Andrew M. Strassman

Behavioral evidence of trigeminal neuropathic pain following chronic constriction injury to the rat's infraorbital nerve

Video recordings of free behavior and responses to mechanical facial stimulation were analyzed to assess whether chronic constriction injury (CCI) to the rats infraorbital nerve (IoN) results in behavioral alterations indicative of neuropathic pain. A unilateral CCI was produced by placing loose chromic gut ligatures around the IoN. After CCI to the IoN, rats exhibited changes in both non-evoked and evoked behavior. Behavioral changes developed in two phases. Early after CCI (postoperative days 1–15), rats showed increased face-grooming activity with face-wash strokes directed to the injured nerve territory, while the responsiveness to stimulation of this area was decreased. Later after CCI (postoperative days 15–130), the prevalence of asymmetric face grooming was reduced but remained significantly increased compared to control rats. The early hyporesponsiveness was abruptly replaced by an extreme hyperresponsiveness: all stimulus intensities applied to the injured nerve territory evoked the “maximal” response (brisk head withdrawal, avoidance behavior plus directed face grooming). This response was never observed in control rats. Concurrently, IoN ligation rats showed a limited increase in the responsiveness to stimulation of the contralateral IoN territory, and around postoperative days 30–40 the responsiveness to stimulation of facial areas outside the IoN territories also increased. The hyperresponsiveness to stimulation of the ligated IoN territory slightly decreased from 60 d postoperative. Throughout the study, IoN ligation rats showed decreased exploratory behavior, displayed more freezing-like behavior, had a slower body weight gain, and a higher defecation rate, compared to control rats. The behavioral alterations observed after CCI to the IoN are indicative of severe sensory disturbances within the territory of the injured nerve: mechanical allodynia develops after a period of relative hypo- /anesthesia during which behavioral signs of recurrent spontaneous, aversive (possibly painful) sensations (paresthesias/dysesthesias) are maximal.

Anatomical and physiological characteristics of vestibular neurons mediating the vertical vestibulo-ocular reflexes of the squirrel monkey.

The morphology of 35 vestibular neurons whose firing rate was related to vertical eye movements was studied by injection of horseradish peroxidase intracellularly into physiologically identified vestibular axons in alert squirrel monkeys. The intracellularly injected cells were readily classified into four main groups. One group of cells, down position‐vestibular‐pause neurons (down PVPs; N = 12), increased their firing rate during downward eye positions, paused during saccades, and were located in the medial vestibular nucleus (MV) and the adjacent ventrolateral vestibular nucleus (VLV). They had axons that crossed the midline and ascended in the medial longitudinal fasciculus (MLF) to terminate in the trochlear nucleus, the lateral aspect of the caudal oculomotor nucleus, and the dorsal aspect of the rostral oculomotor nucleus. A second group of cells (N = 15) were also located in the MV and VLV, but increased their firing rate during upward eye positions, and paused during saccades. These cells had axons that crossed the midline and ascended in the contralateral MLF to terminate in the medial aspect of the oculomotor nucleus. A third group of cells (N = 4) were located in the superior vestibular nucleus, generated bursts of spikes during upward saccades, and increased their tonic firing rate during upward eye positions. These cells had axons that ascended laterally to the ipsilateral MLF to terminate in regions of the trochlear and oculomotor nuclei similar to those in which down PVPs terminated. A fourth group of cells (N = 4), located in the VLV, had axons that projected to the spinal cord, although they had firing rates that were significantly correlated with vertical eye position. Electrical stimulation of the vestibular nerve evoked spikes at monosynaptic latencies in each of the above classses of cells, six of which were injected with horseradish peroxidase.

Mast cell degranulation activates a pain pathway underlying migraine headache.

Abstract Intracranial headaches such as that of migraine are generally accepted to be mediated by prolonged activation of meningeal nociceptors but the mechanisms responsible for such nociceptor activation are poorly understood. In this study, we examined the hypothesis that meningeal nociceptors can be activated locally through a neuroimmune interaction with resident mast cells, granulated immune cells that densely populate the dura mater. Using in vivo electrophysiological single unit recording of meningeal nociceptors in the rat we observed that degranulation of dural mast cells using intraperitoneal administration of the basic secretagogue agent compound 48/80 (2 mg/kg) induced a prolonged state of excitation in meningeal nociceptors. Such activation was accompanied by increased expression of the phosphorylated form of the extracellular signal‐regulated kinase (pERK), an anatomical marker for nociceptor activation. Mast cell‐induced nociceptor interaction was also associated with downstream activation of the spinal trigeminal nucleus as indicated by an increase in c‐fos expression. Our findings provide evidence linking dural mast cell degranulation to prolonged activation of the trigeminal pain pathway believed to underlie intracranial headaches such as that of migraine.

Somatotopic Activation in the Human Trigeminal Pain Pathway

Functional magnetic resonance imaging was used to image pain-associated activity in three levels of the neuraxis: the medullary dorsal horn, thalamus, and primary somatosensory cortex. In nine subjects, noxious thermal stimuli (46°C) were applied to the facial skin at sites within the three divisions of the trigeminal nerve (V1, V2, and V3) and also to the ipsilateral thumb. Anatomical and functional data were acquired to capture activation across the spinothalamocortical pathway in each individual. Significant activation was observed in the ipsilateral spinal trigeminal nucleus within the medulla and lower pons in response to at least one of the three facial stimuli in all applicable data sets. Activation from the three facial stimulation sites exhibited a somatotopic organization along the longitudinal (rostrocaudal) axis of the brain stem that was consistent with the classically described “onion skin” pattern of sensory deficits observed in patients after trigeminal tractotomy. In the thalamus, activation was observed in the contralateral side involving the ventroposteromedial and dorsomedial nuclei after stimulation of the face and in the ventroposterolateral and dorsomedial nuclei after stimulation of the thumb. Activation in the primary somatosensory cortex displayed a laminar sequence that resembled the trigeminal nucleus, with V2 more rostral, V1 caudal, and V3 medial, abutting the region of cortical activation observed for the thumb. These results represent the first simultaneous imaging of pain-associated activation at three levels of the neuraxis in individual subjects. This approach will be useful for exploring central correlates of plasticity in models of experimental and clinical pain.

Calcitonin gene–related peptide does not excite or sensitize meningeal nociceptors: Implications for the pathophysiology of migraine

Migraine is among the most common types of pain, but its mechanisms are poorly understood. A growing body of evidence points to a critical role of calcitonin gene–related peptide (CGRP) in the pathophysiology of migraine headache. During migraine, CGRP is thought to be released from peripheral endings of perivascular meningeal nociceptors primary and to promote vasodilatation. A current hypothesis suggests that peripheral CGRP and its related meningeal vasodilatation results in activation and sensitization, leading to the generation of migraine headache. However, direct evidence supporting this idea is lacking. Here, using electrophysiological, extracellular, single‐unit recording combined with laser–Doppler flowmetry measurements of dural blood flow (DBF), we examined whether CGRP and meningeal vasodilatation promote activation or sensitization of meningeal nociceptors. Changes in (DBF), ongoing discharge, and responsiveness to mechanical stimulation of the dura were studied after either topical administration or intravenous infusion of rat α‐CGRP in anesthetized rats. Both topical and systemic administration of CGRP caused a significant increase in dural blood flow; however, neither method of CGRP administration resulted in activation or sensitization of meningeal nociceptors. The results of this study suggest that CGRP effects in the meninges, including meningeal vasodilatation, are not sufficient to activate or sensitize meningeal nociceptors. Ann Neurol 2005;58:698–705

Response of brainstem trigeminal neurons to electrical stimulation of the dura

The extracellular response of medullary trigeminal neurons to electrical stimulation of the dura was studied in anesthetized cats. Fifty-six medullary trigeminal units were excited by stimulation sites near major dural vessels with an average latency of 11.0 ms. Many units also responded to infraorbital nerve shock and had cutaneous receptive fields that included the ipsilateral periorbital region. These cutaneous responses were either wide dynamic range or nociceptive specific in type. Electrical stimulation of the midbrain periaqueductal gray region suppressed the response of medullary trigeminal units to either dural stimulation or infraorbital nerve shock. Medullary trigeminal neurons that receive convergent inputs from dura and facial skin may provide a physiological substrate for the cutaneous referral of dural sensation.

Is the jaw-opening reflex a valid model of pain?

Tooth pulp shock does not produce only pain; low intensity stimulation results in a non-painful sensation that is termed pre-pain. In animals low intensity tooth pulp shock does not evoke escape behavior; the similarity of the animal escape/detection threshold ratio with the human pain/pre-pain threshold ratio is evidence that pre-pain and pain may be present in animals as in humans. Both pre-pain and pain may arise from the activation of a common afferent modality. The TP-JOR does not correlate with the degree of pain experienced under all conditions. The TP-JOR threshold is at or near the sensory detection threshold, at stimulation intensities which evoke pre-pain. Under normal conditions both the magnitude of the TP-JOR response and the degree of pain experienced increase with increasing stimulation intensity. The TP-JOR and the tooth pulp-evoked pain are affected in parallel by sensory habituation and both appear to relay in the rostral trigeminal complex. There are no cases where the TP-JOR is suppressed and pain is still experienced from tooth pulp shock; the suppression of the TP-JOR may therefore be an accurate index of analgesia. However, in humans treatments that produce analgesia have not been shown to produce suppression of the TP-JOR. Thus, the TP-JOR that persists following analgesic treatments is not a reliable index of either analgesia or pain.

The NMDA antagonist memantine blocks pain behavior in a rat model of formalin-induced facial pain

&NA; Recent studies have provided evidence that excitatory amino acid antagonists can exert analgesic effects in animals. These studies, however, have focused primarily on phasic pain or hyperalgesia rather than tonic pain. The present study evaluates the effects of systemic administration of Memantine (1‐amino‐3,5‐dimethyl‐adamantane), a clinically used N‐methyl‐d‐aspartate (NMDA) receptor antagonist, on formalin‐induced phasic and tonic pain behavior in the rat. Memantine (2.5, 5.0, 10.0 and 20.0 mg/kg) or normal saline was injected i.p. l h prior to a s.c. injection of formalin (5%, 50 &mgr;l) into the vibrissal pad of adult rats (n = 5/group). Pain behavior was measured by the number of seconds of formalin‐induced face grooming during a 42‐min post‐injection observation period. Saline‐injected animals displayed a biphasic face‐grooming response, consisting of an early, phasic phase (0–6 min) and a delayed, prolonged tonic phase (12–42 min). Memantine at doses of 2.5–10 mg/kg produced a significant dose‐related inhibition of the second phase (65–93%) and a much smaller inhibition of the first phase (up to 52%). A higher dose (20 mg/kg) further inhibited both phases but also produced other motor effects (increased exploratory and decreased freezing behavior, hind‐paw weakness and gait ataxia) which were not observed at the lower doses. These results suggest that the NMDA receptor antagonist Memantine can block formalin‐induced tonic and, to a lesser extent, phasic pain, at doses that do not alter other observed motor behaviors.

Sensitization and Activation of Intracranial Meningeal Nociceptors by Mast Cell Mediators

Intracranial headaches such as migraine are thought to result from activation of sensory trigeminal pain neurons that supply intracranial blood vessels and the meninges, also known as meningeal nociceptors. Although the mechanism underlying the triggering of such activation is not completely understood, our previous work indicates that the local activation of the inflammatory dural mast cells can provoke a persistent sensitization of meningeal nociceptors. Given the potential importance of mast cells to the pain of migraine it is important to understand which mast cell-derived mediators interact with meningeal nociceptors to promote their activation and sensitization. In the present study, we have used in vivo electrophysiological single-unit recording of meningeal nociceptors in the trigeminal ganglion of anesthetized rats to examine the effect of a number of mast cell mediators on the activity level and mechanosensitivity of meningeal nociceptors. We have found that that serotonin (5-HT), prostaglandin I2 (PGI2), and to a lesser extent histamine can promote a robust sensitization and activation of meningeal nociceptors, whereas the inflammatory eicosanoids PGD2 and leukotriene C4 are largely ineffective. We propose that dural mast cells could promote headache by releasing 5-HT, PGI2, and histamine.

Anatomy and physiology of intracellularly labelled omnipause neurons in the cat and squirrel monkey

SummarySaccadic omnipause neurons (OPNs) were intracellularly labelled with horseradish peroxidase (HRP) in alert cats and squirrel monkeys. The somas of OPNs were located on or near the midline in the caudal pons and their axons projected to regions of the pontomedullary reticular formation that contain the excitatory and inhibitory burst neurons.