The retrograde tracer Fluoro-Gold interferes with the infectivity of herpes simplex virus

Fluoro-Gold has been used previously to identify those trigeminal ganglion cells that innervate the central cornea. To examine the effects of Fluoro-Gold treatment on infection and spread of HSV in vivo, we measured the number of plaque forming units recovered from trigeminal ganglia 3 or 5 days after corneal scratch and inoculation with Fluoro-Gold and HSV. Treatment with Fluoro-Gold reduced the amount of virus recovered after retrograde transport 63% at 3 days and 28% at 5 days after inoculation. When we examined trigeminal ganglion sections from animals treated with HSV and Fluoro-Gold, we found the number of neurons double labeled with antibodies that recognize HSV and Fluoro-Gold was only 13% of all Fluoro-Gold labeled neurons. This was significantly fewer cells that we had anticipated, on the basis of double labeling experiments with wheat germ agglutinin combined with Fluoro-Gold. The effects of varying doses of the retrograde tracer, Fluoro-Gold on Herpes simplex virus (type 1) (HSV) infectivity were also assayed in vitro using a standard viral plaque assay. At 1 × 10⁻³ mg/ml Fluoro-Gold there was no effect on the number of plaque forming units. At 5 × 10⁻¹ mg/ml the number of plaques was reduced about 67%. We conclude that Fluoro-Gold interferes with productive HSV infection in vivo and in vitro after retrograde transport of HSV by neurons.     

Calcitonin gene-related peptide in nucleus ambiguus motoneurons in rat: Viscerotopic organization

Calcitonin gene-related peptide has been reported in the rat nucleus ambiguus. This nucleus comprises a dorsal division that is the source of special visceral efferents innervating the striated muscle of the upper alimentary tract and a ventral division supplying general visceral efferents primarily to the heart. The distribution of caldtonin gene-related peptide immunoreactive neurons in the two divisions was determined by using a combination of immunocytochemical techniques and fluorescent retrograde tracing. In 22 rats, injections of Fluoro-Gold were made into either the supranodosal vagus nerve, palatopharynx, larynx, esophagus, or heart. Following colchicine injection, medullary sections were processed immunocytochemically for calcitonin gene-related peptide. Injection of Fluoro-Gold into the supranodosal vagus resulted in prominent labeling of neurons in the dorsal and ventral divisions of the nucleus ambiguus. The majority of fluorescent labeled neurons in the dorsal division were found to be immunoreactive for calcitonin gene-related peptide, while those labeled neurons in the ventral division were unreactive for the peptide. With esophageal, and palatopharyngeal and cricothyroid injections, many fluorescent labeled neurons that were immunoreactive for calcitonin gene-related peptide were found respectively in the compact and semicompact formations of the dorsal division. In contrast, injections of the heart resulted in fluorescent labeled neurons, which were unreactive for calcitonin gene-related peptide, localized to the external formation. The results demonstrate that calcitonin gene-related peptide immunoreactive neurons are localized entirely to the dorsal division of the nucleus ambiguus and that all striated muscular areas of the alimentary tract are innervated by calcitonin gene-related peptide containing motoneurons. The localization of calcitonin gene-related peptide to vagal motoneurons also known to contain acetylcholine and the increase in acetylcholine receptor synthesis caused by this peptide suggest that calcitonin gene-related peptide acts as a cotransmitter with acetylcholine in special visceral efferent vagal motoneurons.     

A procedure to deliver herpes simplex virus to the murine trigeminal ganglion

Although initial herpes simplex virus (HSV) infections of the cornea are relatively easily treated, recurrent infections following reactivation of latent virus in the sensory ganglion cells are more difficult to treat. Untreated infections may result in severe consequences, including corneal scarring, glaucoma, and encephalitis. To develop such treatments, an experimental in vivo model was needed in which HSV can be applied directly to trigeminal ganglion cells. We have previously developed such a model to examine the mechanisms of HSV spread from trigeminal neurons to corneal epithelial cells. The current paper describes in detail the technical steps required for implementation of that model. Immunocytochemistry and electron microscopy have been used to validate the efficacy of the described procedures. This technique will be useful for future in vivo studies of neurotrophic viral infections of trigeminal ganglion cells.     

Growth of trigeminal neurites and interactions with corneal cells in embryonic chick organ cultures

The investigation of interactions between growing neurites and target cells during the development of the sensible corneal innervation is of crucial importance for understanding certain corneal diseases which are related to abnormal patterns of innervation. The purpose of the present work was to establish a culture system of cornea and trigeminal neurons and to examine interactions between these tissues. The responses of neurons derived from explanted embryonic chick trigeminal ganglia to co-explanted slices prepared from embryonic cornea were monitored over several days in culture. The growth of trigeminal fibers, but not of neurites derived from control tissues such as trigeminal mesencephalic nucleus or ciliary ganglion, was preferentially directed towards the co-cultured corneal slices. The ingrowth of trigeminal axons into the cornea was followed by formation of elaborate axonal terminal branches. Individual dissociated trigeminal neurons of pseudo-unipolar or bipolar classes developed their typical morphologies in culture. In co-cultures with corneal slices, they reacted to the corneal co-explant by frequently retracting some branches and forming or elongating other ones, which were predominantly directed towards the target tissue. In addition, the presence of a co-explanted trigeminal ganglion increased the rate of growth in the dissociated trigeminal neurons. The effect was not additive when cornea was present. Antibodies against nerve growth factor (NGF) and the low-affinity p75-NGF receptor (LANGFR) revealed that trigeminal ganglion cells support neuritic growth by secreting NGF, whereas corneal cells secrete additional factor(s) which act via the LANGFR.     

HSV (Type 1) infection of the trigeminal complex

Following corneal inoculation with herpes simplex virus (Type 1) (HSV), virus spreads to the CNS by axonal transport in the central branches of trigeminal ganglion cell neurons. Although this mode of viral entry to the CNS is rare for humans, it appears to be the principal route of entry into the CNS in animal models of herpetic corneal disease. In this study, the corneas of BALB/c mice were unilaterally inoculated with HSV, and the distribution of HSV-immunoreactive label was studied to identify the central branches of the axons of infected trigeminal ganglion cells. Virus was first noted in the brainstem trigeminal complex 4 days after corneal inoculation, when HSV-labeled afferents were found throughout the course of the descending tract of V as well as in interstitial neurons in the tract. By 5 days labeled neurons were also found not only in the n. caudalis and portions of the n. interpolaris of the trigeminal complex but also in laminae I–IV of the dorsal horn of the upper cervical levels of the spinal cord. No immunoreactivity was seen in other regions of the complex, including the n. oralis or the main sensory n. of V. By 6 days, however, the infection had spread to the main sensory division of V.     

Peripheral and central distribution of TRPV1, substance P and CGRP of rat corneal neurons

The rat corneal neurons expressing vanilloid receptor TRPV1, substance P (SP) and calcitonin-gene-related peptide (CGRP) were examined. In the cornea, some TRPV1-immunoreactive nerve fibers displayed either SP- or CGRP immunoreactivity also. For observing corneal neuronal elements in the trigeminal ganglion (TG) and in the medulla oblongata, retrograde and anterograde cholera toxin subunit B (CTB) tracing methods combining with triple immunofluorescence technique were performed. The corneal neuronal somata were located in the ophthalmic division of the TG; 37% of them were immunoreactive for TRPV1. One third and three quarters of the corneal TRPV1-immunoreactive neurons co-expressed SP and CGRP, respectively. All of SP-immunoreactive corneal neurons exhibited TRPV1 immunoreactivity. They were predominantly medium-sized (mean +/- SE = 638.2 +/- 49.5 microm(2)) and significantly larger than SP-immunoreactive and TRPV1-immunonegative neurons in the ophthalmic division of the TG. The central projection fibers of corneal neurons co-expressing TRPV1 with SP and CGRP were observed at the subnucleus interpolaris/caudalis transition within trigeminal nucleus. The present study suggests that TRPV1 of the corneal neurons works in close relation to SP and CGRP both in the cornea and CNS for healing and nociceptive transduction.     

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.     

Distribution of a splice variant of choline acetyltransferase in the trigeminal ganglion and brainstem of the rat: comparison with calcitonin gene-related peptide and substance P

Rat trigeminal ganglion neurons have been shown to contain a splice variant of choline acetyltransferase (pChAT). Here we report the distribution pattern of pChAT-containing afferents from the trigeminal ganglion to the brainstem, compared with that of calcitonin gene-related peptide (CGRP) and substance P (SP), by use of the immunohistochemical techniques in the rat. Most of CGRP(+) SP(+) ganglion cells contain pChAT, whereas half of the pChAT(+) ganglion cells possess neither CGRP nor SP. In the brainstem, pChAT(+) nerve fibers are found exclusively in the trigeminal and solitary systems, although the distribution pattern differs from that of CGRP(+) or SP(+) fibers. First, the ventral portion of the principal sensory nucleus contains many pChAT(+) fibers, with few CGRP(+) or SP(+) fibers. Because this portion receives projections of nociceptive corneal afferents, a subpopulation of pChAT(+) CGRP(-) SP(-) primary afferents is most probably nonpeptidergic nociceptors innervating the cornea. Second, the superficial laminae of the medullary dorsal horn, the main target of nociceptive afferents, contain dense CGRP(+) and SP(+) fibers but sparse pChAT(+) fibers. Because pChAT occurs in most CGRP(+) SP(+) ganglion cells, such sparseness of pChAT(+) fibers implies poor transportation of pChAT to axon branchlets. Another important finding is that pChAT(+) axons are smooth and nonvaricose, whereas CGRP(+) or SP(+) fibers possess numerous varicosities. Our confocal microscopy suggests colocalization of these three markers in the same single axons in some brainstem regions. The difference in morphological appearance, nonvaricose or varicose, appears to reflect the difference in intraaxonal distribution between pChAT and CGRP or SP.     

Coexistence of calcitonin gene-related peptide and substance P-like peptide in single cells of the trigeminal ganglion of the rat: immunohistochemical analysis

The localization of calcitonin gene-related peptide (CGRP) and substance P (SP) in the rat trigeminal ganglion was examined by means of the indirect immunofluorescent method. About 40% of neurons in the ganglion contained CGRP-like immunoreactivity (CGRPI), while about 20% of neurons showed SP-like immunoreactivity (SPI). In serial sections, nearly all the SPI neurons contained CGRPI.     

A quantitative correlation of substance P-, calcitonin gene-related peptide- and cholecystokinin-like immunoreactivity with retrogradely labeled trigeminal ganglion cells innervating the eye

To evaluate the intraganglionic organization of ocular sensory neurons in the guinea pig, we studied the retrograde axonal transport from the eye to the trigeminal ganglion of cholera toxin B subunit and then applied immunohistochemistry for substance P, calcitonin gene-related peptide and cholecystokinin. Retrogradely labeled cells were observed only in the anteromedial portion of the ipsilateral ganglion. We observed no somatotopical organization to trigeminal neurons containing any of these three peptides, either for cells projecting to the eye or for the ganglion as a whole. The relative proportion of neurons immunoreactive for each of these three peptides was similar among the population of neurons retrogradely labeled with cholera toxin B and among the population of neurons without direct projections to the eye.     

Parvalbumin- and calretinin-immunoreactive trigeminal neurons innervating the rat molar tooth pulp

Calcium-binding proteins and neuropeptides were examined in trigeminal neuronal cell bodies retrogradely labeled with Fast blue (FB) from the maxillary molar tooth pulp of the rat. FB-labeled cells were located in the maxillary division of the trigeminal ganglion. 30 and 50% of the labeled cells were immunoreactive for parvalbumin and calcitonin gene-related peptide (CGRP), respectively. The coexpression of these substances was observed in 9.5% of FB-labeled cells. On the other hand, 2.4% of FB-labeled cells exhibited calretinin-immunoreactivity (CR-ir) and 20% tachykinin (TK)-ir. The coexpression of CR and TK was observed in 1.9% of FB-labeled cells, i.e., most of CR-ir FB-labeled neurons coexpressed TK-ir. An immuno-EM method revealed that all parvalbumin-ir nerve fibers in the root pulp were myelinated and that CGRP-ir nerve fibers were both myelinated (15%) and unmyelinated (85%). The present study indicated that primary nociceptors innervating the rat molar both pulp contained parvalbumin and CR and coexpressed these calcium-binding proteins and neuropeptides. It was suggested that peripheral axons of parvalbumin-ir tooth pulp primary neurons are all myelinated. Most peripheral CR-ir axons are probably unmyelinated because TK-ir myelinated axons have never been demonstrated in any peripheral organ.     

Central projections and trigeminal ganglion location of corneal afferent neurons in the monkey, Macaca fascicularis

The method of transganglionic transport of horseradish peroxidase-wheat germ agglutinin conjugate (HRP-WGA) was used to determine the location within the monkey trigeminal ganglion of the primary afferent neurons that innervate the cornea, and the brainstem and spinal cord termination sites of these cells. In each of four animals. Gelfoam pledgets were saturated with 2% HRP-WGA in saline and applied to the scratched surface of the central cornea for 30 minutes. Postmortem examination of the corneal whole mounts revealed that the tracer solution remained confined to approximately the central one-fourth of the cornea with no spread into the peripheral cornea or limbus. Seventy-two to 96 hours after tracer application, 126-242 labeled cell bodies were observed in the medial region of the ipsilateral trigeminal ganglion. The majority of neurons were concentrated in an area of the ganglion that lay directly caudal to the entering fibers of the ophthalmic nerve, but smaller numbers of cells lay somewhat more laterally, near the region where the ophthalmic and maxillary nerves come together. A very small number of neurons in one animal innervated the cornea by sending their fibers into the maxillary nerve. HRP-WGA-labeled terminal fields were present to some extent in all four major rostrocaudal subdivisions of the ipsilateral trigeminal brainstem nuclear complex (TBNC), but the size of the terminal fields and the intensity of labeling differed markedly from one level of the TBNC to the next. Labeled fibers projected heavily to the transitional zone between caudal pars interpolaris and rostral pars caudalis (i.e., the "periobex" region of the TBNC) and moderately to the trigeminal main sensory nucleus, pars oralis, and caudal pars caudalis at the level of the pyramidal decussation. Remaining areas of the TBNC, including rostral pars interpolaris and the midlevel of pars caudalis, received few, if any, corneal afferent projections. Occasional labeled fibers were observed in the dorsal horn of C1 and in the rostral half of C2. It is hoped that data generated in the current investigation of nonhuman primates will contribute to a better understanding of the neural substrates that subserve corneal sensation and the blink reflex in humans.     

Trigeminal collaterals in the nasal epithelium and olfactory bulb: a potential route for direct modulation of olfactory information by trigeminal stimuli

The nasal epithelium is richly invested with peptidergic (substance P and calcitonin gene-related peptide [CGRP]) trigeminal polymodal nociceptors, which respond to numerous odorants as well as irritants. Peptidergic trigeminal sensory fibers also enter the glomerular layer of the olfactory bulb. To test whether the trigeminal fibers in the olfactory bulb are collaterals of the epithelial trigeminal fibers, we utilized dual retrograde labeling techniques in rats to identify the trigeminal ganglion cells innervating each of these territories. Nuclear Yellow was injected into the dorsal nasal epithelium, and True Blue was injected into the olfactory bulb of the same side. Following a survival period of 3-7 days, the trigeminal ganglion contained double-labeled, small (11.8 x 8.0 microm), ellipsoid ganglion cells within the ethmoid nerve region of the ganglion. Tracer injections into the spinal trigeminal complex established that these branched trigeminal ganglion cells also extended an axon into the brainstem. These results indicate that some trigeminal ganglion cells with sensory endings in the nasal epithelium also have branches reaching directly into both the olfactory bulb and the spinal trigeminal complex. These trigeminal ganglion cells are unique among primary sensory neurons in having two branches entering the central nervous system at widely distant points. Furthermore, the collateral innervation of the epithelium and bulb may provide an avenue whereby nasal irritants could affect processing of coincident olfactory stimuli.     

Gene expression of histamine H1 receptor in guinea pig primary sensory neurons: a relationship between H1 receptor mRNA-expressing neurons and peptidergic neurons

Pharmacological studies have suggested that a subgroup of primary sensory neurons is responsive to histamine via the histamine H1 receptor. We addressed this issue using in situ hybridization histochemistry with a cRNA probe for the guinea pig H1 receptor gene. About 15% of the trigeminal and lumber dorsal root ganglion (DRG) neurons, but none of nodose ganglion neurons, were intensely labeled with this probe. The H1 receptor mRNA-positive neurons were exclusively small in size, and were demonstrated to give rise to unmyelinated fibers by ultrastructural analysis of isolectin B4-labeling. However, the H1 receptor mRNA-expressing DRG neurons were not immunoreactive to substance P (SP) and calcitonin gene-related peptide (CGRP). A marked increase in the number of mRNA-positive DRG neurons were observed 1-5 days after a crush injury of the sciatic nerve (3-4-fold of the control value). These neurons turned mRNA-positive after the nerve crush were also mainly small-sized. The mRNA signals were detected in many peptidergic (SP/CGRP) neurons, in contrast to the normal state. On the other hand, in the neurons which showed intense labeling in the normal condition, the mRNA signals were down-regulated. These results suggest that primary sensory neurons include two kinds of H1 receptor-expressing sensory neurons, one expressing H1 receptor mRNAs in the normal state and the other up-regulating the mRNAs following the peripheral nerve damage.     

GDNF family ligand receptor components Ret and GFRalpha-1 in the human trigeminal ganglion and sensory nuclei

The occurrence of Ret and GFRalpha-1 receptors is shown by immunohistochemistry in the human trigeminal sensory system at pre-, postnatal and adult age. Receptor-labeled neurons occur in both trigeminal ganglion and mesencephalic nucleus. In adult trigeminal ganglion, about 75% of Ret- and 65% of GFRalpha-1-labeled neurons are small- and medium-sized. The proportion of Ret+ and GFRalpha-1+ trigeminal ganglion neurons in the adult is about 25 and 60%, respectively. The majority of Ret+ are double labeled for GFRalpha-1 and glial cell line-derived neurotrophic factor (GDNF). Most of the GFRalpha-1+ cells contain GDNF and about 50% of them contain Ret. Triple labeling shows many Ret+/GDNF+/GFRalpha-1+ neurons, but also a number of Ret-/GDNF+/GFRalpha-1+ and Ret+/GDNF-/GFRalpha-1+ cells. Both Ret+ and GFRalpha-1+ neuronal subpopulations overlap with that containing calcitonin gene-related peptide. Ret+ pericellular basket-like nerve fibers occur in the adult trigeminal ganglion. Centrally, immunoreactivity is restricted to the spinal nucleus pars caudalis and pars interpolaris and to the mesencephalic nucleus. In adult specimens, Ret+ nerve fibers and puncta gather in the inner substantia gelatinosa. Ret+ neurons occur in the spinal nucleus and are more frequent in newborn than in adult subjects. Central GFRalpha-1+-labeled neurons and punctate elements are sparse. These findings support the involvement of GDNF and possibly other cognate ligands in the trophism of human trigeminal primary sensory neurons from prenatal life to adulthood, indicating a selective commitment to cells devoted to protopathic and proprioceptive sensory transmission. They also support the possibility that receptor molecules other than Ret could be active in transducing the ligand signal.     

Cerebral arterial innervation by nerve fibers containing calcitonin gene-related peptide (CGRP): I. Distribution and origin of CGRP perivascular innervation in the rat

The origin, density and distribution of calcitonin gene-related peptide (CGRP) immunoreactivity in cerebral perivascular nerves and the trigeminal ganglion of rats were examined in this study. CGRP immunoreactive axons were abundant on the walls of the rostral circulation of the major cerebral arteries in the circle of Willis. The fibers form a grid- or meshwork of longitudinal and circumferential axons studded with numerous varicose swellings. The density of CGRP fibers was particularly high at the bifurcation of major arteries. A few CGRP fibers cross the midline to innervate arteries on the contralateral side of the arterial tree. The arteries of the caudal circulation were sparsely innervated by CGRP fibers. In the trigeminal ganglion, about 30% of the ganglion cells had CGRP immunoreactivity. The cell size of most (75%) of CGRP neurons was less than 30 micron in diameter. There was no significant difference in staining density between small and large CGRP neurons. Unilateral transection of the maxillary and mandibular divisions of the trigeminal nerve caused a substantial decrease of CGRP immunoreactivity in the ipsilateral dorsal two-thirds of the trigeminal nucleus and cervical spinal cord but did not noticeably change the diameter of the vascular lumen or the densities of CGRP fibers in the walls of the cerebral arteries. In contrast, unilateral transection that included the ophthalmic division eliminated CGRP fibers on the ipsilateral cerebral arteries and eliminated CGRP immunoreactivity throughout the trigeminal nucleus in the brainstem and rostral cervical cord. In addition, these lesions caused a significant reduction in the diameter of the denervated arteries. The present study demonstrates that CGRP, a putative neurotransmitter/neuromodulator, is especially abundant in the rostral cerebral circulation and is derived from the ipsilateral ophthalmic division of the trigeminal nerve. In addition, the loss of CGRP perivascular nerves is associated with a reduction of the arterial lumen. This suggests that CGRP is a strong candidate as a nerve-derived trophic factor at trigeminal terminals and provides additional evidence that CGRP is a component in the trigeminovascular system influencing vascular diameter.     

Implications of Transient Receptor Potential Cation Channels in Migraine Pathophysiology

Migraine is a common and debilitating headache disorder. Although its pathogenesis remains elusive,abnormal trigeminal and central nervous system activity is likely to play an important role. Transient receptor potential(TRP) channels, which transduce noxious stimuli into pain signals, are expressed in trigeminal ganglion neurons and brain regions closely associated with the pathophysiology of migraine. In the trigeminal ganglion,TRP channels co-localize with calcitonin gene-related peptide, a neuropeptide crucially implicated in migraine pathophysiology. Many preclinical and clinical data support the roles of TRP channels in migraine. In particular,activation of TRP cation channel V1 has been shown to regulate calcitonin gene-related peptide release from trigeminal nerves. Intriguingly, several effective antimigraine therapies, including botulinum neurotoxin type A, affect the functions of TRP cation channels. Here, we discuss currently available data regarding the roles of major TRP cation channels in the pathophysiology of migraine and the therapeutic applicability thereof.     

Sympathetic innervation of the rat cornea as demonstrated by the retrograde and anterograde transport of horseradish peroxidase-wheat germ agglutinin

The sympathetic innervation of the rat cornea was studied by using the method of intraaxonal transport of horseradish peroxidase-wheat germ agglutinin conjugate (HRP-WGA). In the first set of experiments, the relative number of superior cervical ganglion neurons that innervate the rat central cornea was estimated by the method of retrograde HRP-WGA transport. Following tracer application to the scarificed central corneal surface, 49-198 labeled neurons were observed in the ophthalmic region of the ipsilateral trigeminal ganglion and zero to four cells in the rostral pole of the ipsilateral superior cervical ganglion. In the second set of experiments, the three-dimensional distribution and termination sites of the corneal sympathetic nerve fibers were investigated by the technique of HRP-WGA anterograde transport from the superior cervical ganglion. HRP-WGA-labeled axons in corneal whole mounts were identified by the presence within their axoplasm of linear arrays of HRP-TMB reaction product, and their distribution was plotted faithfully onto line drawings made with a drawing tube attachment. Large numbers of HRP-labeled fibers were found in all animals within the corneoscleral limbus where the majority were associated with blood vessels. Fewer fibers (zero to 14 per animal) entered the cornea proper. The latter fibers entered the peripheral cornea in the deep to middle layers of the stroma and ascended into progressively more superficial layers as they coursed centrally. The majority of fibers branched infrequently in the peripheral cornea and increased in branching complexity near the central cornea. HRP-labeled axonal varicosities suggestive of terminal and preterminal expansions were located preferentially in the subepithelial layer of the corneal stroma and in the basal epithelium. Approximately 75% of the axonal varicosities were located in the central half of the cornea. The results of the current investigation reveal that the rat cornea is innervated sparsely by sympathetic nerve fibers derived from the superior cervical ganglion. These data provide additional support to current theories that corneal sympathetic nerve fibers may influence select aspects of corneal physiology, including ion transport and hydration, mitogenesis and wound healing, and sensitivity.     

c-Jun expression after axotomy of corneal trigeminal ganglion neurons is dependent on the site of injury

The proto-oncogene c-Jun has been implicated in the control of neuronal responses to injury and in axonal growth during regenerative processes. We have investigated the expression of c-Jun during normal terminal remodelling in trigeminal ganglion neurons innervating the cornea and after acute injury of epithelial nerve terminals or parent axons. Remodelling and rearrangement, or damage limited to corneal epithelium endings, was not a trigger for activation of c-Jun expression. However, injury of parent axons in the stroma or in the orbital ciliary nerves induced c-Jun expression in 50% of the population of corneal neurons, which included all of the large myelinated and 20% of the small neuropeptide-containing corneal neurons. This suggests that c-Jun expression in trigeminal ganglion neurons is not associated with normal remodelling or regeneration of peripheral nerve terminals, and that it takes place only when parent axons are injured. A substantial number of damaged neurons do not express c-Jun, indicating that in primary sensory neurons, injury and regeneration may not always be coupled to the expression of this proto-oncogene.