Calcitonin gene-related peptide receptors in rat trigeminal ganglion do not control spinal trigeminal activity

Calcitonin gene-related peptide (CGRP) is regarded as a key mediator in the generation of primary headaches. CGRP receptor antagonists reduce migraine pain in clinical trials and spinal trigeminal activity in animal experiments. The site of CGRP receptor inhibition causing these effects is debated. Activation and inhibition of CGRP receptors in the trigeminal ganglion may influence the activity of trigeminal afferents and hence of spinal trigeminal neurons. In anesthetized rats extracellular activity was recorded from neurons with meningeal afferent input in the spinal trigeminal nucleus caudalis. Mechanical stimuli were applied at regular intervals to receptive fields located in the exposed cranial dura mater. α-CGRP (10(-5) M), the CGRP receptor antagonist olcegepant (10(-3) M), or vehicle was injected through the infraorbital canal into the trigeminal ganglion. The injection of volumes caused transient discharges, but vehicle, CGRP, or olcegepant injection was not followed by significant changes in ongoing or mechanically evoked activity. In animals pretreated intravenously with the nitric oxide donor glyceryl trinitrate (GTN, 250 μg/kg) the mechanically evoked activity decreased after injection of CGRP and increased after injection of olcegepant. In conclusion, the activity of spinal trigeminal neurons with meningeal afferent input is normally not controlled by CGRP receptor activation or inhibition in the trigeminal ganglion. CGRP receptors in the trigeminal ganglion may influence neuronal activity evoked by mechanical stimulation of meningeal afferents only after pretreatment with GTN. Since it has previously been shown that olcegepant applied to the cranial dura mater is ineffective, trigeminal activity driven by meningeal afferent input is more likely to be controlled by CGRP receptors located centrally to the trigeminal ganglion.     

Response properties of trigeminal brain stem neurons with input from dura mater encephali in the rat

The responsiveness of trigeminal brain stem neurons to selective local mechanical and chemical stimulation of the cranial dura mater was examined in a preparation in the rat. The dura mater encephali was exposed and its surface stimulated with electrical pulses through bipolar electrodes. Extracellular recordings were made from neurons in the subnucleus caudalis of the spinal trigeminal nucleus. Single neurons driven by meningeal input were identified by their responses to electrical stimulation and to probing their receptive fields on the dura. Facial receptive fields were defined mechanically. Chemical stimuli (a combination of inflammatory mediators, bradykinin, prostaglandin E2, serotonin, capsaicin and acidic Tyrode's solution) were applied topically to the dura and by injection through a catheter into the superior sagittal sinus. All neurons with input from the parietal dura mater had convergent input from the facial skin, with preponderance of the periorbital region. Proportions of units were activated by the combination of inflammatory mediators (55%), bradykinin (64.5%), acidic Tyrode's solution (64.1%) and capsaicin (78.6%). We conclude that, among the chemical mediators of inflammation, bradykinin and low pH are the most effective chemical stimuli in activating meningeal nociceptors. These stimuli may be important during meningeal inflammatory processes that lead to the generation of headaches.     

Trigeminohypothalamic and reticulohypothalamic tract neurons in the upper cervical spinal cord and caudal medulla of the rat

Sensory information that arises in orofacial organs facilitates exploratory, ingestive, and defensive behaviors that are essential to overall fitness and survival. Because the hypothalamus plays an important role in the execution of these behaviors, sensory signals conveyed by the trigeminal nerve must be available to this brain structure. Recent anatomical studies have shown that a large number of neurons in the upper cervical spinal cord and caudal medulla project directly to the hypothalamus. The goal of the present study was to identify the types of information that these neurons carry to the hypothalamus and to map the route of their ascending axonal projections. Single-unit recording and antidromic microstimulation techniques were used to identify 81 hypothalamic-projecting neurons in the caudal medulla and upper cervical (C(1)) spinal cord that exhibited trigeminal receptive fields. Of the 72 neurons whose locations were identified, 54 were in laminae I-V of the dorsal horn at the level of C(1) (n = 22) or nucleus caudalis (Vc, n = 32) and were considered trigeminohypothalamic tract (THT) neurons because these regions are within the main projection territory of trigeminal primary afferent fibers. The remaining 18 neurons were in the adjacent lateral reticular formation (LRF) and were considered reticulohypothalamic tract (RHT) neurons. The receptive fields of THT neurons were restricted to the innervation territory of the trigeminal nerve and included the tongue and lips, cornea, intracranial dura, and vibrissae. Based on their responses to mechanical stimulation of cutaneous or intraoral receptive fields, the majority of THT neurons were classified as nociceptive (38% high-threshold, HT, 42% wide-dynamic-range, WDR), but in comparison to the spinohypothalamic tract (SHT), a relatively high percentage of low-threshold (LT) neurons were also found (20%). Responses to thermal stimuli were found more commonly in WDR than in HT neurons: 75% of HT and 93% of WDR neurons responded to heat, while 16% of HT and 54% of WDR neurons responded to cold. These neurons responded primarily to noxious intensities of thermal stimulation. In contrast, all LT neurons responded to innocuous and noxious intensities of both heat and cold stimuli, a phenomenon that has not been described for other populations of mechanoreceptive LT neurons at spinal or trigeminal levels. In contrast to THT neurons, RHT neurons exhibited large and complex receptive fields, which extended over both orofacial ("trigeminal") and extracephalic ("non-trigeminal") skin areas. Their responses to stimulation of trigeminal receptive fields were greater than their responses to stimulation of non-trigeminal receptive fields, and their responses to innocuous stimuli were induced only when applied to trigeminal receptive fields. As described for SHT axons, the axons of THT and RHT neurons ascended through the contralateral brain stem to the supraoptic decussation (SOD) in the lateral hypothalamus; 57% of them then crossed the midline to reach the ipsilateral hypothalamus. Collateral projections were found in the superior colliculus, substantia nigra, red nucleus, anterior pretectal nucleus, and in the lateral, perifornical, dorsomedial, suprachiasmatic, and supraoptic hypothalamic nuclei. Additional projections (which have not been described previously for SHT neurons) were found rostral to the hypothalamus in the caudate-putamen, globus pallidus, and substantia innominata. The findings that nonnociceptive signals reach the hypothalamus primarily through the direct THT route, whereas nociceptive signals reach the hypothalamus through both the direct THT and the indirect RHT routes suggest that highly prioritized painful signals are transferred in parallel channels to ensure that this critical information reaches the hypothalamus, a brain area that regulates homeostasis and other humoral responses required for the survival of the organism.     

Sensory processing in a thermal afferent pathway

Extracellular recordings were made from cold-receptive afferent fibers in the trigeminal ganglion of rats anesthetized with halothane. By applying a standardized series of steady or changing temperatures to the receptive fields, we recorded the static and dynamic responses of the afferents. Comparable recordings were made from neurons in the marginal layer of the caudal trigeminal nucleus onto which the cold fibers synapse. The static and dynamic responses of the afferent fibers were reproduced faithfully by the second-order neurons, but at a much higher level of activity. Ganglionectomy silenced the second-order cells. Their continuous high level of activity appears to depend on the tonic input from the afferent fibers and not on any intrinsic circuits in the medulla.     

Modulation of trigeminal spinal subnucleus caudalis neuronal activity following regeneration of transected inferior alveolar nerve in rats

Modulation of trigeminal spinal subnucleus caudalis neuronal activity following regeneration of transected inferior alveolar nerve in rats. To clarify the neuronal mechanisms of abnormal pain in the face innervated by the regenerated inferior alveolar nerve (IAN), nocifensive behavior, trigeminal ganglion neuronal labeling following Fluorogold (FG) injection into the mental skin, and trigeminal spinal subnucleus caudalis (Vc) neuronal properties were examined in rats with IAN transection. The mechanical escape threshold was significantly higher at 3 days and lower at 14 days after IAN transection, whereas head withdrawal latency to heat was significantly longer at 3, 14, and 60 days after IAN transection. The number of FG-labeled ganglion neurons was significantly reduced at 3 days after IAN transection but increased at 14 and 60 days. The number of wide dynamic range (WDR) neurons with background (BG) activity was significantly higher at 14 and 60 days after IAN transection compared with na?ve rats, and the number of WDR and low-threshold mechanoreceptive (LTM) neurons with irregularly bursting BG activity was increased at these two time points. Mechanically evoked responses were significantly larger in WDR and LTM neurons 14 days after IAN transection compared with na?ve rats. Heat- and cold-evoked responses in WDR neurons were significantly lower at 14 days after transection compared with na?ve rats. Mechanoreceptive fields were also significantly larger in WDR and LTM neurons at 14 and 60 days after IAN transection. These findings suggest that these alterations may be involved in the development of mechanical allodynia in the cutaneous region innervated by the regenerated IAN.     

Jaw-muscle spindle afferent feedback to the cervical spinal cord in the rat

Putative synaptic contacts between masticatory-muscle spindle afferents and brainstem neurons which project to the cervical spinal cord were studied in rats by combining retrograde and intracellular neuronal labeling. Spinal cord projecting neurons were retrogradely labeled via injection of horseradish peroxidase unilaterally or bilaterally into cervical spinal cord segments C2 through C5. Twenty-four hours after the injection of horseradish peroxidase, one to five jaw-muscle spindle afferent axons were physiologically identified and intracellularly stained with biotinamide on each side of the brainstem. Horseradish-peroxidase-labeled neurons were found bilaterally in the supratrigeminal region, trigeminal principal sensory nucleus, parvicellular reticular nucleus including its alpha division, spinal trigeminal subnuclei oralis and interpolaris and the medullary reticular formation. Retrogradely labeled neurons were most numerous in the spinal trigeminal subnucleus oralis, parvicellular reticular formation and the ventral part of the spinal trigeminal subnucleus interpolaris. A small number of horseradish-peroxidase-labeled neurons were also present in the trigeminal mesencephalic nucleus and spinal trigeminal subnucleus caudalis. Appositions between jaw-muscle spindle afferent boutons and spinal projecting neurons were found in the supratrigeminal region, dorsomedial portions of the trigeminal principal sensory nucleus and spinal trigeminal subnuclei oralis and interpolaris, and the parvicellular reticular formation including its alpha division. Putative synaptic contacts were most frequent in the parvicellular reticular formation and the dorsomedial portion of the trigeminal subnucleus oralis. These results indicate that some orofacial proprioceptive feedback transmitted via the mesencephalic trigeminal nucleus reaches the cervical spinal cord directly and suggests that jaw-muscle spindle afferent feedback reaches the cervical spinal cord predominately via relays in the dorsomedial part of the spinal trigeminal subnucleus oralis and the parvicellular reticular formation. It is hypothesized that these pathways are primarily involved in the coordination of jaw and neck movement during mastication and biting. Materials and methods 27 January 1999 / Accepted: 9 May 1999     

The contribution of the principal and spinal trigeminal nuclei to the receptive field properties of thalamic VPM neurons in the rat

Primary sensory information from neurons innervating whisker follicles on one side of a rat's face is relayed primarily through two subnuclei of the brainstem trigeminal complex to the contralateral thalamus. The present experiments were undertaken to separate the contribution of the principal trigeminal nucleus (PrV) from that of the spinal trigeminal nucleus (SpV) to whisker evoked responses in the ventral posterior medial (VPM) nucleus in the adult rat thalamus. Extracellular single-unit responses of VPM neurons to controlled stimulation of the contralateral whiskers under urethane anesthesia were quantified in terms of receptive field size, modal latency, response probability and response magnitude. The SpV contribution to VPM cell responses was isolated by making kainic acid lesions of the PrV. The PrV contribution was ascertained by cutting the trigeminothalamic axons arising from SpV just before they cross the midline. After destruction of the PrV, the SpV pathway alone produced large receptive fields (mean: 9.04 whiskers) and long latency (mean: 11.07 ms) responses from VPM neurons. In contrast, PrV input alone (SpV disconnected) generated small receptive fields (mean: 1.06 whiskers) and shorter latency (mean: 6.74 ms) responses. With both pathways intact the average receptive field size was 2.4 whiskers and peak (modal) response latency was 7.33 ms. The responses with both pathways intact were significantly different from either pathway operating in isolation. Response probability and magnitude followed the same trend. We conclude that normal responses of individual VPM neurons represent the integration of input activity transmitted through both PrV and SpV pathways.     

5-HT-immunoreactive fibers in the trigeminal nuclear complex of the rat

Summary The distribution of serotonin immunoreactive (5-HT-IR) fibers in the trigeminal nuclear complex of the rat was mapped. In the sensory nuclei, innervation appeared to be dense in areas primarily related to nociceptive afferent activity, and sparse in areas primarily related to nonnociceptive afferent activity. Specifically, the marginal and gelatinosa layers of the spinal subnucleus caudalis contained many 5-HT-IR fibers while few labeled fibers were seen in the magnocellular portion of subnucleus caudalis or in the principal sensory nucleus. The spinal subnuclei oralis and interpolaris were sparsely innervated except for a few areas which contained more 5-HT-IR fibers. The motor nucleus contained as many immunoreactive fibers as the subnucleus caudalis, although fibers in the motor nucleus were thicker and varicosities more irregularly spaced than in caudalis.This research was supported by grant BNS-79-06474 from the National Science Foundation awarded to Efrain Azmitia     

Localization of P2X2 and P2X3 receptors in rat trigeminal ganglion neurons

Purine receptors have been implicated in central neurotransmission from nociceptive primary afferent neurons, and ATP-mediated currents in sensory neurons have been shown to be mediated by both P2X3 and P2X2/3 receptors. The aim of the present study was to quantitatively examine the distribution of P2X2 and P2X3 receptors in primary afferent cell bodies in the rat trigeminal ganglion, including those innervating the dura. In order to determine the classes of neurons that express these receptor subtypes, purine receptor immunoreactivity was examined for colocalization with markers of myelinated (neurofilament 200; NF200) or mostly unmyelinated, non-peptidergic fibers (Bandeiraea simplicifolia isolectin B4; IB4). Forty percent of P2X2 and 64% of P2X3 receptor-expressing cells were IB4 positive, and 33% of P2X2 and 31% of P2X3 receptor-expressing cells were NF200 positive. Approximately 40% of cells expressing P2X2 receptors also expressed P2X3 receptors and vice versa. Trigeminal ganglion neurons innervating the dura mater were retrogradely labeled and 52% of these neurons expressed either P2X2 or P2X3 or both receptors. These results are consistent with electrophysiological findings that P2X receptors exist on the central terminals of trigeminal afferent neurons, and provide evidence that afferents supplying the dura express both receptors. In addition, the data suggest specific differences exist in P2X receptor expression between the spinal and trigeminal nociceptive systems.     

Trigeminal nociceptive neurons in the subnucleus reticularis ventralis. I. Response properties and afferent connections.

Trigeminal nociceptive neurons within the subnucleus reticularis ventralis medullae oblongatae (SRV), which lies ventral to the trigeminal subnucleus caudalis and subnucleus reticularis dorsalis medullae oblongatae, were studied in urethane/chloralose-anesthetized cats and monkeys. These neurons were called 'SRV neurons'. They were almost regularly excited by pressure to the ipsilateral cornea or to both corneas at a strength well above the human corneal pain threshold. Most of them were activated by noxious mechanical stimulation of the pinna, face and/or tongue. A significant fraction of SRV units was responsive to tapping of the ipsilateral dorsum of the nose and/or electrical stimulation of tooth pulp afferents. Evidence was obtained that responses to tapping of the dorsum of the nose were due to mechanical distortion of the nasal mucosa. Intracellular injection of HRP into SRV neurons demonstrated that injected neurons were large neurons characteristic of the SRV. Trigeminal tractotomy just rostral to the obex did not eliminate responses of SRV units to trigeminal inputs. Neurons relaying trigeminal inputs to SRV neurons were electrophysiologically identified in the nucleus reticularis parvocellularis which is ventromedially adjacent to the subnuclei oralis and interpolaris of the trigeminal spinal tract nucleus. These findings were supported by HRP injection into the SRV. Units having receptive fields similar to those of SRV neurons were found in lamina VII of the first cervical cord, suggesting that SRV neurons may be trigeminal lamina VII neurons.     

Excitatory amino acid binding sites in the trigeminal principal sensory and spinal trigeminal nuclei of the rat.

Quantitative autoradiography was used to examine the density and distribution of excitatory amino acid (EAA) binding site subtypes in the principal sensory and spinal trigeminal nuclei of the rat trigeminal complex. The highest densities of N-methyl-D-aspartate (NMDA), alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA), kainate and metabotropic receptors were found in the superficial laminae (I and II) of subnucleus caudalis, a region known to be densely innervated by primary afferent nociceptive terminals. Lower densities of EAA binding sites were observed in spinal subnuclei interpolaris and oralis and within the principal sensory nucleus. These results are consistent with the hypothesis that EAAs are involved in primary afferent nociceptive neurotransmission.     

Corneal representation within the trigeminal subnucleus caudalis and adjacent bulbar lateral reticular formation of the cat.

Corneal units in the trigeminal subnucleus caudalis and adjacent bulbar lateral reticular formation were studied in urethane-chloralose anesthetized cats. Corneal units were categorized into four classes: low-threshold corneal (LTC) units, high-threshold corneal (HTC) units, wide dynamic range (WDR) units with corneal input, and subnucleus reticularis ventralis (SRV) units with corneal input. Corneal receptive fields of these four classes of corneal afferent units consisted of 3-6 spots. Mechanical thresholds of LTC units were lower than 30 mg (2.6 g/mm2) and were comparable to the sensory threshold of the human cornea measured in patients with cataract. Mechanical thresholds of the other 3 classes of corneal afferent units were well above the pain threshold in the human cornea. LTC units were located in the magnocellular layer of trigeminal subnucleus caudalis and were intermingled with cutaneous low-threshold mechanoreceptive units. HTC units were coexistent with nociceptive specific units in the marginal layer and in the outer zone of substantia gelatinosa. WDR units with corneal input were found in the lateral part of trigeminal lamina V equivalent, which corresponds to the lateral part of subnucleus reticularis dorsalis. These 3 classes of corneal units were found at a level 2.7-3.5 mm caudal to the obex. SRV units were found in the dorsolateral part of SRV along the entire length of the medulla oblongata caudal to the obex. These results support the suggestion that either nonpainful sensation or pain can be evoked from the cornea.     

Mechanical response properties of A and C primary afferent neurons innervating the rat intracranial dura

The intracranial dura receives a small-fiber sensory innervation from the trigeminal ganglion that is thought to be involved in some types of headaches, including migraine. Mechanical response properties of dural afferent neurons were examined to investigate variation across the population in the properties of threshold, slope, adaptation, and incidence of mechanosensitivity. Dural afferent neurons were recorded in the trigeminal ganglion of urethan-anesthetized rats and were identified by their constant-latency response to dural shock. Neurons were classified as fast A (>5 m/s), slow A (5 >or= conduction velocity (CV) >or= 1.5 m/s), or C (<1.5 m/s), based on response latency to dural shock. Mechanical receptive fields were identified by stroking or indenting the outer surface of the dura. Stimulus-response curves were obtained from responses to 2-s constant-force indenting stimuli of graded intensities delivered to the dural receptive field with a servo force-controlled mechanical stimulator. The slow A population had the highest percentage of mechanosensitive units (97%) as well as the highest slopes and the lowest thresholds. Thus by all three criteria, the slow As had the highest mechanosensitivity. Conversely, the fast A population had the lowest mechanosensitivity in that it had the lowest percentage of mechanosensitive units (66%), the lowest slopes, and the highest thresholds. The C population was intermediate with respect to all three properties but was much more similar to the slow As than to the fast As. All three fiber classes showed a negative correlation between slope and threshold. The majority of neurons showed a slowly adapting response to a maintained 2-s stimulus. Adapting neurons could be subdivided based on whether the fitted exponential curve decayed to zero or to a nonzero plateau; the latter group contained the most sensitive neurons in that they had the lowest thresholds and highest slopes. Nonadapting neurons generally had lower initial firing rates than adapting neurons. Fast A neurons exhibited greater and more rapid adaptation than slow A and C neurons. Neurons with the lowest slopes, regardless of CV, had relatively rapid adaptation. The more slowly conducting portion of the C population was distinguished from the other C neurons by a number of properties: more mechanically insensitive neurons, higher thresholds, and more nonadapting neurons. These differences in mechanical response properties may be related in part to differences in membrane currents involved in impulse generation that have been described in subpopulations of dorsal root ganglion cells.     

Biphasic response to nitric oxide of spinal trigeminal neurons with meningeal input in rat--possible implications for the pathophysiology of headaches

Nitric oxide (NO) is suggested to play a causative role in the pathogenesis of primary headaches. Infusion of NO donors can trigger headache attacks, and products of NO metabolism are found to be increased in the cranial circulation in patients suffering from such headaches. To examine if NO is involved in mediating and maintaining spinal trigeminal neuronal activity, an animal model of meningeal nociception was used. In barbiturate-anesthetized rats, a cranial window was made to expose the parietal dura mater. An access to the medullary brain stem allowed extracellular action potentials to be recorded from neurons in the spinal trigeminal nucleus that received afferent input from the exposed dura. Slow intravenous infusion of the NO donor, sodium nitroprusside (SNP, 50 microg/kg), transiently increased spontaneous activity in a subset of neurons and, with a latency of 50 min, caused a progressive increase in impulse activity across the entire sample of neurons. A similar pattern of delayed activation was seen after topical application of the same dose of SNP onto the exposed medulla. Slow injection of the nonspecific inhibitor of NO synthase, N(omega)-nitro-l-arginine methyl ester (20 mg/kg), reduced the spontaneous activity in all neurons within 15 min. The results suggest that NO can induce delayed, slowly developing activation of central trigeminal neurons and that endogenous release of NO may contribute to the ongoing activity of these neurons. The delayed changes in neuronal activity may include gene expression of pro-nociceptive mediators. These mechanisms may be relevant for the pathogenesis of chronic headaches.     

Dependence of electrical activity of neurons in rostral parts of spinal trigeminal nucleus on functional state of trigeminal Gasser ganglion

Unilateral injection of 100 μl 1% lidocaine into the trigeminal Gasser ganglion of narcotized rats produced a long-term moderation of the discharge rate of neurons in the ipsilateral (relative to the side of injection) rostral area of the spinal trigeminal nucleus. Activity of neurons in the contralateral rostral area of the spinal trigeminal nucleus was not blocked. Functional state of neurons in the trigeminal ganglion determines discharge activity of ipsilateral neurons of the spinal trigeminal nucleus. Activity of neurons in the contralateral rostral area of spinal trigeminal nucleus was not inhibited. Functional state of the cells in the trigeminal ganglion determines the character of electrical activity of neurons in the ipsilateral rostral area of spinal trigeminal nucleus. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 144, No. 7, pp. 4–6, July, 2007     

Activity during mastication of periodontal mechanosensitive neurons of the trigeminal subnucleus oralis of the rabbit

1. The activity of mechanosensitive neurons was examined before and during mastication. One hundred and seventy-eight neurons were recorded in the rostral parts of the trigeminal sensory nuclei of 20 rabbits anesthetized with urethan. Twenty-eight neurons received inputs from the periodontal mechanoreceptors, all on the ipsilateral side. Nineteen had receptive fields that were restricted to one tooth; 2 could be activated from more than 1 tooth, and 6 included parts of the mucosa. Only the latter were spontaneously active. 2. All periodontal neurons with a mandibular input responded to graded electrical stimulation of the inferior alveolar nerve at minimum latencies of less than or equal to 3.4 ms, and approximately half had inputs from the sensorimotor cortex. 3. Almost all periodontal units recorded were found to lie in, or just outside, the dorsal part of the most rostral subdivision of the spinal trigeminal nucleus (subnucleus oralis, pars gamma). None projected to the ipsi- or contralateral thalamus. 4. All periodontal neurons fired during mastication. Those without mucosal receptive fields fired during jaw closure, with almost all activity confined to the slow-closing phase when pressure is applied to the teeth. Injections of local anesthetic showed that input from mucosal fields was responsible for activating neurons in other phases of the cycle. 5. Possible roles in the control of mastication for these periodontal interneurons were discussed.     

Tooth pulp input to the shell region of nucleus ventralis posteromedialis of the cat thalamus

A population of neurons in the somatosensory part of the nucleus ventralis posteromedialis (VPM proper) that responded to electrical stimulation of the tooth pulp were studied in cats under urethan-chloralose anesthesia. Two classes of units responsive to electrical stimulation of the contralateral canine tooth pulp were identified. One class was responsive only to tooth pulp stimulation and these units were designated as tooth pulp specific (TPS) units. The other class of units responded to mechanical stimulation of the contralateral trigeminal integument in addition to tooth pulp stimulation. Their receptive field characteristics identified them as wide dynamic range (WDR) units responsive to tooth pulp stimulation. Both classes of units were located in the shell region of the caudal VPM proper; TPS units were coexistent with trigeminal nociceptive specific (NS) units and were found in the dorsomedial as well as ventromedial parts of the NS zone. WDR units responsive to electrical stimulation of the tooth pulp were located in the dorsomedial as well as ventromedial parts of WDR zone, a narrow band, approximately 300 micron wide, just in front of the NS zone. Tooth pulp units in the dorsomedial shell region of the VPM proper responded to the maxillary canine tooth pulp, whereas those in the ventromedial shell region responded to the mandibular canine tooth pulp. Some tooth pulp units in these two regions were responsive to stimulation of both maxillary and mandibular canine teeth. Both TPS and WDR units were antidromically excited by electrical stimulation of the SI area of the somatosensory cortex. Cooling the dorsolateral surface of the caudal medulla oblongata reversibly blocked tooth pulp evoked responses of TPS and WDR units. Trigeminal tractotomy just above the level of the obex irreversibly abolished tooth pulp-evoked responses of TPS and WDR units. These findings suggested that TPS neurons in the marginal layer of the trigeminal subnucleus caudalis and WDR neurons in the lateral part of the subnucleus reticularis dorsalis relay afferent impulses derived from the tooth pulp to the shell region of the VPM proper.     

Peripheral and spinal components of the sensitization of spinal neurons during an acute experimental arthritis

In cats the injections of kaolin and carrageenan into the knee joint lead to an acute arthritis which develops within 1–3 hours. In parallel articular afferents (low, high threshold and unresponsive ones) are becoming (more) sensitive to movements in the working range of the joint and many show (enhanced) ongoing discharges. Consequently spinal nociceptive-specific and wide dynamic range neurons with afferent input from the inflamed knee develop (increased) responsiveness to gentle stimulation of the joint. But in addition most of these neurons display enhanced reactions to non-inflamed parts of their receptive fields, too, and some neurons show enlargement of their total receptive fields. These latter findings indicate that the sensitization of spinal neurons is not simply reflecting the increased afferent input from the inflamed knee but that intrinsic spinal mechanisms may participate in the sensitization process.     

Diffuse noxious inhibitory controls in anti-nociception produced by acupuncture and moxibustion on trigeminal caudalis neurons in rats

Numerous studies have demonstrated that acupuncture and moxibustion induce analgesic effects. This study examined whether diffuse noxious inhibitory controls (DNIC) participated in acupuncture and moxibustion induced-analgesia. Single unit extracellular recordings from neurons in the trigeminal nucleus caudalis of urethane-anesthetized Wistar rats were obtained with a glass micropipette. A total of 52 single units, including 36 wide dynamic range (WDR), 5 nociceptive specific (NS) and 11 low-threshold mechanoreceptive (LTM) units were examined. During noxious test stimulation (cutaneous pinch or electrical stimulation), acupuncture, moxibustion or pinch stimulation was applied as the conditioning stimulus to the remote area of the receptive fields. When the conditioning stimulation induced rapid suppression of noxious receptive field stimulation response, examination revealed that various areas of the entire body were affected and suppression increased in an intensity-dependent manner. These features resemble DNIC phenomena. The suppression was observed on both WDR and NS neurons but not on LTM neurons. Eight of 16 WDR neurons examined were inhibited by acupuncture, five of 14 by moxibustion, and seventeen of 21 by pinching stimulation. Of the NS neurons, one of 2 units examined was suppressed by acupuncture, one of 2 by moxibustion, and two of 3 by pinch stimulation. Pinch stimulation induced the most profound suppression followed by manual acupuncture. Moxibustion induced moderate suppression with a long induction time. These results suggest that DNIC may be involved in the analgesic mechanism of acupuncture and moxibustion.     

Ontogeny of the calcium binding protein parvalbumin in the rat nervous system

Summary In the adult rat brain, the calcium-binding protein parvalbumin is preferentially associated with spontaneously fast-firing, metabolically active neurons and coexists with gamma-amino-butyric acid (GABA) in cortical inhibitory interneurons. Whether this is so in developing neurons has not been explored. To this end, we have used parvalbumin immunohistochemistry to study expression of this protein in the rat nervous system during pre- and postnatal life. Our results indicate that parvalbumin first appears at embryonic day 13 in sensory system of the spinal cord, in the vestibular (VIII), the trigeminal (V) and the visuomotor (III, IV VI) systems, and develops rapidly during the following days. In these locations the expression of parvalbumin coincides with the beginning of physiological activity in nerve cells. In the gamma-aminobutyric acid (GABA)-containing interneurons of the cerebral cortex and the hippocampus, as well as in the Purkinje cells of the cerebellum, parvalbumin only appears postnatally. It lags behind the development of GABA-immunoreactivity by 1 to 2 weeks. The beginning of its expression, in the cerebellum at least, coincides with the arrival of excitatory synaptic input and the onset of spontaneous activity. Thus, during the development of the nervous system, the expression of parvalbumin is subordinate to the establishment of physiological activity.Abbreviations 3 oculomotor nucleus - 4 trochlear nucleus - 4n trochlear nerve - 6 abducens nucleus - 12 hypoglossal nucleus - 3n oculomotor nerve - 4V 4th ventricle - 5g trigeminal ganglion - 5n trigeminal nerve - 5mx trigeminal nerve, maxillary branch - 8c1 cochlear ganglion - 8g vestibular ganglion - 8n vestibular nerve - 10n vagal nerve - Amb ambiguus nucleus - CaBP calcium-binding protein - Ce cerebellum - ChP choroid plexus - cl cochlea - CPu caudate putamen - Cu cuneate nucleus - Cx cerebral cortex - df dorsal funiculus spinal cord - dr dorsal root spinal nerve - E15 embryonic day 15 of gestation - ECN external cuneate nucleus - Fr formatio reticularis - GABA gamma-amino-butyric acid - GAD glutamate decarboxylase - gl granular layer cerebellum - Gr gracile nucleus - Hip hippocampus - H heart - inc inferior colliculus - IOK inferior olive, kap cooy medial nucleus - Li liver - LMol lacunosum moleculare layer hippocampus - Lu lung - LV lateral ventricle - LVe(v) lateral vestibular nucleus (ventral) - LVe(d) lateral vestibular nucleus (dorsal) - me5 mesencephalic trigeminal tract - Me5 mesencephalic trigeminal nucleus - Mes mesencephalon - ml molecular layer cerebellum - MVe medial vestibular nucleus - Or oriens layer hippocampus - P 2 postnatal day 2 - Pu Purkinje-cell layer, cerebellum - Py pyramidal cell layer, hippocampus - R red nucleus - Rhom rhombencephalon - Rt reticular thalamic nucleus - sk skin - sn substantia nigra - spgl spinal ganglion - sp5 spinal trigeminal tract - SpVe spinal vestibular nucleus - Sp5I spinal trigeminal nucleus, interpolar - suc superior colliculus - SuVe superior vestibular nucleus - TBS tris-buffered saline - tch tactile hair sinus - ve vestibular epithelium - vb vertebral body - vh ventral horn spinal cord - VL ventrolateral thalamic nucleus - VP ventral pallidum - vr ventral root spinal nerve