Innervation of rat and human dura mater and pericranial tissues in the parieto-temporal region by meningeal afferents.

OBJECTIVE: To reinvestigate the innervation pattern of the dura mater of rat and human middle cranial fossa, the morpho-functional substrate of headache generation, and adjacent extracranial tissues with neuronal in vitro tracing. BACKGROUND: This study was initiated by recent structural and functional findings of meningeal afferent fibers which innervate the cranial dura mater and may project to extracranial tissues. METHODS: Anterograde and retrograde neuronal in vitro tracing was made in formaldehyde fixed hemisected rat and human skulls. The fluorescent tracer DiI was applied to proximally cut meningeal nerves in rat and to distal branches of the spinosus nerve in human calvaria lined by dura mater. After several weeks, the dura mater and deep extracranial tissues were examined with fluorescence microscopy. RESULTS: In addition to a network of meningeal nerve fibers, several fiber bundles were observed, leaving the skull through emissary canals and fissures to innervate the pericranial temporal, parietal, and occipital periosteum. Traced fibers were seen spreading into deep layers of the temporal and upper neck muscles. Retrograde neuronal tracing revealed labeled cell bodies exclusively in the mandibular and maxillary division of the rat trigeminal ganglion, and centrally projecting fibers were identified in the spinal trigeminal tract. Electron microscopy of the cross-sected spinosus nerve showed myelinated and unmyelinated axons with similar numbers in human and rat. CONCLUSIONS: We conclude that a proportion of meningeal afferents innervates extracranial tissues like periosteum and pericranial muscles via collaterals projecting through the skull. These afferents may be nociceptive, some may subserve proprioceptive functions. The finding of extracranial projections of meningeal afferents may be important for our understanding of extracranial impacts on headache generation and therapy.

Analysis of the sensory innervations of the greater trochanter for improving the treatment of greater trochanteric pain syndrome.

In medical practice, greater trochanteric pain syndrome has an incidence of 5.6 per 1,000 adults per year, and affects up to 25% of patients with knee osteoarthritis and low back pain in industrialized nations. It also occurs as a complication after total hip arthroplasty. Different etiologies of the pain syndrome have been discussed, but an exact cause remains unknown. The purpose of this study was to obtain a better understanding of the sensory innervations of the greater trochanter in attempt to improve the treatment of this syndrome. Therefore, we dissected the gluteal region of seven adult and one fetal formalin fixed cadavers, and both macroscopic and microscopic examination was performed. We found a small sensory nerve supply to the periosteum and bursae of the greater trochanter. This nerve is a branch of the n. femoralis and accompanies the arteria and vena circumflexa femoris medialis and their trochanteric branches to the greater trochanter. This nerve enters the periosteum of the greater trochanter directly caudal to the tendon of the inferior gemellus muscle. This new anatomical information may be helpful in improving therapy, such as interventional denervation of the greater trochanter or anatomically guided injections with corticosteroids and local anesthetics.

Sensory Innervation of Human Bone: An Immunohistochemical Study to Further Understand Bone Pain.

Skeletal diseases and their surgical treatment induce severe pain. The innervation density of bone potentially explains the severe pain reported. Animal studies concluded that sensory myelinated A∂-fibers and unmyelinated C-fibers are mainly responsible for conducting bone pain, and that the innervation density of these nerve fibers was highest in periosteum. However, literature regarding sensory innervation of human bone is scarce. This observational study aimed to quantify sensory nerve fiber density in periosteum, cortical bone, and bone marrow of axial and appendicular human bones using immunohistochemistry and confocal microscopy. Multivariate Poisson regression analysis demonstrated that the total number of sensory and sympathetic nerve fibers was highest in periosteum, followed by bone marrow, and cortical bone for all bones studied. Bone from thoracic vertebral bodies contained most sensory nerve fibers, followed by the upper extremity, lower extremity, and parietal neurocranium. The number of nerve fibers declined with age and did not differ between male and female specimens. Sensory nerve fibers were organized as a branched network throughout the periosteum. The current results provide an explanation for the severe pain accompanying skeletal disease, fracture, or surgery. Further, the results could provide more insight into mechanisms that generate and maintain skeletal pain and might aid in developing new treatment strategies. PERSPECTIVE: This article presents the innervation of human bone and assesses the effect of age, gender, bone compartment and type of bone on innervation density. The presented data provide an explanation for the severity of bone pain arising from skeletal diseases and their surgical treatment.

Identifying spinal afferent (sensory) nerve endings that innervate the marrow cavity and periosteum using anterograde tracing.

While sensory and sympathetic neurons are known to innervate bone, previous studies have found it difficult to unequivocally identify and characterize only those that are of sensory origin. In this study, we have utilized an in vivo anterograde tracing technique to selectively label spinal afferent (sensory) nerve endings that innervate the periosteum and marrow cavity of murine long bones. Unilateral injections of dextran-biotin (anterograde tracer; 20% in saline, 50-100 nl) were made into L3-L5 dorsal root ganglia. After a 10-day recovery period to allow sufficient time for selective anterograde transport of the tracer to nerve terminal endings in bone, the periosteum (whole-mount) and underlying bone were collected, processed to reveal anterograde labeling, and immuno-labeled with antibodies directed against protein gene product (pan-neuronal marker; PGP9.5), tyrosine hydroxylase (sympathetic neuron marker; TH), calcitonin gene-related protein (peptidergic nociceptor marker; CGRP), and/or neurofilament 200 (myelinated axon marker; NF200). Anterograde-labeled nerve endings were dispersed throughout the periosteum and marrow cavity and could be identified in close apposition to blood vessels and at sites distant from them. The periosteum and the marrow cavity were each innervated by myelinated (NF200+) sensory neurons, and unmyelinated (NF200-) sensory neurons that were either peptidergic (CGRP+) or nonpeptidergic (CGRP-). Spinal afferent nerve endings did not express TH, and lacked the cylindrical morphology around blood vessels characteristic of sympathetic innervation. This approach to selective labeling of sensory nerve terminal endings will help to better identify how different sub-populations of sensory neurons, and their peripheral nerve terminal endings, interact with bone.

Innervation is higher above Bone Remodeling Surfaces and in Cortical Pores in Human Bone: Lessons from patients with primary hyperparathyroidism.

Mounting evidence from animal studies suggests a role of the nervous system in bone physiology. However, little is known about the nerve fiber localization to human bone compartments and bone surface events. This study reveals the density and distribution of nerves in human bone and the association of nerve profiles to bone remodeling events and vascular structures in iliac crest biopsies isolated from patients diagnosed with primary hyperparathyroidism (PHPT). Bone sections were sequentially double-immunostained for tyrosine hydroxylase (TH), a marker for sympathetic nerves, followed by protein gene product 9.5 (PGP9.5), a pan-neuronal marker, or double-immunostained for either PGP9.5 or TH in combination with CD34, an endothelial marker. In the bone marrow, the nerve profile density was significantly higher above remodeling surfaces as compared to quiescent bone surfaces. Ninety-five percentages of all nerve profiles were associated with vascular structures with the highest association to capillaries and arterioles. Moreover, vasculature with innervation was denser above bone remodeling surfaces. Finally, the nerve profiles density was 5-fold higher in the intracortical pores compared to bone marrow and periosteum. In conclusion, the study shows an anatomical link between innervation and bone remodeling in human bone.

Cutaneous innervation of lower eyelid.

The loss of skin sensation or numbness after lower blepharoplasty is not uncommon. The aim of this study is to elucidate the infraorbital nerve (ION) and zygomaticofacial nerve (ZFN) in detail. Twenty-one hemifaces of 14 fresh Korean adult cadavers were dissected. Infraorbital nerve and ZFN came out of infraorbital foramen and zygomaticofacial foramen. They ran along superficial to the periosteum within and beneath the epimysium of the orbicularis oculi muscle and then through orbicularis oculi muscle perpendicularly and distributed to the skin. The distal branch approached to the lower border of the tarsal plate. Most terminal branches (93.8%) of ION were distributed medial to the lateral canthus. Only a few branches (6.2%) were lateral to the lateral canthus. Most (99.4%) terminal branches of ZFN were distributed lateral to the lateral canthus. Very few (0.6%) branches were medial to the lateral canthus. We conclude that the skin-muscle flap infringes less than the skin flap on the terminal branches of ION and ZFN in exposure of the orbital floor as well as in lower blepharoplasty.

A quantitative analysis of the nerve fibres of the acetabular periosteum of dogs.

There are many techniques for the treatment of hip dysplasia, and novel research is currently being undertaken in the hope of obtaining more efficient and less traumatic techniques. The denervation of the hip joint capsule is a simple and effective technique that allows recovery of the functional activity of the affected limbs in significantly less time than other techniques. This surgical procedure consists of removing the acetabular periosteum, thus eliminating the nerve fibres with consequent analgesia. The aim of this investigation was to quantify the number of nerve fibres present in different regions of the acetabular periosteum. The knowledge of regional differences is potentially valuable for the refining of the denervation technique of the hip joint capsule. Thirty canine acetabular fragments were used to compare the nerve fibre density of the periosteum. The results showed a significant difference between the mean density of nerve fibres at the cranial and dorsal-lateral portion (approximately 75 fibres/mm2) and caudal lateral portion (approximately 60 fibres/mm2) of the acetabulum. Those fibres at the periosteum are almost positioned in a sagittal plane, pointing towards the joint capsule, suggesting the same density in the latter region. These results indicate a new approach to the articular denervation technique, thus obtaining even better results for the treatment of hip dysplasia in dogs.

The transient receptor potential cation channel subfamily V members 1 and 2, P2X purinoceptor 3 and calcitonin gene-related peptide in sensory neurons of the rat trigeminal ganglion, innervating the periosteum, masseter muscle and facial skin.

OBJECTIVE: Distribution of the transient receptor potential cation channel subfamily V members 1 (TRPV1) and 2 (TRPV2), and P2X purinoceptor 3 (P2 × 3) was investigated in rat trigeminal ganglion neurons innervating the periosteum, masseter muscle and facial skin. DESIGN: Double immunofluorescence method for TRPV1 and TRPV2 ion channels or ATP receptor P2 × 3 with calcitonin gene-related peptide (CGRP) was performed on trigeminal ganglion neurons retrogradely labeled from the mandibular periosteum, masseter muscle, or facial skin in 15 male Wistar rats. RESULTS: The cell size of periosteum neurons (mean ± S.D. = 810.7 ± 36.1 µ m2) was smaller than that of masseter muscle neurons (927.0 ± 75.6 µ m2), and larger than that of facial skin neurons (661.3 ± 82.2 µ m2). Periosteum neurons contained TRPV1- (26.7%), TRPV2- (47.1%) and P2 × 3-immunoreactivity (50.1%). Expression of TRPV2-immunoreactivity was more abundant among periosteum neurons than among facial skin neurons (16.1%). Regarding to TRPV1 and P2 × 3 expression, however, there was no significant difference between periosteum neurons and, masseter muscle and facial skin neurons. TRPV1- immunoreactive trigeminal ganglion neurons which innervated the periosteum, masseter muscle and facial skin mostly had small and medium-sized cell bodies, whereas TRPV2- and P2 × 3-immunoreactive trigeminal ganglion neurons innervating those tissues were of various cell body sizes. Approximately 20% of periosteum (19.2%), masseter muscle (19.2%) and facial skin (21.5%) neurons contained both TRPV1- and CGRP-immunoreactivity. Some periosteum neurons also co-expressed CGRP-immunoreactivity with TRPV2- (10.9%) or P2 × 3- immunoreactivity (11.1%). Distributions of perivascular and free nerve fibers containing CGRP and either TRPV1, TRPV2, or P2 × 3 were occasionally very similar in the mandibular periosteum. CONCLUSIONS: The present study indicated that trigeminal ganglion nociceptors innervating the periosteum as well as those innervating the masseter muscle and facial skin have vanilloid, acidic, thermal, mechanical and ATP sensors. In some periosteum neurons, CGRP may act as inflammatory mediator through activation of TRPV1, TRPV2 and P2 × 3.

Post-traumatic orbital reconstruction: anatomical landmarks and the concept of the deep orbit.

Dissection deep within the orbit is a cause for concern to surgeons because of the perceived risks of injuring critical structures such as the contents of the superior orbital fissure and the optic nerve. Although "safe distances" (those distances within which it is considered safe to dissect within the orbit) have been described, these are of limited value if the orbit is severely disrupted or is congenitally shallow. In addition, traumatic defects in the orbital floor, in particular, often extend beyond these distances. Reliable landmarks based on the relations between anatomical structures within the orbit, rather than absolute distances, are described that permit safe dissection within the orbit. We present the concept of the deep orbit and describe its relevance to repair of injuries.

How many types of cholinergic sympathetic neuron are there in the rat stellate ganglion?

Sympathetic cholinergic postganglionic neurons are present in many sympathetic ganglia. Three classes of sympathetic cholinergic neuron have been reported in mammals; sudomotor neurons, vasodilator neurons and neurons innervating the periosteum. We have examined thoracic sympathetic ganglia in rats to determine if any other classes of cholinergic neurons exist. We could identify cholinergic sudomotor neurons and neurons innervating the rib periosteum, but confirmed that cholinergic sympathetic vasodilator neurons are absent in this species. Sudomotor neurons contained vasoactive intestinal peptide (VIP) and calcitonin gene-related peptide (CGRP) and always lacked calbindin. Cholinergic neurons innervating the periosteum contained VIP and sometimes calbindin, but always lacked CGRP. Cholinergic neurons innervating the periosteum were usually surrounded by terminals immunoreactive for CGRP. We conclude that if any undiscovered populations of cholinergic neurons exist in the rat thoracic sympathetic chain, then they are indistinguishable in size, neurochemistry and inputs from sudomotor or cholinergic neurons innervating the periosteum. It may be that the latter two populations account for all cholinergic neurons in the rat thoracic sympathetic chain ganglia.

Biodegradable nanofiber-membrane for sustainable release of lidocaine at the femoral fracture site as a periosteal block: In vitro and in vivo studies in a rabbit model.

The aim of this study was to evaluate the efficacy of a biodegradable, lidocaine-embedded, nanofibrous membrane for the sustainable analgesic release onto fragments of a segmental femoral fracture site. Membranes of three different lidocaine concentrations (10%, 30%, and 50%) were produced via an electrospinning technique. In vitro lidocaine release was assessed by high-performance liquid chromatography. A femoral segmental fracture, with intramedullary Kirschner-wire fixation and polycaprolactone stent enveloping the fracture site, was set-up in a rabbit model for in vivo assessment of post-operative recovery of activity. Eighteen rabbits were randomly assigned to three groups (six rabbits per group): group A comprised of rabbits with femoral fractures and underwent fixation; group B comprised of a comparable fracture model to that of group A with the implantation of lidocaine-loaded nanofibers; and group C, the control group, received only anesthesia. The following variables were measured: change in body weight, food and water intake before and after surgery, and total activity count post-surgery. All membranes eluted effective levels of lidocaine for more than 3 weeks post-surgery. Rabbits in group B showed faster recovery of activity post-operatively, compared with those in group A, which confirmed the pain relief efficacy of the lidocaine-embedded nanofibers. Nanofibers with sustainable lidocaine release have adequate efficacy and durability for pain relief in rabbits with segmental long bone fractures.

Sympathetic nervous system does not mediate the load-induced cortical new bone formation.

UNLABELLED: The contribution of the SNS to bone's response to mechanical loading is unclear. Using a noninvasive model of axial loading of the murine tibia, we found that sciatic neurectomy enhances load-induced new cortical bone formation and that pharmacological blockade of the SNS does not affect such responses, indicating that the SNS does not mediate the osteogenic effects of loading in cortical bone. INTRODUCTION: There is increasing evidence that the sympathetic nervous system (SNS) contributes to the regulation of bone mass and may influence remodeling by modulating bones' response to mechanical load-bearing. The aim of this study was to examine the effect of sciatic neurectomy (SN) on the changes in cortical bone formation induced in response to mechanical loading and to investigate whether the SNS is directly involved in such load-induced responses. MATERIALS AND METHODS: Accordingly, load-induced responses were compared in tibias of growing and adult control C57Bl/J6 mice and in mice submitted to unilateral SN; noninvasive axial loading that induced 2,000 microstrain on the tibia lateral midshaft cortex was applied cyclically, 5 or 100 days after surgery, for 7 minutes, 3 days/week for 2 weeks, and mice received calcein on the third and last days of loading. Tibias were processed for histomorphometry, and transverse confocal images from diaphyseal sites were analyzed to quantify new cortical bone formation. Chemical SNS inactivation was achieved by prolonged daily treatment with guanethidine sulfate (GS) or by the introduction of propranolol in drinking water. RESULTS: Our results show that new cortical bone formation is enhanced by loading in all tibial sites examined and that load-induced periosteal and endosteal new bone formation was greater in the SN groups compared with sham-operated controls. This SN-related enhancement in load-induced cortical bone formation in tibias was more pronounced 100 days after neurectomy than after 5 days, suggesting that longer periods of immobilization promote a greater sensitivity to loading. In contrast, the increases in new bone formation induced in response to mechanical loading were similar in mice treated with either GS or propranolol compared with controls, indicating that inactivation of the SNS has no effect on load-induced cortical new bone formation. CONCLUSIONS: This study shows that SN, or the absence of loading function it entails, enhances loading-related new cortical bone formation in the tibia independently of the SNS.

Temporal summation of pain evoked by mechanical stimulation in deep and superficial tissue.

UNLABELLED: Temporal summation of deep tissue pain has been suggested to be facilitated in chronic musculoskeletal pain syndromes. This study aimed to test whether temporal summation of mechanical induced pressure pain is (1) more pronounced at short (1 second) interstimulus intervals (ISIs) compared with long ISI (30 seconds), (2) more potent than summation elicited by pure skin stimulation, and (3) attenuated in women compared with men. Twelve age-matched men and 12 women were included. A computer-controlled pressure stimulator with a probe surface of 1 cm2 was used to give 10 stimulations to the tibialis anterior, tibia periosteum, and the first web of the hand. Sequential stimulation at pressure pain threshold intensity was applied with different ISIs (1, 3, 5, 10, and 30 seconds). The pain intensity was assessed on a visual analog scale (VAS) after each individual stimulus. The VAS scores after the 10th stimulation with 1-second ISI were increased (P < .05) by 418% +/- 77%, 378% +/- 89%, and 234% +/- 66% compared with the first stimulation for tibia, tibialis anterior, and web, respectively. Temporal summation of pain was observed for all ISIs in tibialis anterior and tibia, eg, 30-second ISI evoked a VAS increase of 192% +/- 71 % (tibia) and 117% +/- 42% (tibialis anterior) compared with the first stimulation. The VAS score after the 10th web stimulation was smaller (P < .05) than that of the 10th tibialis anterior or tibia stimulation. A regression analysis between stimulation number and VAS score showed that the pain intensity increased progressively (1) more for 1-second ISIs compared with longer ISIs (P < .01) and (2) faster in deep tissue compared with skin (P < .01). No gender difference was observed. The temporal summation might be related to both central and peripheral mechanisms. PERSPECTIVE: Pain originating in deep tissue influences central pain processing systems more than superficial tissue. This might be of importance in patients with musculoskeletal pain.

Distribution of galanin in bone and joint tissues.

The article describes the distribution of galanin in normal bone and joint tissues. Periosteum, cortical bone, bone marrow, and synovial membrane of normal rats were analyzed. Immunoelectron microscopy (iEM) was used to analyze the distribution of galanin, and radioimmunoassay (RIA) was used to determine its concentration. Immunoelectron microscopy showed that galanin is abundant in nerve fibers and endothelial cells in the periosteum and also in macrophage-like-cells and nerve fibers of the synovial membrane. The concentration of galanin measured by RIA showed the highest concentration in bone marrow, followed by periosteum and cortical bone. This study demonstrates that galanin is present and can be quantified in different compartments of bone and joint tissues and illustrates the possible role of galanin under physiological conditions.

Chemical sympathectomy impairs bone resorption in rats: a role for the sympathetic system on bone metabolism.

The possibility that the nervous system may control bone metabolism has been raised, as neuromediators physiologically conveyed by sympathetic fibers (eg, vasoactive intestinal peptide) influence bone resorption in vitro. In this study, the sympathetic system was inactivated by treating rats with guanethidine (40 mg/kg/day), a sympathetic neurotoxic, for 21 days, after which a wave of osteoclastic resorption was induced along the mandibular buccal cortex. The effects of denervation were assessed 4 days later (corresponding to the peak of resorption in this model). The rats exhibited ptosis soon after starting guanethidine, proving the success of the sympathectomy. This was associated with a significant increase in calcitonin gene-related peptide- (+54%, p < 0.02) and substance P-immunoreactive sensory fibers (+29%,p < 0.02), a known effect of sympathectomy. For the quantitation of the bone parameters, the study zone was divided into a juxta-osseous alkaline phosphatase-positive osteogenic compartment and a nonosteogenic compartment. In the osteogenic compartment, the resorption surface was reduced by 56% (p < 0.001) in the treated animals, together with a fall in the number of osteoclasts (-25%,p < 0.05) and impaired osteoclast access to the bone surface. Tartrate-resistant acid phosphatase-positive (TRAP+) mononuclear preosteoclasts were found only in this compartment; they were reduced by 43% (p < 0.05) by the sympathectomy. No change in non-specific esterase (NSE)+ osteoclast precursors was found. In the nonosteogenic compartment, vasodilation was the only effect of sympathectomy (+80%,p < 0.05); in particular, the number of NSE+ cells was not modified. Our results indicate that: (1) interactions of NSE+ precursors with osteogenic cells are required for their differentiation into TRAP+ preosteoclasts; (2) the sympathetic nervous system is not involved in osteoclast precursor recruitment; but (3) has a significant effect on resorption by inhibiting preosteoclast differentiation and disturbing osteoclast activation. These data suggest that depletion of sympathetic mediators may disturb osteogenic cell-mediated osteoclast differentiation.

Origins of skeletal pain: sensory and sympathetic innervation of the mouse femur.

Although skeletal pain plays a major role in reducing the quality of life in patients suffering from osteoarthritis, Paget's disease, sickle cell anemia and bone cancer, little is known about the mechanisms that generate and maintain this pain. To define the peripheral fibers involved in transmitting and modulating skeletal pain, we used immunohistochemistry with antigen retrieval, confocal microscopy and three-dimensional image reconstruction of the bone to examine the sensory and sympathetic innervation of mineralized bone, bone marrow and periosteum of the normal mouse femur. Thinly myelinated and unmyelinated peptidergic sensory fibers were labeled with antibodies raised against calcitonin gene-related peptide (CGRP) and the unmyelinated, non-peptidergic sensory fibers were labeled with the isolectin B4 (Bandeira simplicifolia). Myelinated sensory fibers were labeled with an antibody raised against 200-kDa neurofilament H (clone RT-97). Sympathetic fibers were labeled with an antibody raised against tyrosine hydroxylase. CGRP, RT-97, and tyrosine hydroxylase immunoreactive fibers, but not isolectin B4 positive fibers, were present throughout the bone marrow, mineralized bone and the periosteum. While the periosteum is the most densely innervated tissue, when the total volume of each tissue is considered, the bone marrow receives the greatest total number of sensory and sympathetic fibers followed by mineralized bone and then periosteum. Understanding the sensory and sympathetic innervation of bone should provide a better understanding of the mechanisms that drive bone pain and aid in developing therapeutic strategies for treating skeletal pain.

Distribution of substance P and calcitonin gene-related peptide-like immunoreactive nerve fibers in the rat temporomandibular joint.

The density and distribution of substance P-like immunoreactive (SP-LI) and calcitonin gene-related peptide-like immunoreactive (CGRP-LI) nerve fibers in rat temporomandibular joint (TMJ) were investigated in whole-mount preparations and frozen sections by immunohistochemistry with the avidin-biotin-peroxidase complex method. Both types of immunoreactive nerves were observed primarily in the joint capsule, the peripheral articular disc, the synovial membrane, and the periosteum. The distribution of CGRP-LI nerves was similar to that of SP-LI nerves. The anterior portion of the joint capsule and disc was most densely innervated, followed by the posterior, lateral, and medial portions. In addition, CGRP-LI nerves were more numerous and more dense in immuno-intensity than SP-LI nerves. In the synovial membrane, many SP- and CGRP-LI nerves terminated in the subsynovial layer, but some branches extended into the superficial synovial lining layer close to the joint cavity. Immunolabeled nerves were prominently located in the disc attachment and peripheral portion of the disc, and occasional nerves were located in the dense collagenous disc band as an actual disc. However, no fibers were detected in the central disc band. Thus, most of the disc was not innervated by any nerves. The present study provides a morphological basis for the possible roles of neuropeptides in endocytosis by synoviocytes, regulation of blood flow in the synovial membrane, nociception mechanisms of the TMJ, and modulation of the inflammatory response in the TMJ.

Fine structure of calcitonin gene-related peptide-immunoreactive nerve fibres in the rat temporomandibular joint.

The distribution and fine structure of these nerve fibres was examined by immunoelectron microscopy. CGRP-immunoreactive fibres were seen in the nerve bundles, blood vessels and periosteum around the condyle as well as in the disc. These nerve fibres were unmyelinated and had diameters varying from 200 to 600 nm. They were completely or partially enclosed by Schwann cell cytoplasm and did not form synaptic contact with any cells. CGRP-immunoreactive nerve fibres may be sensory nature and this peptide could be involved in pain transmission and neurogenic inflammation.

The effect of tooth extraction on periodontal ligament mechanoreceptors represented in the mesencephalic nucleus of the cat.

When a force is applied to a tooth, mechanoreceptors in the periodontal ligament are stimulated. When teeth are extracted the remnants of the periodontal ligament break down and disappear, but it is not known what happens to the mechanoreceptor neurones that innervated it. The present study seeks to determine the effect of tooth extraction on the population of periodontal ligament mechanoreceptor neurones represented in the mesencephalic nucleus of the fifth cranial nerve. The incisor and canine teeth were extracted from adult cats; terminal experiments were performed between 7.5 months and 2 yr later. Recordings were made in the mesencephalic nucleus with microelectrodes, and neurones were identified in the inferior alveolar nerve that previously innervated the periodontal ligament of one of the extracted mandibular teeth. The majority of these neurones responded only to electrical stimuli applied to the edentulous ridge of the mandible in the area where the incisor or canine teeth had previously been. It was not possible to stimulate them mechanically, despite the use of large forces. A small number had reinnervated new soft-tissue sites. They could be mechanically stimulated and were found adjacent to the area in which the mandibular incisor and canine teeth had been. Thus the population of periodontal ligament mechanoreceptor neurones represented in the mesencephalic nucleus do not all degenerate after tooth extraction. As the majority of those still present do not appear to reinnervate new tissues in which they can be mechanically stimulated, it is unlikely that they have any functional role after tooth loss.

Segmental distribution and morphometric features of primary sensory neurons projecting to the tibial periosteum in the rat.

Previous reports have demonstrated very rich innervation pattern in the periosteum. Most of the periosteal fibers were found to be sensory in nature. The aim of this study was to identify the primary sensory neurons that innervate the tibial periosteum in the adult rat and to describe the morphometric features of their perikarya. To this end, an axonal fluorescent carbocyanine tracer, DiI, was injected into the periosteum on the medial surface of the tibia. The perikarya of the sensory fibers were traced back in the dorsal root ganglia (DRG) L1-L6 by means of fluorescent microscopy on cryosections. DiI-containing neurons were counted in each section and their segmental distribution was determined. Using PC-assisted image analysis system, the size and shape of the traced perikarya were analyzed. DiI-labeled sensory neurons innervating the periosteum of the tibia were located in the DRG ipsilateral to the injection site, with the highest distribution in L3 and L4 (57% and 23%, respectively). The majority of the traced neurons were of small size (area < 850 microm2), which is consistent with the size distribution of CGRP- and SP-containing cells, regarded as primary sensory neurons responsible for perception of pain and temperature. A small proportion of labeled cells had large perikarya and probably supplied corpuscular sense receptors observed in the periosteum. No differences were found in the shape distribution of neurons belonging to different size classes.