Kanako Ochi

The Ruffini Ending as the Primary Mechanoreceptor in the Periodontal Ligament: Its Morphology, Cytochemical Features, Regeneration, and Development

The periodontal ligament receives a rich sensory nerve supply and contains many nociceptors and mechanoreceptors. Although its various kinds of mechanoreceptors have been reported in the past, only recently have studies revealed that the Ruffini endings--categorized as low-threshold, slowly adapting, type II mechanoreceptors--are the primary mechanoreceptors in the periodontal ligament. The periodontal Ruffini endings display dendritic ramifications with expanded terminal buttons and, furthermore, are ultrastructurally characterized by expanded axon terminals filled with many mitochondria and by an association with terminal or lamellar Schwann cells. The axon terminals of the periodontal Ruffini endings have finger-like projections called axonal spines or microspikes, which extend into the surrounding tissue to detect the deformation of collagen fibers. The functional basis of the periodontal Ruffini endings has been analyzed by histochemical techniques. Histochemically, the axon terminals are reactive for cytochrome oxidase activity, and the terminal Schwann cells have both non-specific cholinesterase and acid phosphatase activity. On the other hand, many investigations have suggested that the Ruffini endings have a high potential for neuroplasticity. For example, immunoreactivity for p75-NGFR (low-affinity nerve growth factor receptor) and GAP-43 (growth-associated protein-43), both of which play important roles in nerve regeneration/development processes, have been reported in the periodontal Ruffini endings, even in adult animals (though these proteins are usually repressed or down-regulated in mature neurons). Furthermore, in experimental studies on nerve injury to the inferior alveolar nerve, the degeneration of Ruffini endings takes place immediately after nerve injury, with regeneration beginning from 3 to 5 days later, and the distribution and terminal morphology returning to almost normal at around 14 days. During regeneration, some regenerating Ruffini endings expressed neuropeptide Y, which is rarely observed in normal animals. On the other hand, the periodontal Ruffini endings show stage-specific configurations which are closely related to tooth eruption and the addition of occlusal forces to the tooth during postnatal development, suggesting that mechanical stimuli due to tooth eruption and occlusion are a prerequisite for the differentiation and maturation of the periodontal Ruffini endings. Further investigations are needed to clarify the involvement of growth factors in the molecular mechanisms of the development and regeneration processes of the Ruffini endings.

Alterations in Ultrastructural Localization of Growth-associated Protein-43 (GAP-43) in Periodontal Ruffini Endings of Rat Molars during Experimental Tooth Movement

It is known that orthodontic forces induce discomfort and/or abnormal sensation after application of an orthodontic appliance in patients, suggesting the adaptation of periodontal neural elements to environmental changes. However, no morphological data have been provided. The present study investigated, by immunoelectron microscopy, the localization of growth-associated protein-43 (GAP-43) in periodontal Ruffini endings in rat molars during experimental tooth movement. In the untreated control group, immunoelectron microscopy demonstrated that GAP-43-like immunoreactivity in the Ruffini endings was confined to the Schwann sheaths around the axon terminals, and was in neither the cell bodies of terminal Schwann cells nor the axon terminals themselves. Immunoelectron microscopic observation revealed alterations in the localization of GAP-43-like immunoreactivity in the periodontal Ruffini endings during experimental tooth movement. After 1 day of treatment, the cell bodies of the terminal Schwann cells associated with Ruffini endings appeared to contain immunoreaction products for GAP-43, and retained GAP-43-like immunoreactivity during tooth movement. From 5 to 7 days, a major population of the axoplasm of the periodontal Ruffini endings, which was immunonegative in control, filled the GAP-43 immunoreactions, showing a tendency to decrease in number later, and disappeared completely at 14 days. These findings suggest that orthodontic forces easily induce the remodeling of the mechanoreceptive Ruffini endings as well as the active tissue remodeling in a close relationship. Since the ultrastructural localization of GAP-43-like immunoreactivity was drastically changed in the Ruffini endings during tooth movement, GAP-43 functions as one of the key molecules in the remodeling of mechanoreceptive Ruffini endings during tooth movement.

EXPRESSION OF TRKB-LIKE IMMUNOREACTIVITY IN NON-NEURAL CELLS OF RAT PERIODONTAL LIGAMENT

The Trk family, a group of high-affinity neurotrophin receptors, is divided into three subtypes, TrkA, TrkB and TrkC. These were originally found in neural elements, and are involved in neural development, maintenance and survival. Recent studies have shown that non-neural cells in vitro also express mRNA encoding some neurotrophin receptors. In our preliminary study, TrkB-like immunoreactivity (LI) was found in the various non-neural cells in the rat periodontal ligament. The present study was undertaken to clarify which cell types express Trk-LI, in particular two types of TrkB-LI, in the periodontal ligament of mature rats, using an immunocytochemical technique with polyclonal antibodies. Intense full-length TrkB-LI was clearly recognized in non-neural cells such as fibroblasts, osteoclasts, odontoclasts and cementoblasts as well as in neural elements. Relatively large cells with many cytoplasmic processes were also frequently immunopositive for full-length TrkB. Immunocytochemistry for TrkB[TK-], a truncated type, also demonstrated a similar immunostaining pattern to that of full-length TrkB in non-neural periodontal cells, and intense positive reactions in endothelial cells. Some non-neural cells were positive for TrkA and TrkC. These findings suggest that neurotrophic factors, the ligands of the Trk family, have certain effects on the proliferation and/or differentiation of non-neural cells, as well as on their neurotrophic functions.

Calretinin-like immunoreactivity in the Ruffini endings, slowly adapting mechanoreceptors, of the periodontal ligament of the rat incisor

The distribution and ultrastructural localization of calretinin (CR)-like immunoreactivity (-LI) were investigated in the lingual periodontal ligament of rat incisors. Some thick nerve fibers within the nerve bundle displayed CR-LI; these CR-like immunoreactive (-IR) nerve fibers entered the alveolar half of the lingual periodontal ligament of the incisor where dendritic terminal arborization was exhibited. Thin and beaded CR-IR nerve fibers were rarely observed in the periodontal ligament. Observations of adjacent sections immunostained with protein gene-product 9.5 (PGP 9.5) revealed that most, if not all, PGP 9.5-IR nerve terminals showing a dendritic arborization expressed CR-LI. Immunoelectron microscopic observations showed that electron-opaque immunoreaction products were localized in the axoplasm of the axon terminals, except for the mitochondria, which were surrounded by Schwann sheaths and multiple-layered basal lamina. Neither cell bodies, the cytoplasmic extension of terminal Schwann cells, nor other cellular elements such as periodontal fibroblasts exhibited CR-LI. The present findings suggest that Ruffini endings, an essential mechanoreceptor in the periodontal ligament and categorized as a slowly adapting mechanoreceptor, express CR-LI, and that CR may participate in the Ca2+ homeostasis against external stimuli in the periodontal Ruffini endings.

Immunohistochemical localization of calbindin D28k in the periodontal Ruffini endings of rat incisors

It was examined whether calbindin D28k (CB) might be located in the rat incisor periodontal Ruffini ending, an essential mechanoreceptor in periodontal ligament, by light- and electron-microscopic immunohistochemistry. Some thick nerve fibers showing CB-like immunoreactivity (LI) entered the lingual half of the periodontal ligament of the incisor and showed the dendritic terminal arborization. Electron-dense immunoreaction products indicating CB-LI were distributed diffusely in axoplasm of the axon terminals, no mitochondria, however, were not labeled. Neither cell bodies nor cytoplasmic extensions of the terminal Schwann cells exhibited CB-LI. CB was presumed to be involved in the maintenance of Ca2+ homeostasis in the mechano-electric transduction in mechanoreceptors in the periodontal ligament.

Craniofacial morphology and inclination of the posterior slope of the articular eminence in female patients with and without condylar bone change

Abstract This study investigated the association of craniofacial and glenoid fossa shapes and temporomandibular joint (TMJ) pathology in 39 orthodontic patients with signs and symptoms of TMJ disorders, using helical CT scans. Cephalometric measurements showed that 21 subjects with bilateral condylar bone change (BBC) had significantly smaller SNB angles, ramus heights and S-Ar/N-Ba ratios, as well as larger mandibular plane angles and lower anterior facial height than the 18 subjects with no condylar bone change (NBC). The average posterior slope of the left and right articular eminence in their central and lateral sections was significantly steeper in NBC than in BBC. Condylar bone change might, therefore, not only be related to the morphology of the mandible, but also of the glenoid fossa and cranial base. This appears to reflect adaptive changes in the condyle, articular eminence and cranial base in response to changes in loading.

Immunocytochemical demonstration of ß1-subunit of Na+/K+-ATPase in the mechanoreceptive Ruffini-like endings of the rat incisor ligament

The localization of one of the isoforms of Na+/K(+)-ATPase, the beta 1-subunit, was investigated in the periodontal Ruffini endings of rat incisors by light- and electron-microscopic immunocytochemistry. Immunoreactivity for the rat beta 1-subunit followed the pattern of dendritic terminal arborization in the alveolar half of the lingual periodontal ligament. Ultrastructurally, the reaction products were localized in dilatations of axons, possibly the terminals of Ruffini-like endings in the periodontal ligament. No immunoreactivity was seen in Schwann cells. The immunostaining results support the view that the beta 1-subunit of Na+/K(+)-ATPase is the predominant isoform in sensory neurones, and that this protein is a useful marker for periodontal Ruffini-like endings.

Calbindin D28K-like immunoreactive nerve fibres in the predentine of rat molar teeth

Immunoelectron-microscopy was applied to reveal the existence of nerve fibres and terminals showing calbindin D28k (CB)-like immunoreactivity (IR) in the rat molar tooth pulp. In the root pulp, thick, smooth-surfaced CB-IR nerve fibres were in bundles accompanying the blood vessels. In the coronal pulp, the fibres arborized repeatedly and extensively. CB-IR nerve fibres had a predominantly thick, smooth-surfaced appearance, though parts appeared thin and beaded. Occasionally some thin, varicose CB-IR nerve fibres ran along the odontoblasts, penetrating into the predentine alongside the dentinal tubules. They could be traced for approx. 10-20 microns into the predentine from the pulp-predentine border. Immunoelectron-microscopy revealed that only some of the nerve terminals in the predentine showed CB-IR, and that predentinal CB-IR nerve terminals were located close to the odontoblast processes. No synaptic structures were observed between them. The presence of CB-IR nerve terminals in the predentine suggests that many, if not all, CB-IR nerve fibres could be nociceptors. The CB could be involved in Ca2+ homeostasis during the activation of nociceptors.

Carbonic anhydrase isozyme II immunoreactivity in the mechanoreceptive Ruffini endings of the periodontal ligament in rat incisor

The present study describes the distribution of carbonic anhydrase isozyme II (CA II) in the lingual periodontal ligament of the rat incisor. Some thick nerve fibers in the nerve bundle displayed CA II-like immunoreactivity (LI) as well as non-neuronal elements such as osteoclasts. At the alveolar half of the lingual periodontal ligament of the incisor, thick CA II-like immunoreactive (-IR) nerve fibers showed a tree-like raminification, but thin and beaded CA II-IR nerve fibers were rare. Under the electron microscope, CA II-LI were diffusely localized in the axoplasm of the axon terminals surrounded by Schwann sheaths which were immunonegative for CA II. The cell bodies of the terminal Schwann cells associated with the periodontal Ruffini endings did not exhibit CA II-LI. The present immunohistochemical evidence indicates that CA II may participate in the regulation of the intra-neuronal ion in the periodontal Ruffini endings which are thought to be in a state of high neuronal activity.