Reflex responses of single salivatory neurons to stimulation of trigeminal sensory branches in the cat

Responses of 71 single salivatory neurons, identified by antidromic spikes evoked by stimulation of the chorda tympani, were tested to stimulation of the ipsilateral infraorbital (IO), inferior alveolar (IA) and lingual nerves (LN) in the cat. Fifty-one neurons responded with spike potentials to stimulation of one or more of these nerves (responsive type, R), while the remaining 20 neurons did not respond to stimulation of any of them (non-responsive type, NR). Thirty-three R neurons activated by stimulation of all of the 3 trigeminal afferent branches, while 12 neurons responded with spikes to stimulation of only one branch, usually of LN. Reflex spike responses appeared with a latency of 5.6–14.6 ms to LN stimulation, 6.4–15.7 ms to IO stimulation and 6.0–26.0 ms to IA stimultion. Impulses of both Aβ and Aδ afferent fibres of the trigeminal nerve were found to be effective for activation of salivatory neurons.     

Infrared sensory neurons in the trigeminal ganglia of crotaline snakes: transganglionic HRP transport

Trigeminal neurons were labeled by inserting HRP into holes cut in the pit receptor membranes of a crotaline snake, Agkistrodon blomhoffi brevicaudus. Neurons were labeled in the ophthalmic ganglion and the maxillary division of the maxillo-mandibular ganglion, and the HRP was further transported across the ganglia and through the lateral descending trigeminal tract (dlv) to label axon terminals exclusively in the dlv nucleus (DLV). In 6 successful preparations, 7.1-19.3% of totals of 5568-5986 cells in the maxillary division of the ganglion were labeled, but none at all were labeled in the mandibular division. Only a few or none at all were labeled in the ophthalmic ganglion. Cells in the two ganglia ranged in size from 10 to 55 micrometers, but large cells (greater than or equal to 40 micrometers) were scarce (4.9% of the total population). All HRP-labeled neurons fell in the median range of 20-39 micrometers. We concluded that these ganglion cells were infrared neurons, and were therefore the origin of the A delta fibers in the pit membrane. There were no HRP-labeled neurons above or below this range, in spite of the fact that smaller cells (less than or equal to 19 micrometers) made up 35.8% of the total population. In normal Nissl preparations we found both light- and dark-staining cells, but the size range of neither corresponded to the size range of infrared neurons.     

Retrograde axonal transport of asialoglycoproteins in mouse trigeminal neurons in vivo and in rat dorsal root ganglia neurons in vitro

Different terminal sugars of the glycoprotein orosomucoid were exposed by sequential glycosidase digestions. The orosomucoid and its different derivatives were conjugated to horseradish peroxidase by a two-step glutaraldehyde coupling procedure, injected into the snout of 12-day-old mice or exposed to dorsal root ganglia neurons from embryonic rats, cultivated in a two chamber system. A marked increase in transport of the conjugates in the trigeminal and dorsal root ganglia neurons was observed histochemically after removal of sialic acid, exposing galactose as the terminal sugar. Quantitative hydrolysis of galactose residues resulted in reduced uptake. The data suggest the presence of a galactose-recognition molecule in the axon-terminal membrane, involved in retrograde axonal transport.     

Substance P-containing trigeminal sensory neurons project to the nucleus of the solitary tract

Intense substance P-like immunoreactivity (SPLI) was identified in fiber bundles coursing between the spinal nucleus of the trigeminal nerve and the ventrolateral nucleus of the solitary tract at the level of the area postrema. These bundles were apparent only when tissue was stained for substance P immunoreactivity and were not visible in preparations treated with antisera to somatostatin or neurotensin. Following unilateral section of the trigeminal nerve, the SPLI-containing fiber bundles were absent ipsilateral to the nerve section. The fibers were absent bilaterally in rats which were previously injected with capsaicin. Unilateral removal of the nodose ganglion did not diminish the intensity or apparent number of SPLI fibers. These data indicate the presence of a trigeminosolitary projection which is composed of primary trigeminal sensory neurons containing substance P. The results provide an anatomical route by which substance P of trigeminal origin may modulate vagal or glossopharyngeal sensory information.     

The morphology of trigeminal nociceptive neurons in the caudal bulbar lateral reticular formation of the cat

Two types of trigeminal nociceptive neurons, i.e. subnucleus reticularis ventralis (SRV) and wide dynamic range (WDR) neurons were identified in the caudal bulbar lateral reticular formation (LRF) and intracellularly stained with horseradish peroxidase. SRV neurons were large neurons characteristic of the subnucleus reticularis ventralis. Their dendrites were confined to the LRF. WDR neurons were situated in the subnucleus reticularis dorsalis. Their dendrites penetrated into the magnocellular layer, but did not reach the substantia gelatinosa.     

Mechanosensitivity of voltage-gated K+ currents in rat trigeminal ganglion neurons

We investigated the mechanosensitivity of voltage-gated K+ channel (VGPC) currents by using whole-cell patch clamp recording in rat trigeminal ganglion (TG) neurons. On the basis of biophysical and pharmacological properties, two types of VGPC currents were isolated. One was transient (I(K,A)), the other sustained (I(K,V)). Hypotonic stimulation (200 mOsm) markedly increased both I(K,A) and I(K,V) without affecting their activation and inactivation kinetics. Gadolinium, a well-known blocker of mechanosensitive channels, failed to block the enhancement of I(K,A) and I(K,V) induced by hypotonic stimulation. During hypotonic stimulation, cytochalasin D, an actin-based cytoskeletal disruptor, further increased I(K,A) and I(K,V), whereas phalloidin, an actin-based cytoskeletal stabilizer, reduced I(K,A) and I(K,V). Confocal imaging with Texas red-phalloidin showed that actin-based cytoskeleton was disrupted by hypotonic stimulation, which was similar to the effect of cytochalasin D. Our results suggest that both I(K,A) and I(K,V) are mechanosensitive and that actin-based cytoskeleton is likely to regulate the mechanosensitivity of VGPC currents in TG neurons.     

Response characteristics of primary periodontal mechanoreceptive neurons in the trigeminal mesencephalic nucleus to trapezoidal mechanical stimulation of a single tooth in the rat

The response characteristics of primary periodontal mechanoreceptive neurons in the trigeminal mesencephalic nucleus (MeNV) were studied by changing the rate and magnitude of trapezoidal pressure applied to the upper incisor in very lightly anesthetized rats.Using a metal microelectrode in the MeNV either for recording or for stimulation, the projection site of primary afferents in the anterior superior alveolar nerve innervating the upper incisor was determined. Its stereotaxic coordinates were 1.0–2.0 mm posterior to the interaural plane and 1.2–1.3 mm lateral to the mid-sagittal plane, corresponding to the caudal part of the MeNV.From this site of 19 animals, single unitary activity from 41 primary periodontal mechanoreceptive neurons was recorded, which were identified by: (1) the constant, brief latency of 1.0–4.0 ms (2.0 ± 0.6ms, mean±S.D.) from the onset of single tooth tapping; (2) the wave forms of their unitary spike responses; and (3) the ability to follow faithfully trains of repetitive stimuli applied to the anterior superior alveolar nerve at rates of more than 100 Hz for 2 s. The responses of the identified primary mechanoreceptive neurons in the MeNV were tested for repetitive tapping and trapezoidal pressure to the ipsilateral upper incisor. The highest frequency of one-to-one following was observed in repetitive tapping at 100 Hz. All of the 27 tested neurons exhibited such very rapid adaptation as to show only on-off responses to trapezoidal pressures, and did not respond at all unless the pressures were applied more rapidly than 0.6 Newton/s (N/s). In most of these neurons, the number of evoked spikes was greater in on-responses than in off-responses, and the number and frequency of spikes were increased with an increase in the rate of pressure application. But in several neurons only one spike was triggered even when the pressures were applied at the rate of more than 83.1 N/s.     

Organization of HRP-labeled trigeminal mandibular primary afferent neurons in the rat

Horseradish peroxidase (HRP) applied to the transected mandibular division of the trigeminal (V) ganglion was transported anterogradely to pri-mary afferent terminal zones in the dorsal and dorsomedial trigeminal brain-stem nuclear complex (TBNC). Primary V afferents of ganglionic origin were also visible in the ipsilateral cerebellar cortex (crus I and II, paraflocculus) and the dentate, cuneate, solitary, supratrigeminal, and dorsal motor vagal nuclei, parvicellular reticular formation, area postrema and C1–C6 dorsal horn, laminae I–V. Contralateral subnucleus caudalis and C1–C2 dorsal horn were also innervated by primary afferents which crossed in the spinal gray to terminate medially, primarily in laminae I, II, and V. Almost all of these projections were also labeled in various combinations when HRP was applied to individual sensory branches of the mandibular nerve: lingual, infe-rior alveolar, mylohyoid, and auriculotemporal. Transganglionic transport of HRP in the latter four cases revealed strong evidence for mtradivisional somatotopy among the four branches in both the ganglion and TBNC. Cell bodies innervating posterior and/or lateral portions of the head and face (i.e., auriculotemporal and mylohyoid) were found with greater frequency in dor-sal mandibular ganglion regions, while somata supplying more rostral oral-perioral regions (i.e., lingual and inferior alveolar) were predominant ventrally. Components of the mandibular projection to the TBNC were organized topographically in at least some portion of all of its three dimen-sions. Subnuclear preferences were not clear-cut; all four nerves innervated at least some portion of principalis, oralis, interpolaris, and caudalis, save for mylohyoid, which did not project to caudalis. Lingual fibers were most prominent in principalis and oralis, occupied medial portions of the mandib-ular projection to the TBNC, and descended only to rostral caudalis, most notably laminae I-III. Inferior alveolar afferents were ubiquitous in the mandibular component of the TBNC and C1–C2, save for its far lateral bor-der. Mylohyoid terminals were sparse, most prominent in interpolaris, and occupied only dorsolateral TBNC regions and laminae III and IV of C1–C3. The auriculotemporal innervation of the mandibular TBNC was heaviest in interpolaris and was restricted to mostly ventrolateral regions. Its primary focus, however, was laminae III and IV of C1–C4. The clinical implications of this topographical organization are discussed, particularly with respect to the rostrocaudal intradivisional lamination in caudalis and the cervical dorsal horn.     

Inhibition of hyperactive trigeminal subnucleus caudalis neurons after experimental trigeminal rhizotomy in response to thalamic sensory relay nucleus stimulation*

The effects of thalamic sensory relay nucleus stimulation on the single neuron activity and field potentials within the trigeminal subnucleus caudalis, which is a trigeminal equivalent of the dorsal horn were investigated in intact cats as well as in cats subjected to retro-Gasserian rhizotomy 1-6 months before the experiments. inhibition of nociceptive neural activity and a positive field potential were evoked by thalamic stimulation in the dorsal horn of the intact animals. A positive field potential followed double pulse stimulation with frequencies ranging up to approximately 50 Hz. The inhibitory periods ranged from 60 to 100 ms. Train pulse stimulation with frequencies ranging from 30 to 50 Hz produced long-lasting inhibition of nociceptive neural activity and a positive shift of the field potentials. Essentially identical inhibition of abnormal neural hyperactivity occurring in the rhizotomized dorsal horn was observed. The positive field potential corresponding to this inhibition also displayed characteristics similar to the field potential seen in the intact dorsal horn. These data indicate that the pathways involved in the inhibitory responses induced by thalamic. sensory relay nucleus stimulation, unlike some other pain inhibitory systems, can exert their influence even to the dorsal horn which has undergone reorganization of neural circuits after rhizotomy.     

Effects of chronic descending tractotomy on the response patterns of neurons in the trigeminal nuclei principalis and oralis.

Unilateral, descending, trigeminal tractotomy was carried out on adult cats using aseptic technique. Unit activity, evoked by bipolar noxious stimulation of the tooth pulp and by innocuous mechanical stimulation of other receptive fields, was recorded from neurons in the trigeminal principalis and oralis nuclei, 7 to 13 weeks after tractotomy. Seventy neurons from the lesion side of the brain stem and 59 from the control side were studied. All units responded to pulpal stimuli and the great majority to mechanical stimuli. Three general response configurations were produced by each of the two forms of stimulation: brief bursts of 1 to 5 spikes, trains of 6 to 20 spikes, and multiple component discharges consisting of sequences of brief bursts. No statistically significant differences between the two populations were found for peripheral field size or location, for discharge characteristics such as latency, configuration, or mean density, or for responsiveness to the two types of stimuli. However, a statistically greater proportion of control units fired dense impulse trains (more than eight spikes) to intense pulpal stimuli. The results provide evidence that the response patterns of most somatosensory neurons in nuclei principalis and oralis are unaffected by descending trigeminal tractotomy and that stimulus modality is not coded by the response characteristics of polymodal units in these nuclei. Although some modulatory influence is exerted on polymodal neurons in the rostral nuclei by the trigeminal nucleus caudalis, it appears doubtful that this is a significant factor in determining whether a peripheral stimulus is perceived as noxious or innocuous.     

Satellite glial cell proliferation in the trigeminal ganglia after chronic constriction injury of the infraorbital nerve

We have examined satellite glial cell (SGC) proliferation in trigeminal ganglia following chronic constriction injury of the infraorbital nerve. Using BrdU labeling combined with immunohistochemistry for SGC specific proteins we positively confirmed proliferating cells to be SGCs. Proliferation peaks at approximately 4 days after injury and dividing SGCs are preferentially located around neurons that are immunopositive for ATF‐3, a marker of nerve injury. After nerve injury there is an increase GFAP expression in SGCs associated with both ATF‐3 immunopositive and immunonegative neurons throughout the ganglia. SGCs also express the non‐glial proteins, CD45 and CD163, which label resident macrophages and circulating leukocytes, respectively. In addition to SGCs, we found some Schwann cells, endothelial cells, resident macrophages, and circulating leukocytes were BrdU immunopositive. GLIA 2013;61:2000–2008     

痛刺激后大鼠三叉神经尾侧亚核星形细胞和神经元相互关系的电镜观察

目的：观察大鼠上唇皮下注射福尔马林后，三叉神经尾侧亚核（Sp5C)内反应性星形细胞和神经元之间相互关系的超微结构。方法：用DAB染色的抗胶质原纤维酸性蛋白（GFAP）、抗connexin43(Cx43）和金颗粒标记抗Cx32双标记免疫电镜方法。结果：电镜下观察到Sp5C内反应性星形细胞和神经元之间存在4种联系结构：第一种是突触样结构；第二种是三种成分的突触复合体，第三种是缝隙连接；第四种是由Cx32和Cx43构成的异源性缝隙连接（HGJ）。HGJ表现为两侧膜增厚，Cx43阳性物质和Cx32阳性金颗粒分别位于星形细胞和神经元一侧，痛刺激后HGJ数明明显增加。结论：神经元和星形细胞之间有多种信息通道，HGJ可能是一种快速，适应性信息通道。Sp5C星形细胞可能通过HGJ调节神经元的活动，共同参与中枢神经系统对刺激反应的调节。     

Physiological properties of neurons in different parts of the cat trigeminal sensory complex

The following points emerge from a systematic investigation of the 4 divisions of the cat trigeminal sensory complex.(1) The subnucleus oralis receives a large representation from the oral cavity, a region also represented in the 3 other divisions of the trigeminal sensory complex.(2) Nucleus principalis cells project heavily to the contralateral and to the ipsilateral ventroposterior thalamus. Ipsilateral projections are only from the oral cavity representation.(3) Units responding to noxious mechanical stimulation have been found at two different loci: the subnucleus caudalis for the entire trigeminal area, and subnucleus oralis for the oral cavity alone.(4) The dental pulp projects to the 4 divisions of the trigeminal sensory complex, but the heaviest projection was found in the rostral part (nucleus principalis and subnucleus oralis).(5) Three distinct types of post-synaptic responses were found to be evoked by dental pulp stimulation: (a) short latency, consistent and synaptically secure, (b) strongly variable latency, inconstant and easily fatigued and (c) a class showing progressive enhancement by progressive increase in stimulus intensity and repetition.     

Ultrastructural observations of the migration and early development of trigeminal motoneurons in chick embryos

The migration of immature chick trigeminal (V) motoneurons from a miclline medial column to a lateral nucleus was studied between days 3 and 7 of incubation. The somatic translocation of these cells is preceded by the lateral outgrowth of their axons. At days 3–4 the axons grow within extracellular spaces dorsal to pial end feet at the external limiting membrane. At later ages the axons grow deeper within the intermediate zone. Upon reaching the medial border of the V sensory tract the axons turn ventrally, exit from the neural tube, and grow toward the mandibular arch. The medial column somas rotate from their original radial orientation to lie parallel to the pial surface. They migrate laterally in close association with fiber bundles composed of axonal and trailing processes of medial column and other migrating V motoneurons. Several punctate contacts are made between the migrating cells and the axon bundles; these contacts may stabilize the somas while the growing tips of their axons elongate. During migration the cytoplasm of the V motoneuron matures, acquiring rough endoplasmic retic-ulum, lysosomes, and multivesicular bodies. Immature synapses contact premigratory, migratory, and postmigratory cells on day 4. Their maturation on day 5 coincides with the onset of active jaw movements. Some V motoneurons secondarily migrate dorsally from the lateral nucleus while their trailing axons are in the head mesenchyme. Few punctate contacts are observed between these cells and their substrate, the longitudinal fibers of the lateral longitudinal fasciculus. Postmigratory dorsal V motoneurons are organelle poor and have few synaptic contacts by day 7. This undifferentiated state may reflect a later onset of the spontaneous movement of their target, the levator palpebrae muscle.     

Birth dates of trigeminal motoneurons and metamorphic reorganization of the jaw myoneural system in frogs

Drastic alterations in oral behavior characterize metamorphosis of anuran amphibians. Changes cascade through all components of the jaw apparatus from bone to muscle to nerve. In this investigation, tritiated thymidine autoradiography was used to determine the production schedule of the trigeminal motoneurons in the leopard frog, Rana pipiens. The time of origin of these neurons and their subsequent fate are of special interest given the breakdown of the larval jaw muscles and the de novo generation of adult muscle fibers during metamorphosis. Specifically, we wanted to learn whether trigeminal motoneurons are added, deleted, or reused during metamorphic climax. The entire complement of trigeminal motoneurons was produced over a 4-day span commencing at embryonic stage 13 and terminating at stage 20. Newly formed neurons are added to the primordial trigeminal nucleus in an orderly pattern. Firstborn neurons settle in the ventrorostral region of the nucleus; cells with progressively later birth dates were added in a posterodorsal direction. No additional trigeminal motoneurons are generated during larval maturation or at metamorphosis, thus indicating that the same population of neurons is present throughout the lifespan of the animal. From these observations we suggest that, during metamorphosis, the trigeminal motoneurons that supply the larval muscles switch their allegiance to the newly formed adult jaw muscles. This change of peripheral targets can be viewed as a respecification of the trigeminal motoneurons.     

Re-evaluation of acute neurotoxic effects of Cd on mesencephalic trigeminal neurons of the adult rat

The mechanism of Cd²⁺ neurotoxicity, which is considered to be secondary to changes in blood vessels, was re-evaluated in dissociated mesencephalic trigeminal (Me5) neurons of the adult rat. Cd²⁺ induced morphological changes in Me5 neurons at 0.1 and 1 mM but not at 0.01 mM. The changes appeared predominantly in the cytoplasm: destruction of the cytoplasmic organelles, swelling and vacuolization of the cell body, and finally resulted in cell lysis. These observations indicate necrosis rather than apoptosis, and no sign of degraded nuclear DNA, characteristic to apoptosis, was detected by the TUNEL technique. Using a Ca²⁺-sensitive dye Indo-1, Cd²⁺ was found to elevate the intracellular Ca²⁺ concentration [Ca²⁺]_i (both in the cytoplasm and the nucleus). Both the elevation in [Ca²⁺]_i and the morphological alteration were inhibited either by removing Ca²⁺ from the bathing medium or by the application of BAPTA/AM (10 μM), a membrane-permeable intracellular Ca²⁺ chelator. Furthermore, neither morphological changes nor elevation in [Ca²⁺]_i by Cd²⁺ occurred in the presence of Zn²⁺. It is concluded that (1) Cd²⁺ can directly affect nerve cells, (2) toxicity of Cd²⁺ on Me5 neurons is mediated by continuous elevation in [Ca²⁺]_i, (3) Cd²⁺ induces necrotic cell death, and (4) Cd²⁺ neurotoxicity can be antagonized by Zn²⁺.     

Somatotopic organization of the primary sensory trigeminal neurons in the hagfish, Eptatretus burgeri

Primary sensory trigeminal projections were investigated in the hagfish following application of horseradish peroxidase (HRP) to the sensory branches. In our control preparations we were able to distinguish five sensory ganglia and their respective nerves. HRP application confirmed the almost exclusive relation of each of these nerves to their respective ganglia, with very little overlap. In normal frontal sections of the medulla oblongata, five columns of fibers surrounded by neuronal cell bodies could be clearly distinguished, but the number is probably fortuitous, for there was no one-on-one relationship with the five trigeminal ganglia. From their peripheral connections, we surmised that columns 1 and 3 handle general cutaneous sensation, columns 2, 4, and 5 handle taste sensation, and column 5 handles general mucous cutaneous sensation conveyed by utricular ganglion cells. Dorsally located columns received projections from nerves with dorsal peripheral connections, and more ventrally located columns received projections from nerves with ventral peripheral connections. This relation is the reverse of that seen in other vertebrates.     

Suppression of neurokinin-1 receptor in trigeminal ganglia attenuates central sensitization following inflammation

This study examined whether local application of a neurokinin-1 (NK1) receptor antagonist into the trigeminal ganglia (TRGs) modulates hyperexcitability of trigeminal spinal nucleus caudalis (SpVc) wide-dynamic range (WDR) neuron activity innervating both the temporomandibular joint (TMJ) region and facial skin following TMJ inflammation. Extracellular single unit recording combined with multibarrel electrodes was used. TMJ inflammation was induced by the injection of complete Freund's adjuvant (CFA). WDR neurons responding to electrical stimuli of the TMJ region and facial skin were recorded from the SpVc in anesthetized rats. The spontaneous and mechanical stimulation-induced discharge frequencies of WDR neurons were significantly larger in inflamed rats than in control rats. The spontaneous WDR activities were current-dependently decreased by local iontophoretic application of an NK1 receptor antagonist into the TRGs after 1 and 2 days of inflammation. The firing frequency of WDR neurons and threshold evoked by mechanical stimulation of facial skin returned to control levels by application of the NK1 receptor antagonist into TRGs after 1 day, but not 2 days, of inflammation. These results suggest that in the early stages of inflammation suppression of the NK1 receptor mechanism in TRGs may prevent central sensitization of SpVc nociceptive neurons.     

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.     

Trigeminal ganglion neurons which project by way of axon collaterals to both the caudal spinal trigeminal and the principal sensory trigeminal nuclei

Employing a combination of fluorescent retrograde double labeling and immunofluorescence histochemistry, we found that some single neurons in the trigeminal ganglion of the rat projected by way of axon collaterals both to the caudal spinal trigeminal nucleus and to the principal sensory trigeminal nucleus, and that about 40% or 57% of these neurons showed respectively substance P- or calcitonin gene-related peptide-like immunoreactivity.