral STN DBS implantation surgery. Root mean square (RMS) and power spectral density (PSD) of the MERs were used to train and test an HMM in identifying the dorsolateral oscillatory region (DLOR) and nonoscillatory subterritories within the STN. The HMM demarcations were compared to the decisions of a human expert. The HMM identified STN‐entry, the ventral boundary of the DLOR, and STN‐exit with an error of ?0.09 ± 0.35, ?0.27 ± 0.58, and ?0.20 ± 0.33 mm, respectively (mean ± standard deviation), and with detection reliability (error < 1 mm) of 95, 86, and 91%, respectively. The HMM was successful despite a very coarse clustering method and was robust to parameter variation. Thus, using an HMM in conjunction with RMS and PSD measures of intraoperative MER can provide improved refinement of STN entry and exit in comparison with previously reported automatic methods, and introduces a novel (intra‐STN) detection of a distinct DLOR‐ventral boundary. © 2009 Movement Disorder Society     

Simultaneous recordings of wind-up of paired spinal dorsal horn nociceptive neuron and nociceptive flexion reflex in rats

To study the sensory-motor interaction of spinal processing underlying the neuronal mechanisms of the nociceptive flexion reflex (NFR) and its temporal facilitation, 16 spinal dorsal horn (DH) wide-dynamic-range (WDR) neurons and paired 16 single motor units (SMU) from the gastrocnemius soleus muscle (GS) were simultaneously recorded using extracellular single unit and electromyographic techniques in spinal, halothane-anesthetized rats. The paired DH WDR neuron and GS SMU showed a parallel increase in the firing rate and duration of spike responses to noxious pinch stimuli applied to their common cutaneous receptive field (cRF) on the ipsilateral hind paw skin. Innocuous brush or pressure evoked no, or less, firing in the SMU but evoked a graded increase in spike responses in the simultaneously-recorded WDR neuron. Moreover, both pressure and noxious pinch stimuli evoked a short-lasting after-discharge (for several min) in the WDR neuron but without any after-discharge in the simultaneously-recorded SMU. The paired WDR neuron and SMU also showed a parallel basal response (termed as early and late components according to latency), after-discharge and wind-up of the late response to repetitively applied supra-threshold electrical stimulation (intensity: >1.5 T, duration: 1 ms and frequency: 1 Hz for 15 s). Linear regression and cross-correlation histogram analyses showed that the DH WDR neuron had a significant correlation with the simultaneously-recorded SMU and they were functionally located in the spinally-organized NFR circuitry via polysynaptic connections. Systemic administration of fentanyl, an opioid receptor agonist, resulted in a parallel, naloxone-reversible suppression of both basal late response component and wind-up response in both WDR neuron and SMU paired; however, fentanyl suppressed only the early response of the SMU without any effect on that of the DH WDR neuron. The present results provide new direct evidence showing an essential role of spinal DH WDR neurons in the mediation of spinally-organized NFR as well as its temporal facilitation (wind-up). Based on these data, the spinal DH WDR neuron seems to function as a signal discriminator or frequency encoder of multireceptive primary afferent impulses that may determine excitable level of motor output and the occurrence of a behavioral NFR via polysynaptic connections. Consequently, the spinal WDR neuron-mediated NFR and its temporal facilitation are likely to be modulated by spinal endogenous opioid peptides via opioid receptors on the nociceptive sensory components of the spinally-organized NFR circuitry.     

Changes in synaptic populations in the spinal dorsal horn following a dorsal rhizotomy in the monkey

Studies in monkeys have shown substantial neuronal reorganization and behavioral recovery during the months following a cervical dorsal root lesion (DRL; Darian‐Smith [2004] J. Comp. Neurol. 470:134–150; Darian‐Smith and Ciferri [2005] J. Comp. Neurol. 491:27–45, [2006] J. Comp. Neurol. 498:552–565). The goal of the present study was to identify ultrastructural synaptic changes post‐DRL within the dorsal horn (DH). Two monkeys received a unilateral DRL, as described previously (Darian‐Smith and Brown [2000] Nat. Neurosci. 3:476–481), which removed cutaneous and proprioceptive input from the thumb, index finger, and middle finger. Six weeks before terminating the experiment at 4 post‐DRL months, hand representation was mapped electrophysiologically within the somatosensory cortex, and anterograde tracers were injected into reactivated cortex to label corticospinal terminals. Sections were collected through the spinal lesion zone. Corticospinal terminals and inhibitory profiles were visualized by using preembedding immunohistochemistry and postembedding γ‐aminobutyric acid (GABA) immunostaining, respectively. Synaptic elements were systematically counted through the superficial DH and included synaptic profiles with round vesicles (R), pleomorphic flattened vesicles (F; presumed inhibitory synapses), similar synapses immunolabeled for GABA (F‐GABA), primary afferent synapses (C‐type), synapses with dense‐cored vesicles (D, mostly primary afferents), and presynaptic dendrites of interneurons (PSD). Synapse types were compared bilaterally via ANOVAs. As expected, we found a significant drop in C‐type profiles on the lesioned side (∼16% of contralateral), and R profiles did not differ bilaterally. More surprising was a significant increase in the number of F profiles (∼170% of contralateral) and F‐GABA profiles (∼315% of contralateral) on the side of the lesion. Our results demonstrate a striking increase in the inhibitory circuitry within the deafferented DH. J. Comp. Neurol. 518:103–117, 2010. © 2009 Wiley‐Liss, Inc.     

Chronic-pain-associated astrocytic reaction in the spinal cord dorsal horn of human immunodeficiency virus-infected patients

Studies with animal models have suggested that reaction of glia, including microglia and astrocytes, critically contributes to the development and maintenance of chronic pain. However, the involvement of glial reaction in human chronic pain is unclear. We performed analyses to compare the glial reaction profiles in the spinal dorsal horn (SDH) from three cohorts of sex- and age-matched human postmortem tissues: (1) HIV-negative patients, (2) HIV-positive patients without chronic pain, and (3) HIV patients with chronic pain. Our results indicate that the expression levels of CD11b and Iba1, commonly used for labeling microglial cells, did not differ in the three patient groups. However, GFAP and S100β, often used for labeling astrocytes, were specifically upregulated in the SDH of the "pain-positive" HIV patients but not in the "pain-negative" HIV patients. In addition, proinflammatory cytokines, TNFα and IL-1β, were specifically increased in the SDH of pain-positive HIV patients. Furthermore, proteins in the MAPK signaling pathway, including pERK, pCREB and c-Fos, were also upregulated in the SDH of pain-positive HIV patients. Our findings suggest that reaction of astrocytes in the SDH may play a role during the maintenance phase of HIV-associated chronic pain.     

Anatomical changes in cat dorsal horn cells after transection of a single dorsal root

Adult cats were subjected to unilateral L6, L7, or S1 rhizotomy. After survival times of 3 to 224 days each cat's spinal cord segments L1 to S2 were serially sectioned and stained with either Golgi or Nissl stains. The Nissl-stained material was used to determine whether or not significant cell death occurred as a result of transneuronal degeneration. The Golgi material was used to determine if changes in dendritic structure occurred. Laminae IV to VI were used for these analyses. There were no statistically significant changes in cell counts when the operated side was compared with the normal side, suggesting that cell death was rare or absent. However, dendritic complexity (mean number of branches of different orders per cell) and total dendritic length decreased with time after L7 rhizotomy in segments L3 to S2. There were no significant changes in segments L1 and L2. Atrophy of dendrites was most rapid and severe in the L7 segment, decreasing as a function of distance from L7. Two hypotheses are advanced, on the basis of these data: (a) The rapidity and degree of atrophy of a dendrite is a function of severity of deafferentation of that dendrite; and (b) rapid dendritic atrophy without cell death, as is seen in L6 to S1 after L7 rhizotomy, is possible because some dendrites are totally or largely deprived of their synaptic complement, but enough of the neuron's total synaptic input is left intact to sustain the cell and prevent cell death.     

Direct catecholaminergic innervation of spinal dorsal horn neurons with axons ascending the dorsal columns in cat

Previous ultrastructural studies have shown that catecholamine-containing nerve terminals in the spinal dorsal horn form synaptic junctions with dendrites and somata, but the identity of the neurons giving rise to these structures is largely unknown. In this study we have investigated the possibility that spinomedullary neurons, which project through the dorsal columns to the dorsal column nuclei, are synaptic targets for descending catecholaminergic axons. Neurons with axons ascending the dorsal columns were retrogradely labelled after uptake of horseradish peroxidase by their severed axons in the thoracic (T10–T12) or cervical (C2–C3) dorsal columns. After the retrogradely labelled neurons were visualized, the tissue was immunocytochemically stained with antisera raised against tyrosine hydroxylase or dopamine-β-hydroxylase. Three hundred forty-three retrogradely labelled neurons within laminae III–V of the lumbosacral dorsal horn were examined under high power with the light microscope. In Triton X-100 treated material, over 60% of cells were found to have dopamine-β-hydroxylase-immunoreactive varicosities closely apposed to their somata and proximal dendrites. The number of contacts per cell varied from 1 to 22, with a mean number of 4.5. Fewer cells (34%) received contacts from axons immunoreactive for tyrosine hydroxylase as a consequence of the weaker immunoreaction produced by this antiserum. Correlated light and electron microscopic analysis confirmed that many of these contacts were regions of synaptic specialization and that immunostained boutons contained pleomorphic (round to oval) agranular vesicles together with several dense core vesicles. These observations suggest that catecholamines regulate sensory transmission through this spinomedullary pathway by a direct postsynaptic action upon its cells of origin. Such an action would be predicted to suppress transmission generally through this pathway. © 1993 Wiley-Liss, Inc.     

ATP excites a subpopulation of rat dorsal horn neurones

1. Intracellular recordings were made from dorsal horn neurones grown in dissociated cell culture for periods of 4–12 weeks.

2. Results showed that a subpopulation of dorsal horn neurones that are likely to be involved in the processing and regulation of sensory stimuli were potently and selectively excited by ATP.

3. In dissociated cell culture, the identification and origin of dorsal hown neurones that respond to ATP were unclear.     

Changes in dorsal horn synaptic disc numbers following unilateral dorsal rhizotomy

The present study estimates the numbers of synaptic discs and numbers of degenerating synaptic terminals in laminae I-IV of the rat S2 dorsal horn ipsi- and contralateral to unilateral dorsal rhizotomy. These data allow us to estimate the loss of synapses of primary afferents and to correlate this loss with the rate of axon disappearance in the proximal stump of a transected S2 dorsal root. Our first findings are that 47% of the ipsilateral synapses and 27% of the contralateral synapses disappear within a day following unilateral rhizotomy. Conclusions are that the predominant synaptic population in this part of the rat spinal cord is of primary afferent origin and that there is an extensive bilateral projection of the dorsal root fibers. The contralateral projection is confirmed by the appearance of numerous degenerating terminals on the contralateral side. We also find that synaptic loss and appearance of degenerating terminals occur relatively synchronously in laminae I-IV. Finally we find that the time course of the synaptic loss correlates primarily with the disappearance of unmyelinated fibers in the proximal stump of the transected dorsal root.     

Cell death in the superficial dorsal horn in a model of neuropathic pain

The aims of this study were to investigate the occurrence of apoptotic cell death in the dorsal horn of the adult rat spinal cord following chronic constriction injury (CCI) to the sciatic nerve and to correlate this with behavioural responses. Six groups of six rats were used as follows: 1) CCI, 2) CCI, 3) MK801 + CCI, 4) axotomy, 5) sham, and 6) naive. Group 1 animals were behaviourally tested for thermal hyperalgesia 8 days following surgery and sacrificed and the spinal cords removed and frozen. The rest of the groups underwent the same procedure 14 days following surgery. The lumbar region of the spinal cord was cryosectioned and the incidence of apoptotic cells investigated using the TUNEL technique plus Hoechst double labelling. By 8 days post-CCI, hyperalgesia had developed in the ipsilateral paw, which was still present 14 days after the injury compared to the contralateral paw and naive and sham animals. Preemptive MK-801 prevented the onset of hyperalgesia. Significant numbers of apoptotic cells were present in the ipsilateral dorsal horn of the spinal cord 8 and 14 days following CCI compared to the contralateral side and to naive and sham animals. Preemptive treatment with MK-801 reduced the extent of apoptosis resulting from CCI to the level seen in control animals. This study demonstrates that cells undergo apoptosis as a result of CCI simultaneous with the occurrence of hyperalgesia. Furthermore, MK-801 prevents the onset of hyperalgesia and reduces the extent of apoptotic cell death, suggesting, perhaps, that apoptosis contributes to the initiation/maintenance of hyperalgesia.     

Ureteral ligation induces Fos expression in the dorsal horn

To describe a sympathetic afferent circuit, the left ureter was ligated in anesthetized rats for 1.5–2 h followed by immunocytochemical processing to localize expression of either the immediate early gene (IEG) c-fos or Krox-24 in the spinal cord or dorsal root ganglia (DRG). No IEG expression was detected in DRG. Both Fos and Krox-24 expression was found in the dorsal horn. More Fos immunocytochemically stained cells were found in the dorsal hom both ipsi- and contralateral to the ligated ureter at spinal segments T₁₀–T₁₃ after ureteral ligation than after either sham ligation or anesthesia control procedures. More Fos stained cells were in the dorsal horn ipsilateral to the ligated ureter than on the contralateral side. The Fos staining patterns in the dorsal horn of ligated and sham-ligated animals were similar with most labeled cells in dorsomedial portions of laminae I and II. In contrast, the Fos staining pattern in the dorsal horn in anesthetized animals (unoperated controls) was noticeably different from operated animals with the most Fos cells in the ventrolateral part of laminae I-II. These results indicate that (1) Fos immunocytochemistry may be useful for tracing sympathetic afferent pathways, (2) the sensory pathway activated by ureteral ligation enters the spinal cord at lower thoracic levels, where renal and upper ureteral afferents are terminating, and (3) some of this sympathetic afferent pathway is located contralateral to the stimulated kidney. Neurons activated by ureteral ligation in the contralateral dorsal horn may mediate reno-renal reflexes.     

Extension of dorsal horn neurons into the severed and implanted dorsal root

After dorsal root ganglionectomy in adult rats, the dorsal root was cut close to the spinal cord and implanted into the dorsal horn. Outgrowth from neurons in he dorsal horn of the spinal cord into the implanted dorsal root could be demonstrated after 3 months by means of retrograde HRP labeling. Double-labeling experiments showed that some of these neurons had retained their central projections while extending new processes into the implanted root. The possibility to reconstruct the sensory pathway by replacing the damaged primary sensory neuron with peripheral outgrowth from secondary sensory neurons is discussed.     

Increase in substance P in the dorsal horn during a chemogenic nociceptive stimulus

Substance P has been implicated as a neuromediator of nociception in the dorsal horn of the spinal cord. This is the first report of quantitative increases in substance P following an acute nociceptive stimulus. Female Wistar rats were injected subcutaneously in the plantar aspect of the right hindpaw with 5% formalin. Substance P levels were determined by quantitative immunohistochemistry 1 h after the injection. The present results show that substance P levels in the upper two laminae of the ipsi-and contralateral dorsal horn were significantly increased 1 h after the formalin injection. These results suggest a direct functional relationship between substance P levels in the upper dorsal horn and an acute chemogenic nociceptive stimulus.     

Dendritic changes in the spinal dorsal horn following transection of a peripheral nerve

Microinjections of different doses of bicuculline methiodide (BM) were performed into the mesencephalic central gray (CG), the medial hypothalamus (MH) and lateral hypothalamus (LH). Flight reactions could be induced by microinjections of BM into either the CG or the MH. However, the type of flight behavior was different whether the injection was made in the CG or the MH. Furthermore, microinjections of 35 ng of BM in either structure produced an increase in locomotor activity whose time course differed according to the injected structure, and an increase in rearings was induced at MH but not at CG sites. At lateral hypothalamic sites, BM produced an increase in locomotor activity and rearings but no jump. These effects were antagonized in a dose-dependent manner by a local pretreatment with 4,5,6,7-tetrahydroisoxazolo(5,4-c)pyridin-3-ol (THIP), a GABA agonist. These results suggest that (1) at the level of both the MH and the CG, a GABAergic link is involved in the inhibition of a substrate whose activation produces aversive effects, and (2) the aversive effect induced by CG BM microinjection seems to be different from that induced by MH BM microinjection.     

Intracellular recordings from lateral horn cells fo the spinal cord in vitro

Intracellular recording was used in studies of the preganglionic neurons of the autonomic nervous system. These were carried out on isolated segments of the cat spinal cord. It was found that the lateral horn cells have electrical membrane characteristics similar to postganglionic neurons, but many of them have a much longer afterhyperpolarization. 5-Hydroxytryptamine, noradrenaline and aspartate induce depolarizations in lateral horn cells which are characteristically associated with an increased membrane resistance. EPSPs in lateral horn cells are not cholinergic in nature, though many cells are endowed with excitatory nicotinic receptors. Some interneurons also appear to have excitatory muscarinic receptors. Glutamate can depolarize many lateral horn cells. This excitatory amine does not seem to be responsible for the production of an EPSP, since the EPSP persisted during the continued presence of glutamate in the superfusing medium. All the neurons examined in the lateral horn are susceptible to the hyperpolarizing and shunting actions of GABA and glycine. In a small group of neurons, noradrenaline caused a hyperpolarization.     

Bilateral subthalmic nucleus deep brain stimulation with microelectrode recordings in the setting of mild inherited hemophilia B: a case report

Abstract

Hemophilia B is an X linked recessive deficiency of factor IX that presents with a range of clinical severity that co-relates with factor levels. Although guidelines exist to guide perioperative hemostasis in such patients, there is scarce data on elective high-risk neurosurgeries, resulting in a reluctance to offer these patients elective neurosurgeries. These patients thus rarely if ever undergo such procedures. We report a unique case of undiagnosed mild hemophilia B in a gentleman that was found incidentally at age 64 during pre-operative workup. This gentleman had intractable Parkinson’s disease for which subthalmic deep brain stimulation was indicated. He was found to have a prolonged APTT on initial lab testing. After subsequent workup, and having excluded the presence of inhibitors, he was diagnosed with Hemophilia B. With the use of Factor IX concentrates (AlphaNine®) and close clinical, laboratory, and radiological monitoring a plan was made for this patient to undergo this procedure. Our patient successfully underwent subthalmic deep brain stimulation with microelectrode recordings and intraoperative test stimulation in a two-step procedure, followed by single channel implantable neurostimulator and extension wire implantations 2?weeks later. The successful peri-operative course of this patient using this novel approach is described, and the need for future data in this regard is emphasized.     

Single-unit analysis of the spinal dorsal horn in patients with neuropathic pain.

Despite the key role played by the dorsal horn of the spinal cord in pain modulation, single-unit recordings have only been performed very rarely in this structure in humans. The authors report the results of a statistical analysis of 64 unit recordings from the human dorsal horn. The recordings were done in three groups of patients: patients with deafferentation pain resulting from brachial plexus avulsion, patients with neuropathic pain resulting from peripheral nerve injury, and patients with pain resulting from disabling spasticity. The patterns of neuronal activities were compared among these three groups. Nineteen neurons were recorded in the dorsal horns of five patients undergoing DREZotomy for a persistent pain syndrome resulting from peripheral nerve injury (i.e., nondeafferented dorsal horns), 31 dorsal horn neurons were recorded in nine patients undergoing DREZotomy for a persistent pain syndrome resulting from brachial plexus avulsion (i.e., deafferented dorsal horns), and 14 neurons were recorded in eight patients undergoing DREZotomy for disabling spasticity. These groups were compared in terms of mean frequency, coefficient of variation of the discharge, other properties of the neuronal discharge studied by the nonparametric test of Wald-Wolfowitz, and the possible presence of bursts. The coefficient of variation tended to be higher in the deafferented dorsal horn group than in the other two groups. Two neurons displaying burst activity could be recorded, both of which belonged to the deafferented dorsal horn group. A significant difference was found in term of neuronal behavior between the peripheral nerve trauma group and the other groups: The brachial plexus avulsion and disabling spasticity groups were very similar, including various types of neuronal behavior, whereas the peripheral nerve lesion group included mostly neurons with "nonrandom" patterns of discharge (i.e., with serial dependency of interspike intervals).