Trigeminal Innervation Research Articles

Desmoplastic melanoma: patterns of recurrence.

Desmoplastic melanoma is a rare type of malignant melanoma, recognized since 1971. Other variants of desmoplastic melanoma include neural transforming melanoma and neurotropic melanoma. The pathology and clinical features of 58 patients whose tumor had the features of desmoplastic melanoma, neural transforming melanoma, and neurotropic melanoma, either separately or in combination, were examined to assess patterns of recurrent disease. The tumor was situated on the head and neck in 41% of patients and was amelanotic in 71% of patients. There was an associated superficial melanoma in 48% of patients. There was a combination of the 3 histologic patterns, commonly found in the 1 melanoma. Local recurrence occurred in 29% of patients and malignant cranial neuropathies were documented in 4 patients. Nineteen percent of patients have died from disseminated disease. Neurotropic melanomas had a lower incidence of visceral recurrence. Desmoplastic and neural transforming melanomas had similar rates of local and visceral recurrence. When this specific variant of melanoma is compared with larger series of malignant melanoma in general, they appear to be more advanced locally, with a higher incidence of local recurrence. When considered in relation to the thicker nondesmoplastic melanomas, the survival is no worse and may be more favorable. Surgeons should excise the primary tumor and local recurrences with wide margins and adopt close follow-up. On the head and neck, symptoms and signs relating to trigeminal or facial nerve innervation may herald a developing malignant cranial neuropathy.

Evidence for prenatal competition among the central arbors of trigeminal primary afferent neurons

Previous studies have shown that damage to vibrissa follicles in newborn rats and mice does not alter the brainstem representations of the remaining vibrissa as demonstrated by staining for mitochondrial enzymes such as cytochrome oxidase (CO) succinic dehydrogenase. This study asked whether this lack of effect might be due to the fact that the trigeminal primary afferents in rodents are already quite well developed at birth. We assessed this possibility by using CO staining the evaluate patterns in the brainstems of pre- and postnatal rats. A vibrissa-related pattern began to emerge in trigeminal nucleus principalis and subnucleus interpolaris (Spl) by embryonic day (E-) 19 and appeared fully developed by the day of birth (P-0). We also made partial lesions of the vibrissa pad on E-15-20 and on P-0, killed pups on P-5-7, and measured the size of the CO-stained patches in Spl on both sides of the brainstem. The correspondence between CO patches and clusters of primary afferent terminal arbors was verified in some animals by combining transganglionic horseradish peroxidase tracing and CO staining. Vibrissa pad damage on E-15-18 resulted in significant (20.1-36.9%) increases in the average area of the remaining CO patches in Spl ipsilateral to the lesion. Vibrissa pad damage on E-19, E-20, and P-0 produced small (6.2-8.9%), but insignificant, increases in patch size in Spl ipsilateral to the lesion. We used anatomical and electrophysiological methods to determine whether our lesions altered the trigeminal innervation of surviving vibrissa follicles. We recorded single trigeminal ganglion cells from 12 rats that sustained vibrissa pad lesion on E-17. As in normal rats, all of the 49 vibrissa-sensitive ganglion cells isolated in the lesioned animals were responsive to deflection of one and only one vibrissa. We also dissected 11 deep vibrissal nerves from intact follicles in adult rats that sustained fetal vibrissa pad damage on E-17, and counted numbers of myelinated axons in 1 microns plastic sections. These data were compared with counts from corresponding follicles on the intact side of the face. The average number of myelinated axons innervating follicles in the damaged vibrissa pads was 196.8 +/- 27.9, and that for the corresponding contralateral nerves was 194.6 +/- 25.7. These data suggest that competitive interactions among the central arbors of trigeminal primary afferents in fetal life may influence the development of central vibrissa representations and, further, that lesion-induced central changes need not be correlated with alterations in the peripheral innervation of undamaged follicles.

Ophthalmologic Aspects of Headache

Pain around the eye can be caused by local ophthalmic disorders or by disease of other structures sharing trigeminal nerve sensory innervation. In general, most ocular causes for pain also cause the eye to be red, thus alerting the examiner to the focality of the problem. However, conditions like eyestrain, intermittent angleclosure glaucoma or neovascular glaucoma, and low-grade intraocular inflammation can be painful and not be associated with obvious redness. Ocular signs and symptoms also occur with numerous other causes of headache. Double vision in association with periocular pain can result from orbital lesions, isolated cranial neuropathies, and cavernous sinus lesions. Pupillary abnormalities like Horner's syndrome may result from a variety of painful conditions, including cluster headache, parasellar neoplasms or aneurysms, internal carotid dissection or occlusion, and Tolosa-Hunt syndrome. Pain with a dilated and unreactive pupil may reflect a benign condition like Adie's syndrome or ophthalmoplegic migraine, or it may herald the presence of a life-threatening posterior communicating artery aneurysm. Headache and transient visual loss can be manifestations of classic migraine, or be symptoms of ocular hypoperfusion from ipsilateral internal carotid occlusion or increased intracranial pressure from pseudotumor cerebri. In a young patient, head pain with a fixed visual deficit may result from optic neuritis, in an older adult, temporal arteritis may be the culprit. Ophthalmologic aspects of headache thus encompass problems that range from simple and benign to complex and formidable.

Chronic Trigeminal Ganglionectomy or Topical Capsaicin Application to Pial Vessels Attenuates Postocclusive Cortical Hyperemia but Does Not Influence Postischemic Hypoperfusion

Marked hyperemia accompanies reperfusion after ischemia in the brain, and may account for the propensity of cerebral hemorrhage to follow embolic stroke or carotid endarterectomy, and for the morbidity that follows head injury or the ligation of large arteriovenous malformations. To evaluate the contribution of trigeminal sensory fibers to the hyperemic response, CBF was determined in 12 symmetrical brain regions, using microspheres with up to five different isotopic labels, in four groups of cats. Measurements were made at 15-min intervals for up to 2 h of reperfusion after global cerebral ischemia induced by four-vessel occlusion combined with systemic hypotension of either 10- or 20-min duration. In normal animals, hyperemia in cortical gray matter 30 min after reperfusion was significantly greater after 20 min (n = 10) than after 10 min (n = 7) of ischemia (312 ml/100 g/min versus 245 ml/100 g/min; p less than 0.01). CBF returned to preischemic levels approximately 45 min after reperfusion and was reduced to approximately 65% of basal CBF for the remaining 75 min. In cats subjected to chronic trigeminal ganglionectomy (n = 15), postocclusive hyperemia in cortical gray matter was attenuated by up to 48% on the denervated side (249 versus 150 ml/100 g/min; p less than 0.01) after 10 min of ischemia. This effect was maximal in the middle cerebral artery (MCA) territory, and was confined to regions known to receive a trigeminal innervation. In these animals, substance P (SP) levels in the MCA were reduced by 64% (p less than 0.01), and the density of nerve fibers containing calcitonin gene-related peptide (but not vasoactive intestinal polypeptide or neuropeptide Y) was decreased markedly on the lesioned side. Topical application of capsaicin (100 nM; 50 microliters) to the middle or posterior temporal branch of the MCA 10-14 days before ischemia decreased SP levels by 36%. Postocclusive hyperemia in cortical gray matter was attenuated throughout the ipsilateral hemisphere by up to 58%, but the cerebral vascular response to hypercapnia (PaCO2 = 60 mm Hg) was unimpaired. The duration of hyperemia and the severity of the delayed hypoperfusion were not influenced by trigeminalectomy, capsaicin application, or the intravenous administration of ATP. These data demonstrate the importance of neurogenic mechanisms in the development of postischemic hyperperfusion, and suggest the potential utility of strategies aimed at blocking axon reflex-like mechanisms to reduce severe cortical hyperemia.

Carnosine-, calcitonin gene-related peptide- and tyrosine hydroxylase-immunoreactivity in the mouse olfactory bulb following peripheral denervation

We report the effects of olfactory peripheral deafferentation by intranasal irrigation with ZnSO 4 on carnosine and CGRP immunoreactivities in the mouse olfactory system. In the normal rodent olfactory epithelium carnosine immunoreactivity is associated with the olfactory receptor neurons. Conversely, CGRP immunoreactivity appears to be associated with the trigeminal innervation of the nasal cavity. Following lesion the magnitude of carnosine immunoreactivity in the olfactory epithelium is strongly reduced while CGRP immunoreactivity is unaffected. In the olfactory bulb, deafferentation causes a strong reduction of carnosine immunoreactivity in the glomerular layer and, concurrently, of TH immunoreactivity in the juxtaglomerular neurons. CGRP immunoreactive fibers in the olfactory bulb are abundant in the glomerular layer both before and after deafferentation. These data demonstrate that, in the adult mouse, the immunocytochemically detectable levels of CGRP are not altered following lesion and indicate CGRP is not released directly from the olfactory neurons to induce TH production.

The trigeminal sensory innervation to the middle ear, eustachian tube, and pharynx: A study by the horseradish peroxidase tracer method

The trigeminal sensory innervations to the middle ear, the eustachian tube, and the pharynx were examined using the horseradish peroxidase (HRP) method in cats. After HRP application to the middle ear and the eustachian tube, HRP-labeled cells were found mainly in the posterolateral portion and the anteromedial portion of the trigeminal ganglion, respectively. In the case of injection into the epipharynx and the mesopharynx, HRP-labeled cells were found in the anteromedial portion and the posterolateral portion, respectively. HRP-labeled cells were hardly observed when HRP was applied to the hypopharynx. These results indicate that trigeminal nerve sensory fibers branch to the middle ear, including the eustachian tube, epipharynx, and mesopharynx.

Detection of CCK mRNA in the motor nucleus of the rat trigeminal nerve with in situ hybridization histochemistry

The regional distribution of dorsal pontine tegmental neurons expressing cholecystokinin (CCK) messenger RNA (mRNA) was examined using in situ hybridization histochemical techniques. A distribution of neurons containing CCK mRNA was observed in the motor nucleus of the trigeminal nerve, a region which has not been shown to contain CCK by immunocytochemical means. In addition, CCK mRNA was seen in the caudal portion of the dorsal raphe nucleus which correlates with prior immunocytochemical observations. The presence of CCK mRNA in the cholinergic motor trigeminal nucleus suggests that these cells express the CCK gene and have gone undetected in previous immunocytochemical studies. It is also suggested that CCK may have an influence on muscular contraction at sites of trigeminal nerve innervation (e.g. muscles of mastication).

THE NERVOUS CONTROL OF INTRALUMINAL PRESSURE IN THE VOMERONASAL ORGAN OF THE DOMESTIC RAM

In order to examine whether an aspiration mechanism exists in the vomeronasal organ (VNO) of the domestic ram, experiments were conducted in chloralose-anaesthetized animals using acute electromanometry and electrophysiological techniques. Stimulation of the ipsilateral cervical sympathetic nerve activated a mechanism producing a negative (with respect to atmospheric pressure) intraluminal pressure which drew fluid into the VNO from the nasopalatine canal. The mechanism was under alpha-adrenergic control and did not seem to involve the carotid blood supply. An ipsilateral increase in intraluminal pressure occurred following electrical excitation of the maxillary trigeminal innervation. Thus a mechanism was demonstrated in the ram for conveying odours to the putative olfactory receptors in the VNO.

Effect of fetal infraorbital nerve transection upon trigeminal primary afferent projections in the rat

Transganglionic tracing with a combination of horseradish peroxidase (HRP) and wheat germ agglutinin-conjugated HRP (WGA-HRP) was employed to compare the trigeminal (V) innervation of the brainstem in adult rats that sustained transection of the infraorbital nerve (ION) on either the day of birth or just prior to the beginning of the 17th embryonic day (E-17). The same methods were also employed to assess the effects of such lesions upon the innervation of the brainstem by the lingual, inferior alveolar, mylohyoid, and auriculotemporal V branches. Previous experiments (Chiaia et al.: Dev. Brain Res. 36:75-88, '87) showed that application of HRP and WGA-HRP to the ION in normal adult rats (N = 3) labelled 12,553 +/- 1,455 (mean +/- s.d.) V ganglion cells while application of these tracers to the regenerated ION after neonatal transection (N = 9) labelled 5,001 +/- 1,287 ganglion cells. Application of HRP and WGA-HRP to the regenerated ION in adulthood (N = 6) after fetal transection labelled 5,476 +/- 3,056 ganglion cells. Thus, the numbers of ganglion cells giving rise to the regenerated ION after fetal and neonatal transection were equivalent (P greater than .05). The central projections of the ION after fetal transection were qualitatively different from those observed after neonatal injury. After neonatal transection, the central terminal field of regenerated ION fibers in adulthood is almost completely restricted to layers I and II of subnucleus caudalis (SpC; Jacquin and Rhoades: Brain Res. 269:137-144, '83; Chiaia et al.: Dev. Brain Res. 36:75-88, '87). After fetal transection, regenerated ION axons terminate heavily in all portions of the V brainstem complex. After neonatal ION transection, we (Jacquin and Rhoades: J. Comp. Neurol. 235:129-143, '85) have been unable to detect central sprouting of undamaged V mandibular axons by means of transganglionic tracing with HRP and WGA-HRP. Such sprouting was evident in both V subnucleus interpolaris (SpI) and SpC after fetal ION transection. We carried out one additional experiment to determine whether ION ganglion cells that survived fetal axotomy were more resistant to axonal damage than the population of neurons that normally contribute to this nerve on the day of birth. Rats (N = 5) sustained transection of the ION on E-17 and again on the day of birth. The regenerated ION was then labelled with HRP and WGA-HRP when the animals reached adulthood.(ABSTRACT TRUNCATED AT 400 WORDS)

Enrichment of a vasoactive neuropeptide (calcitonin gene related peptide) in the trigeminal sensory projection to the intracranial arteries

Trigeminal sensory innervation of cerebral vessels and the surrounding dura is responsible for most intracranial head pain. Small-diameter fibers containing substance P (Sub P) have been observed in the periadventitia around feline cerebral blood vessels, and it has been suggested that these fibers are the trigeminal substrate for vascular pain associated with cluster and migraine headaches. Calcitonin gene related peptide (CGRP) coexists with Sub P in some of these fibers and with some Sub P containing neurons in the trigeminal ganglion. In addition, a population of trigeminal neurons containing CGRP but not Sub P has been observed. We now report that the population of trigeminal ganglion cells projecting to the cerebral vasculature is enriched in CGRP-containing neurons, and especially in the population of neurons containing CGRP and not Sub P. Using retrograde tracing of fluorescent tracers combined with immunocytochemistry after explant culture, we found approximately 32% of trigeminal ganglion cells projecting to the cerebral vasculature contained CGRP. Approximately 18 and 17% of these cells contained Sub P and cholecystokinin (CCK), respectively. The 32% of ganglion cells projecting to the cerebral vasculature that contain CGRP stands in contrast to the 12% CGRP positive seen in the population of ganglion cells projecting out to another target (the forehead), and the 21 and 23% CGRP positive observed in the mandibular branch and entire ganglion, respectively. Sub P and CCK are not enriched in the trigeminal innervation of the vasculature compared with their presence in cells throughout the ganglia.(ABSTRACT TRUNCATED AT 250 WORDS)

Pattern of intracranial and extracranial projections of trigeminal ganglion cells

The trigeminal sensory innervation of the major cerebral vessels is thought to carry the nociceptive information during a migraine headache, and this pain is usually referred to the forehead area. Using retrograde tracing techniques, we have described the distribution of sensory trigeminal cells that innervate the middle cerebral artery (MCA) and the forehead. Nearest-neighbor analysis of the ophthalmic division of the trigeminal ganglion revealed that cells innervating the forehead tend to be clumped around individual cells that innervate the MCA. An average of less than 1 cell per animal was found to project divergent collaterals to both areas. The close association of ganglion cell bodies innervating the cerebral vasculature and those innervating forehead areas may underlie the convergence of their central processes onto common brain-stem trigeminal nucleus cells, and thus the referral of headache pain. In contrast to the lack of ganglion cells with axonal collaterals to the cerebral vasculature and forehead, a significant population of cells that innervate the MCA also have collateral projections to other cerebral arterial branches (branches of the middle meningeal artery), as well as the surrounding dura. Thus, the innervation targets of individual trigeminal cells are very widespread intracranially (including arteries and dura), but separate cells in the ophthalmic division innervate extracranial targets.

Cell death organizes the postnatal development of the trigeminal innervation of the cerebral vasculature

In the adult trigeminal ganglion single cell bodies that innervate the middle cerebral artery (MCA) are different from but situated near to one or more cell bodies that innervate the forehead (O'Connor and Van der Kooy, submitted). Multiple fluorescent retrograde axonal tracing in postnatal day 3–90 rats was employed to describe the development of this adult pattern of trigeminal projections. We found that close to 90% of the cells that innervate the MCA at postnatal day 5 (PND 5) are eliminated by PND 90. Less than 20% of the ganglion cells innervating the forehead die over the same postnatal period. Subpopulations of cells in the ganglion were observed to have a maximal rate of death during different postnatal periods. First, 15–20% of the cells throughout the ophthalmic division die between PND 5 and PND 10. Second, a small population of cells that had early projections to the contralateral MCA die out completely by PND 22. Third, cells with a projection only to the MCA die primarily between PND10 and PND 54. Fourth, during the first postnatal week there are many cells that project to both the MCA and the forehead; however, 90% of this population dies by PND 90. This elimination is observed latest in the postnatal period, with these cells exhibiting their greatest rate of cell death between PND 22 and PND 90. Thus, cell death is the primary postnatal mechanism that produces this organization in the ophthalmic division of the trigeminal ganglion and retraction of axonal collaterals is a minor mechanism. We suggest that the latest period of death in cells with divergent artery and forehead projections as well as the ultimate persistence of some artery projecting cells beyond PND 90, may be due to the larger peripheral fields of innervation of these trigeminal ganglion cells.

An experimental study of timing and topography of early tooth development in the mouse embryo with an analysis of the role of innervation

The putative involvement of the innervation in determining the sites of dental development was investigated by intra-ocular homografting and organ culture methods. Mandibular arches were dissected from 9 and 10-day-old (E9–E10) mouse embryos and grafted to the anterior eye chambers of homologous adult mice for 12–14 days. There was no significant difference in the incidence of tooth formation between grafts in which the trigeminal ganglion was included ( n = 82, 68 per cent) or excluded ( n = 72, 65 per cent). A parallel in vitro study in which E9 and E10 mouse embryo mandibular arches were cultured in the absence of trigeminal innervation showed that definitive tooth germs were formed during the 7–9 day culture period. It is concluded that innervation plays no part in determination of tooth development. In further experiments with E9 and E10 material, the complete mandibular arch, the hemimandibular arch and the ventral midline region of the mandibular arch (including the median epithelial isthmus) were each grafted for a period of 21 days. Alizarin-red whole-mount staining of recovered grafts revealed that bone had been deposited and mineralized in the majority of grafts of all types. Incisor and molar teeth with near normal crown shapes developed in grafts of complete mandibular arches. Hemimandibular arches gave rise almost exclusively to molars. Grafts of the E10 ventral midline region gave rise exclusively to incisors, but the same graft performed at E9 did not produce teeth. It is concluded that incisor primordia are localized in or near the median epithelial isthmus (and are thus destroyed or damaged when the mandibular arch is hemisected) and that incisors are not determined until E10. At E9 the odontogenic neural crest has yet to complete its ventrad migration into proximity with presumptive incisor epithelium. By this time, however, odontogenic crest and presumptive molar epithelium have already reached juxtaposition and molar primordia are fully competent. In grafts of the frontonasal and maxillary processes made at E10, prior to merging of their respective mesenchymes, frontonasal processes gave rise exclusively to incisors, whereas maxillary processes gave rise exclusively to molars.

Innervation of the feline cerebral vasculature by nerve fibers containing calcitonin gene-related peptide: Trigeminal origin and co-existence with substance P

The presence of a population of nerve fibers containing immunoreactive calcitonin gene-related peptide (CGRP) has been demonstrated around cerebral arteries of the cat with immunocytochemistry and radioimmunoassay. CGRP immunoreactivity in the feline cerebral vasculature, as characterized by high-performance liquid chromatography, is similar to authentic rat CGRP. Numerous perikarya containing CGRP are present in the trigeminal ganglia, and surgical lesions of the trigeminal ganglia significantly reduce the levels of CGRP in the cerebral vasculature, suggesting that this cranial nerve is the principal origin of these cerebrovascular nerve fibers. As demonstrated by sequential immunocytochemistry, CGRP coexists with substance P both in the trigeminal ganglion and nerve fibers around cerebral blood vessels. The presence of CGRP in the cerebrovascular trigeminal innervation provides further versatility and complexity for this sensory afferent system putatively involved in the transmission of intracranial pain.

Parasympathetic, sympathetic, and sensory interactions in the iris: nerve growth factor regulates cholinergic ciliary ganglion innervation in vivo

Interactions between peptidergic sensory nerves, noradrenergic sympathetic nerves, and cholinergic parasympathetic fibers were examined in the rat iris. The putative peptide neurotransmitter, substance P (SP), was used as an index of the trigeminal sensory innervation, tyrosine hydroxylase (TH) activity served to monitor the sympathetic fibers, and choline acetyltransferase (CAT) activity was used as an index of the parasympathetic innervation. Destruction of the sympathetic innervation by neonatal administration of 6-hydroxydopamine resulted in increased SP development and a smaller increase in CAT activity in the iris. Moreover, trigeminal ablation resulted in an increase in both TH and CAT activities. Finally, ciliary ganglionectomy resulted in increased SP and a smaller increase in TH activity in the iris. Administration of nerve growth factor (NGF) into the anterior chamber substantially increased both SP and TH activity in the iris and also increased CAT activity to a lesser extent. Moreover, administration of anti-NGF into the anterior chamber prevented both the sympathectomy-induced increases in SP and CAT, and the increases in TH and CAT activities after trigeminal ablation, suggesting that NGF mediated these increases. These observations suggest that the sympathetic, sensory, and parasympathetic innervations of the iris interact by altering availability of NGF elaborated by the iris. Regulation of iris CAT activity was examined in greater detail. Injection of the cholinergic toxin, AF64A, into the anterior chamber concurrently with ablation of the sympathetic and sensory innervations paradoxically increased CAT activity, whereas AF64A alone decreased CAT activity.(ABSTRACT TRUNCATED AT 250 WORDS)

Trigeminal and facial innervation of cirri in three teleost species

Innervation of the cirri in three teleost species (Hypsoblennius gilberti, Hypsoblennius gentilis, Oxylebius pictus) was investigated with the use of HRP- and cobalttracing techniques. All projections were found to be ipsilateral. Labeled cells were demonstrated in both portions of the trigeminal ganglion and in the facial ganglion. Cirrus nerve fibers running in the trigeminal nerve project to terminal fields in an isthmic sensory trigeminal nucleus, to areas adjacent to the descending trigeminal root in the brainstem, and to the medial funicular nucleus in the medulla. Distribution of labeled cells in the trigeminal ganglion complex suggests a functional distinction of the two ganglion portions. Cirrus nerve fibers belonging to the facial nerve terminate in a circumscribed part of of the facial lobe, indicating a somatotopic projection. Pathways were principally the same in all three species investigated. Findings of facial innervation of teleost cirri suggest a suspected gustatory function of teleost head appendages.

A Tactile Fovea in the <i>Scolopacidae?</i>

In the Scolopacidae, which includes the sandpipers and snipe, there is a significant enlargement of the neural apparatus subserving the behaviour of probing which is the preferred feeding strategy of this family of birds. Along with a sophisticated motor arrangement which allows fine manipulation by the tip of the bill without abduction of the mandibles, there is an obvious expansion of the forebrain area devoted to the trigeminal innervation of receptor-rich bill tip. This 'trigeminal expansion' forms a visible bulge on the surface of the brains of the Scolopacidae but not on the brains of related shorebirds in the Charadriidae; this difference would be evident in fossil endocast material and could provide new information on the evolution of the avian brain.

Substance P levels differ in sympathetic target organ terminals and ganglion perikarya

Regulation of the putative neurotransmitter, substance P (SP), was examined in the rat iris and pineal, targets of the peptide-containing superior cervical sympathetic ganglion (SCG). Decentralization (denervation) of the SCG, which is known to increase ganglion SP, did not alter the peptide in either target. Conversely, surgical or pharmacologic (6-hydroxydopamine) ablation of the SCG actually increased SP in both targets. Moreover, destruction of the trigeminal sensory innervation reduced iris SP to virtually blank levels. Finally, trigeminal ablation abolished the rise in iris SP subsequent to sympathetic destruction with 6-hydroxydopamine. Our observations suggest that sympathetic terminals in targets, in contrast to ganglion perikarya and processes, contain negligible quantities of SP. Consequently, SP may not be transported from ganglion to periphery, but may serve an intraganglionic function. Our studies also suggest that sympathetic terminals modulate sensory peptidergic innervation of the iris.

A re-evaluation of the retrograde axonal transport of horseradish peroxidase isoenzymes by semi-quantitative cytochemical analysis

Retrograde axonal transport of 2 horseradish peroxidase (HRP) isoenzymes, the cationic isoenzyme C (HRP-C) and anionic isoenzyme A (HRP-A), was compared using the trigeminal innervation of the cornea in the rabbit. The effects of various fixatives, cytochemical reaction solutions and intracellular decay on the enzymatic activity and cytochemical visualization of both HRP-C and HRP-A were first evaluated quantitatively. Spectrophotometric assays showed that HRP-C and HRP-A respectively retained 92% and 84% of their initial activity after exposure to a fixative containing 1% paraformaldehyde and 0.5% glutaraldehyde for 1 h at 20°. The specific activity of HRP-A was 26% that of HRP-C when assayed in a solution containing 0.5 mg/ml diaminobenzidine and 0.01% H 2O 2 in 0.15 m citrate buffer, pH 5.1. By killing the rabbits at different times following injection of HRP into the corneas, the intracellular half-life of HRP-C was estimated to be 40–52 h and HRP-A 8–12 h. After taking all of these differences into consideration, corrected concentrations of the 2 isoenzymes were compared for retrograde axonal transport. Results showed that both label intensity (scored according to 4 classes) and total number of labeled cell profiles in animals injected with HRP-C was significantly greater than in animals injected with HRP-A. These data are consistent with evidence from previous in vivo and in vitro studies demonstrating a certain selectivity of both uptake and retrograde axonal transport of HRP isoenzyme C that does not appear to be simple ‘fluid-phase’ internalization. The methodology and precautionary steps taken in this study in obtaining a semi-quantitative evaluation of HRP cytochemical visualization should be of general use in a variety of histochemical studies.

Representation of head and face in postcentral gyrus of the macaque.

The receptive field and submodality characteristics of individual neurons within the cytoarchitectural and topographic subdivisions of the head and face areas of the postcentral gyrus (SI) were determined with the technique of extracellular recording. Correlation of the single-unit data with the intracortical location of the recording electrode provided a detailed description of the functional organization within each of the several cytoarchitecturally distinct regions contributing to the representation of the head and face in SI. The data indicate that the functional organization of the SI cortex which receives its principal input from trigeminal mechanoreceptors is comparable to the organization within those SI regions which receive their input from the mechanoreceptors of the limbs, trunk, and tail. In each topographic subdivision of the SI cortex 1) a single region in the periphery is represented several times in widely separated locations, each time in a context of different submodalities and peripheral receptive fields; and 2) neurons belonging to the different submodality classes are segregated so that projections from cutaneous afferents terminate mainly in cytoarchitectural area 3 in the adjacent anterior portion of area 1, while projections from the afferents innervating deep tissues terminate mainly in cytoarchitectural area 3a, area 2, and the posterior part of each 1. Although the mechanoreceptor input to SI is segregated according to submodality and the mechanoreceptors from most body regions project to multiple widely separated regions within SI, neurons with receptive fields confined to the ophthalmic division of the trigeminal peripheral innervation field are found within a restricted region of the anterior postcentral gyral crown which is positioned symmetrically about the junction of cytoarchitectural areas 1 and 3. Neurons with receptive fields confined to the maxillary division of the trigeminal innervation field are found within a ring of cortex which a) completely surrounds the representation of the ophthalmic field, and b) includes parts of cytoarchitectural area 2, 1, 3, and 3a. SI neurons with receptive fields restricted to the mandibular division of the trigeminal innervation field occupy the largest portion of the SI face area and form a ring of cortical cell columns which completely surrounds that cortical region which receives its input from the maxillary peripheral innervation field.