Cell size-specific appearance of neuropeptide Y in the trigeminal ganglion following peripheral axotomy of different branches of the mandibular nerve of the rat

The effect of peripheral axotomy of the mental nerve (MN) and the cutaneous branch of the mylohyoid nerve (MhN) on the appearance of neuropeptide Y-like immunoreactivity (NPY-IR) in cells in the trigeminal ganglion of the rat was examined with combined retrograde-tracing and immunohistochemistry. Retrograde-tracing with True Blue (TB) revealed that the cell-size spectrum of the trigeminal cells sending peripheral processes to the MN (TB MN cells) ranged from 75.9 to 1560.5 μm² (or from 9.8 to 44.6 μm in diameter); 53% of TB MN cells were 300–600 μm². TB MhN cells ranged from 47.7 to 1261.5 μm² (or from 7.8 to 40.1 μm in diameter); 56% of TB MhN cells were <300 μm². In the normal trigeminal ganglion, there were no NPY-IR cells. 14 days after MN transection, 35% of TB MN cells displayed NPY-IR. The distribution of the cross-sectional areas of NPY-IR cells after MN transection was very similar to that of TB MN cells. Transection of MhN also induced the appearance of NPY-IR in the trigeminal ganglion but to a lesser extent (17% of TB MhN cells). The distribution of the cross-sectional areas of NPY-IR cells after MhN transection was similar to that of NPY-IR cells after MN transection. These results indicate that injury-evoked NPY-IR is specific for the medium- and large-sized ganglion cells.     

Partial coexistence of neuropeptide Y and calbindin D28k in the trigeminal ganglion following peripheral axotomy of the inferior alveolar nerve in the rat

Immunohistochemistry was applied to examine the correlation between neuropeptide Y (NPY) and the two calcium binding proteins (CaBPs) parvalbumin (PV) and calbindin D28k (CB) in the trigeminal ganglion following peripheral axotomy of the inferior alveolar nerve (IAN) in the rat. Five days following transection and application of FluoroGold (FG) to the cut end of the IAN, approximately 14.8% (80/539) and 18.6% (90/483) of FG-labeled IAN neurons in the trigeminal ganglion showed PV-like immunoreactivity (-LI) and CB-LI, respectively. The mean ± S.D. area of FG-labeled PV-like immunoreactive (-IR) cells (FG/PV-IR cells) and FG/CB-IR cells were 835.9 ± 303.1 μm² and 712.7 ± 246.0 μm², respectively. FG/PV-IR cells were significantly larger than FG/CB-IR cells. Fourteen days following peripheral axotomy of the IAN, NPY-LI appeared in the medium- to large-sized cells. Double immunostaining revealed that approximately 3.3% (52/1569) of NPY-IR cells in the axotomized trigeminal ganglion displayed PV-LI, while approximately 26.7% (371/1392) of NPY-IR cells displayed CB-LI. The mean ± S.D. cross-sectional areas of PV-IR and CB-IR trigeminal ganglion cells displaying NPY-LI were 819.5 ± 265.6 μm² and 766.5 ± 279.7 μm², respectively. There were no significant differences in the cross-sectional areas either between NPY/PV-IR cells and NPY/CB-IR cells, or between FG/PV-IR cells and NPY/PV-IR cells, or between FG/CB-IR cells and NPY/CB-IR cells. The present results indicate that injury-evoked medium- to large-sized NPY neurons were a different population from large-sized PV neurons, and NPY was partly co-localized with CB.     

Immunoelectron microscopic study on neuropeptide Y in the periodontal ligament of the incisor following peripheral nerve injury to the inferior alveolar nerve in the rat

Immunoelectron microscopic analysis was carried out to examine whether neuropeptide Y (NPY)-like immunoreactivity (-LI) is localized in mechanoreceptors in the lingual periodontal ligament of the rat incisor following peripheral nerve injury to the inferior alveolar nerve (IAN). In the lingual periodontal ligament of normal animals, no NPY-like immunoreactive (-IR) primary afferents were observed, except for a very few sympathetic perivascular nerve fibers which showed NPY-LI. Fourteen days following chronic constriction injury to the IAN combined with sympathectomy of the superior cervical ganglion, thick NPY-IR nerve fibers showing tree-like raminifications were detected in the shear zone between the tooth-related part and alveolus-related part as well as in the alveolus-related part. Immunoelectron microscopy revealed that expanded NPY-IR nerve terminals were covered with several Schwann sheaths and that a part of the axoplasm expanded to the surrounding tissues. These ultrastructural features of NPY-IR structures were identical to those of periodontal Ruffini endings, categorized as slowly adapting mechanoreceptors. Thick (6–8 μm in diameter) NPY-IR axons were also observed without any apparent myelin sheath. The present results provide further evidence that NPY is closely associated with thick axons, probably myelinated nerves and Ruffini endings, following peripheral nerve injury.     

Effects of peripheral nerve injuries and tissue inflammation on the levels of neuropeptide Y-like immunoreactivity in rat primary afferent neurons

Changes in neuropeptide Y-like immunoreactivity (NPYir) in the rat L₄ and L₅ spinal cord and dorsal root ganglia (DRG) were examined after different sciatic nerve injuries (transection, loose ligation, and crush) and a localized, painful inflammation of the hind paw. Inflammation had no effect on NPYir. All the nerve injuries produced comparable increases in NPYir in ipsilateral laminae III–V axons and varicosities, and induction of NPYir in many DRG cells. Most NPYir DRG cells were medium to large (mean diameters: 40–45 μm); less than 2% of the cells had diameters of 25 μm or less. We conclude that the nerve injury-evoked increase in NPYir occurs mostly in the somata and intraspinal arbors of low-threshold mechanoreceptors; very few, if any, C-fiber afferents are involved. Nerve injury, rather than a painful condition, appears to be the stimulus for the induction of NPYir synthesis.     

Neuropeptide Y-immunoreactive primary afferents in the dental pulp and periodontal ligament following nerve injury to the inferior alveolar nerve in the rat

The distribution of neuropeptide Y (NPY)-immunoreactive (IR) primary afferents in the dental pulp and periodontal ligament of the rat mandible were examined following combined chronic constriction injury (CCI) of the inferior alveolar nerve (IAN) and sympathectomy of the superior cervical ganglion (SCG). NPY-IR nerve fibers were observed around the blood vessels in the trigeminal ganglion, dental pulp and periodontal ligament in normal animals. Following combined CCI of the IAN and sympathectomy of SCG (SCGx), perivascular NPY-IR nerve fibers originating from SCG disappeared completely, but many NPY-IR nerve fibers coming from the trigeminal ganglion appeared in the dental pulp and periodontal ligament. In the molar dental pulp, thick NPY-IR nerve fibers were observed within the nerve bundle, and some thin NPY-IR nerve fibers ran towards the odontoblast layer; very few NPY-IR nerve fibers were observed in the incisor pulp. In the periodontal ligament of molar, thick NPY-IR nerve fibers appeared at the alveolar part following combined CCI of IAN and SCGx. In the lingual portion of the periodontal ligament of the incisor, many thick NPY-IR nerve fibers were observed. These occasionally showed a tree-like appearance, resembling immature Ruffini endings; slowly adapting mechanoreceptors. The present results indicate that periodontal mechanoreceptors are among the main targets of injury-evoked NPY following IAN injury.     

Osteopontin-immunoreactivity in the rat trigeminal ganglion and trigeminal sensory nuclei

Osteopontin-immunoreactivity (OPN-ir) was examined in the oro-facial tissues and trigeminal sensory nuclei (principal sensory nucleus and spinal trigeminal nucleus) to ascertain the peripheral ending and central projection of OPN-containing primary sensory neurons in the trigeminal ganglion (TG). No staining was observed using mouse monoclonal anti-OPN antibody preabsorbed with recombinant mature OPN. OPN-immunoreactive (ir) peripheral endings were classified into two types: encapsulated and unencapsulated types. Unencapsulated endings were subdivided into two types: simple and complex types. Simple endings were characterized by the thin neurite that was usually devoid of ramification. These endings were seen in the hard plate and gingiva. The complex type was characterized by the thick ramified neurite, and observed in the vibrissa, hard palate, and molar periodontal ligament. Encapsulated endings were found only in the hard palate. The trigeminal sensory nuclei contained OPN-ir cell bodies and neuropil. The neuropil was devoid of ir in laminae I and II of the medullary dorsal horn (MDH), and had various staining intensities in other regions of the trigeminal sensory nuclei. Transection of the infraorbital and inferior alveolar nerves caused an increase of OPN-ir intensity in ipsilateral TG neurons. The staining intensity of the neuropil also increased in the trigeminal sensory nuclei ipsilateral to the neurotomy excepting laminae I and II of the MDH. The present study indicates that OPN-ir primary sensory neurons in the TG innervate encapsulated and unencapsulated corpuscular endings. Such neurons probably project their central terminals to the trigeminal sensory nuclei except for the superficial laminae of the MDH.     

Involvement of nitric oxide in re-innervation of rat molar tooth pulp following transection of the inferior alveolar nerve

The aim of this study was to elucidate whether nitric oxide (NO) is involved in re-innervation of rat molar tooth pulp following transection of the inferior alveolar nerve. The inferior alveolar nerves (IAN) of rats were transected unilaterally under anesthesia with chloral hydrate. The animals received horseradish peroxidase (HRP) application to mandibular molar tooth pulps on both sides and were fixed by transvascular perfusion. The average number of labeled cells on each side of the trigeminal ganglion was not significantly different [101±11 (mean±S.E.M.; n=6, left) and 89±11 (n=6, right)]. With HRP application on postoperative day 3, the ratio of the number of labeled neurons in the transected vs. non-transected (contralateral) sides was 31.5±5.8% (n=11). The i.p. administration of N^ω-nitro-

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-arginine methyl ester (

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-NAME; 100 mg/kg, once a day for a period of 4 days), but not

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-NAME, significantly decreased the ratio of the number of labeled neurons (10.1±7.0%, n=10).

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-Arginine (300 mg/kg, i.p., once a day for a period of 4 days) slightly increased the number of labeled neurons on the transected side. Clonidine (25 μg/kg, i.p., once a day for a period of 4 days) failed to exhibit any significant effect on nerve regeneration. In the trigeminal ganglion ipsilateral to the transected IAN on postoperative day 4, NADPH-diaphorase (NADPH-d)-positive neurons had significantly increased. On the other hand, no changes in NADPH-d were observed in the superficial layers of the subnucleus caudalis of the spinal trigeminal nucleus from where primary neurons innervating the mammalian tooth pulp project. These results suggest that NO is involved in several mechanisms related to neuronal regeneration.     

Immunohistochemical evidence for enkephalin and neuropeptide Y in rat inferior colliculus neurons that provide ascending or commissural fibers

Enkephalin- and neuropeptide Y-expressin neurons which offer commissural axons or axons toward auditory thalamus were identified in the rat inferior colliculus. These neurons exhibited a differential distribution pattern. The results provide evidence for regional specificity and chemical heterogeneity of neurons in the auditory midbrain.     

Ca(2+)/calmodulin-protein kinase IIalpha in the trigeminal subnucleus caudalis contributes to neuropathic pain following inferior alveolar nerve transection

Calcium-calmodulin protein kinase IIalpha (CaMKIIalpha) is mainly found in brain cells, and the mRNA concentrates highly in the postsynaptic density. CaMKIIalpha is an effector of calcium and calmodulin mediated functions, and the phosphorylated CaMKIIalpha (pCaMKIIalpha) activates glutamate receptors, such as the AMPA receptor, and enhances its function. In the present study, we examined whether CaMKIIalpha in trigeminal brainstem neurons contributed to the neuropathic pain induced by inferior alveolar nerve (IAN) transection. Using immunohistochemistry and in situ hybridization, we found that the expression of CaMKIIalpha and pCaMKIIalpha increased in the trigeminal subnucleus caudalis (Vc) after IAN transection. The significant increase in the protein of CaMKIIalpha peaked at 30 min after IAN transection, and the mRNA of CaMKIIalpha increased from 2 to 14 days. Double immunofluorescent staining for CaMKIIalpha and MAP2, a marker of dendrite, revealed a significant increase in the overlapping area at 30 min after injury. This suggests that CaMKIIalpha protein is synthesized from the local mRNA pool in the dendrite 30 min after IAN transection and may quickly transmit information after nerve injury. In the behavioral test in which the escape threshold from mechanical stimulation to the lateral face was measured, intrathecal administration of KN-93, a CaMKII inhibitor, for 7 days significantly inhibited mechano-allodynia induced by IAN transection, as compared with administration of a control peptide. These data suggest that CaMKIIalpha in the trigeminal subnucleus caudalis may be involved in neuropathic pain caused by IAN transection.     

The effect of substance P on the spontaneous discharge from injured inferior alveolar nerve fibres in the ferret

Previous studies in our laboratory have shown that, at a site of inferior alveolar nerve (IAN) injury, there is a close association between the development of spontaneous neural activity and the accumulation of neuropeptides in the damaged axons. As this ectopic activity may contribute to the development of sensory disturbances after injury, we have examined further this relationship by determining the potential role of one neuropeptide, substance P (SP), in the initiation or modulation of the spontaneous discharge. Thirty-six adult ferrets were anaesthetised, the IAN sectioned, and the animals allowed to recover for 3-4 days. In terminal experiments under general anaesthesia, electrophysiological recordings were made from axons in fine filaments dissected from the nerve, central to the injury site. The effect of SP (2 x 10(-12), 10(-6) and 10(-4) M) and SP-antagonist (10(-4) M) applied either close-arterially or topically to the injury site was determined. Of the 101 units studied, 59% were spontaneously active. Close-arterial administration of SP increased the level of spontaneous discharge in a dose-dependent manner, with higher concentrations affecting more units (2 x 10(-12) M, 14%; 10(-6) M, 58%; 10(-4) M, 85%). SP also initiated spontaneous discharge in some previously silent units. Activity in 46% of units also increased in response to the SP-antagonist. None of the units responded to topical application of either SP or SP-antagonist. This study shows that SP can both initiate and modulate the spontaneous discharge from damaged axons, and this mechanism may be a potential therapeutic target in the management of sensory disturbances after nerve injury.     

The effects of peripheral nerve injury of the masseteric nerve on the levels of calcium binding proteins and neuropeptide Y, and their correlation in the mesencephalic trigeminal nucleus of the rat

Combined retrograde neuronal tracing with FluoroGold (FG) and a double immunofluorescence method was performed to examine the effects of peripheral nerve injury of the masseteric nerve (MassN) on the levels of two calcium binding proteins (CaBPs), parvalbumin (PV) and calbindin D28k (CB), and neuropeptide Y (NPY) in the mesencephalic trigeminal nucleus (MesV) in the rat. In the normal MesV, many medium- to large-sized unipolar PV-like immunoreactive (-IR) cells were detected through the entire rostrocaudal extent, but CB-IR cells were rarely observed. No NPY-IR cells were observed in the normal MesV. The distributions of these three neurochemical markers in the MesV contralateral to the transection of Mass were almost identical to those observed in the normal MesV. Four days following transection and application of FG to the MassN, approximately 52% (572/1104) and 38% (414/1104) of FG-labeled cells (FG cells) in the MesV displayed PV-like immunoreactivity (-LI) and NPY-LI, respectively; Approximately 24% (265/1104) of FG cells showed both PV-LI and NPY-LI. Approximately 47% (265/572) of FG cells with PV-LI showed NPY-LI or 64% (265/414) of FG cells with NPY-LI displayed PV-LI. Fourteen days following transection and application of FG, the percentage of FG cells with PV-LI significantly decreased to 36% (365/1024) compared to that observed 4 days post-injury; approximately 44% (448/1024) of FG cells displayed NPY-LI; approximately 38% (141/365) of FG cells with PV-LI showed NPY-LI and approximately 31% (141/448) of FG cells with NPY-LI displayed PV-LI. In contrast, FG cells showing CB-LI were very rare on 4 days (1%; or 14 days (1%; 16/1085) following MassN transection. The present results indicate that the levels of PV in the MesV decreased 14 days following the MassN injury compared to those observed 4 days post-injury and rapid induction of NPY in the injured MesV neurons, and that the correlation between CaBP and NPY in the MesV following the MassN transection is different from that observed in the trigeminal ganglion, which is equivalent to the MesV, following peripheral nerve injury of the inferior alveolar nerve.     

Parvalbumin, calretinin and carbonic anhydrase in the trigeminal and spinal primary neurons of the rat

The cell-body size of parvalbumin-immunoreactive (-ir) primary neurons was measured in the trigeminal (TG) and lumber dorsal root ganglia (DRG). In the DRG, parvalbumin-ir was mostly detected in large cells (94% in the range of 600–2800 μm²). Parvalbumin-ir TG cells were smaller than similar DRG cells and yet parvalbumin-ir TG cells of < 400 μm² (2.86%) were rare. Trichrome stains for parvalbumin, calretinin (CR) and carbonic anhydrase (CA), and for parvalbumin, calcitonin gene-related peptide (CGRP) and CA were performed to estimate possible overlap of these substances. Virtually all parvalbumin-ir DRG cells contained CA activity while a small subpopulation (28.5%) of CR-ir DRG cells lacked CA activity. All the CR-ir DRG cells that exhibited CA were also ir for parvalbumin. 31.1% of parvalbumin-ir DRG cells exhibited CR-ir while 71.5% of CR-ir DRG cells showed parvalbumin-ir. All the CR-ir DRG cells of < 400 μm² lacked CA activity and parvalbumin-ir while all those of > 800 μm² exhibited both activities. 30% of CR-ir DRG cells in the size range of 400–800 μm² co-expressed CA. DRG cel co-expressing parvalbumin and CGRP were rare (1%). As was the case for the DRG, most of parvalbumin-ir TG cells exhibited CA activity (89.24%) and lacked CGRP-ir (96.6%). CR-ir TG cells were also subdivided into two groups; one with and the other without co-expression of CA. Unlike in the DRG, however, co-expression of parvalbumin and CR could never be detected in the TG.     

Direct excitatory effects of neuropeptide Y (NPY) on rat hippocampal neurones in vitro

Intracellular recordings from granule cells of the rat dentate gyrus show neuropeptide Y (NPY) applied by pressure ejection from pipettes containing 1.2-12 microM by pressures of less than 200 kPa for 1-5 s in duration to consistently evoke membrane depolarisations accompanied by a reduction in membrane resistance. The depolarisations were accompanied by an increase in excitability. Since the depolarisations evoked by NPY were not attenuated by either tetrodotoxin or kynurenic acid a direct excitatory action of NPY is postulated.     

Cholecystokinin and neuropeptide Y receptors on single rabbit vagal afferent ganglion neurons: site of prejunctional modulation of visceral sensory neurons

A [¹²⁵I]cholecystokinin (CCK) analog and [¹²⁵I]peptide YY (PYY) were used to localize and characterize CCK and neuropeptide Y (NPY) receptor binding sites in the rabbit vagal afferent (nodose) ganglion. High concentrations of CCK and NPY binding sites were observed in 10.6% and 9.2% of the nodose ganglion neurons, respectively. Pharmacological experiments using CCK or NPY analogs suggest that both subtypes of CCK (CCK-A and CCK-B) and NPY (Y1 and Y2) receptor binding sites are expressed by discrete populations of neurons in the nodose ganglion. These results suggest sites at which CCK or NPY, released in either the nucleus of the solitary tract or a peripheral tissue, may modulate the release of neurotransmitters from a select population of visceral primary afferent neurons. Possible functions mediated by these receptors include modulation of satiety, opiate analgesia, and the development of morphine tolerance.     

Characteristics of periodontal mechanoreceptors reinnervated after freezing the inferior alveolar nerve

Previous electrophysiological investigations have shown that reinnervated periodontal mechanoreceptors, 12 weeks after crushing or sectioning the inferior alveolar nerve, have altered discharge properties. These alterations may have resulted from the regeneration of axons along inappropriate endoneurial sheaths distal to the injury site and this possibility was investigated by comparing the properties of reinnervated mechanoreceptors after nerve crush with those after nerve freeze (after which endoneurial sheath guidance is not lost). In 4 cats, 67 periodontal mechanosensitive units supplying the lower canine were examined 12 weeks after freezing the inferior alveolar nerve. When compared with controls, they were found to have significantly smaller arcs of sensitivity, raised force thresholds, lower discharge frequencies and reduced dynamic indices. These characteristics were, however, similar to those of units examined after nerve crush. These results suggest that the alterations in receptor properties after nerve injury cannot be attributed to a mismatch between the regenerating axons and their receptors.     

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.     

Neurophysiological assessment of trigeminal nerve reflexes in disorders of central and peripheral nervous system.

The trigeminal nerve and nuclei (the trigeminal complex) are unique in the human body with regard to their anatomical and physiological characteristics. They are also special regarding the lesions in which they are involved, both at the peripheral level because of the susceptibility of some terminal branches, and at the nuclei because of their large size and the large amount of connections with other centers. Conventional magnetic resonance imaging studies are often not sufficiently informative to demonstrate very tiny lesions that could be responsible for an important damage in the brainstem. Therefore, clinical neurophysiology and specifically, the techniques used in the study of the trigeminal functions, remain as convenient diagnostic and research tools to document clinically evident lesions or uncover subclinical abnormalities. This review is focussed on the clinical applicability of the study of trigeminal reflexes, including methods employed in the documentation of focal lesions of peripheral branches, trigeminal involvement of peripheral neuropathies, specific lesions of the trigeminal ganglia, central nervous dysfunctions causing abnormalities in the excitability of trigeminal neurons, and the possible use of trigeminal nerve reflexes in the study of facial pain syndromes and headache.     

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.     

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.     

Regulation of neuronal and glial galectin-1 expression by peripheral and central axotomy of rat primary afferent neurons

Galectin-1 (Gal1) is an endogenously-expressed protein important for the embryonic development of the full complement of primary sensory neurons and their synaptic connections in the spinal cord. Gal1 also promotes axonal regeneration following peripheral nerve injury, but the regulation of Gal1 by axotomy in primary afferent neurons has not yet been examined. Here, we show by immunohistochemistry and in situ hybridization that Gal1 expression is differentially regulated by peripheral nerve injury and by dorsal rhizotomy. Following peripheral nerve injury, the proportion of Gal1-positive DRG neurons was increased. An increase in the proportion of large-diameter DRG neurons immunopositive for Gal1 was paralleled by an increase in the depth of immunoreactivity in the dorsal horn, where Gal1-positive terminals are normally restricted to laminae I and II. Dorsal rhizotomy did not affect the proportions of neurons containing Gal1 mRNA or protein, but did deplete the ipsilateral dorsal horn of Gal1 immunoreactivity, indicating that it is transported centrally by dorsal root axons. Dorsal rhizotomy also resulted in an increase in Gal1 mRNA the nerve peripheral to the PNS-CNS interface (likely within Schwann cells and/or macrophages), and to a lesser extent within deafferented spinal cord regions undergoing Wallerian degeneration. This latter increase was notable in the dorsal columns and along the prior trajectories of myelinated afferents into the deeper dorsal horn. These results show that neuronal and glial expressions of Gal1 are tightly correlated with regenerative success. Thus, the differential expression pattern of Gal1 following peripheral axotomy and dorsal rhizotomy suggests that endogenous Gal1 may be a factor important to the regenerative response of injured axons.