Innervation of periodontal ligament and dental pulp in the rat incisor: An immunohistochemical investigation of neurofilament protein and glia-specific S-100 protein

Summary Nervous elements in the periodontal ligament and dental pulp of rat incisors were investigated by means of immunohistochemistry for neurofilament protein (NFP) and glia-specific S-100 protein. The periodontal ligament in the incisors was densely innervated by NFP-immunoreactive nerve fibers; the distribution of the nerve fibers and their terminations differed markedly from those in molars. NFP-positive, thick nerve bundles entered the lingual periodontal ligament through slits located in the mid-region of the alveolar socket, and immediately formed numerous Ruffini-like corpuscles. In the labial periodontal ligament, all of the NFP-immunoreactive nerve fibers terminated in free endings. The restricted location of the stretch receptor, Ruffini-like corpuscle, in the lingual periodontal ligament appears to be an essential element, because this region is regularly extended during mastication. The nervous elements were restricted to the alveolar half of the periodontal ligament in every region; they avoided the dental half of the periodontal ligament, which presumably moves continuously with the tooth. Pulpal nerve fibers in incisors also showed a characteristic distribution different from those in molars; individual nerve fibers with beaded structures ran in the center of the pulp toward the incisai edge, and did not form the subodontoblastic nerve plexus of Raschkow.Immunostaining for S-100 protein revealed a distribution pattern of nervous elements similar to that for NFP, suggesting that the nerves supplying the periodontal ligament and dental pulp were mostly covered by a Schwann sheath.     

Expression of Vesicular Glutamate Transporters VGLUT1 and VGLUT2 in the Rat Dental Pulp and Trigeminal Ganglion following Inflammation

Background

There is increasing evidence that peripheral glutamate signaling mechanism is involved in the nociceptive transmission during pathological conditions. However, little is known about the glutamate signaling mechanism and related specific type of vesicular glutamate transporter (VGLUT) in the dental pulp following inflammation. To address this issue, we investigated expression and protein levels of VGLUT1 and VGLUT2 in the dental pulp and trigeminal ganglion (TG) following complete Freund’s adjuvant (CFA) application to the rat dental pulp by light microscopic immunohistochemistry and Western blot analysis.

Results

The density of VGLUT2− immunopositive (+) axons in the dental pulp and the number of VGLUT2+ soma in the TG increased significantly in the CFA-treated group, compared to control group. The protein levels of VGLUT2 in the dental pulp and TG were also significantly higher in the CFA-treated group than control group by Western blot analysis. The density of VGLUT1+ axons in the dental pulp and soma in the TG remained unchanged in the CFA-treated group.

Conclusions

These findings suggest that glutamate signaling that is mediated by VGLUT2 in the pulpal axons may be enhanced in the inflamed dental pulp, which may contribute to pulpal axon sensitization leading to hyperalgesia following inflammation.     

Substance P dans les neurones sensitifs primaires innervant la pulpe dentaire,chez le cobaye

Primary sensory trigeminal neurons supplying the dental pulp of incisors in guinea pigs were labelled by retrograde axonal transport. Using an autometallographic intensification procedure, 48 h after injection of wheat germ agglutinin/colloidal gold in the pulp, gold particles were detected in the cytoplasm of the neurons as black granulations. A morphometric study showed a bimodal repartition of the labelled neurons of the ganglion. By submitting ganglion slices to an anti-substance P immunserum revealed by immunocytochemistry, it could be observed that, among the neurons supplying the dental pulp of incisors, the majority of the largest were substance P immunopositive while the smallest were substance P immunonegative. These observations suggest that there could be at least two different populations of nerve fibres supplying the guinea pig incisor dental pulp. Substance P negative neurons could express different neurotransmitters.     

Dental pulp cells produce neurotrophic factors, interact with trigeminal neurons in vitro, and rescue motoneurons after spinal cord injury.

Interactions between ingrowing nerve fibers and their target tissues form the basis for functional connectivity with the central nervous system. Studies of the developing dental pulp innervation by nerve fibers from the trigeminal ganglion is an excellent example of nerve-target tissue interactions and will allow specific questions regarding development of the dental pulp nerve system to be addressed. Dental pulp cells (DPC) produce an array of neurotrophic factors during development, suggesting that these proteins might be involved in supporting trigeminal nerve fibers that innervate the dental pulp. We have established an in vitro culture system to study the interactions between the dental pulp cells and trigeminal neurons. We show that dental pulp cells produce several neurotrophic factors in culture. When DPC are cocultured with trigeminal neurons, they promote survival and a specific and elaborate neurite outgrowth pattern from trigeminal neurons, whereas skin fibroblasts do not provide a similar support. In addition, we show that dental pulp tissue becomes innervated when transplanted ectopically into the anterior chamber of the eye in rats, and upregulates the catecholaminergic nerve fiber density of the irises. Interestingly, grafting the dental pulp tissue into hemisected spinal cord increases the number of surviving motoneurons, indicating a functional bioactivity of the dental pulp-derived neurotrophic factors in vivo by rescuing motoneurons. Based on these findings, we propose that dental pulp-derived neurotrophic factors play an important role in orchestrating the dental pulp innervation.     

Dexamethasone effects on Na<Subscript>v</Subscript>1.6 in tooth pulp,dental nerves,and alveolar osteoclasts of adult rats

Dexamethasone causes extensive physiologic reactions including the reduction of inflammation and pain. Here, we asked whether it also affected dental or periodontal cells or dental innervation by altering voltage-gated sodium channel Na_v1.6 immunoreactivity (IR) or neural synaptophysin. Daily dexamethasone (0.2 mg/kg) given for 1 week to rats caused 12-fold increased intensity of Na_v1.6-IR in dendritic pulpal cells of normal molars and incisors compared with vehicle treatment. These cells also co-localized monocyte (ED-1) or dendritic cell (CD11b/Ox42) markers, and their location in molars expanded during dexamethasone treatment to include deeper pulp. Furthermore, dexamethasone caused a 10-fold decrease in the number of Na_v1.6-immunoreactive multinucleate osteoclasts along the alveolar bone of molar root sockets. No changes occurred for neural Na_v1.6 at axonal nodes of Ranvier, even though IR for calcitonin gene-related peptide was greatly decreased, as expected, and neural synaptophysin-IR was decreased 59% by dexamethasone. At 4 days after tooth injury, pulpal vasodilation and increased Na_v1.6-immunoreactive pulp cells were similar for all groups. Thus, dexamethasone changes dental pulp cell and alveolar osteoclast Na_v1.6-IR in normal teeth, but different mechanisms occur after tooth injury when tissue reactions were similar for dexamethasone- and vehicle-treated rats. Steroid-induced alterations of dental pain and inflammation coincide with altered exocytic capability in dental nerve fibers as shown by synaptophysin-IR and with altered pulp cell Na_v1.6-IR and osteoclast number, but not with any changes in Na_v1.6-IR for nodes of Ranvier in myelinated dental axons.     

S100 protein-immunoreactive trigeminal neurons innervating the rat molar tooth pulp

S100-immunoreactivity (ir) was examined in tooth pulp primary neurons of the rat. An immunofluorescence method demonstrated that the molar tooth pulp contained S100-immunoreactive (ir) nerve fibers. In the root pulp, pulp horn and roof of the pulp chamber, S100-ir smooth and varicose fibers ramified and formed subodontoblastic nerve plexuses. All the fibers became varicose at the base of the odontoblastic layer and extended to the odontoblastic layer. Some varicose endings could be traced into the dentin. The trigeminal neurons retrogradely labeled with fluorogold (FG) from the first and second maxillary molar tooth pulps exhibited S100- and parvalbumin-ir. Approximately 60% and 24% of the labeled cells were ir for S100 and parvalbumin, respectively. Virtually all parvalbumin-ir FG-labeled cells showed S100-ir, while 40% of S100-ir ones coexpressed parvalbumin-ir. An immunoelectron microscopic method revealed that all myelinated axons and half of the unmyelinated axons in the root pulp contained S100-ir. In the odontoblastic layer, predentin and dentin, S100-ir neurites lost the Schwann cell ensheathment and made close contact with cell bodies and processes of odontoblasts. The odontoblastic layer also contained parvalbumin-ir neurites. These neurites were devoid of the Schwann cell ensheathment and in close apposition to cell bodies and processes of odontoblasts. S100-ir pulpal axons seemed to be insensitive to repeated neonatal capsaicin treatment. This study suggests that S100-ir tooth pulp primary neurons are mostly myelinated and that S100-ir unmyelinated axons in the root pulp are preterminal segments of myelinated stem axons.     

Galanin-immunoreactive nerves in the rat iris: alterations induced by denervations

Summary The iris and choroid membrane of the adult rat contain nerve fibers expressing immunoreactivity to the neuropeptide galanin. The density and distribution of galanin-positive nerve fibers varied from iris to iris and, particularly, among animals. Smooth, non-terminal axons were seen running in nerve bundles consisting of otherwise negative fibers. From the choroid membrane these bundles reached the iris via the ciliary body. Axons were frequently seen to branch giving rise to a sparse system of varicose, single fibers in the dilator plate and sphincter area. Galanin-positive fibers were sometimes also seen outlining blood vessels.Capsaicin, in a dose that causes permanent depletion of substance P- and cholecystokinin-immunoreactive fibers in the iris, caused no change in amount of galanin-positive fibers. Removal of the superior cervical ganglion caused a rapid and pronounced increase in the number of galanin-immunoreactive nerve fibers. Similarly, removal of the ciliary ganglion appeared to increase galanin immunoreactivity, while removal of the pterygopalatine ganglion was less effective. Lesioning of the trigeminal ganglion caused a disappearance of galanin immunoreactivity. The sympathetectomy-induced increase was counteracted by capsaicin.Galanin-positive nerve cell bodies were present in both the superior cervical and the trigeminal ganglia. In the superior cervical ganglion, immunoreactive galanin did not seem to coexist with neuropeptide Y-positive cells; in the trigeminal ganglion, some galanin-positive cells also contained calcitonin gene-related peptide (CGRP) immunoreactivity, while most cells did not. In the iris, double-staining suggested that CGRP and galanin immunoreactivities were contained in different fiber populations.We conclude that the rat iris and choroid membrane contain a sparse plexus of nerve fibers expressing galanin-like immunoreactivity. It is suggested that these fibers are derived from the trigeminal ganglion. The iris is able to respond with a pronounced increase in number of galanin-immunoreactive nerve fibers to certain denervation procedures.     

Sema3A chemorepellant regulates the timing and patterning of dental nerves during development of incisor tooth germ

Semaphorin 3A (Sema3A) axon repellant serves multiple developmental functions. Sema3A mRNAs are expressed in epithelial and mesenchymal components of the developing incisor in a dynamic manner. Here, we investigate the functions of Sema3A during development of incisors using Sema3A-deficient mice. We analyze histomorphogenesis and innervation of mandibular incisors using immunohistochemistry as well as computed tomography and thick tissue confocal imaging. Whereas no apparent disturbances in histomorphogenesis or hard tissue formation of Sema3A ^?/? incisors were observed, nerve fibers were prematurely seen in the presumptive dental mesenchyme of the bud stage Sema3A ^?/? tooth germ. Later, nerves were ectopically present in the Sema3A ^?/? dental papilla mesenchyme during the cap and bell stages, whereas in the Sema3A ^+/+ mice the first nerve fibers were seen in the pulp after the onset of dental hard tissue formation. However, no apparent topographic differences in innervation pattern or nerve fasciculation were seen inside the pulp between postnatal and adult Sema3A ^+/+ or Sema3A ^?/? incisors. In contrast, an abnormally large number of nerves and arborizations were observed in the Sema3A ^?/? developing dental follicle target field and periodontium and, unlike in the wild-type mice, nerve fibers were abundant in the labial periodontium. Of note, the observed defects appeared to be mostly corrected in the adult incisors. The expressions of Ngf and Gdnf neurotrophins and their receptors were not altered in the Sema3A ^?/? postnatal incisor or trigeminal ganglion, respectively. Thus, Sema3A is an essential, locally produced chemorepellant, which by creating mesenchymal exclusion areas, regulates the timing and patterning of the dental nerves during the development of incisor tooth germ.     

Innervation of the incisors and periodontal ligament in several rodents: an immunohistochemical study of neurofilament protein and glia-specific S-100 protein

The present immunohistochemical study by use of antisera against neurofilament protein (NFP) and S-100 protein dealt with the innervation of the upper incisors and periodontal ligament in five species of rodents including the guinea pig, hamster, Mongolian gerbil (Meriones unguicularis), mouse and squirrel (Tamias sibiricus). The innervation pattern of the periodontal ligament and dental pulp in the incisors of five rodents was fundamentally identical to that in the rat, which we have previously demonstrated by the same method. The NFP-positive Ruffini-like corpuscles were concentrated in the middle region of the lingual periodontal ligament in all the species examined, suggesting that this particular arrangement of Ruffini-like corpuscles, possibly stretch receptors, was essential to the rodent incisor. The labial periodontal ligament, on the other hand, contained less numerous NFP-positive nerves, these terminating among collagen fibers as free endings. The gerbil and squirrel in particular possessed only a few nerve fibers in the labial periodontal ligament. It was thus presumed that the labial periodontal ligament might be less significant as a mechanoreceptive site than the lingual periodontal ligament. The NFP-positive pulpal nerves, beaded or smooth in shape, ran parallel to the tooth axis, but never extended to the odontoblastic layer; no subodontoblastic plexus was found in the incisors of any of the rodents. S-100-immunopositive nervous elements were distributed in the periodontal ligament and dental pulp of all the rodent species examined, showing a distribution pattern similar to the NFP-positive nerves. Only in the squirrel did odontoblasts show an intense S-100 immunoreactivity.     

Expression and localization of reelin in human odontoblasts.

Reelin is a large extracellular matrix (ECM) glycoprotein strongly expressed during embryonic development in the central nervous system and involved in architectonic brain development. It could participate in axon plasticity processes or adhesion-recognition between nerve fibers in adulthood. Previously identified from a subtractive cDNA library of fully differentiated human odontoblasts, reelin might be involved in the relationship between dental nerves and odontoblasts in as so far the latter are in close association with pulpal nerve fibers. Here, we show by in situ hybridization and immunohistochemistry that reelin is specifically expressed by human odontoblasts in vivo and in vitro and that an intense expression of the reelin gene is detected in odontoblasts in comparison with pulpal cells (PC). Co-cultures of rat trigeminal ganglion (TG) and odontoblasts allow to mimic odontoblast innervation and demonstrate that neurites contact these cells with reelin molecules as observed in vivo in human dental pulp. Moreover, by RT-PCR, we show that both reelin receptors (namely apolipoprotein E receptor [ApoER-2], very low density lipoprotein receptor [VLDLR] and cadherin-related neuronal receptor [CNR]) and the cytoplasmic adapter Disabled-1 implicated in the reelin signal transduction, were expressed by trigeminal ganglion. On the basis of these data, we suggest that reelin might be an extracellular matrix molecule involved in the terminal innervation of the dentin-pulp complex, promoting adhesion between dental nerve endings and odontoblasts.     

Pulp sensibility test in elderly patients

doi: 10.1111/j.1741‐2358.2012.00623.x
Pulp sensibility test in elderly patients Background: The ageing process transforms the histological composition of the dental pulp and may affect the response to pulp sensibility tests. Objectives: The aim of this study was to assess the influence of age on pulp response time and on pain intensity. Material and methods: Fifty elderly patients and 50 young patients were selected. Different classes of teeth were evaluated. The pulp sensibility test was performed with a refrigerant spray. The pulp response time was measured in seconds and the pain intensity was assessed by visual analogue scale. Results: The Spearman coefficient was calculated and detect a positive correlation between age and pulp response time for maxillary incisors, premolars, mandibular incisors, and mean (p < 0.05). On the contrary, there was a negative correlation between age and pain intensity for maxillary incisors, mandibular incisors, and mean (p < 0.05). Also, the results of elderly and young groups were compared by Mann–Whitney test. Significant difference was noted regarding the pulp response time for maxillary incisors, premolars, mandibular incisors, and mean (p < 0.05). Significant difference was detected regarding the pain intensity for mandibular incisors only (p < 0.05). Conclusions: Pulp response time increases when people get older while pain intensity decreases. There were variations among the classes of teeth.     

Evidence for GABAergic fibers entering the superior cervical ganglion of rat from the preganglionic nerve trunk

The origin of gamma-aminobutyric acid immunoreactive (GABA-IR) nerve fibers present in the superior cervical ganglion (SCG) of rat was investigated. With immunocytochemical techniques many nerve fibers showed GABA-like positivity in the cervical sympathetic trunk, whereas similar staining could not be revealed in the internal carotid nerve or in the external carotid nerve. Ligation of the cervical sympathetic trunk for 24 h resulted a dramatic reduction in the staining density in the ganglion and in the cervical sympathetic trunk distal to the ligature. After transection of the preganglionic nerve fibers for eleven days or more, very few fibers staining for GABA were seen in the ganglion. The immunohistochemical results suggest that a major source of GABA within the SCG is a population of GABAergic axons entering from the preganglionic trunk.     

Evidence for GABAergic fibers entering the superior cervical ganglion of rat from the preganglionic nerve trunk

Summary The origin of gamma-aminobutyric acid immunoreactive (GABA-IR) nerve fibers present in the superior cervical ganglion (SCG) of rat was investigated. With immunocytochemical techniques many nerve fibers showed GABA-like positivity in the cervical sympathetic trunk, whereas similar staining could not be revealed in the internal carotid nerve or in the external carotid nerve. Ligation of the cervical sympathetic trunk for 24 h resulted a dramatic reduction in the staining density in the ganglion and in the cervical sympathetic trunk distal to the ligature. After transection of the preganglionic nerve fibers for eleven days or more, very few fibers staining for GABA were seen in the ganglion. The immunohistochemical results suggest that a major source of GABA within the SCG is a population of GABAergic axons entering from the preganglionic trunk.     

Electron microscopic studies of the innervation of the human spleen

Summary The innervation of four normal human spleens was investigated by electron microscopy. Unmyelinated nerve fibers accompanied the arterial vascular system up to the arterioles of the red pulp. Neither myelinated nerve fibers nor ganglion cells were seen in the splenic hilum or in the splenic tissue itself. The nerve fibers terminated against the smooth muscle cells of the blood vessels in a manner that is typical of the autonomic nervous system. The terminal axons contained small and large granular vesicles and thus were adrenergic nerve fibers. In contrast to the results of previous studies using silver impregnation methods innervation of the red or white pulp could not be demonstrated. The findings on human spleens agree with those on mammalian spleens obtained by other authors using ultrastructural and fluorescence histochemical methods.We are indebted to Prof. Dr. K. Unsicker for his help in discussing the results     

Immunohistochemical analysis of two stem cell markers of α-smooth muscle actin and STRO-1 during wound healing of human dental pulp

Recent studies have employed two markers, alpha-smooth muscle actin (α-SMA) and STRO-1, to detect cells with mesenchymal stem cell properties in dental pulp. The present study aimed to explore the expression profile of α-SMA and STRO-1 in intact dental pulp as well as during wound healing in adult dental pulp tissue. Healthy pulps were mechanically exposed and capped with the clinically used materials MTA (ProRoot White MTA) or Ca(OH)(2) to induce a mineralized barrier at the exposed surface. After 7-42?days, the teeth were extracted and processed for immunohistochemical analysis using antibodies against α-SMA, STRO-1 and nestin (a neurogenic cytoskeletal protein expressed in odontoblasts). In normal pulp, α-SMA was detected in vascular smooth muscle cells and pericytes. Double immunofluorescent staining with STRO-1 and α-SMA showed that STRO-1 was localized in vascular smooth muscle cells, pericytes and endothelial cells, in addition to nerve fibers. During the process of dental pulp healing, numerous α-SMA-positive cells emerged at the wound margin at 14?days, and the initially formed mineralized barrier was lined with α-SMA-positive cells similar in appearance to reparative odontoblasts, some of which co-expressed nestin. STRO-1 was abundant in nerve fibers. In the advanced stage of mineralized barrier formation at 42?days, cells lining the barrier were stained with nestin, and no staining of α-SMA was detected in those cells. These observations indicate that α-SMA-positive cells temporarily appear along the wound margin during the earlier phase of mineralized barrier formation and STRO-1 is confined in vascular and neuronal elements.     

Normal and abnormal pathfinding of facial nerve fibers in the chick embryo.

Development of the facial nerve was studied in normal chicken embryos and after surgical disruption of ingrowing sensory facial nerve fibers at 38-72 h of incubation. Disruption of facial nerve fibers by otocyst removal often induced a rostral deviation of the facial nerve and ganglion to the level of the trigeminal ganglion. Cell bodies of the geniculate ganglion trailed their deviating neurites and occupied an abnormal rostral position adjacent to the trigeminal ganglion. Deviating facial nerve fibers were labeled with the carbocyanine fluorescent tracer DiI in fixed tissue. Labeled fibers penetrated the cranium adjacent to the trigeminal ganglion, but they did not follow the trigeminal nerve fibers into the brain stem. Rather, after entering the cranium, they projected caudally to their usual site of entrance and proceeded towards their normal targets. This rostral deviation of the facial nerve was observed only after surgery at 48-72 h of incubation, but not in cases with early otocyst removal (38-48 h). A rostral deviation of the facial nerve was seen in cases with partial otocyst removal when the vestibular nerve was absent. The facial nerve followed its normal course when the vestibular nerve persisted. We conclude that disruption of the developing facial pathway altered the routes of navigating axons, but did not prevent pathfinding and innervation of the normal targets. Pathfinding abilities may not be restricted to pioneering axons of the facial nerve; later-developing facial nerve fibers also appeared to have positional information. Our findings are consistent with the hypothesis that navigating axons may respond to multiple guidance cues during development. These cues appear to differ as a function of position of the navigating axon.     

Normal and abnormal pathfinding of facial nerve fibers in the chick embryo

Development of the facial nerve was studied in normal chicken embryos and after surgical disruption of ingrowing sensory facial nerve fibers at 38–72 h of incubation. Disruption of facial nerve fibers by otocyst removal often induced a rostral deviation of the facial nerve and ganglion to the level of the trigeminal ganglion. Cell bodies of the geniculate ganglion trailed their deviating neurites and occupied an abnormal rostral position adjacent to the trigeminal ganglion. Deviating facial nerve fibers were labeled with the carbocyanine fluorescent tracer Dil in fixed tissue. Labeled fibers penetrated the cranium adjacent to the trigeminal ganglion, but they did not follow the trigeminal nerve fibers into the brain stem. Rather, after entering the cranium, they projected caudally to their usual site of entrance and proceeded towards their normal targets. This rostral deviation of the facial nerve was observed only after surgery at 48–72 h of incubation, but not in cases with early otocyst removal (38–48 h). A rostral deviation of the facial nerve was seen in cases with partial otocyst removal when the vestibular nerve was absent. The facial nerve followed its normal course when the vestibular nerve persisted. We conclude that disruption of the devloping facial pathway altered the routes of navigating axons, but did not prevent pathfinding and innervation of the normal targets. Pathfinding abilities may not be restricted to pioneering axons of the facial nerve; later-developing facial nerve fibers also appeared to have positional information. Our findings are consistent with the hypothesis that navigating axons may respond to multiple guidance cues during development. These cues appear to differ as a function of position of the navigating axon. © 1992 John Wiley & Sons, Inc.     

Tooth pulp tissue promotes neurite outgrowth from rat trigeminal ganglia in vitro

The mammalian tooth pulp becomes innervated by nociceptive and sympathetic axons relatively late during development, when part of the root has formed. In the adult, regenerating axons from an injured tooth nerve or sprouting axons from uninjured nerves in the vicinity rapidly reinnervate denervated tooth pulps. These observations indicate that tooth pulp tissue can use molecular factors to attract pulpal axons from local nerve trunks. The present study examines the hypothesis that these factors include nerve growth factor (NGF), brain derived neurotrophic factor (BDNF) and glial cell line derived neurotrophic factor (GDNF). Explants of trigeminal ganglia from neonatal rat pups showed a distinct neurite outgrowth when co-cultured with pulpal explants collected from molar teeth of 12-day old pups, or after application of a pulpal extract. Control cultures, containing single ganglionic explants, or explants co-cultured with heat-treated pulpal tissue, exhibited a sparse neurite outgrowth. Exogenous NGF and/or GDNF, but not exogenous BDNF, stimulated neurite outgrowth from ganglionic explants. Unexpectedly, application of antibodies against NGF, BDNF and/or GDNF to co-cultures of ganglionic and pulpal explants did not inhibit neuritogenesis. Control experiments showed that IgG molecules readily penetrate the gel used for culture and that even very high concentrations of NGF and GDNF antibodies in combination failed to block neurite growth. On the basis of these data we suggest that other as yet unknown neurite-promoting factors might be present and active in TG/pulpal co-cultures.     

Neurokinin A-like immunoreactivity in feline dental pulp: its distribution,origin and coexistence with substance P-like immunoreactivity

Summary The distribution and origin of neurokinin A (NKA)-like immunoreactivity were investigated in feline dental pulp by an indirect immunofluorescence method. NKA-containing nerve fibres with varicosities, which entered the dental pulp via apical foramen, were distributed throughout this tissue. Many NKA-containing nerve fibres were localized around blood vessels, but some were observed apart therefrom. At the odontoblastic layer, thin NKA-containing nerve fibres were observed running straight toward the pulp-predentinal border between odontoblasts. After inferior alveolar nerve section, all NKA-containing nerve fibres disappeared in the dental pulp, while the removal of the superior cervial ganglion resulted in no change in the distribution of these fibres. The correlation of NKA-like immunoreactivity and substance P (SP)-like immunoreactivity was also investigated by double-immunofluorescence technique. The distribution of NKA-containing nerve fibres was very similar to that of SP-containing nerve fibres; it appeared that all NKA-containing nerve fibres contained SP.     

Neuropeptide Y and vasoactive intestinal peptide coexist in rat thyroid nerve fibers emanating from the thyroid ganglion

Neuropeptide Y (NPY) and vasoactive intestinal peptide (VIP) occur in nerve fibers around blood vessels and between follicles in the thyroid gland of the mouse and rat. VIP-immunoreactive fibers are numerous, while NPY-immunoreactive fibers are fewer. Most of the latter fibers contain noradrenaline (NA) as well as NPY, while a subpopulation was found to contain VIP instead of NA. We have determined the origins of rat thyroid nerve fibers containing NPY, VIP or NPY/VIP by investigating 3 conceivable sources, i.e. the superior cervical ganglion, the nodose ganglion and the thyroid ganglion. Chemical sympathectomy or removal of the superior cervical ganglion did not affect the frequency of VIP-immunoreactive fibers but eliminated most of the NPY-immunoreactive fibers as well as all NA-containing nerve fibers (recognized by antibodies to dopamine-beta-hydroxylase). The NPY-immunoreactive fibers that remained after sympathectomy occurred around blood vessels and between follicles and contained VIP. Cervical vagotomy (removal of the nodose ganglion including the adjacent vagus) did not overtly affect the frequency of NPY/VIP-, VIP-, or NPY/NA-containing fibers in the thyroid. In contrast, extirpation of the thyroid ganglion, which is situated immediately outside the thyroid capsule, greatly reduced the number of VIP- and NPY/VIP-containing fibers in the rat thyroid. On the whole, the results of radioimmunoassay of NPY and VIP agreed well with the immunocytochemical findings. High performance liquid chromatography confirmed the identity of NPY and VIP. The present findings suggest the existence in the rat thyroid of one NPY-containing nerve fiber population that harbours NA and emanates from the superior cervical ganglion; one NPY-containing fiber population that is non-adrenergic, harbours VIP and originates in the thyroid ganglion; and a second VIP-containing fiber population that is devoid of NPY and appears to derive from the thyroid ganglion.