Toshiyuki Itota

Osteocalcin-immunoreactive primary sensory neurons in the rat spinal and trigeminal nervous systems.

1200 micrometer(2) and 9% of those in the range 600-1200 micrometer(2) showed the immunoreactivity (ir). DRG neurons <600 micrometer(2)800 micrometer(2) showed the ir and 21% of those in the range 400-800 micrometer(2) were immunoreactive for this protein. TG neurons <400 micrometer(2) were mostly devoid of OPN-ir (2%). Virtually all (99%) Mes5 primary sensory neurons exhibited the ir. Muscle spindles in the soleus and masseter muscles contained OPN-ir spiral axon terminals. In the hard palate and incisor periodontal ligament, unencapsulated corpuscular endings exhibited the ir. The co-expression of OPN with parvalbumin and calcitonin gene-related peptide (CGRP) was also examined in the DRG and TG. In the DRG, virtually all (97%) OPN-ir neurons exhibited parvalbumin-ir. Conversely, 66% of parvalbumin-ir DRG neurons co-expressed OPN-ir. In the TG, 81% of OPN-ir neurons exhibited parvalbumin-ir and 69% of parvalbumin-ir ones showed OPN-ir. Virtually all OPN-ir DRG and TG neurons were devoid of CGRP-ir. The present study indicates that OPN-ir primary sensory neurons in the DRG and Mes5 are spinal and trigeminal proprioceptors. OPN-ir TG neurons appear to include low-threshold mechanoreceptors.

Alteration of odontoblast osteonectin expression following dental cavity preparation

Cavity preparation can increase the active synthesis and secretion of non-collagenous proteins by odontoblasts, thus resulting in the deposition of tertiary dentine. In this study, the effect of cavity preparation on osteonectin expression was examined in odontoblasts of the rat tooth pulp. A class V cavity was prepared in rat first molars to stimulate odontoblastic secretory activity, and the animals were killed at various intervals. In the normal pulp, osteonectin immunoreactivity was detected in odontoblasts but not other cells. At 1 day after cavity preparation, immunoreactivity had diminished beneath the cavity. At 3 days, strong immunoreactivity could be detected in odontoblasts beneath the cavity. Numerous round cells underlying the odontoblastic layer also demonstrated immunoreactivity. Thereafter, the intensity of osteonectin immunoreactivity in odontoblasts beneath tertiary dentine decreased gradually, and at 30 and 60 days, it was weaker than in normal pulp. These findings suggest that osteonectin is actively synthesized by odontoblasts underlying a cavity in the initial stage of tertiary dentine formation.

Effects of fluoride release from bis-GMA/TEGDMA resin regulated by γ-methacryloxypropyltrimethoxysilane on demineralization of bovine enamel

We previously demonstrated that fluoride release from resins could be regulated by the polysiloxane coating of the fluoride additives. The present study investigated the effects of regulated fluoride release from resin on enamel demineralization in vitro. Bovine enamel cavities were restored with bis-GMA/TEGDMA resins containing 50 wt% NaF powders treated with or without gamma-methacryloxypropyltrimethoxysilane. Specimens were immersed in distilled water that was changed daily to measure the amount of fluoride released over 40 days, and thereafter subjected to pH-cycling. Microradiographic observations were performed to determine total mineral loss (AZ) and lesion depth (Ld) on the enamel. In addition, fluorine distribution was analyzed using EPMA. The resin containing untreated NaF exhibited high-rate and short-term fluoride release, whereas the resin containing treated NaF released low concentrations of fluoride over a longer period. The former showed high fluorine uptake in the adjacent enamel. In contrast, the latter showed high fluorine uptake not only in the adjacent enamel, but also in a wider area of enamel surface. The latter also showed lower AZ and Ld values in the surrounding enamel, indicating a high inhibitory effect on caries formation. Therefore, it is suggested that regulated fluoride release from the resin based on polysiloxane coating is effective in preventing caries formation.

Calbindin D-28k distribution in odontoblasts underneath tertiary dentine in human carious teeth.

OBJECTIVE The aim of this study was to examine the calbindin D-28k immunoreactivity in carious teeth to know whether this protein may have a function in tertiary dentine formation. METHODS Human extracted teeth with or without carious lesions were immersion-fixed with Zamboni fixative, demineralized in 4.13% EDTA solution (pH 7.4), frozen-sectioned, and processed for calbindin immunoreactivity and hematoxylin-eosin stain. The intensity of the immunostaining was evaluated by quantitative densitometry. RESULTS In intact teeth, numerous odontoblasts were aligned underneath the secondary dentine and their cell bodies showed the immunoreactivity. In carious teeth, tertiary dentine had poor- or rich tubular patterns under the carious lesion. Underneath the tubule-poor tertiary dentine, distinct odontoblasts could not be seen at the central site. However, some cells with a flat appearance were located at this site and were immunonegative for calbindin D-28k. On the other hand, columnar odontoblasts were seen at the peripheral site, and their cell bodies and processes showed strong immunoreactivity. Underneath the tubule-rich tertiary dentine, columnar odontoblasts were abundantly distributed, and the strong immunoreactivity was observed in their cell bodies and processes. The immunoreactivity in odontoblasts underneath the tertiary dentine with poor or rich tubular pattern was more intense than that for the secondary dentine in intact teeth (P<0.05). On the other hand, the intensity of the immunoreactivity in odontoblasts was similar underneath the secondary dentine in intact and carious teeth. CONCLUSIONS The present study demonstrated that calbidin D-28k was actively synthesised by odontoblasts under the carious lesion. These findings may suggest that this protein plays an important role in the tertiary dentine formation.

Dopamine Receptor Presence in the Rat Area Postrema Identified by RT-PCR, Immunohistochemistry, and In Situ Hybridization

Abstract Dopamine (DA) is a major central nervous system (CNS) neurotransmitter with many important physiological activities. Investigations into the neuroanatomy and neurologic functions of the dopaminergic neural systems have generated much debate. Regarding neuroanatomy, physiological and pharmacological criteria have divided DA receptors into D1 and D2 subtypes. The genes encoding these subtypes have been cloned and classified into a D1 subfamily encompassing D1 and D5 receptors and a D2 subfamily with D2, D3, and D4. Based on the sequences of the cloned receptors, we prepared antibodies and riboprobes to elucidate the expression of the corresponding proteins and mRNAs in the rat area postrema (AP) by immunohistochemistry and in situ hybridization (ISH). The AP was obtained from adult male Sprague-Dawley rats undergoing brain surgery, and tissue samples were used for RT-PCR, immunohistochemistry, and ISH. The results showed that D2 and D5 receptors and their mRNAs exist in the rat AP. On the other hand, D1, D3, and D4 receptors and their mRNAs were not detected.